CAROTENOIDS

CHEMISCHE REIHE
BAND 23

LEHRBUCHER UND MONOGRAPHIEN

AUS DEM GEBIETE DER EXAKTEN WISSENSCHAFTEN
CAROTENOIDS
Edited by
OTTO ISLER
Chemical Research Department
F. Hoffmann-La Roche & Co. Ltd., Basle, Switzerland

Co-editors Huoo GuTMANN and ULRICH SoLMS

Chemical Research Department
F. Hoffmann-La Roche & Co. Ltd., Basle, Switzerland

SPRINGER BASEL AG 1971

 ISBN 978-3-0348-5832-8 ISBN 978-3-0348-5831-1 (eBook)
DOI 10.1007/978-3-0348-5831-1

©Springer Basel AG 1971

Urspriinglich erschienen bei Birkhiiuser Verlag Basel, 1971
Softcover reprint of the hardcover 1st edition 1971

All rights reserved

No part of this book may be reproduced in any form, by photostat, microfilm,
or any other means, without written permission from the publisher
 In memory of
PAUL KARRER
Nobel Laureate

Dedicated to
All Scientists in the Field of Carotenoids
 LIST OF CONTRIBUTORS

J.C. BAUERNFEIND, Nutley, New Jersey, U.S.A.

G. B. BRUBACHER, Basle, Switzerland
G. ENGLERT, Basle, Switzerland
T. W. GooDWIN, Liverpool, England
0. ISLER, Basle, Switzerland
H. M. KL.AUI, Basle, Switzerland
N.l. KRINSKY, Boston, Massachusetts, U.S. A.
S. LIAAEN-JENSEN, Trondheim, Norway
W.L. MARUSICH, Nutley, New Jersey, U.S.A.
H. MAYER, Basle, Switzerland
G. A. J. PITT, Liverpool, England
N. RIGASSI, Basle, Switzerland
U. SCHWIETER, Basle, Switzerland
0. STRAUB, Basle, Switzerland _
H. THOMMEN, Basle, Switzerland
W. VETTER, Basle, Switzerland
B. C. L. WEEDON, London, England
 CONTENTS

List of Contributors . . 7
I. Introduction by 0. ISLER 11
II. Occurrence by B. C. L. WEEDON 29
III. Isolation, Reactions by S. LIAAEN-JENSEN 61
IV. Spectroscopic Methods by W. VETTER, G. ENGLERT, N. RIGASSI and
U. SCHWIETER . . . . . . . . . . 189
V. Stereochemistry by B. C. L. WEEDON 267
VI. Total Syntheses by H. MAYER and 0. ISLER 325
VII. Biosynthesis by T. W. GooDWIN 577
VIII. Metabolism by H. THOMMEN 637
IX. Function by N.l. KRINSKY 669
X. Vitamin A by G. A J. PITT 717
XI. Use of Carotenoids by J. C. BAUERNFEIND, G. B. BRUBACHER, H. M.
KLAUI and W. L. MARUSICH . . . . . . . 743
XII. Lists of Natural Carotenoids by 0. STRAUB . . . . . . . . . 771
Appendix. Tentative Rules for the Nomenclature of Carotenoids 851
Author Index 865
Subject Index 899

Colour plates showing occurrence and applications of carotenoids, as

well as crystal forms, appear between pages 32 and 33.
 11

I. Introduction
0. ISLER
Chemical Research Department, F. Hoffmann-LaRoche&Co. Ltd., Basle, Switzerland

A. General Remarks . . . 12
B. Historical Development 13
C. Scope and Limitations 15
D. Vitamins A and Provitamins A . 15
E. Carotenoids as Natural Colouring Matters . 19
F. Other Natural Pigments and Related Compounds. 21
G. Nomenclature . . . 22
H. Lists of Carotenoids 24
I. Acknowledgments 25
References . . . . . . 25
12 0. ISLER

A. General Remarks

The brightly coloured carotenoid pigments have aroused the curiosity

of scientists since the beginning of organic chemistry. Indeed some of the
oldest studies were published during the early 19th century. Research on
carotenoids can be separated into four broad periods according to the selection
of problems and the methods of attacking them. During the 19th century,
the emphasis was on isolation of the pigments and their characterization by
measurements of light absorption. The second period (1900-1927) centred on
the determination of empirical formulae and on tentative efforts to discover
a role in photosynthesis. The third period (1928-1949) was dominated by the
provitamin A concept, by establishing structural formulae and developing
synthetic methods. The latest period (1950 to the present) has seen an expo-
nential increase in the number of known carotenoids accompanied by notable
advances in total synthesis and in the determination of absolute configura-
tions.
The recent explosive growth in knowledge has in no small part been due
to new separation methods (e.g. thin layer chromatography) whereby the
number of individual carotenoids has increased from about 80 in 1948 to
the present total of more than 300. Moreover, techniques of structure determina-
tion have changed fundamentally so that today spectroscopic measurements
such as proton magnetic resonance and mass spectra have become factors of
prime importance. Partial and total syntheses are usually effected and serve
to confirm the structures.
Great advances, especially in stereochemistry, have been made in recent
years (e.g. in X-ray crystallography and optical rotatory dispersion combined
with structural chemical correlations) and have reached a climax in the com-
plete elucidation of the absolute configuration of many carotenoids. This
progress has for the first time been summarized (Chapter V).
The present monograph is to be regarded as arising from and developing
the standard work 'Carotenoids' of Karrer and Jucker [1]. It also serves in
part as an extension of Goodwin's 'The Comparative Biochemistry of the
Carotenoids' [2], a work shortly to appear in its second edition. In 1962,
Zechmeister published his specialized monograph 'Cis-trans Isomeric Ca-
rotenoids, Vitamins A and Arylpolyenes' [3], which remains indispensable in
its field.
When Karrer and Jucker planned their survey, it was natural simply to
proceed from one compound to another. Today the multiplicity of carotenoids
makes it imperative to deal with related members of subgroups. This is reflected
in the separate chapters, whether the emphasis is on natural occurrence,
structure or synthesis.
The very rapid growth in the number of known carotenoids and in the
pertinent literature has made it necessary to summarize the information in
a new way. A characteristic of the book of Karrer and Jucker was that it
noted all the then-known carotenoids and recounted their history and proper-
 I. Introduction 13

ties. Today, a more selective function is exercised by a 'Subcommittee on

Carotenoids' of the U.S. National Academy of Sciences, which publishes at
intervals' Specifications and Criteria' of selected carotenoids [ 4]. In the present
work, all the carotenoids described up to mid-1970 have been listed, and the
information about them has been displayed and its provenance indicated,
according to a systematic procedure (Chapter XII).

B. Historical Development

The 19th century saw the accumulation of information on the occurrence

and detection of carotenoids [cf. 5-7]. The isolation of carotene by Wacken-
roder in 1831 [6] and the naming of the yellow pigment of autumn leaves as
xanthophyll by Berzelius in 1837 [7] are worthy of mention. In 1902, Kohl
published [8] a monograph with about 800 literature references, but despite
this very few pure crystalline pigments were then known.
Between 1900 and 1927, Tswett and the Willstatter School worked out
procedures for the separation and purification of carotene, lycopene, xantho-
phyll (lutein), fucoxanthin and bixin, and this was followed by success in
determining many empirical formulae. Tswett [9] invented column chromato-
graphy for the separation of leaf pigments into chlorophylls, xanthophylls
(oxygen containing carotenoids) and carotenes (less strongly adsorbed). To him
must be credited the concept of a carotenoid family. Willstatter and Mieg
recognized in 1907 [10] a formal connection between carotenoids and isoprene.
The monographs of Willstatter and Stoll [11] on chlorophyll in 1913 and on
the assimilation of carbon dioxide in 1918 still merit study, although in the
light of subsequent developments their hypotheses concerning the roles of
carotenoids and chlorophylls in photosynthesis were less than adequate.
In 1922 Palmer contributed to the American Chemical Society Monograph
Series a volume on carotenoids and related pigments [12], which presented
to chemists and biologists a wealth of challenging information which no doubt
stimulated wide interest in the topic and revealed how much had yet to be
done.
Investigations of structure by the schools of Karrer, Kuhn, Zechmeister
and Heilbron began to appear in 1928. The polyene concept was first advanced
by Zechmeister, and the occurrence of conjugation in the chain of double
bonds was shown spectroscopically to be necessary for colour in carotenoids
(Kuhn). The isoprene rule (Ruzicka) had considerable influence on the process
of postulating structures. In 1930-1931 Karrer recognized the symmetrical
nature of the structures of /3-carotene, lycopene and zeaxanthin, and the
constitution of vitamin A was shown to be closely related to half the /3-carotene
molecule.
Indeed, as will be seen, /3-carotene was shown to be the main precursor of
vitamin A in the animal. This provitamin A concept was entirely new, and it
proved to have great scientific and economic importance.
14 0. ISLER

Adsorption chromatography on columns was developed as a widely useful

separation method which in this field was applicable, for example, to the
isolation of ex-, P- and y-carotenes. Oxidative degradation (e.g. by ozone or
permanganate), complete hydrogenation and the synthesis of perhydro
compounds were exploited in structural investigations. Thus it was incidentally
recognized that the familiar pigments crocetin and P-citraurin might be
formed by oxidative degradation at both sides and on one side respectively
of C40 -carotenoids.
In 1934 Zechmeister [13] published his book entitled 'Carotinoide, ein
biochemischer Bericht tiber pflanzliche und tierische Polyenfarbstoffe', which
acted as a bridge between the survey by Palmer and that of Karrer and Jucker.
Between 1934 and 1938 Lederer [14, 15, 15a] reviewed carotenoids of plants
and animals. Later, Karrer and his colleagues isolated mutatochrome and
aurochrome and by means of partial synthesis elucidated the structures of
5,6- and 5,8-epoxides [16].
The number of known naturally occurring carotenoids increased between
1933 and 1948 from 15 to about 80, and the structures of some 35 of these
pigments were definitely established. At this stage research on the synthesis
of vitamin A inevitably overlapped with that of carotenoids, and after a tech-
nical process for vitamin A had been developed at Roche, the schools of
Karrer and Inhoffen achieved almost simultaneously the total synthesis of
P-carotene in 1950. Inhoffen's synthesis was improved and developed by
Roche on a technical scale. Further total syntheses followed, and an im-
portant stimulus was provided by Wittig, whose olefin synthesis, first
exploited in the carotenoid field, has now become a standard method in
organic chemistry.
A notable broadening of the chemistry of natural carotenoids is due
especially to workers in the laboratories of Liaaen-Jensen [17-19] and
Weedon [20, 21]. This expansion is to be seen in the summary by Davis
[22], in the symposium volume 'Carotine und Carotinoide' edited by
Lang [23] and in the review on carotenoproteins by Cheesman et al. [24].
Important surveys were made at the two IUPAC Symposia on 'Carotenoids
other than Vitamin A' held in 1966 at Trondheim and in 1969 at Las Cruces
[25,26].
Significant progress was made in 1966 by MacGillavry [27], who showed
that the polyene chains of carotenoids are almost planar with an S-type
distortion. In another advance Weedon elucidated the absolute configurations
of carotenoids possessing asymmetric centres. In yet another direction, great
advances were achieved by systematic studies on biosynthesis so that a consist-
ent pattern of pathways emerged.
It has become evident that all the more important purely chemical objectives
as regards structure, synthesis and stereochemistry will soon be attained.
The outstanding challenge is biochemical, and the remaining task is to elucidate
modes of action in living cells.
 I. Introduction 15

C. Scope and Limitations

Chemical advances in the last two decades have been so comprehensive

that the present monograph had to be written as a multi-author volume.
To this end eminent University research workers and a group of Roche
scientists have compiled accounts in Chapters II-VI which include all chemical
findings of the period 1950-1970 and added (Chapters VII-XI) reviews on
biochemical aspects; these may well become the main research areas in the
future for workers on carotenoids. The literature has been surveyed up to
mid-1970.

•
Acetyl-Co A

Isopentenyl pyrophosphate

•
,-------> Terpenoid metabolites
(C 5 -unit)

•
Tetraprenyl pyrophosphate ,-----> Vitamins A
(C 20 -unit)

r
Colourless caroten~o'ds IApo-carotenoids I
• i
Carotenoid, , . ,,..-H-ig-h-er-c-ar-o-te-n-oi-ds-,1

Fig. 1. Biosynthesis and metabolism

Fig. 1 illustrates the genesis, classification and breakdown of carotenoids.

Among the metabolites are important substances like vitamin A, trisporic acid
and possibly abscisic acid. The four main types of carotenoids-en closed in the
rectangles-are treated fully in the text; vitamin A is included in the chapters
on spectroscopic methods and total syntheses because there is unavoidable
overlapping. Included in the chapter on total syntheses are numerous
carotenoids not known to occur naturally.
Chapter XII contains lists of the naturally occurring carotenoids compiled
so as to display in a standard manner as much information as possible.
In the subject index there are no references to carotenoid-containing plant
and animal species. For these, reference should be made to Goodwin's book [2],
which, in the first edition, is planned around distribution in nature.

D. Vitamins A and Provitamins A

In 1909 Stepp described a fat-soluble principle obtained from egg yolk,

which proved essential for life. Shortly afterwards other nutrition researchers
16 0. ISLER

found the same factor in butter fat, egg yolk, and cod liver oil. The active
substance was subsequently designated vitamin A by McCollum. It promotes
growth when fed to young rats maintained on a fat-free diet and prevents eye
damage (e.g. xerophthalmia and night blindness). The elapse of 20 years then
saw the differentiation of vitamin A from the other fat-soluble biologically
active substances, namely the antirachitic vitamin D, the anti-oxidant vitamin E
and vitamin K essential for blood clotting [28]. Fish oils were shown to be
the most important natural sources. In animal products the vitamin A activity
increased with the intensity of the ultraviolet absorption at 325-328 nm and
the Carr-Price reaction (blue colouration at 620 nm with antimony trichloride
in chloroform). These observations made it possible to control enrichment
procedures which led to highly active preparations from fish liver oils. In
plant products, however, the vitamin A activity increased parallel to the
intensity of the yellow carotene colour. In 1928 von Euler showed that crys-
talline carotene possesses high vitamin A activity, and in 1930 Moore demon-
strated by very elegant experiments in the rat that absorbed carotene is meta-
bolized to vitamin A and is stored as such in the liver. Karrer's structural
elucidation, in the following year, of P-carotene and vitamin A then provided
the simple chemical explanation that vitamin A has the structure of half of the
P-carotene molecule with an added molecule of water in the end position.
The carrot pigment P-carotene is the most important provitamin A, while
oc- and y-carotene and cryptoxanthin are also provitamins. Their potential
vitamin A activity is, however, only half that of P-carotene. The P-apo-
carotenals, as might have been expected, are very active. Quite generally, all
carotenoids with an unsubstituted P-carotene half are provitamin A com-
pounds, in that they can be converted more or less efficiently in vivo and in vitro
by oxidative degradation into vitamin A via vitamin A aldehyde.
All provitamin A compounds are produced by plants or microorganisms.
Animals possess enzymes occurring mainly in the intestinal mucosa which
convert ingested provitamins to vitamin A. So far this conversion has not
been observed to occur in plants.
Wald showed in 1935 and 1937 that, in denaturing the visual purple of the
ox and frog retina, vitamin A or retinene is formed depending on the conditions.
The latter showed an ultraviolet peak at about 380 nm and in the Carr-Price
reaction an absorption band at 664 nm. Morton demonstrated in 1944 the
identity of retinene with vitamin A aldehyde. In doing so he oxidized vitamin A
with manganese dioxide, discovering thereby a generally useful method of
oxidizing allylic alcohols. Vitamin A acid was synthesized in 1946 by van Dorp
and Arens. It possesses full growth-promoting activity but, unlike vitamin A
and vitamin A aldehyde, is not stored in the body. In 1960 Dowling and Wald
showed that vitamin A acid cannot replace vitamin A in the visual process,
and the Liverpool workers found later that the acid cannot overcome mam-
malian sterility induced by vitamin A deficiency. It is now known that the
transformations shown in Fig. 2 starting from P-carotene and other pro vitamins
occur in the animal organism.
 I. Introduction 17

P-Carotene (Provitamin A)

l
Vitali A •ldohydo ~Vit~i• A ocid ~ '
Vital\" A ~11-d•Vtmin
A •ldohydo

Vitamin A ester Visual purple

(Storage in the liver) (Storage in the eye)
Fig. 2. In vivo transformations in the vitamin A series

Provitamin A compounds are broken down to vitamin A aldehyde by an

oxidation system in the intestinal mucosa. Alcohol dehydrogenase reduces
the aldehyde to the all-trans-vitamin A, which is transported to the liver
or to sites of action. For storage vitamin A is enzymically esterified and
laid down in the liver, chiefly as palmitate. The esters can be reconverted
to vitamin A and vitamin A aldehyde. On the other hand, the oxidation to
vitamin A acid by an oxidase is irreversible. A specific isomerase present in
the retina effects isomerization of vitamin A aldehyde to the compound of
11-cis configuration. As Wald and Oroshnik established, this is a sterically
hindered spatial form, which by condensation with a specific protein (opsin)
forms visual purple or rhodopsin, the pigment concerned in low-intensity
vision. This is cleaved on illumination, liberating all-trans-vitamin A aldehyde.
The recognition of the simple relationships depicted in Fig. 2 was rendered
difficult by the highly sensitive nature of the vitamin A compounds, which
give, under unsuitable treatment, stereoisomers (e. g. 13-cis- and 9-cis-vitamin A)
and artifacts (e.g. retroanhydro- and isoanhydro-vitamin A). Crystalline all-
trans-vitamin A (1), called retinol according to the new nomenclature [29],
was obtained by isolation in 1942 and by synthesis in 1947. The usual com-
mercial forms are vitamin A acetate and vitamin A palmitate, also designated
as vitamin A 1 compounds.

11 13
CH 2 0H
"<::::, "<::::, "<::::,
10 12 14

Vitamin A {I) Vitamin A aldehyde (2)

(Retinol) (Retinal)

11
C02 H
"<::::, "<::::, "<::::,

Vitamin A acid (3)

(Retinoic acid) CHO
11-cis-Vitamin A aldehyde
(11-cis-Retinal)

Carotenmds 2
18 0. ISLER

3-Dehydroretinol, with a Carr-Price maximum at 693 nm, was recognized

in 1931 as a vitamin A congener in the unsaponifiable fraction of sea-fish
liver-oils and in 1937 was prepared in purer state from the unsaponifiable
fractions of the liver oils or eye lipids of fresh-water fish and designated vita-
min A 2 (Morton). Retinene1 and retinene 2 proved more readily separable
than vitamin A 1 and vitamin A 2 and purified retinene 2 was then reduced to
vitamin A 2 • The structure was demonstrated by partial synthesis from vita-
min A aldehyde (2, retinal) and from vitamin A acid (3, retinoic acid). Crys-
tallization followed in 1962. The in vivo and in vitro transformations in the
vitamin A 1 and vitamin A 2 series are completely analogous, but the vitamin A 2
compounds are significantly less stable and less active in animals and therefore
are unlikely to attain economic importance. The active compounds of the
vitamin A 1 and vitamin A 2 series are characterized in Table 1 by physical data
(melting point, ultraviolet absorption maxima and Carr-Price reaction).

Table 1. Melting points and absorption maxima of vitamin A compounds

Vitamin A compounds M.p. (oC) U. v. absorption Carr-Price

(in ethanol) reaction

A. .... E!% Amax E!%

!em !em
(nm) (nm)

Vitamin A 1 62-64 325 1832 620 4800

Vitamin A 1 aldehyde 61-62 381 1530 664 3470
11-cis-Vitamin A 1 aldehyde 63.5-64.4 376.5 878 664 3470
Vitamin A 1 acid 179-180 350 1510
VitaminA 2 63-65 350 1455 693 4100
Vitamin A 2 aldehyde 78-79 401 1470 741 4200
11-cis-Vitamin A 2 aldehyde Oil 393 882 741 4200
Vitamin A 2 acid 183-184 370 1395

Reviews of the chemical and biochemical aspects of the vitamin A field

appeared in 1960, 1967 and 1970 as standards works [28, 30-32]. The excellent
monograph of Moore [33] was the first comprehensive account.
The carotenoid field is very wide but there is no doubt that, according to
international nomenclature, vitamin A falls _outside the category of carotenoids.
Nevertheless, the provitamin A concept is an integral part of the history of
carotenoid research.
Vitamin A, as the most significant carotene metabolite, cannot be left out
of this book. The spectroscopic properties of carotenoids and of vitamin A
derivatives are closely interrelated. The synthetic methods first elaborated for
vitamin A are used in the synthesis of carotenoids, and indeed vitamin A
derivatives are themselves intermediates. Wherever therefore it seemed
appropriate, information about vitamin A has been included in the different
chapters. All the same, the subject of vitamin A is itself too large to be given
systematic coverage here. An exception has been made of visual pigments
 I. Introduction 19

because they, like the carotenoproteins, are important conjugated proteins

with significant differences as well as resemblances.
The technical processes of Roche for production of P-carotene and vita-
min A are based on the Grignard reaction and those of the Badische Anilin-
& Soda-Fabrik on Wittig's olefin synthesis. The syntheses of P-carotene are
considerably more costly than those of the vitamin A esters. Synthetic P-
carotene can therefore never compete, as regards vitamin A activity, with the
more active and better absorbed vitamin A. Since the introduction of synthetic
vitamin A the production of concentrates from fish oils has decreased rapidly.
The production of high potency concentrates is no longer economic.
International committees have been concerned with standards for vitamin A
and provitamin A [34]. The advent of pure all-trans-vitamin A acetate and
pure all-trans-p-carotene provided the needed reference substances and
permitted precise definitions for international units of vitamin or provitamin
activity.
Evaluation of natural products is, however, still somewhat difficult because
bio-assays have a considerable margin of error and spectroscopic methods
ideally require a determination of the proportions of cis and trans isomers,
which differ in physical constants and intrinsic potencies. The analytical
methods recommended in the Pharmacopoeias are therefore, to a minor
extent, based on approximations.

E. Carotenoids as Natural Colouring Matters [35]

The carotenoids form one of the most important groups of natural pigments
and are to be found in all families of the vegetable and animal kingdoms. It is
estimated that more than 100 million tons are produced annually in nature. The
diversity of natural carotenoids is discussed in Chapter II, and many important
carotenoid extracts are now used to colour foods and feed-stuffs.
Table 2 illustrates some of the compounds and their uses. The significant
aspect of this development is that innocuous pigments, some of which have
always been present in natural food and are readily metabolized, have been
made available as safe colouring agents.
Crystalline synthetic P-carotene (3) * was introduced commercially by
Roche in 1954 as a food colourant. Its main application is in the colouring
and fortification of margarine. In 1960 synthetic P-apo-8' -carotenal (248), in
1962 P-apo-8' -carotenoic acid (250) ester and in 1964 canthaxanthin (193) were
introduced for the colouring of foods and feed-stuffs. In 1968 the Badische
Anilin- & Soda-Fabrik brought citranaxanthin (237) on to the market as a
feed additive.
So far all industrial production processes for carotenoids are based on
P-ionone, an important intermediate of the perfumery industry. This can be
obtained by total synthesis from acetone and acetylene via dehydrolinalool, or
by partial synthesis from P-pinene via citral. Production from lemongrass oil,
* Arabic numerals in bold-face type refer to Section A of Chapter XII, as explained on p. 24.
 20 0. ISLER

Table 2. Carotenoid preparations used as yellow and red colourants

Colour Natural source Main pigments Use Vitamin A Pigmentation

activity of eggs and
broiler

Yellow Carrots P-Carotene, IX-carotene Food ++

Palm oil Carotenes, lutein Food ++ +
Alfalfa, grass meal Lutein, P-carotene Feed + ++
Leaves Lutein, P-carotene Feed + ++
Tagetes Lutein, P-carotene Feed (+) ++
Yell ow maize Cryptoxanthin, zeaxanthin Feed (+) ++
Saffron Crocetin, P-carotene, Food and + +
zeaxanthin feed
Annatto seeds Bixin and decomposition Food and ++
products feed
Peels of oranges Esters of lutein Food (+)
and tangerines
Red Paprika (red pepper) Capsanthin, capsorubin Feed ++
Rhodotorula rubra Torularhodin, P-carotene, Food + ++
(red yeast) torulene

which contains 75% citral, is no longer economic. Fig. 3 gives a survey of the
industrial synthetic procedures.
P-CwAldehyde and P-C 19 -aldehyde are important key products of the
Roche processes. From two moles of P-C19 -aldehyde and acetylene, symmetrical

Acetone P-Pinene

1 1
Do~Citnl < - - - - Lemongrass oil

P-Ionone - - - - - - - - - - - - - . . .

1
P-C 14-Aldehyde ---------+

1
1
P-C 19 -Aldehyde ----+------>
c____,---

1 P-Carotene (3) I

1
Canthaxanthin (193)

Fig. 3. Survey of the industrial synthetic processes

 I. Introduction 21

p-carotene (3) is synthesized and from the latter the similarly symmetrical
canthaxanthin (193). The P-apo-carotenoids (248, 250) are prepared from
p-C19 -aldehyde by successive chain extensions. Citranaxanthin (237) is formed
from the P-apo-carotenal (248) and acetone.
Also based on P-ionone are all industrial processes for the production of
vitamin A. More recent procedures, using Wittig's olefin synthesis, also lead
to p-carotene and to the P-apo-carotenoids via vitamin A. At the present
time two further carotenoids, the tomato pigment lycopene (19) and an ester
of the saffron pigment crocetin (269), are undergoing trials as red and yellow
food colourants, respectively. In this use the non-toxic carotenoids are replacing,
to an increasing extent, yellow and red azo dyes which are being prohibited by
food legislation. The carotenoid pigments, which have been present in man's
food throughout the whole of his evolution, are readily metabolized.

F. Other Natural Pigments and Related Compounds

Important pigments of the plant and animal kingdoms which may accom-
pany the carotenoids belong to the following main classes: flavonoids (e.g.
anthocyanins, flavones, flavonols), quinones (e.g. terpenoid quinones, anthra-
quinones, naphthoquinones), pteridines (e.g. flavins, pterins), pyrrole pigments
(e.g. chlorophylls, porphyrins, bile pigments) and other nitrogen-containing
pigments (e.g. phenazines, phenoxazines, melanins, indigo). Most of the yellow
and red colours in the vegetable kingdom, which contrast with the dominant
green of chlorophyll, originate from water-soluble flavonoids or fat-soluble
carotenoids. In the animal world the striking colours are mostly due to nitroge-
nous pigments or carotenoids. The latter are frequently present in the form of
carotenoproteins [24], a class of compounds so far insufficiently investigated.
Natural pigments have been described in monographs or reviews by W. Kar-
rer [36], Goodwin [37], Fox and Vevers [38], Bentley [39], Bielig [40],
Gore et al. [41].
Fig. 4 places carotenoids in a wider biosynthetic perspective. Attention is
drawn to the fact that two Cwunits combine tail to tail to give squalene, a
key compound in the elaboration of sterols. Similarly, two C 20 -units com-
bine to give the colourless C40 -materials from which the coloured carotenoids
are derived. Important metabolites occur in both the C 30 - and the C 40 -
families.
Squalene is one member of a sequence of compounds all arising in the
animal organism by endogenous enzymic reactions. A very different picture
is seen for carotenoids since animals seem unable to synthesize the C 40 -
precursors and are dependent on exogenous carotenoids which they can,
however, utilize by enzymic degradation. In the polyprenes and polyprenyl
quinones the isoprenoid chain is lengthened normally to give a different set
of biologically important compounds. Recent symposia (Goodwin 1970, 1971)
develop this theme [ 42].
22 0. ISLER

•
Acetate

•
C 5 -umt

•
C 10 -umt ~ Monoterpenes

~
Sesqmterpenes

•
C 15 -umt

~
DJterpenes

C 20 -umt

J ~
Vttamms K 1 and E

C -umt
30
VttammsK2

J
C40-umt
Ubtqumones
Carotenmds
Ubtchromenols

I
C 50 -umt
Plastoqumones

Plastochromenols

I ==:I
C 511 -umt Polymer tsoprenmds I
Fig. 4. Biogenesis of terpenoids

G. Nomenclature

At the suggestion of Karrer [43] 'Definitive Rules for Nomenclature of

Carotenoids' were worked out [ 44]. This involves 11 rules, which, because of
the development of carotenoid chemistry, had to be modified and transformed
into 13 more differentiated rules. These new 'Tentative Rules' are printed in
the Appendix of this book; most of the contributions to this volume use,
mainly, the old rules.
Common to both nomenclatures is the definition of carotenoids as a class
of compounds built from 8 isoprene units in such a manner that the linking
of the isoprene units is reversed in the middle of the molecule. As a result, the
two methyl groups near the centre of the polyene chain are separated by
6 C-atoms and the other methyl groups by 5 C-atoms. This is illustrated by
the structural formula of the tomato pigment lycopene (19), from which almost
all other carotenoids can be formally derived by hydrogenation, dehydrogena-
tion, cyclization or oxidation or any combination of these processes. The
similarly common numbering of the carotenoid halves from the ends to the
centre as 1-15 and 1'-15' and the additional numbering of the methyl groups
16-20 and 16'-20' are drawn in the structure of the carrot pigment {3-caro-
tene (3).
Formerly, the end groups were compared with the structures of oc-, {3- and
pseudoionone (t/1), and in asymmetric molecules primed numbers were assigned
in the first place to the t/1 configuration and in the second to the oc configuration
(old rules). Now the configurations at both ends are indicated by Greek letters.
 I. Introduction 23

~
i' ~ "i::: ~ "i:::

Lycopene (19)

c.

4'

11 13 15 14' 12' 10' 8'

~ ~ ~ ~ ~ ~ ~ ~ ~
10 12 14 15' 13' 11' 9'
20' 19'

P-Carotene (3)

The use of the letters rt. and y was abandoned, and new designations were
introduced, namely £ (epsilon) for the halves of £-carotene, K (kappa) for the
five-ring end group of the paprika carotenoids, t/1 (psi) for the lycopene half as
well as q> (phi) and x (chi) for the aromatic end groups. This is illustrated by
the carotenes (5), (4) and (5). The Greek letters appear in alphabetical order
({3, £, K, q>, x, t/J), and the formulae are written correspondingly from left to right,
whereby the right side with the later cited letter is given the primed numbers
(new rule 3).
4'

p p,e-Carotene = IX-Carotene (5)

4'

K K,rp-Carotene (4)

6' 4' 2'

~,. 3' ~1'

X x,t/1-Carotene (5)
 24 0. ISLER

Most carotenoids contain oxygen functions and belong to the class of the
C40 -xanthophylls. Hitherto for such compounds trivial names ending in
'xanthin' were usual (old rule 7). In the new semi-systematic nomenclature the
root of each compound is '-carotene-' (new rule 2). The nomenclature of the
apo-carotenoids had to be broadened (new rule 10). New rules were created
for the designation of the retro compounds (new rule 9), the higher carotenoids
(new rule 11) and stereochemistry (new rule 12).
The formula and semi-systematic name are given for the saffron pigment
crocetin (269) which provide an example of an old apo-carotenoid, and for
decaprenoxanthin (226), one of the higher carotenoids discovered in 1966.

8,8'-Diapocarotene-8,8'-dioic acid (269)

CH 2 0H
HOH 2 C ~

2,2'-Bis(4-hydroxy-3-methyl-2-butenyl)-E,E-carotene (226)

The new rule 13 reads: 'Trivial names will be of value in natural product
and biochemical work, but their use in organic or systematic work should be
restricted. If trivial names are used in a paper, the semi-systematic name
should always be given in parantheses or in a footnote at the first mention'.

H. Lists of Carotenoids

Chapter XII contains three lists of naturally occurring carotenoids, namely:

A. List of nearly 300 compounds of known structure.
B. Alphabetical list of carotenoids of partly or completely unknown
structure.
C. Alphabetical list of trivial names no longer in use.
In the first and most important list the compounds are systematically arranged
in the following order:
1. C40 -Hydrocarbons.
2. C 40-Xanthophylls, sub-divided into mono-, di- and poly-hydroxy com-
pounds, ethers, aldehydes, ketones and acids.
3. Carotenoids with more than 40 C-atoms in the carbon framework.
4. Apo-carotenoids with less than 40 C-atoms in the carbon framework.
Each of these naturally occurring carotenoids of known structure has an
arabic numeral in bold-face type, which is used by the authors throughout this
 I. Introduction 25

book. Examples: /)-carotene (3), lycopene (19), /),a-carotene or ()(-carotene (5),

crocetin (269) and decaprenoxanthin (226), as presented above.
Above each structural formula are the trivial name or synonyms and the
semisystematic name according to the new nomenclature, and under the
formulae are given literature references grouped according to physical data,
chemical reactions, syntheses, etc.
The literature references at the end of Chapter XII are arranged alpha-
betically (according to the name of the first author) and have not been repeated
in the author index of the whole book.
Carotenoids not occurring naturally, intermediates and reaction products
are numbered as they occur in each chapter by arabic numerals in italics.
Examples: K,<p-carotene (4) and x,t/1-carotene (5).

I. Acknowledgments

Thanks are due to the contributors, who have succeeded in presenting

the state of knowledge in their own fields in a most stimulating manner.
The main aim of the authors has been to provide scientists, active in the study
of carotenoids, with the best possible survey.
The late Professor Paul Karrer has followed with interest the planning of
the book and has expressed his satisfaction that so authoritative a set of authors
(academic and industrial) has been assembled as advisers and contributors.
We are grateful to the authorities of IUPAC and IUB for permission to
print the 'Tentative Rules for Nomenclature' as an Appendix.
We are indebted to Professor R. A. Morton of Liverpool for his invaluable
help and advice, particularly in the co-ordination of the biochemical chapters.
Special thanks are due to Professor S. Liaaen-J ensen and to Professor B. C. L.
Weedon as well as to Professor T. W. Goodwin for fruitful discussions in the
planning phase of the book. Dr. A. Cohen and Dr. A.L. Morrison of Welwyn
Garden City, England, read the manuscripts prepared in Switzerland, and
have made valuable suggestions on language difficulties.
The Management of Roche have provided generous financial support so
that the advanced state of knowledge concerning carotenoids could be dis-
played properly and in a manner matching the fascination of the subject.

References

a) Complementary monographs

[1] P. Karrer and E. Jucker, Carotinoide (Birkhiiuser, Basle 1948); English translation by
E. A. Braude (Elsevier, Amsterdam 1950).
[2] T. W. Goodwin, The Comparative Biochemistry of the Carotenoids (Chapman and Hall,
London 1952); 2nd Ed. in preparation.
[3] L. Zechmeister, Cis-trans Isomeric Carotenoids, Vitamins A and Arylpolyenes (Springer,
Vienna 1962).
26 0. ISLER

[4] National Academy of Sciences, Specifications and Criteria for Biochemical Compounds,
2nd Ed., Nat. Acad. Sci.-Nat. Res. Counc., Pub!. No.1344 (Washington D.C. 1967); Sup-
plements in the press.

b) Some old references

[5] H. Braconnot (paprika), Ann. Chim. Phys. [2] 6, 122 (1817); Aschoff (crocin), Berlin. Jahrb.
Pharm. 19, 142 (1818).
[6] H. Wackenroder, Mag. Pharm. 33, 144 (1831).
[7] J. J. Berzelius, Ann. Pharm. 21, 257 (1837).
[8] F. G. Kohl, Untersuchungen uber das Carotin und seine physiologische Bedeutung in der
Pflanze (Borntraeger, Leipzig 1902).
[9] M. Tswett, Ber. Deut. Botan. Ges. 24, 316, 384 (1906); 26, 88 (1908); 29, 630 (1911).
[10] R. Willstiitter and W. Mieg, Ann. Chern. 355, 1 (1907).
[11] R. Willstiitter and A. Stoll, Untersuchungen uber Chlorophyll, Methoden und Ergebnisse
(Springer, Berlin 1913); Untersuchungen uber die Assimilation der Kohlensiiure (Springer,
Berlin 1918).
[12] L. S. Palmer, Carotenoids and Related Pigments, Amer. Chern. Soc. Monograph Ser. (Chemical
Catalog, New York 1922).
[13] L. Zechmeister, Carotinoide (Springer, Berlin 1934).
[14] E. Lederer, Les Carotenoides des Plantes (Hermann, Paris 1934).
[15] E. Lederer, Les Carotenoides des Animaux (Hermann, Paris 1935).
[15a] E. Lederer, Recherches sur les Carotenoides des Animaux lnferieurs et des Cryptogames,
These (Paris 1938).
[16] P. Karrer, Fortschr. Chern. Org. Naturst. 5, 1 (1948).

c) Some new references

[17] S. Liaaen-Jensen, The Constitution of Some Bacterial Carotenoids and their Bearing on
Biosynthetic Problems, Kgl. Norske Videnskab. Selskabs Skrifter 1962, Nr. 8.
[18] S. Liaaen-Jensen and A. Jensen, Progr. Chern. Fats Other Lipids 8, 129 (1965).
[19] S. Liaaen-Jensen, Experientia 26, 697 (1970).
[20] B. C. L. Weedon, in Terpenoids in Plants, ed. by J. B. Pridham (Academic Press, London 1967),
p.119.
[21] B. C. L. Weedon, Fortschr. Chern. Org. Naturst. 27, 81 (1969).
[22] J. B. Davis, in Rodd's Chemistry of Carbon Compounds, 2nd Ed., ed. by S. Coffey, Vol. 2B
(Elsevier, Amsterdam 1968), p. 231.
[23] K. Lang (ed.), Carotine und Carotinoide, Wiss. Veroff. Deut. Ges. Erniihr., Vol. 9 (Steinkopff,
Darmstadt 1963).
[24] D. F. Cheesman, W.L. Lee and P.F. Zagalsky, Bioi. Rev. Cambridge Phil. Soc. 42, 131 (1967);
see also R. Kuhn and H. Kiihn, Eur. J. Biochem. 2, 349 (1967) (last carotenoid publication
of past-master Kuhn).
[25] Pure Appl. Chem.14, 215-278 (1967).
[26] Pure Appl. Chern. 20, 365-553 (1969).
[27] C. H. MacGillavry, Chern. Weekbl. 62, 78 (1966); J. C. J. Bart and C. H. MacGillavry, Acta
Crystallogr., Sect. B 24, 1587 (1968).

d) Vitamins A and provitamins A

[28] R.A. Morton (ed.), Fat-Soluble Vitamins, International Encyclopaedia of Food and Nutrition,
Vol. 9 (Pergamon Press, Oxford 1970).
[29] IUPAC Commission on the Nomenclature of Biological Chemistry, J. Amer. Chern. Soc. 82,
5581 (1960).
[30] W.H.Sebrell,Jr., and R.S.Harris (eds.), The Vitamins, 2nd Ed., Vol.l (Academic Press,
New York 1967).
 I. Introduction 27

[31] U. Schwieter and 0. Isler, in Ullmanns Encyklopiidie der technischen Chemie, 3rd Ed., ed. by
W. Foerst, Vol.18 (Urban & Schwarzenberg, Munich 1967), p. 225.
[32] Vitamins Hormones 18, 289-571 (1960).
[33] T. Moore, Vitamin A (Elsevier, Amsterdam 1957).
[34] IUP AC Applied Chemistry Section, Food Division, Vitamin Assay Subdivision, The Vitamin A
Potency of Beta-Carotene (Butterworths, London 1959); World Health Organ. Tech. Rep. Ser.
No.187, 10 (1960); No. 222, 10, 51 (1961); No. 362, 15, esp. 24 (1967).

e) Natural colouring matters

[35] 0. Isler, R. Riiegg and U. Schwieter, Pure Appl. Chern. 14,245 (1967); 0. Isler, M. Montavon
and R. Riiegg, C. R. 36e Congr. Internat. Chim. Ind., Bruxelles 1966, publ. in Ind. Chim.
Beige, 32, Spec. No., Vol. 3, 853 (1967).
[36] W. Karrer, Konstitution und Vorkommen der organischen Pflanzenstoffe (Birkhiiuser, Basle
1958); 2nd Ed. in preparation.
[37] T. W. Goodwin (ed.), Chemistry and Biochemistry of Plant Pigments (Academic Press, London
1965).
[38] H. M. Fox and G. Vevers, TheN ature of Animal Colours (Sidgwick and Jackson, London 1960).
[39] K.W. Bentley, The Natural Pigments (Interscience, New York 1960).
[40] H.-J. Bielig, in Ullmanns Encyklopiidie der technischen Chemie, 3rd Ed., ed. by W. Foerst,
Vol. 7 (Urban & Schwarzenberg, Munich 1956), p. 84.
[41] T. S. Gore, B.S. Joshi, S.V. Sunthankar and B.D. Tilak (eds.), Recent Progress in the Chemistry
of Natural and Synthetic Colouring Matters and Related Fields (Academic Press, New York
1962).
[42] T. W. Goodwin (ed.), Natural Substances Formed Biologically from Mevalonic Acid, Biochem.
Soc. Symp. No. 29 (Academic Press, London 1970); T. W. Goodwin (ed.), Aspects of Terpenoid
Biochemistry (Academic Press, London 1971).

f) Nomenclature
[43] C. R.14th Conf. IUPAC, London 1947, 142; C. R.16th Conf IUPAC, New York 1951, 110.
[44] IUPAC Commission on the Nomenclature of Biological Chemistry, J. Amer. Chern. Soc. 82,
5583 (1960).
 29

II. Occurrence
B. C. L. WEEDON
Department of Chemistry, Queen Mary College, Mile End Road, London, E.1, England

A. Introduction. . . . . 30
B. Acyclic Carotenes . . 31
C. Acyclic Xanthophylls . 33
D. Alicyclic Carotenes . . 34
E. Alicyclic Xanthophylls 36
1. Oxygen substituent at C-3 . 36
2. Oxygen substituent at C-4 . 37
3. Oxygen substituent at both C-3 and C-4 . 38
4 Unsubstituted P-ring . 40
F. Aromatic Carotenoids ~u
G. Epoxides (5,6 and 5,8) . 41
H. Cyclopentyl Ketones . 42
I. Allenic Carotenoids . 43
1. Acetylenic Carotenoids 44
K. Methyl-oxidized Carotenoids 45
L. Higher Carotenoids (C 45 and C 50 ) 46
M. Degraded Carotenoids 48
1. Seco-carotenoids. 48
2. Apo-carotenoids . 48
3. Related terpenes . 49
N. General Comments. 52
References . . . . . . 53
30 B. C. L. WEEDON

A. Introduction

Man has always derived a great deal of aesthetic pleasure from the rich
variety of colour in nature. It is therefore not surprising that, since the earliest
days of their subjects, biologists, organic chemists and biochemists should
have been intrigued by the pigments present in living organisms. Though the
structures of many of these substances have now been solved by the chemist,
and a large number of them have been synthesized, major problems remain
concerning their formation and function.
Of the various classes of pigments in living organisms, there can be no
doubt that the carotenoids are among the most widespread and important.
They are found throughout the plant kingdom, though their presence is often
masked by chlorophyll. They are responsible for many of the brilliant yellow
and red colours in flowers and fruits. They also occur in insects, birds and other
animals.
The ability to produce carotenoids seems to have been developed at an
early stage in evolution. Some bacteria, the algae and the higher plants preserve
this capacity, but animals, certainly the higher orders, seem to be dependent
for their carotenoids on those present in their diet. However, subsequent
transformation of carotenoids obtained from the diet sometimes leads to
characteristic animal pigments which are not normally found in organisms
capable of carotenogenesis de novo.
Most carotenoids are brightly coloured due to the presence in the molecule
of a chromophore consisting mainly, or entirely, of a chain of conjugated
double bonds. However a few carotenoids have polyene chromophores which
are too short for detection by the human eye. A number of these substances are
now believed to be important intermediates in the biosynthesis of other
carotenoids (Chapter VII).
The majority of carotenoids are tetraterpenes and, as such, can be formally
regarded as resulting from the joining together of eight isoprene units. The
linkage of these units is in the normal head-to-tail manner, except at the centre
of the molecule where the order is reversed so that the C 40 -skeleton, viewed as
a whole, is symmetrical.
In many ways the acyclic compound lycopene (19) can be taken as the
prototype of the carotenoids. Other carotenoids can then be considered as
related to it by means of structural changes in one or both halves of the mole-
cule. Thus the well known P-carotene (3) can be regarded as a bicyclic lycopene.
Though it is often convenient to stress such structural relationships to identify
different classes of carotenoids, it is not intended to suggest that in nature the
corresponding transformations necessarily involve the specific precursor men-
tioned, or that the transformations themselves are as simple as a superficial
inspection of the gross structures might seem to imply.
Though the numbers of basic structural modifications are comparatively
few, their occurrence in different combmations accounts for the remarkable
variety of natural carotenoids. About 300 are now known and the number is
 II. Occurrence 31

increasing rapidly. Most of these are oxygenated carotenoids, xanthophylls,

and the number of hydrocarbons, or carotenes, is relatively small.
The concentrations in which carotenoids occur are low, but vary enormously
from one source to another. The record for carotenes is held by the red fringe
of the corona of the pheasant's-eye narcissus, Narcissus majalis. Here fJ-carotene
(3) can constitute up to 16% of the dry weight. Furthermore the daily rate of
fJ-carotene formation reaches 70 lJ.g/mg dry matter; this is over 10,000 times
the rate observed in carrot roots [1].
The total carotenoid production in nature has been conservatively esti-
mated at about 108 tons a year [2]. Most of this output is in the form of four
major carotenoids: fucoxanthin (190), the characteristic pigment of many
marine algae and without question the most abundant natural carotenoid, and
the three main carotenoids in green leaves, lutein (73), violaxanthin (135) and
neoxanthin (122). By comparison, all othet carotenoids are produced in quite
small amounts, though some, like fJ-carotene (3) and zeaxanthin (67), occur
very widely, and others, such as lycopene (19), capsanthin (170), bixin (265) and
spirilloxanthin (108), constitute the principal pigment in a particular organism.
A great deal of information has been published concerning the carotenoids
present in various plant and animal species, and no attempt will be made here
to repeat the excellent compilations and reviews already available t3-6].
Instead attention will be directed to the different types of carotenoids, and the
principal organisms in which they are encountered. For this purpose it will be
convenient to divide carotenoids into a dozen main classes. In the following
account, unsymmetrical carotenoids are normally considered as representatives
of the second of the two main classes which they illustrate.

B. Acyclic Carotenes

Lycopene (19) together with its hydro and dehydro derivatives form a
structurally well defined class of acyclic carotenoids.
The decahydro derivative (34), lycopersene, has been reported to occur in
the mycelia of Neurospora crassa [7, 8] and in carrots [9], but many workers
have been unable to detect lycopersene in carotenogenic systems [10-12]. If
it occurs at all in nature, it seems likely that it is an artifact formed, perhaps,
from geranylgeranyl pyrophosphate as the result of some lack of specificity of
squalene-synthetase for farnesyl pyrophosphate. Certainly few authorities
would attribute to lycopersene a role in carotenogenesis comparable to that
known to be played by its C 30 -analogue, squalene, in the case of the triterpenes
(Chapter VII).
The occurrence of the octahydrolycopene, phytoene (32), of the hexahydro-
lycopene, phytofluene (30), of the tetrahydrolycopene, C-carotene (26), and of
the dihydrolycopene, neurosporene (22), is however well authenticated [13].
It is now generally accepted that these substances are the key C 40-intermediates
in the biosynthesis of other carotenoids (Chapter VII), each being converted
32 B. C. L. WEEDON

into the next member of the series by a dehydrogenation which introduces an

extra double bond and extends the conjugated system by two double bonds
(Scheme 1). As might be expected, representatives of this group of hydrolyco-

.....~.--···
Scheme 1

penes have been detected in most types of organisms capable of carotenogenesis

de novo. Thus they have been observed in bacteria [14, 15], algae [6], fungi
[16, 17] and higher plants [18].
The hydrolycopenes, though widespread, are normally found in only small
amounts. Carrots [19, 20], in particular carrot oil [21, 22], and tomatoes [23]
have provided the main sources of phytoene, phytofluene and neurosporene.
Several instances are known of strains or mutants of carotenogenic organisms
in which the ability to perform some of the final stages in the dehydrogenation
sequence seems to have been impaired with the result that the production of the
main carotenoids is reduced and one or more hydrolycopenes accumulate.
Thus neurosporene (22), rather than spheroidene (99), accumulates in a green
mutant of the photosynthetic bacterium Rhodopseudomonas spheroides [24].
Phytoene (32) and other hydrolycopenes are formed rather than the normal
carotenoids when a mutant of Chiarella vulgaris is grown in the dark [25].
Phytoene (32) and phytofluene (30) have been obtained in relatively large
amounts from hybrid tomatoes [26]. Even in normal strains it is sometimes
possible to inhibit chemically the enzymes responsible for some steps in the
dehydrogenation sequence. Thus the production of rx-, /3- and y-carotene, and
of lycopene, is almost completely inhibited, and the concentration of phytoene,
phytofluene, (-carotene and neurosporene is considerably increased, when the
fungus Phycomyces blakesleeanus is grown in the presence of diphenylamine [ 4].
Similarly phytoene and other hydrolycopenes accumulate in place of spirilla-
xanthin (108) when the purple bacterium Rhodospirillum rubrum is cultured
in the presence of the same inhibitor [14, 27].
It is now known that the conjugated heptaene from diphenylamine-
inhibited cultures of R. rubrum is not (-carotene (26), as previously assumed,
but its unsymmetrical isomer, 7,8,11,12-tetrahydrolycopene (25) [28]. There
is reason to believe that even in purple bacteria that have not been inhibited
with diphenylamine the heptaene involved in the biosynthesis of carotenoids
is also the unsymmetrical compound (25) [28-30]. However, with the gram-
positive bacterium Flavobacterium dehydrogenans there is evidence for the
occurrence of both (-carotene (26) and its isomer (25) [15, 31].
Recently a series of acyclic carotenes has been reported which are charac-
terized by a terminal isopropyl group (20, 21, 23, 24, 27-29, 31, 33) [32]. These
1,2-dihydro compounds only occur in trace amounts, and the possibility
cannot at present be excluded that they result from some adventitious reduction
of the terminal isopropylidene group in the more common hydrolycopenes, or
in lycopene itself.
 Plates I-VII

Carotenoids

1 Flamingo
2 Cock of the rock (Rupico/a peruviana)
3 Cock of the rock (Rupicola rupicola)
4 OccJirrence in plants and animals
5 Occurrence in plants and animals
6 Eschscholtzia
7 Rose hips
8 8-Carotene

1
2

3
5
6

7
8
10 ~
 II. Occurrence 33

The hydrogenation level of most carotenoids is exemplified in the acyclic

series by lycopene (19). This well known pigment of red tomatoes also occurs
in a number of other fruit and in many berries [3-5]. Its presence as a minor
pigment in bacteria [14, 15] provides an important clue to the pathway by
which carotenoids are formed in these organisms (Chapter VII).
Though the lycopene molecule still contains two positions at which a
dehydrogenation formally analogous to that shown in Scheme 1 can occur,
very few organisms seem to have the ability to carry out these reactions.
However 3,4-didehydrolycopene (18) has been isolated from a microorganism
[33] and from fungi [34], and traces of 3,4,3',4'-tetradehydrolycopene (17), the
end product of the dehydrogenation sequence, have been observed in Spanish
oranges (Valencia) [33].

C. Acyclic Xanthophylls

Over the last ten to fifteen years, extensive studies on the pigments of the
so-called purple photosynthetic bacteria have revealed the existence of a major
class of acyclic xanthophylls [35-38]. These bacteria are primitive organisms
of the sub-order Rhodobacteriineae (order Eubacteriales) and comprise two
families: the Thiorhodaceae which accumulate sulphur globules within the
cells when grown in the presence of sulphide, and the Athiorhodaceae which
do not. All species contain bacteriochlorophyll, and it is interesting to note
that the accompanying xanthophylls are quite distinct from those which are
associated with chlorophyll a and b in higher plants.
Some two dozen of these acyclic bacterial xanthophylls have now been
identified, and consideration of their structures suggests that they are formed
from lycopene (19), or one of its more highly hydrogenated precursors, as the
result of transformations of the types summarized in Schemes 2 to 5.

H20~_ ...··----+ HO~.....-

H0 Scheme2

HO~.. / ----+CH,O~ ....-

Scheme 3

RO~.--···-RO~...···
Scheme4

RO~/~R~/
(R=H or CH 3 )
Scheme 5

Carotenouls 3
34 B. C. L. WEEDON

Spirilloxanthin (108) is one of the commonest carotenoids of the Athio-

rhodaceae and has also been reported in several speCies of ThiMhodaceae
[36, 39, 40]. Such minor pigments as rhodopin (56), 3,4-dehydrorhodopin (55),
anhydrorhodovibrin (97), rhodovibrin (106) and OH-spirilloxanthin (105),
which have also been isolated from R. rubrum, are probably intermediates in
the conversion of lycopene (19) into spirilloxanthin (108) in these bacteria.
Some, if not all, of the transformations summarized by Schemes 2--4
apparently occur with phytoene (32), 7,8,11,12-tetrahydrolycopene (25) and
neurosporene (22), since the pigments in diphenylamine-inhibited cultures of
R. rubrum include 1-hydroxy-1,2-dihydrophytoene (61) [41], 1-hydroxy-1,2-
dihydrophytofluene (60) [29], 1-hydroxy-1,2,7',8',11',12'-hexahydrolycopene
(59) [29], 3,4,11',12'-tetrahydrospheroidene (102), 11',12'-dihydrospheroidene
(101) and spheroidene (99) [28-30a]. The tetrahydrospirilloxanthin (110) is
present in a photolithotropic bacterium [ 42], and the corresponding glycol (81)
is a minor pigment in Rhodomicrobium vannielii [ 43].
In some species of Athiorhodaceae there seems to be little if any dehydro-
genation of neurosporene (22) to lycopene (19) before the onset of transfor-
mations of the types shown in Schemes 2-5. Thus spheroidene (99) is the main
carotenoid when Rhodopseudomonas spheroides is grown anaerobically, and
spheroidenone (182) when it is grown aerobically [39, 44]. The green mutant
of Rps. spheroides which produces neurosporene (22) also contains chloro-
xanthin (58) which can be regarded as an intermediate in the formation of
spheroidene (99) [39, 45]. Minor pigments from Rhodopseudomonas include
OH-spheroidene (107), OH-spheroidenone (185) and 2,2'-diketospirilloxanthin
(208) [46].
The Sarcoscyphaceae fungi synthesize carotenoids with structures similar
to those of the keto series in Athiorhodaceae bacteria, except that tertiary
alcohols or their esters are produced rather than tertiary methyl ethers. Thus
the major pigment in Phillipsia carminea is a diester of phillipsiaxanthin (207)
[47]. Another important development is the discovery that oscillaxanthin, a
characteristic pigment of blue-green algae, is the dirhamnoside (95) of the cor-
responding tetrol [ 48, 49]. The related bis(O-methyl-5-C-methylpentoside)
(96) has been isolated from one batch of Oscillatoria limosa [49a].
Recently a minor xanthophyll in tomatoes was identified as phytoene
1,2-epoxide (116) [50]. There is, however, no evidence that this, or other 1,2-
epoxides, are involved in the biosynthesis ofthe 1-hydroxy or the 1,2-dihydroxy
carotenoids.

D. Alicyclic Carotenes

An oc- or a /3-ionone ring (1 and 2 respectively) is a common feature of many

natural carotenoids not only in higher plants but also in some bacteria, fungi
and algae. The ability to effect a cyclization of the type represented by Schemes
6oc and 6/3 must again have developed at an early evolutionary stage.
 II. Occurrence 35

There is no good evidence that cyclization occurs before neurosporene (22)

in carotenogenesis [5]. Though it has been suggested that 17-carotene from the
berries of Lonicera japonica is a bicyclic form (4) of (-carotene (26), and that
another constituent is a cyclic analogue of phytofluene (30) [51], these struc-
tures lack proof. The monocyclic forms of neurosporene are, however, well
known. Both ex- and P-zeacarotene, (12) and (9) respectively, are found in
maize [52], and P-zeacarotene is present in certain Rhodotorula species of
fungi [5]. P-Zeacarotene has also been identified in Phycomyces blakesleeanus
after P-carotene synthesis has been blocked by diphenylamine [53].

(2)
Scheme 6P

The main cyclic carotenes are P-carotene (3) and its ex- (5), y- (8), b- (11) and
e- (10) isomers. Dehydrogenation of ex- and P-zeacarotene probably accounts
for the formation of b-andy-carotene respectively (cf. Scheme 1). With ex-, P-
and e-carotenes a further cyclization must also be involved. The possibility
cannot be excluded that in some organisms cyclization of lycopene (19) may
also occur (Chapter VII).
P-Carotene (3) is one of the most widespread natural carotenoids [3-6].
In higher plants it is often associated with smaller amounts of ex-carotene (5)
and traces of y-carotene (8). P-Carotene is not nearly as common in bacteria
and fungi, although it is well distributed in the Mucorales [ 4, 5].
The + and - forms of certain Mucorales, e. g. Choanephora conjuncta and
Blakeslea trispora, synthesize different small amounts of P-carotene, but when
grown in mixed culture synthesize up to 15 to 20 times as much as do cultures
of either mating type alone [54-56]. This effect is associated with the production
of trisporic acid (see Chapter VII). In another member of the Mucorales,
Phycomyces blakesleeanus, P-carotene synthesis is activated by the addition
to the culture medium of P-ionone or certain other terpenes [57-59].
b-Carotene (11) was first detected in the fruit hulls of Gonocaryum pyri-
forme [60]. It has since been found in trace amounts in carrots, tomatoes and
elsewhere [ 4]. Substantially larger amounts are present in some tomato
hybrids [61].
e-Carotene (10) was first discovered in the green alga Bryopsis corticulans
[62, 63]. It is also found in diatoms (Bacillariophyceae) and in the blue-green
algae (Cyanophyceae) [4, 6].
36 B. C. L. WEEDON

Torulene, a characteristic pigment of red yeasts [64], is now known to be

the didehydro-y-carotene (7) [65]. Both the di- and tetra-dehydro-P-carotene,
(2) and (1) respectively, are stated to be present in the fungus Epicoccum nigrum
if this is grown in the presence of diphenylamine [66]. The red pigment from
the purple leaves of the box Buxus sempervirens is reported to be dianhydro-
eschscholtzxanthin (35) [67].

E. Alicyclic Xanthophylls

1. Oxygen substituent at C-3

The 3-hydroxy end groups (3) and (4) are frequently encountered in natural
carotenoids but, with the possible exception of the fungal pigment aleuria-
xanthin (49), no carotenoid is known with a hydroxyl substituent at C-3 in an
acyclic end group. It therefore seems likely that most, if not all, 3-hydroxy
carotenoids are the result of direct substitution of the C-3 methylene in either
an IX- (1) or a P- (2) end group.

.~/ HO~
~...··'

(3) (4)

Lutein (73) and zeaxanthin (67), the 3,3'-dihydroxy derivatives of IX- and
P-carotene, are extremely common pigments in algae and plants [3-6]. Nor-
mally lutein is present in considerably larger amounts than zeaxanthin, and
this contrasts markedly with the usual relative abundance of the two parent
carotenes. There are, of course, many exceptions to this broad generalization;
thus zeaxanthin is a major pigment in yellow maize [5, 68]. Rather surprisingly
lutein is very uncommon in bacteria and fungi [ 4, 5].
Both lutein and zeaxanthin, doubtless obtained directly from the diet, are
the yellow pigments in many bird feathers; thus (crystalline) lutein has been
isolated from the plumage of the golden auriole, Oriolus auratus [69]. In some
birds (e.g. the canary and budgerigar), the dietary carotenoids are apparently
transformed before deposition in the feathers, and the resulting pigments are
no longer identical with common plant xanthophylls [69].
Small amounts of cryptoxanthin (39), the monohydroxy derivative of P-
carotene, are often found in plants. Zeinoxanthin (42), the corresponding
derivative of IX-carotene, is present in corn [70], in alfalfa [71] and in red
peppers [72, 73], and is probably identical with a pigment from oranges [74].
Rubixanthin (45), the related 3-hydroxy-y-carotene, was first isolated from
rose hips, Rosa rubiginosa [75], and its 5'-cis isomer, gazaniaxanthin (46), is a
major pigment in the petals of Gazania rigens [76-78]. Celaxanthin, from the
red berries of Celastrus scandens, is believed to be the 3',4' -didehydro derivative
(44) of rubixanthin [79].
 II. Occurrence 37

Other compounds with the end group (4) include saproxanthin (75), the
major pigment of the flexibacterium Saprospira grandis [80], myxoxanthophyll
(90), the rhamnoside which is a common pigment of blue-green algae [49, 81]
and the related 0-methyl-5-C-methylpentoside (91) present in the blue-green
alga Oscillatoria limosa [49a].
Eschscholtzxanthin (84), a dehydro derivative of zeaxanthin with end groups
of the type (5), was first isolated from the blooms of Eschscholtzia califor-
nica [82]. The corresponding diketone, rhodoxanthin (209), with end groups
of the type (6), constitutes the principal red pigment in the red fruit of the yew,

.~ (5)

Taxus baccata [83]; the minor pigments include the intermediate hydroxy
ketone, eschscholtzxanthone (192) [84]. Rhodoxanthin (209) is also a charac-
teristic red pigment in the fungus Epicoccum nigrum [85], and in some bird
feathers, such as those of M egaloprepia magnifica [ 69]. These three xanthophylls
with the end groups (5) or (6), together with dianhydroeschscholtzxanthin (35),
are the only natural C40 -carotenoids which possess a retro polyene chromo-
phore [86].
In contrast to the two major bicyclic xanthophylls lutein (73) and zea-
xanthin (67), little is known about the 3,3'-dihydroxy derivative of e-carotene.
Its occurrence has been suggested on a number of occasions, but the evidence is
by no means conclusive. There is, however, reason to believe that a pigment in
the teleost fishes may have this structure (78) [87].

2. Oxygen substituent at C-4

Unlike the 3-hydroxy xanthophylls discussed above, carotenoids with a
hydroxyl group at the 4-position in a terminal ring are rare. There are, however,
grounds for believing that they are intermediates in the biosynthesis of the
4-keto carotenoids (7). It may therefore be significant that 4-hydroxy carot-
enoids have been found in invertebrates (Annelida and Crustacea) [88]. Thus
isocryptoxanthin (40) has been reported in the polychaete Sabella penicillis
[89], in Carcinus maenas [90] and, together with isozeaxanthin (71), in the
marine isopods Idothea granulosa and I. montereyensis [91]. It also occurs in
other anostracan branchiopods, in Branchipus stagnalis and, together with
isozeaxanthin, in Chirocephalus diaphanus [92]. Isozeaxanthin is present in
Tanymastix lacunae [92].

(7)
38 B. C. L. WEEDON

Echinenone (148), the 4-keto derivative of P-carotene, appears to be

widely distributed in algae and marine invertebrates [ 4-6]. The corresponding
derivative (151) of y-carotene, and the related 1'-hydroxy-1',2'-dihydro com-
pound (160), occur as minor carotenoids in Mycobacterium phlei [93]. Struc-
turally related compounds include the 3',4'-didehydro derivative deoxyflexi-
xanthin (158) [94, 97] and its glucoside (159) [95], which occur in some genera
of Myxobacterales. The glycosidic pigment 4-ketophleixanthophyll (172),
from a strain of M. phlei [96], also deserves mention.
Asteroidenone, from star fish, has been tentatively formulated as 3' -hydroxy-
echinenone (154) [98]. A pigment with this structure has been isolated from
blue-green algae [99]. Phoenicopterone, a pigment from the scarlet ibis,
Guara rubra [100], and some flamingoes, e.g. Phoenicopterus chilensis [101],
is regarded as the 4-keto derivative (149) of !X-carotene.
Canthaxanthin, the 4,4' -diketo-P-carotene (193), was first isolated from the
edible mushroom, Cantharellus cinnabarinus [102]. It has since been found in
bacteria [103], blue-green algae [104] and Crustacea [88, 105], and it con-
stitutes one of the chief pigments in the brilliantly coloured feathers of such
birds as the flamingo [101], the scarlet ibis [69, 100] and roseate spoonbill,
Ajaia ajaja [106].
Both canthaxanthin (193) and echinenone (148) are also stated to be pro-
duced in several green algae as secondary carotenoids at the end of the growth
phase instead of, or in addition to, the primary carotenoids [107].

3. Oxygen substituent at both C-3 and C-4

Crustaxanthin, a minor carotenoid which accompanies astaxanthin (203)
in the crustacean Arctodiaptomus salinus, has been formulated as the tetra-
hydroxy-P-carotene (93) [108]. Like other 3,4-dihydroxycarotenes, (64), (85)
and (173), it seems probable that crustaxanthin is either an intermediate in the
biosynthesis of the related 3-hydroxy-4-keto carotenoid, or is formed by
reduction of the latter.
Several organisms are capable of elaborating carotenoids with the 3-
hydroxy-4-keto end group (8). Thus Flexibacteria (which may be regarded as
non-photosynthetic blue-green algae, or as bacteria) produce flexixanthin
(171) [97].

HO
'<:::::,.·····
~/
H 0
(8) (9)

The flowers of Adonis annua contain 3-hydroxyechinenone (153), adonirubin

(196), and adonixanthin which has been tentatively formulated as (167) [109].
Adonirubin is identical with phoenicoxanthin present in flamingoes [100].
The same pigment is also reported to occur, together with echinenone (148)
and canthaxanthin (193), as a secondary carotenoid in green algae [107].
 II. Occurrence 39

Undoubtedly the best known keto xanthophyll is the symmetrical dihydroxy-

diketo-P-carotene, astaxanthin (203). This is a common pigment in Crustacea,
such as the common lobster, Homarus gammarus [3, 4], and in other marine
invertebrates. It is found in the plumage of some birds, e.g. flamingoes [101],
scarlet ibis [100], and the shrike, Laniarius atrococcineus [69], and has also
been reported in insects and fish [3, 4]. It is therefore usually regarded as a
typical animal carotenoid though it appears to be one of the keto carotenoids
formed by green algae at the end ofthe growth phase [107].
Astaxanthin (203) is of special interest as it constitutes the chief prosthetic
group in the carotenoproteins [110]. Carotenoids frequently occur in asso-
ciation with protein, but the term carotenoprotein implies a definite stoichio-
metric relationship between the two components. Many of these complexes
have been reported, so far mainly in invertebrates, but remarkably little is
known about them at the present time [110, 111]. Presumably there must be
specific sites on the protein for the attachment of the carotenoid, but they have
not been identified. The nature of the linkage is also unknown, though theories
have been advanced to explain the marked change in the visible-light absorption
spectrum which often accompanies carotenoprotein formation [112]. The
resulting complex may absorb at longer or shorter wavelengths than the parent
carotenoid: yellow, red, purple, green, blue and even colourless caroteno-
proteins have been reported [110]. Until recently the only prosthetic group to
have been clearly idelltified in any of these complexes was astaxanthin (203), but
it is now necessary to add astaxanthin esters [113] and canthaxanthin (193)
[114].
Very few carotenoproteins have so far been isolated in a state of reasonable
purity. Crustacyanin, the bright blue protein from the lobster carapace, seems
to be a combination of astaxanthin with a simple protein [115]. The caroteno-
proteins from the carapace, mandibles and stomach wall of Aristeus antennatus,
and from the carapaces of a number of other decapod Crustacea, apparently
have a similar composition [116]. Ovoverdin, the green storage protein of
lobster eggs, is a combination of astaxanthin with a lipoprotein particularly
rich in phospholipids [117]. Ovorubin, a red protein in the eggs of the mollusc
Pomacea canaliculata, is a combination of an astaxanthin ester with a glyco-
protein [118].
On exposure to air, particularly in the presence of base, many carotenoids
with the 3-hydroxy-4-keto end group (8) are rapidly converted into the cor-
responding diosphenols with end groups of the type (9). Thus astaxanthin
(203) yields astacene (198), the pigment which gives boiled lobsters and crabs
their attractive colour. The many reports [3, 4] of the natural occurrence of
astacene and other diosphenols must, however, be treated with considerable
reserve. It seems likely that in most, if not all, cases these pigments are artifacts
derived from natural 3-hydroxy-4-keto carotenoids.
In the sea anemone, Actinia equina, the characteristic red pigment is
actinioerythrin [119]. This is a mixture of fatty acid esters (272) of the parent
carotenoid, actinioerythrol, with 2-nor end groups (10) [120, 121]. It has been
40 B. C. L. WEEDON

suggested that the end groups (11) of actinioerythrin arise from those in
astaxanthin by transformations of the type shown in Scheme 7.

m~
~/-~/-.:.~/
~ ~ . .
0 0

HO~/~ RCO,~/
0 0
(10) (II)
Scheme7

Actinioerythrol itself has not yet been reported in nature. Neither has the
beautiful blue pigment, violerythrin, which is formed from it, or from actinio-
erythrin, on treatment with alkali in the presence of oxygen [120-122].
Violerythrin has the cyclopentenedione end groups (12), and its formation from
actinioerythrol represents an autoxidation formally analogous to that involved
in the conversion of astaxanthin (203) into astacene (198) [120, 121, 123].

HO~/
0

(10) (12)

4. Unsubstituted P-ring
The y-carotene derivatives produced by Mycobacterium phlei include the
tertiary hydroxy compound (48) [93] and the tertiary o-glucoside phlei-
xanthophyll (77) [96]. The corresponding glycol, plectaniaxanthin (76), occurs,
together with its mono- and di-ester and the linoleate of dehydroplectania-
xanthin (162), in the fungus Plectania coccinea [124, 125]. Another fungal
pigment, an ester of aleuriaxanthin (49) present in Aleuria aurantia, seems to
represent a further example of a xanthophyll which retains an unsubstituted
P-ring [34].
In addition to the glucoside (159) mentioned earlier, some genera of
Myxobacterales also contain the related compound (47) with a dehydro-P-end
group [95].

F. Aromatic Carotenoids

Though alicyclic carotenoids have long been known, the discovery of their
aromatic counterparts is comparatively recent. Several aromatic compounds
 II. Occurrence 41

are now recognized and are characterized by the presence of the 1,2,5- or
1,2,3-trimethylbenzene end groups (13) or (14) respectively. It has been sug-
gested that these end groups arise in nature from aromatization of the common
alicyclic structures [38, 49, 126].

~/ :()A/
(13) (14)

Aromatic carotenoids were first encountered in the Japanese sea sponge,

Reniera japonica, which contains renieratene (14), isorenieratene (13) and
renierapurpurin (16) [127-129]. Subsequently aromatic carotenoids were
noted in various bacteria. Isorenieratene is now known to be identical with
leprotene [130-132], a pigment isolated many years ago from Mycobacterium
phlei [133, 134]. It is found as well in other Mycobacterium species and in
Phaeobium species [135]. P-Isorenieratene (6) also occurs in Phaeobium
species [136].
The Chlorobia green photosynthetic bacteria contain both chlorobactene
(15), the aryl analogue of y-carotene, and its hydroxydihydro derivative
(53) [137]. The corresponding glucoside (54) is reported to occur in Coryne-
bacterium fascians [138].
The aromatic ketone okenone (181) is the characteristic carotenoid in the
purple photosynthetic bacterium Chromatium okenii [139]. The minor pigments
of 1hiothece gelatinosa include the demethylated okenone (164), and a sub-
stance tentatively formulated as the isomer (180) of okenone with the 1,2,5-
trimethylbenzene end group (13) [139a].
The first phenolic carotenoids, (52) and (79), were recently reported as
occurring in Streptomyces mediolani [140].

G. Epoxides (5,6 and 5,8)

Another major class of pigments in algae and higher plants comp;ises the
5,6-epoxides (15) of zeaxanthin (67) and of the related carotenoids in which one
end group is identical with those of zeaxanthin [3-6, 141]. The symmetrical
diepoxide, violaxanthin (135), is found, together with the isomeric allenic mono-
epoxide neoxanthin (122), in all green leaves. Antheraxanthin (119), the mono-
epoxide of zeaxanthin, occurs in many plants, as does lutein epoxide (120).
However, despite the widespread occurrence of lutein (73), no derivative is
known in which the unconjugated double bond has undergone epoxidation.
Recently it has been suggested that the xanthophyll taraxanthin which was
originally isolated from the dandelion, Taraxacum officinale [142], is identical
with lutein epoxide [143].
42 B. C. L. WEEDON

The 5,6-epoxides (15) rapidly isomerize, on treatment with traces of acid,

into the 5,8-epoxides (16), which are often referred to as furanoid oxides [141].
Many of these 5,8-epoxides have also been isolated from plants, but it is by no
means always certain that they are true natural pigments and not artifacts
formed from the 5,6-epoxides during isolation.

R=OH (15) R=OH (16)

R=H (17) R=H (18)

The 5,6- and 5,8-epoxides, (17) and (18) respectively, derived from a- and
/3-carotene, have also been reported in plants, but are far less common than
the 3-hydroxy compounds [3-6]. Again no derivative is known in which the
isolated double bond of a-carotene has undergone epoxidation.

H. Cyclopentyl Ketones

During the ripening of red peppers, Capsicum annuum, several carotenoids

are formed which are characterized by the presence of an acylcyclopentanol
ring (19) [38, 144-146].
The most abundant of these unusual pigments is capsanthin (170), but its
5,6-epoxide (188), the symmetrical diketone capsorubin (205), and cryptocapsin
(157) are also present.
These compounds appear to be formed from zeaxanthin (67) and crypto-
xanthin (39), via their 5,6-epoxides antheraxanthin (119), violaxanthin (135) and
cryptoxanthin epoxide (117). The key transformation to give the ketones
presumably involves a pinacolic rearrangement of the epoxides, or of a closely
related derivative (Scheme 8) [38, 147-149].

OH
(4) (15) (19)
Scheme 8

In addition to capsanthin, its epoxide and capsorubin, the pollen anthers of

Aesculus hippocastanum are reported to contain the cyclopentanone capsan-
thone (197) [150].
 II. Occurrence 43

I. Allenic Carotenoids

The solution of the long outstanding problem of the structure of the algal
pigment fucoxanthin (190) revealed the first example of an important class of
carotenoids which possess an allenic end group (20) [151-154]. The origin of
this group is not yet known. It is conceivable that it is derived from a related
acetylene (21) [153], but there is no direct evidence to support this view
[155, 156]. It seems more likely that the allenic structure is· formed from a
zeaxanthin end group (4), but whether this involves the intermediate formation
of the 5,6-epoxide (15) [146, 153] or some direct oxidation [156, 157] is again
a matter for speculation.
A_. . .
N~

R=H or Ac ROM~H (20)

The best known sources of fucoxanthin (190) are the brown algae (Phaeo-
phyceae), which include such common marine seaweeds as Fucus vesiculosus
(bladderwrack), F. serratus and Ascophyllum nodosum. However, it is also
present in Chrysophyceae and in Bacillariophyceae (diatoms) [5, 6]. Thus
fucoxanthin can be regarded as the characteristic pigment of all Phaeophyta
except for the Heterokontae. Although the latter do not produce fucoxanthin
they contain several xanthophylls which are regarded as being related to
fucoxanthin. This view has recently received support from studies on vau-
cheriaxanthin, which occurs as a partial ester in species of Vaucheria and
Botrydia and for which structure (124) has been proposed [158].
Two derivatives offucoxanthin, fucoxanthinol (189) and paracentrone (246),
have been isolated from the coelomic epithelium of the sea urchin, Paracentrotus
lividus [159]. The presence of traces of fucoxanthin as well suggests that these
two novel natural allenes are formed in the animals by metabolism of dietary
fucoxanthin [159, 160]. Pentaxanthin, a pigment reported some years ago
in the same organism, may well have been isofucoxanthinol (178) [161].
It is interesting to note that the yellow colour of the yolks of chicken eggs
can be improved by feeding hens with seaweed meal [162]. The pigment
deposited in the yolks, and responsible for the enhanced colour, is probably
identical with isofucoxanthin (179) formed by isomerization of fucoxanthin in
the diet [153, 154]. This isomerization occurs readily on treatment of fuco-
xanthin with weak bases [152-154].
Another important example of an allenic carotenoid is provided by neo-
xanthin (122) [155, 163, 164]. This pigment was first isolated from barley
leaves [165] but is now recognized as a constituent ofEuglenaphyta [166] and
some other algae [5, 6], and as one of the principal xanthophylls in a wide
variety of seed plants (Spermatophyta) and spore-bearing plants (Pterido-
phyta and Bryophyta) [5, 167]. In addition to the 5,6-epoxide, neoxanthin,
44 B. C. L. WEEDON

some plants are said to contain the 5,8-epoxide, neochrome (131) [144]. Trolli-
xanthin, a carotenoid present in the globe flower Trollius europaeus [168], is
now thought to be identical with neoxanthin [153, 156, 156a], whilst a pigment
in the monkey flower M imulus guttatus is believed to be the parent deepoxy-
neoxanthin (86) [169]. The latter may also be identical with trollein [153, 156,
169], which was first observed in orange juice [170], and with a similar
compound from the alga Euglena gracilis [169]. Recently peridinin (sulcato-
xanthin) has been formulated as (273) (see p. 53).

J. Acetylenic Carotenoids

The acetylenic carotenoids were discovered as the result of studies on

pigments from flagellates of the algal class Cryptophyceae. Alloxanthin, the
principal xanthophyll from Cryptomonas ovata var. palustris, Rhodomonas
Strain D3 and Hemiselmis virescens (Droop's Strain), was shown to be the
diacetylenic analogue (65) of zeaxanthin (67) with end groups of the type (21).
Two of the minor xanthophylls, monadoxanthin and crocoxanthin, were
identified as the acetylenic analogues (72) and (41) of lutein (73) and zeinoxan-
thin (42) respectively [171, 172].
Diatoxanthin, a common pigment in diatoms [173] and Chrysophyceae
[174-176], is the monoacetylenic analogue (66) of zeaxanthin [172], whilst
diadinoxanthin from diatoms, such as Nitzschia closterium f. minutissima, and
dinoflagellates [173, 177, 178] is the related 5,6-epoxide (118) [179]. It is
interesting to note that the acetylenes diatoxanthin (66) and diadinoxanthin
(118) often occur together with the allene fucoxanthin (190) in diatoms and
Chrysophyceae. In the Euglenaphyta, such as Euglena gracilis, the principal
xanthophyll is not antheraxanthin (119) (as believed until recently [180]) but
the acetylene diadinoxanthin (118) [179], which occurs in these algae together
with the allene neoxanthin (122) [164, 166] and a related compound thought
to be deepoxyneoxanthin (86) [169].
The acetylene heteroxanthin (94) is closely related structurally to dia-
dinoxanthin (118) [181]. It occurs with the latter and with diatoxanthin (66) in
all Heterokontae so far examined [182-185], and is often accompanied by a
partial ester ofvaucheriaxanthin [181], which has now been formulated as the
allene (124) [158, 182]. Heteroxanthin has not been observed with diadino-
xanthin and diatoxanthin in other organisms such as diatoms [181].
Following the elucidation of the structure of the algal pigment alloxanthin
(65) it was shown that two previously reported carotenoids from marine
animals, pectenoxanthin from the giant scallop Pecten maximus [186-189],
and cynthiaxanthin from the tunicate Halocynthia papillosa [186-190], have
the same diacetylenic structure (65) [191]. Alloxanthin has also been isolated
from the common mussel, Mytilus edulis [191], and there seems little doubt
that the pigment in the californian mussel, M. californianus, which was reported
to be zeaxanthin [192], is also alloxanthin [191].
 II. Occurrence 45

Pectenoxanthin has also been reported in Pectenjacobaeus [193], P. yesso-

ensis [189], the tunicate Botryllus schlosseri [186], and from the red-orange
foot of Andara broughtoni and A. subrenata [189]. A xanthophyll in the mollusc
Aplysia kurodai may again be identical with pectenoxanthin (alloxanthin) [194].
Cynthiaxanthin has also been isolated from Halocynthia roretzi [188].
In addition to pectenoxanthin (alloxanthin), the gonads of the giant scallop
also contain astaxanthin (203) and pectenolone (166), which has one end group
identical with those in alloxanthin (65) and the other with those in astaxanthin
(203) [191]. The same acetylenic ketone (166) has also been detected in the
tunicate Halocynthia papillosa [191], and it is conceivable that glycymerin
from the mollusc Pectunculus glycymeris [187], and hydroxyasteroidenone
from star fish [98], may be related to, or even identical with, pectenolone
(166) [191]. Mytiloxanthin, a pigment in mussels, is also believed to have an
acetylenic end group of the type (21) [156, 191 a].
Further examples of acetylenic keto carotenoids are provided by a recent
re-examination of asterinic acid from the star fish, Asterias rubens. This
xanthophyll, formerly thought to be identical with astaxanthin (203) [195], is
now known to be a mixture of its mono- and di-acetylenic analogues (202) and
(200) which have one and two end groups respectively of the type (22) [49, 196].
It is noteworthy that asterinic acid apparently occurs as a protein complex in
the star fish. As mentioned earlier, the prosthetic group in carotenoproteins is
normally astaxanthin.

A ...
~~
HOY 0
(21) (22)

The acetylenic algal pigments all contain the end group (21). Its origin is
not yet known. Direct dehydrogenation of the common zeaxanthin end group
(4) would involve attack at the sterically hindered 7,8-position. Another possi-
bility is that the acetylenic end group (21) results from dehydration of the allene
(20) [156, 157]. The acetylenic carotenoids in the lower marine animals are
probably derived from their diet. A subsequent transformation of diatoxanthin
(66) into pectenolone (166) and (202), and of alloxan thin (65) into (200), can then
be envisaged by analogy with the postulated biosynthesis of astaxanthin (203).

K. Methyl-oxidized Carotenoids

From the previous sections it will be seen that xanthophylls which can be
regarded as resulting from substitution of a methylene group at C-3 or C-4 are
extremely common. In recent years it has become increasingly apparent that
46 B. C. L. WEEDON

a variety of organisms seem capable of introducing an oxygen function by

substitution of one of the methyl groups in the typical carotenoid skeleton.
This was first realized following the revision of the C 37 -structure proposed for
torularhodin [197], the pigment of the red yeast Torula rubra [198], to the
C40 -formulation (211) [199]. Later the corresponding methyl ester (212) [199a],
aldehyde (142) and alcohol (50) [33] were also reported [200].
Another important revision came when lycoxanthin and lycophyll from
the berries of the woody nightshade, Solanum dulcamara, which had been
formulated as the 3-hydroxy and 3,3'-dihydroxy derivatives of lycopene (19)
[201], were shown to be the primary alcohols (62) and (83) respectively
[202, 203].
Loroxanthin, a pigment from the green algae Scenedesmus obliquus and
Chlorella vulgaris, has recently been identified as a derivative (88) of lutein (73)
in which the C-19 methyl group has been converted into hydroxymethyl [204,
205]. Pyrenoxanthin from C. pyrenoidosa, and provisionally formulated as
(89) [206], is apparently the C-20 analogue of loroxanthin (88). Siphonaxanthin
and siphonein, the two characteristic pigments of the algal class Siphonales,
are now known to be the 19-hydroxy compound (174) and its laurate ester
(175) respectively [205, 207]; the 7-methylene-8-keto structure is a feature
which has previously only been seen in fucoxanthin (190) and some of its
derivatives. Another 19-hydroxy carotenoid is the allenic pigment vaucheria-
xanthin (124) [158], to which reference has already been made.
The photosynthetic purple bacterium Chromatium warmingii contains
several unique carotenoids [208]. The principal pigments, rhodopinal (warm-
ingone) (144) and rhodopinol (warmingol) (80), are now recognized as deriva-
tives of rhodopin (56) substituted at the C-20 methyl [ 49, 209]. Several related
xanthophylls have also been identified in the same bacterium [ 49].

L. Higher Carotenoids (C 45 and C 50)

Until recently no natural carotenoid was known in which the carbon

skeleton contained more than forty carbon atoms. However, studies over the
last few years on the pigments of various non-photosynthetic bacteria have
revealed the occurrence of a number of C 45 - and C 50 -carotenoids.
The first ofthese higher carotenoids to be discovered was decaprenoxanthin
(226), the principal pigment of the gram-positive bacterium Flavobacterium
dehydrogenans [210, 211]. Subsequently the minor pigments in this organism
were shown to include the related monohydroxy C 50 -carotenoid (223) and two
C 45 -carotenoids, nonaprenoxanthin (218) and its 11',12'-didehydro derivative
(217) [212]. Corynexanthin from Corynebacterium species has been identified
as the glucoside (227) of decaprenoxanthin (226) [213].
Sarcinaxanthin from Sarcina lutea, at one time regarded as identical with
decaprenoxanthin [214], is now provisionally formulated as the isomer (224),
and sarcinene as the parent hydrocarbon (222) [ 49].
 II. Occurrence 47

ln Corynebacterium poinsettiae a pigment formerly believed to be spirilla-

xanthin (108) [215, 216] is now regarded as the C 50-diol (228), another
formerly thought to be cryptoxanthin (39) [217] is now represented as the
isomer (220) of decaprenoxanthin, and a third which was at one time thought
to be lycoxanthin (62) [215] is now formulated as (221) [ 49, 217]. In addition
to these C 50 -pigments, C. poinsettiae also contains the C 45 -xanthophyll (219)
[49, 217].
The Halobacteria provide yet another source of C 50 -carotenoids. Bacterio-
ruberin, the principal pigment in many species, though at one time thought to
be the glycol corresponding to spirilloxanthin (108) [218], is now believed to
be (231), or a closely related compound [ 49, 219]. Two minor carotenoids from
H. salinarium are regarded as the mono- and di-anhydro derivatives (230) and
(228) [49].
The biogenesis of these higher carotenoids seems to involve reactions of
the types shown in Schemes 9 and 10 where RED formally represents a dimethyl-
allyl carbonium ion, or a related species [49, 211]. Such a C 5 -unit could
conceivably arise from dimethylallyl pyrophosphate, one of the key inter-
mediates in terpene biosynthesis. The close similarity between Schemes 9 and 6,
and between 10 and 2 are striking. Whereas 6 and 2 involve reactions initiated
by a proton, 9 and 10 indicate analogous reactions which are represented as
being initiated by a C 5 -carbonium ion. The formation of the 2-substituted
cyclohexene ring (Scheme 9) is often accompanied by oxidation of a terminal
methyl group (cf. Section K). The production of the acyclic 2-substituted end
group (Scheme 10) is apparently associated with dehydrogenation and hydra-
tion reactions analogous to those encountered in the C 40-series (cf. Schemes 1,
2 and 4). Whether geranyl and farnesyl pyrophosphates can participate in
reactions of the type envisaged above for dimethylallyl pyrophosphate,
remains to be seen. If so, then we can look forward to the discovery of C 60 -
and C 70 -carotenoids.

-~ HO">l_
H 20 / - , ....- ____.. _.., 'l_...
R00 R
Scheme 10
48 B. C. L. WEEDON

M. Degraded Carotenoids

In the xanthophylls discussed earlier, the oxygen functions seem to have

been introduced by reactions of two main types: (i) substitution of a methyl or
methylene group, and (ii) addition to a carbon-carbon double bond. A further
class of xanthophylls must now be considered, namely those which appear to
be formed by oxidative fission of a carbon-carbon double bond. When this
involves opening of a cyclic double bond, with retention of the C40 -carbon
skeleton, the resulting product is termed a seco-carotenoid. When, as is more
often the case, fission involves rupture of the polyene chain, and loss of one or
both end groups, the product is designated an apo-carotenoid.
On the basis of widely accepted nomenclature rules, a degradation product
which does not retain the C-20 and C-20' methyl groups of the original C40 -
structure is no longer a carotenoid [220]. However, this arbitrary distinction
should not be allowed to mask the important relationship which is known to
exist between carotenoids and many other terpenes, or which seems plausible
from structural considerations. A brief account of some related terpenes is
therefore included in this section.

1. Seco-carotenoids
Both semi-j3-carotenone (213) and P-carotenone (216), which can be
regarded as oxjdation products of P-carotene, have been reported in the citrus
relatives Murraya exotica and 1riphasia trifolia [221]. Semi-a-carotenone
(214) has also been detected in small amounts in M. exotica [222], and tri-
phasiaxanthin from T. trifolia is said to be the related derivative (215) of
cryptoxanthin (39) [223].

2. Apo-carotenoids
Both apo-6'-lycopenal (242) and apo-8'-lycopenal (254) have been detected
in tomatoes [33], and there seems little doubt that they represent breakdown
products of the main pigment, lycopene (19). The major carotenoid present in
the fruits of Shepherdia canadensis, formerly claimed to be the acetate of !yeo-
xanthin [224], has now been shown to be methyl apo-6'-lycopenoate (243)
[225]. The apo-8' glycoside (255) has been isolated from a yellow halophilic
bacterium [226].
The P-apo-10'-carotenal (256) and P-apo-8'-carotenal (248) are minor, but
rather widely distributed, pigments [33, 227]. Like the apo-2' analogue (233)
in citrus fruit [33], they can be regarded as oxidation products of P-carotene.
P-Citraurin (249) is a constituent of orange peel, Citrus aurantium [228],
and is probably derived from zeaxanthin (67). A pigment tentatively formulated
as the apo-10' analogue (257) has been reported in citrus hybrids [229].
Neurosporaxanthin, the P-apo-4'-carotenoic acid (234), is found in the
fungi Neurospora crassa and N. sitophila [230]. Laetiporxanthin, from Laeti-
porus sulphureus, though not yet identified, may well prove to be a structurally
related compound [231].
 II. Occurrence 49

The apo-10' pigment azafrin (261) has long been known as the major
carotenoid in the roots of Escobedia scabrifolia [3, 232, 233] and is associated
with small amounts of the corresponding aldehyde (258) [33]. Like hetero-
xanthin (94), these two compounds contain a ring with an IX-glycol grouping.
In addition to the apo-aldehydes, -acids and -esters, several methyl ketones
are now recognized [221]. Citranaxanthin (237) [234], sintaxanthin (244) [235]
and its 3-hydroxy derivative, reticulataxanthin (238) [236], have all been
isolated from the trigeneric hybrid Sinton citrangequat. Reticulataxanthin has
also been obtained from another citrus relative, Minneola tanger [221]. The
occurrence in Sinton citrangequat of minor pigments believed to be the P-hy-
droxy ketones (239) [237] and (240) [221] suggests that citranaxanthin .and
reticulataxanthin may arise from an aldol-type condensation ofthe appropriate
apo-8' -carotenals with the elements of acetone. Whether sintaxanthin (244) is
formed in a similar fashion, or whether it results from a retro-aldol fission of the
type postulated [160] to explain the formation of paracentrone (246) from
fucoxanthin (190), are matters for conjecture. Tangeraxanthin from tangerines
is tentatively formulated as the methyl ketone (236) with a retro chromophore
[238].
Apparently oxidative degradation can also occur at both ends of the normal
C40 -carbon skeleton. Thus bixin (265), the well known pigment in annatto
which is prepared from the seeds of Bixa orellana [3], can be regarded as a
diapo-carotenoid. A further example is provided by the digentiobioside crocin
(271), which occurs together with small amounts ofthe aglycone, crocetin (269),
in saffron, Crocus sativus [3]. The corresponding dialdehyde (267), and the
half-aldehyde (268), have been isolated from the leaves of Jacquinia angusti-
folia [239].

3. Related terpenes
Under this heading reference must first be made to vitamin A (23), its
aldehyde, retinal (24), and other derivatives [240]. These biologically important
compounds are clearly degradation products of P-carotene (3) and other

(23)

carotenoids which possess one half of the P-carotene structure (Chapter X).
Kitol (25), which occurs in whale liver oil [241], is a naturally occurring dimer
of vitamin A [242, 243]. It is thus a C 40-derivative of P-carotene but is not,
according to the conventional definition, a carotenoid. The trisporic acids, e. g.
Carotenoids 4
50 B.C.L. WEEDON

trisporic acid C (26) [244], which are the principal sexual hormones in the
fungal order Mucorales, are now known to be formed from P-carotene by
way of retinal (24) [245].

OH

(24) (26)

Whether the plant growth regulator abscisic acid (27) is a true sesquiterpene
derived from farnesyl pyrophosphate without further elaboration ofthe carbon
skeleton, or whether it represents a carotenoid degradation product, is a
question that has not yet been fully answered [246]. It may be significant that
both violaxanthin (135) and neoxanthin (122), both widely distributed in
2
HOC~
""::: "":::
OH
0 ~ ~ 2"
(28)

plants, are converted into a product with growth inhibitory properties on

exposure to light [247, 248, 248a]. Natural derivatives of abscisic acid include
abscisyl P-o-glucopyranoside, present in lupins, Phaseus luteus [249], meta-
bolite C (28) [250], and phaseic acid (29) which occurs in the green fruit of
Phaseolus multiflorus [251]. Vomifoliol (30), present in the leaves of Rauwolfia
vomitoria [252], may constitute another member of the series.

~rn~o
(31)

Turning to other compounds where the relationship with carotenoids is

speculative, mention must be made of luciferin (31), the substrate of the bio-
luminescence enzyme system of the freshwater limpet Latia neritoides [253].
The allenic ketone (32) has been identified as a major constituent in an
ant-repellant secretion of the large flightless grasshopper, Romalea microptera
[254]. It seems likely that this ketone is formed by oxidation of the neoxanthin
(122) present in the leaf diet of the insects [255]. It has been suggested that
such compounds as (33) and (34), present in the urine of pregnant mares, also
represent products of carotenoid metabolism [256-258].

N.,.Ao ~ ~
H~H H~ ~ ~
(32) (33) (34)
 II. Occurrence 51

a-Ionone (35) and P-ionone (36) have been encountered in various higher
plants [259, 260], and P-ionone is also present in the alga Trentepohlia iolithus
[260a]. Again it is conceivable that the ionones are derived from a- and P-
carotene [261]. However it must be borne in mind that irone, from the essential
oil and resinoid of iris, is a mixture of methylionones, and that the corre-
sponding 2-methyl substituted carotenoids have not as yet been discovered
in nature [260]. Damascenone (37), from Bulgarian rose oil, Rosa damascena
[262], and 3,4-didehydroionene (38), from, strawberry oil [263], have carbon
skeletons reminiscent of the ionones.

~0 ~0
(35) (36)

m
0

~ (37) (38)

P-Ionone 5,6-epoxide (39) has been identified in tomatoes [260] and in the
flavour of black tea [265]. The latter also contains theaspirone (40) [264, 265]
and dihydroactinidiolide (41) [265-267]. Dihydroactinidiolide [267] is present
in other plants, chiefly cassie [268] and tobacco [269]. It also occurs, together
with actinidiolide (42) and actinidiol (43), in the essential oil of Actinidia poly-
gama [270]. The related loliolide (44) [271-274], also known as digiprolactone,
has been identified in Lolium perenne [268, 271, 275], Fumaria o.fficinalis [276],
Digitalis lanata and D. purpurea [260, 272]. Picrocrocin (45, R = glucosyl) is

~ oi:£A q:ro
(39) (40) (41)

q:ro ~OH ]J:ro 0

HO

ex:
(42) (43) (44)

DCHO

RO
(45) R=H (46)
R=OH (47)
52 B. C. L. WEEDON

the bitter principal of saffron (from Crocus sativus), and the aglycone (45, R =H)
has been reported in unicellular green algae [277]. Finally the trimethylcyclo-
hexanone (46) has been detected in tomatoes [278], and, together with the
hydroxy ketone (47), in black tea [265]. Again further work may show that
some, at least, of these various products are of carotenoid origin.

N. General Comments

Consideration of the main classes of carotenoids, and of the organisms in

which they occur, allows a few generalizations to be made.
The ability to introduce oxygen functions at C-1 or C-2, or at both these
positions, in an acyclic end group of a carotenoid, is a feature of many bacteria,
blue-green algae and Sarcoscyphaceae fungi. None of these substituted acyclic
end groups has been observed as yet in the carotenoids of other classes of algae,
or of the higher plants, and only the photosynthetic purple bacteria seem able
to synthesize the methoxy carotenoids.
Algae, like some bacteria and fungi, produce carotenoids with alicyclic
end groups containing six-membered rings. Such structures are also very
common among the carotenoids of the higher plants. The 5,6-epoxides of these
cyclic end groups, particularly those with a hydroxy substituent at C-3, are also
typical of many algae and plants. However, the alicyclic end group with a
carbonyl function at C-4, though common in some algae, bacteria and animals,
is apparently unknown in higher plants.
The ability of some bacteria to aromatize the cyclic end groups is shared
by at least one sponge (Renierajaponica) and by Streptomyces mediolani. The
production of the C 45 - and C 50-carotenoids with a C 5 -substituent at C-2
and/or C-2' seems to be limited to certain non-photosynthetic bacteria.
Both allenes and acetylenes are produced by algae. All green plants
synthesize the allene neoxanthin, but no example is known of a C 40 -acetylenic
carotenoid from one of the higher plants.
The carotenoids in algae and bacteria provide a wider variety of structural
types than those found in the higher plants. The end groups in the plant
carotenoids are to be seen in the pigments of the simpler organisms-with one
exception. The five-membered ring in capsanthin (170), and related compounds,
seems to represent a novel end group that originated in the higher plants.
The hydroxy carotenoids often occur in nature in the form of derivatives.
Thus xanthophylls in fruit are present mainly as their fatty acid esters, in
contrast to the xanthophylls in leaves [5]. In fungi even the tertiary alcohols
phillipsiaxanthin (207) [ 47], plectaniaxanthin (76) and related pigments [34,
124, 125] occur mainly as fatty acid esters. Recently two pigments, (159) and
(47), in some genera of Myxobacterales have been identified as esters of the
glucosides derived from hydroxy carotenoids [95].
For many years crocin (271), the digentiobiosyl ester of crocetin (269) found
in saffron, provided the sole example of a carotenoid glycoside. Now the
 II. Occurrence 53

bacterial pigments phleixanthophyll (77) and 4-ketophleixanthophyll (172) are

recognized as tertiary n-glucosides [96], corynexanthin (227) provides an
example of a primary glucoside [213], and a pigment (255) from a halophilic
bacterium appears to be a tertiary mannoside [49, 226]. In addition two
characteristic pigments of blue-green algae, myxoxanthophyll (90) and oscilla-
xanthin (95) are now regarded as rhamnosides [ 48, 49], and the corresponding
0-methyl-5-C-methylpentosides have also been observed [49a].
Attention has already been drawn to the carotenoproteins in which there is
a stoichiometric combination between a protein and astaxanthin (203), or
occasionally some related carotenoid. Other carotenoids such as P-carotene (3)
and lutein (73) frequently occur in association with protein without, apparently,
specific interaction [110]. In plants it is assumed that carotenoids are often
located in the grana of the chloroplasts in the form of chromoproteins [5].
However, no reliable information is yet available concerning these complexes.
The widespread occurrence of carotenoids in nature suggests that these
pigments may be of considerable value in chemotaxonomy. A pattern of
evolution of algae, based on a consideration of the distribution of carotenoids
among the various classes, has already been proposed [5], and a similar
approach might be feasible with certain classes of bacteria and fungi. In
adopting carotenoids as a taxon it must, of course, be remembered that there
is a need to confirm many published reports in the light of modern developments
in methods of separation, and in methods of identification. Some characteristic
pigments have no doubt been missed in past surveys, and others wrongly
identified.
Further structural studies on the constituent carotenoids may also be
needed before the relationships between different classes of organisms will
become apparent. Thus elucidation of the structure of peridinin, the charac-
teristic pigment in the Pyrrophyta [3, 279, 280], and of the typical pigments
in the Prasinophyceae [281], might assist studies on the relationship of these
to other types of algae. Similarly the identification of sulcatoxanthin, from the
sea anemone, Anemonia sulcata [282], and of the carotenoids of many other
marine animals, is desirable. These problems are all known to be receiving
attention, but the answers are not available at the time of writing.
Added in proof: The allenic structure (273) has recently been assigned to
peridinin and sulcatoxanthin [283] (see also Chapter III, Section D.4 b).

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54 B. C. L. WEEDON

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[269] W. C. Bailey, Jr., A. K. Bose, R. M. Ikeda, R. H. Newman, H. V. Secor and C. Varsel, J. Org.
Chern. 33, 2819 (1968).
[270] T. Sakan, S. Isoe and S.B. Hyeon, Tetrahedron Letters 1967, 1623.
[271] R. Hodges and A. L. Porte, Tetrahedron 20, 1463 (1964).
[272] D.Satoh, H. Ishii, Y.Oyama, T.Wada and T.Okumura, Pharm. Bull. 4, 284 (1956).
[273] T. Wada and D. Satoh, Chern. Pharm. Bull. 12, 752 (1964).
[274] T. Wada, Chern. Pharm. Bull. 12, 1117 (1964); 13, 43 (1965).
[275] E.P. White, New Zealand J. Agr. Res. 1, 859 (1958).
[276] R.H.F. Manske, Can. J. Res., Sect. B 16,438 (1938).
[277] R. Kuhn and I. Low, Ber. Deut. Chern. Ges. 74, 219 (1941).
[278] Z. Horii, T. Yagami, M. Ito and M. Hanaoka, Chern. Pharm. Bull. 16, 848 (1968).
[279] A. R. Loeblich and V. E. Smith, Lipids 3, 5 (1968).
[280] S. W. Jeffrey and F. T. Haxo, Bioi. Bull. 135, 149 (1968).
[281] T.R. Ricketts, Phytochem. 5, 571 (1966); 6, 1375 (1967).
[282] I. M. Heilbron, H. Jackson and R.N. Jones, Biochem. J. 29, 1384 (1935).
[283] H. H. Strain, W.A. Svec, K. Aitzetmiiller, M.C. Grandolfo, J.J. Katz, H. Kj~sen, S. Norgard, S.
Liaaen-Jensen, F. T. Haxo, P. Wegfahrt and H. Rapoport, J. Amer. Chern. Soc. 93, 1823 (1971).
 61

III. Isolation, Reactions

SYNN0VE LIAAEN-JENSEN

Organic Chemistry Laboratories, Norwegian Institute of Technology,

University of Trondheim, Trondheim, Norway

A. Isolation 63
1. Introduction . 63
2. Extraction . . 63
3. Saponification 65
4. Partition, countercurrent distribution 65
5. Chromatography . . . . . . 66
a) Column chromatography . 66
b) Thin layer chromatography 67
c) Paper chromatography 68
d) Gas chromatography . 69
6. Crystallization . . 69
7. Characterization . 69
a) Purity criteria . 69
b) RF values . . . 70
c) Partition coefficients 71
d) Spectroscopic properties 72
e) Preparation of derivatives . 72
8. Protein complexes 72
B. Group Reactions . . . . . . . 73
1. Introduction . . . . . . . . 73
2. Carbon-carbon multiple bonds 74
a) Addition reactions . . 74
b) Oxidative degradation 74
c) Rearrangements . 75
3. Hydroxy groups . . 76
a) Grignard reaction 76
b) Esterification 77
c) Silylation . . 77
d) Methylation . 77
e) Oxidation. . 78
f) Dehydration 79
g) Character and vicinity 83
4. Carotenol esters . 84
a) Hydrolysis . . 84
b) Hydrogenolysis 85
5. Carbonyl functions 85
a) Reduction . 85
b) Oxidation . . 86
c) Condensation 87
d) Addition . . 87
e) Alkali reactions 87
6. Alkoxy groups . 88
7. Epoxy groups . . 88
a) Colour tests . . 89
b) Epoxide-furanoid rearrangement . 89
62 SYNN0VE LIAAEN-JENSEN

c) Hydrogenolytic cleavage 89
d) De-epoxidation 91
8. Glycosides 91
a) Hydrolysis . . 92
b) Hydrogenolysis 92
c) Elimination . . 92
C. Other Partial Syntheses 93
1. Introduction . . . . 93
2. Dehydrogenation . . 93
3. Introduction of oxygen functions 95
a) Epoxidation. . . . . . . . 95
b) Allylic hydroxy, keto, alkoxy and acyloxy groups . 98
c) Diosphenols . . . . . . . . . . . . . 101
4. Ring contraction . . . . . . . . . . . . . 101
D. Reactions and Structures of Natural Carotenoids 102
1. Introduction . . . . . . . . . . 102
2. Carotenes (C40 ) . • • • • • • • 102
a) Aliphatic and alicyclic carotenes 102
b) Aromatic carotenes . 104
3. C40 -xanthophylls . . . . . . . 105
a) Acetylenes and allenes . . . 106
b) Epoxides and furanoid oxides 113
c) Glycosides . . . . . . . . 116
d) Methyl ethers . . . . . . . 121
e) Carboxylic acids and methyl esters 128
f) Aldehydes . . . . . . . 128
g) Ketones . . . . . . . . . . . 129
h) Alcohols and esters thereof . . . 146
4. Apo-carotenoids and nor-carotenoids with Jess than forty carbon atoms in the skeleton 160
a) Introduction . . . . 160
b) Acetylenes and allenes 161
c) Epoxides . . . . . . 163
d) Glycosides . . . . . 164
e) Carboxylic acids and esters 165
f) Aldehydes . . . . . . . 168
g) Ketones . . . . . . . . 169
5. Carotenoids with more than forty carbon atoms in the skeleton . 173
a) Introduction . 173
b) C45 -carotenoids 173
c) C50 -carotenoids 174
References . . . . . . 177
 III. Isolation, Reactions 63

A. Isolation

1. Introduction
The choice of the method of isolation of carotenoids from natural sources
is mainly determined by the nature of the biological material, the ease of
solvent extraction and the properties and relative quantities of the carotenoids
present. In practice the particular experience of the research worker may
often be the deciding factor in devising the isolation procedure.
For more detailed instruction the reader is referred to publications [1-6].
In this section a general discussion of the various modifications to the general
isolation procedure, see Scheme 1, is given. While isolation invariably involves
solvent extraction and chromatography prior to crystallization, saponification
and partition offer advantages and disadvantages to be considered in each
particular case.
The isolation should be carried out in an inert atmosphere (generally
nitrogen or vacuum), at a low temperature (room temperature to -20° C)
in darkness or diffuse light under acid-free conditions and using pure peroxide-
free solvents [6].

2. Extraction
Details of the extraction of carotenoids from various sources have been
published [3, 4, 7].
To avoid pigment decomposition air-drying of the biological material
should be avoided, but lyophilization or, less satisfactorily, dehydration by
treatment with aqueous methanol [8] are recommended (cf. step a, Scheme 1).
The latter process may, however, result in some undesirable pigment ex-
traction.
Dry material may be extracted with water-immiscible solvents (step b).
Extraction is, however, more easily carried out with a moist sample, and for
this water-miscible solvents such as mixtures of acetone and methanol are
most commonly used (step c) [1-6]. Moistening of dry algal meal [9] or
dried bacteria prior to solvent extraction is often necessary to effect complete
extraction (step a').
To avoid oxidation, isomerization and rearrangements of the carotenoids
by acids sometimes liberated during the extraction process, antioxidants such
as quinol [10] and neutralizing agents like calcium carbonate [11], pyridine
or dimethylaniline [12] may be added.
In cases where solvent extraction may be slow and incomplete [13, 14],
efficient mechanical grinding of the material to be extracted can be usefully
supplemented by, for example, lysing of obligately halophilic bacteria with
water [15] or enzymatic treatment of various bacteria with lysozyme to
break the cell wall [16] before solvent extraction.
Removal of water from the crude extract (step d) is generally carried out
by addition of brine and transfer to an appropriate solvent in a separatory
64 SYNN0VE LIAAEN-JENSEN

(a)

(a')

Water-miscible Water-miscible
solvent or -immiscible
(c)
solvent

Moist crude extract

(d) 1

*
(e) Aliquot for quan-
Pet. ether or ether ex rae --+ tttat1ve spectro-
metric determination

PARTITIOSk
Pet. ether - (g) ONIFICATION
85 %methanol
(e)

Hypophasic Epiphasic Neutral Acidic

carotenoids carotenoids carotenoids carotenoids

1 1 1 1
(g) PARTITION

(e)
Epiphasic Hypophasic
carotenoids carotenoids

1 1
--------------------------------------CHROMATOGRAPHY

l
Creek of chromatographic homogeneity

(
Rechromatography
l
CrYSTALLIZATION -+ Check of purity criteria

(
Recrystallization
l
Pure Carotenoid -+ Characterization

Scheme 1. General isolation scheme

 III. Isolation, Reactions 65

funnel or, when relatively small amounts of water are present, by azeotropic
vacuum distillation with benzene [6]. The former process is, of course, prefer-
able when water-soluble non-carotenoid pigments are present.
Provided chlorophyll is absent, spectrometric estimation (see Chapter IV)
of the total carotenoid content is conveniently carried out at this stage
(step e).

3. Saponification
Information on the alkali-stability of the carotenoids present in the extract
may be obtained by saponifying a small aliquot and comparing the adsorptive
and spectroscopic (visible light) properties of the carotenoid components
before and after alkali treatment.
Providing only alkali-stable carotenoids are present, saponification
(stepf) is included in the purification procedure with great advantage. Standard
conditions have been defined [ 4, 6].
Saponification is the most effective method of removing unwanted lipids
and chlorophylls, the former often abundant in marine invertebrates and the
latter invariably in photosynthetic organisms. Even if some of the carotenoids
present react with alkali (cf. Section B.3) saponification may with advantage
be included, as long as the structural changes this treatment has effected are
allowed for. Sterols, abundant in yeasts and fungi, are of course found in the
unsaponifiable matter together with the carotenoids and may be removed by
fractional crystallization from petroleum ether [ 4] or acetone at low tempera-
ture, or as digitonides [ 4].
The possibility of separating acidic carotenoids from neutral contaminants,
including neutral carotenoids, during the work-up after saponification greatly
facilitate the purification of acidic carotenoids such as diosphenols and
carboxylic acids. Acidic carotenoids are bound as salts in an alkaline hypo-
phase, permitting neutral components to be transferred to ether, and are
themselves transferred to fresh ether after acidification of the hypophase to
approximately pH 4 [17].
When saponification is omitted in the purification procedure, carotenol
esters must be tested for at a later stage.

4. Partition, countercurrent distribution

When carotenoids of greatly differing polarity are present, grouping into
iso-distributive pigments by solvent partition (step g) prior to chromatography
should be considered. As shown in Scheme 1 solvent partition may be carried
out whether saponification is employed or not. If saponification is omitted
solvent partition offers the further advantage of separating chlorophylls
from the xanthophyll fraction. When the xanthophyll fraction contains alkali-
labile carotenoids (e.g. fucoxanthin, 190) this is an alternative way of separating
chlorophylls and lipids from alkali-labile xanthophylls. The chlorophyll-
carotenemonool fraction may thereafter be subjected to saponification.

Carotenmds 5
66 SYNN0VE LIAAEN-JENSEN

Liquid-liquid partition, first introduced to the carotenoid field by Will-

sHitter and Stoll [8], has been described fully, e.g. [3, 4]. Distribution between
petroleum ether and aqueous methanol containing 85-95 per cent methanol
is generally used. The exact distribution between the two phases of carotenoids
with different polarity is discussed in Section A.6. Carotenes are generally
found in the epiphase, monohydroxy xanthophylls are approximately evenly
distributed in the two phases when 90% methanol is used, and diols and
polyols are mainly hypophasic in the latter system. In its simpler form Craig-
distribution is carried out in a separatory funnel, using three to six transfers.
However, Curl [18] in adapting countercurrent distribution to the separation
of carotenoids has developed several solvent systems [18-21] and success-
fully applied the method to the separation of various fruit carotenoids into
iso-distributive groups [19, 22--34]. Each group, containing carotenoids of
very similar polarity, must be separated into individual components by
subsequent column chromatography. Countercurrent distribution has .so far
not found wide application in the carotenoid field.

5. Chromatography
Chromatographic separation of carotenoids, introduced by Tswett [35]
in 1903, rediscovered by Kuhn's school [36, 37] in the early thirties and
further elaborated by the work of Strain [38], is still the method of choice
for the separation of carotenoid mixtures including cis-trans stereoisomeric
sets. However, reports on successful separation of diastereomeric carotenoids
are few [39-41], although some of the furanoid carotenoids described may be
epimeric.

a) Column chromatography
General treatments are given elsewhere [38, 42-44]. Column chromato-
graphy is indispensible for the separation of carotenoid mixtures on a pre-
parative scale, even if better separation may be accomplished by thin-layer
or paper chromatography. Most procedures are based on the principle of
adsorption chromatography. The adsorbents traditionally used are dealt with
in some detail in various monographs [2, 38] and reviews [3, 4]. Other
adsorbents more recently used are cellulose powder [ 45, 46] and magnesium
silicate for strongly polar carotenoids. Generally, carotenes are best separated
on calcium hydroxide or activated alumina columns, monohydroxy xantho-
phylls on deactivated [ 47] alumina and carotenoids of intermediate polarity
on columns of calcium carbonate or magnesium oxide. Strongly polar
carotenoids are best separated on cellulose or sucrose columns. The references
listed above also give information on the solvent mixtures suitable for develop-
ing the chromatograms in each case. Useful comparative studies of various
chromatographic systems have been reported [48, 49].
The practical use of column chromatography of carotenoids by the
adsorption method has been described in detail by several authors [1, 3, 4, 6, 38].
 III. Isolation, Reactions 67

Principally three basic methods are available: (i) The zone chromatogram
where the coloured zones are cut out and eluted after satisfactory development
of the chromatogram. The method is frequently used with calcium carbonate
columns. (ii) The step-wise elution method with sequential elution of the
coloured components in order of increasing adsorbance, using solvent mix-
tures of step-wise increasing polarity. Since the carotenoids form coloured
zones, this fairly primitive method normally gives satisfactory results, and
the more refined (iii) gradient elution method is seldom used. The reliability
of lists which group carotenoids in order of increasing adsorption on various
adsorbents [ 4] decreases with the total number of components listed in
each case.
Although separation of carotenoids by partition chromatography has
not been given sufficient attention, a partition method on silica columns
has been developed [50]. The polyethylene columns used [51] represent
a reversed phase system where the sequence of the carotenoids on the
column is the reverse of that obtained in adsorption systems. The possi-
bility of separating very polar carotenoids by such methods deserves further
attention.

b) Thin layer chromatography

The modern method of thin layer chromatography elaborated by Stahl
[52, 53] has found wide application in the carotenoid field. The popularity
of the method is due to its speed and efficiency in separation, in spite of the
fact that pigment loss on the heavily exposed plates may be considerable.
'Thick layer' separations have not been reported, and the method is still limited
to the semi-micro scale. Comprehensive surveys on thin layer chromato-
graphy and its application to the carotenoid field are published [ 4, 53-58],
as well as a large number of papers describing particular systems and
applications.
Most systems described are based on the adsorption principle, while
more recently partition systems have also been described [59-62]. Systems
described up to 1964 and RF values reported for several carotenes and
xanthophylls have been tabulated [ 4]. A more recent review was published
in 1969 [57].
The various adsorption systems comprise layers of silica gel [53], kiesel-
guhr [63], alumina [4], calcium hydroxide [4], sucrose [64], Mg 2 (0HhC0 3
[62], magnesium oxide [59], magnesium oxide-silica gel [4], calcium hydr-
oxide-silica gel [ 4] and mannitol-starch [65], with appropriate solvent
mixtures. Frequently used are mixtures of petroleum ether-benzene, petroleum
ether-carbon tetrachloride, petroleum ether-ether, benzene, carbon tetra-
chloride and methylene chloride-ethyl acetate [57]. Isomeric carotenoids are
best separated by the adsorption method.
True partition systems are based on paraffin- or triglyceride-impregnated
kieselguhr or cellulose layers as the stationary phase with acetone-methanol-
water as the mobile phase [58, 60, 66]. The partition method has proved
68 SYNN0VE LIAAEN-JENSEN

particularly useful in the separation of carotenol esters with different fatty

acid components [66-71].
Other less clearly classified systems including the polyamide system [61],
which offers particular advantages, have been reviewed [57].
The majority of carotenoids, being strongly coloured compounds, do
not require staining methods. For the detection of uncoloured carotenes
staining with iodine vapour [72] or antimony trichloride [73] and fluorescence
in u. v.light have been described. Rhodanine [74] has been used for the detection
of very small amounts of carotenals.
The practical details of thin layer chromatography are described in
authoritative monographs [53, 56, 75, 76]. The RF values for a large number
of carotenoids are found in several reviews [ 4, 53, 57, 76] and original papers
on carotenes [77, 78], carotenals [79] and carotenoids of photosynthetic
tissues [80-84]. The use of RF values for identification purposes has been
discussed recently [85] (see Section A.7). Although carotenoids may be
eluted successfully from thin layer chromatograms and submitted to spectro-
scopic examination, in practice only sufficient for visible-light and mass
spectrometry is obtained. For the latter purpose the adsorbents should be
pre-treated with the solvents involved in the development of the chromatograms
and elution of the pigments in order to remove contaminants.
c) Paper chromatography
Many reviews on the application of paper chromatography to the carotenoid
field have been published recently [4, 5, 11, 86, 87]. The methods investigated
include one- and two-dimensional as well as circular [88] adsorption and
partition chromatography on unimpregnated or impregnated papers and
papers with special inorganic fillers.
Circular adsorption chromatography on commercial papers with fillers
of kieselguhr, alumina or calcium carbonate (Schleicher & Schiill), silica gel
and aluminium hydroxide (Whatman) have proved very satisfactory for the
separation of carotenes [89] and xanthophylls [90]. Using carefully standard-
ized conditions reproducible RF values are obtained [90]. Pigment recoveries
of 97-100% [90] make the method suitable for quantitative determinations
[91]. Experimental procedures are described [87, 89-91] and reviewed [4-6],
and in these papers RF values for a large number of carotenoids are found,
as in most papers originating from the author's laboratory, where the method
is routinely used for the control of purity and for following chemical reactions.
Various chromatographic systems should be used in combination [85].
Chromatography on kieselguhr paper is normally superior to other micro-scale
methods for the separation of cis-trans-isomeric xanthophylls. It should be
mentioned that eluates from kieselguhr paper chromatograms contain
impurities making such fractions unsuitable for mass-spectrometric exami-
nation.
Quantitative two-dimensional adsorption chromatography of algal pig-
ments on ordinary paper has been described [92], as have more recent studies
 III. Isolation, Reactions 69

of one-dimensional [93], two-dimensional [94] and radial [95] adsorption

chromatography, again on ordinary paper, and the difficulties associated
with these methods.
The partition systems described [96-100] do not generally give satisfactory
resolution.
Finally it may be mentioned that paper chromatography of fatty acids
obtained by oxidative degradation of carotenoids has been reported [101, 102].

d) Gas chromatography
Because of their thermolability and low volatility carotenoids are in
general not amenable to gas chromatography. Only a very limited number
of carotenoid derivatives have been analysed successfully by this method.
Thus gas-chromatographic separation of perhydrosqualene and perhydro-
phytoene at 240-285° C (5% SE-30 on chromosorb W) [103] and of perhydro-
lycopene, perhydro-y-carotene and perhydro-P-carotene have been reported
[104]. Gas-chromatographic identification of methyl esters of fatty acids
derived from natural carotenol esters has also been employed [41].

6. Crystallization
General procedures for obtaining crystalline carotenoids have been out-
lined by various authors [2, 3, 105, 106]. Only fractions chromatographically
pure as to carotenoid content can be used for crystallization, since co-
crystallization otherwise occurs. As pointed out previously [2] crystallization
of carotenoids on the micro scale requires some practice, but the procedures
do not differ essentially from those generally used in other micro-scale
crystallizations, attention to the necessary precautions being obligatory
(Section A.1). Crystallization from suitable solvent pairs on the 1-mg scale
is often successful. Non-carotenoid contaminants are frequently removed
by fractional crystallization in suitable solvent systems prior to crystallization
of the carotenoid. If co-crystallization with non-carotenoid material cannot
be avoided, derivative preparation followed by chromatography may lead
to the pure carotenoid.
The purity of the crystalline material should be checked by the methods
described below (Section A. 7). The number of recommended recrystallizations,
with a maximum of three, depends on the quantity available and the degree
of impurity.

7. Characterization
a) Purity criteria
First of all a pure carotenoid should be chromatographically homogeneous
in at least two paper or thin-layer chromatographic systems. By 'the latter
method colourless impurities may be revealed on spraying with sulphuric
acid. The possibility of extra coloured zones representing cis isomers is checked
70 SYNNf1JVE LIAAEN-JENSEN

by reversible isomerization. The phenomenon of cis-trans isomerism in

carotenoids is dealt with by Zechmeister [107] and discussed in Chapter V.
Ideally a pure carotenoid should exhibit well-defmed crystalline shape
when inspected under the microscope, but frequently in small-scale isolation
work only semi-crystalline solids are obtained.
Provided the melting point (in evacuated capillary tube) is sharp and
correct and there is no depression in melting point when mixed with an
authentic specimen, these criteria can be taken as strong evidence of purity
and identity.
However, if the melting point is unsharp and too low, the degree of impurity
is not revealed by the melting point determination. Extinction coefficient
determination in visible light (of 0.5 mg sample or more) on a molar basis
gives quantitative information on the purity of the compound, when com-
pared with known extinction coefficients for the actual chromophoric type.
Extinction coefficients for most carotenoid chromophores have been reported
(see Chapter IV, various reviews [4, 107] and the original literature). If the
m
molecular weight is unknown comparison of r~ values gives useful in-
dication.
Assuming that the carotenoid at this stage is obtained in the purest possible
form (preferably crystalline, alternatively chromatographically homogeneous
as to other carotenoids) the purpose of further characterization is, in the first
instance, to examine its possible identity with previously described carotenoids.
If the structure is unknown the ultimate aim is to establish the structure of
the compound.
b) RF values
The chromatographic techniques employed have already been described
in Section A.5. The eluent required for a particular carotenoid for a chromato-
graphic column of a given adsorbent gives a rough measure of the polarity
of the compound. RF values measured by thin-layer or paper chromatography
give more precise information. Since the relative polarity of a carotenoid is
primarily determined by the functional groups present, the RF values permit
rough classification into carotene-, monool-, diol- and polyol-type carotenoids.
However, great caution must be shown in drawing conclusions as the chromo-
phore, molecular size and special structural features also have a marked
influence on the RF value. Whereas the chromophore is normally revealed
by the absorption spectrum in visible light, the latter two factors require
special attention. Apo-carotenoids for instance are relatively more strongly
adsorbed than related C40 -carotenoids, C 50-carotenoids less strongly adsorbed
than their C40 -analogues [85], and tX-glycols, for example, less strongly
adsorbed than other diols. Sources for RF values of known carotenoids are
referred to in Section A.5.
RF values are of greatest use in the direct comparison of carotenoids
where their structural relationship is known, as in following the course of a
chemical reaction, and where identity is suspected. RF values are frequently
 III. Isolation, Reactions 71

used as criteria of identity. If two carotenoids are identical they should, of

course, exhibit identical RF values, co-chromatographing, when directly
compared, in all systems. However, the reverse may not be true. Identity,
based on co-chromatography tests alone, is never complete proof but is
better (i) the higher the resolution the system offers (circular chromatography
is frequently superior) and the more systems the identity has been tested
in, (ii) if not only the trans, but also the most abundant cis isomers co-chro-
matograph (direct comparison of iodine-catalysed equilibrium mixtures is
recommended [106, 108]) and finally, (iii) the more derivatives of the com-
pounds co-chromatograph, again preferably by comparison of the stereo-
isomeric sets.
c) Partition coefficients
Use of the liquid-liquid partition method in preparative work has already
been dealt with in Section A.4. Partition ratios of selected carotenoids in the
systems petroleum ether-95% aqueous methanol and petroleum ether-
85% methanol have been determined under standard conditions, and it has
been suggested the method be used as an analytical tool suitable for grouping
carotenoids into carotene-, monool-, diol- and polyol-type. Again the number
of hydroxy groups are of major importance, while more subtle effects are
caused by carbonyl, ester, ether and epoxide functions [109]. Later it was
shown that the chromophore had also to be taken into consideration [106],
that the contribution of the hydroxy groups present depended slightly on
their character [110] and more recently that comparison of carotenoids with
C40 - and C 50-skeletons presented a pit-fall [85]. Consequently comparison
of partition coefficients is only meaningful for carotenoids with closely related
structures.
M 50 values (the M 50 value of a carotenoid is defined as the percentage of
methanol required to give partition ratio 50: 50) have been introduced as a
better measure of polarity [110]. On the basis of the examination of related,
generally bicyclic C40-carotenoids of known structure, relative polarities
were assigned to each carotenoid, the relative polarity being the sum of the
polarities of the functional groups in the molecule, choosing the value 1.00
for a non-allylic hydroxy group as standard. Other functional groups were
assigned the following polarities: allylic secondary hydroxyl 0.89, keto 0.72,
acetoxy 0.47, 5,6-epoxy 0.24 and 5,8-oxide 0.24, thus allowing a linear relation-
ship between the M 50 value and the relative polarity of each carotenoid
examined [110].
Since 1956, carotenoids have been routinely described by their partition
coefficients, and the relative polarity value [110] has been used to suggest
the number and type of functional groups present in an unknown carotenoid
[111-113], disregarding the fact that this figure was worked out for a very
limited group of carotenoids and certainly has no general application [114].
Structural modifications where the suggested polarity values [110] are not
valid, even within the bicyclic C40-series, have been reported [115, 116].
72 SYNNI'JVE LIAAEN-JENSEN

In conclusion, partition coefficients, like RF values, are only meaningful

in the structural sense when comparing structurally closely related carotenoids.
d) Spectroscopic properties
Key information concerning the structure of a carotenoid is obtained
from its spectra. The absorption spectrum in ultraviolet/visible light reveals
the chromophore. The infrared spectrum gives, in a qualitative manner,
information on most of the functional groups present. The proton magnetic
resonance spectrum demonstrates the number and structural surroundings
of the methyl groups present and, less clearly, the number and type of other
hydrogen atoms in the molecule. High-resolution mass spectrometry permits
establishment of the molecular composition, and the fragmentation pattern
gives further structural information. These methods are all treated in detail in
Chapter IV. Whereas the visible light absorption spectrum and the mass
spectrum only require microgram quantities and may be obtained for a
non-crystalline compound, infrared spectra require ca. 0.2 mg and proton
magnetic resonance spectra 5-10 mg pure substance on a 60-MHz instrument,
if no time-averager is available; with a time-averaging device 0.5 mg is
sufficient. For identification purposes co-chromatography and mass spectro-
metry are a satisfactory combination. For full structural elucidation informa-
tion from each type of spectrum must be sought.
e) Preparation of derivatives
Preparation of various derivatives may be carried out in microgram scale
and is useful (i) in making possible chromatographic and mass-spectrometric
comparison at several levels and (ii) confirming, by chemical methods, the
presence of functional groups indicated by spectral data or, in the absence
of infrared, proton magnetic resonance and mass spectra, in indicating the
presence of such groups.
Since the preparation of derivatives depends on the functional groups
present in the molecule, this is treated under group reactions in Section B
below.

8. Protein complexes
Some carotenoids occur in nature as protein complexes, see Chapter II.
The isolation of macromolecular carotenoprotein complexes is of course
based on other principles than those dealt with above.
Carotenoproteins are generally extracted with water or buffer solutions,
fractionally precipitated by ammonium sulphate, and purified by chromato-
graphy on ion-exchange celluloses, Sephadex or by selective adsorption on
calcium phosphate or aluminium hydroxide [117-123].
Isolation of alkali-labile carotenoids such as astaxanthin (203) and asterinic
acid (200 and 202) from lipid-rich sources via protein complexes is a convenient
way of obtaining separation from the lipids without resorting to saponifica-
tion [121, 124].
 III. Isolation, Reactions 73

The topic has been expertly reviewed fairly recently [125], and details
concerning the isolation and characterization of protein complexes are not
included in this chapter.

B. Group Reactions

1. Introduction
The structure of a carotenoid is not fully established until the structure
deduced by physical and chemical methods is proved by total synthesis of the
natural carotenoid or of a suitable derivative, and its stereochemistry deter-
mined. However, by joint use of spectrometry and chemical reactions the
structure of a carotenoid may be established with reasonable certainty. In
structural elucidation, as well as for identification purposes, establishment of
the number and type of the functional groups represents a major step. Elements
other than carbon, hydrogen and oxygen have not yet been encountered in
natural carotenoids. Group reactions in the carotenoid field are therefore
directed towards determination of the oxygen functions in the molecule. In
the present treatment carbon-carbon multiple bond arrangements will also be
considered as functional groups.
The term group reaction is often conceived as derivative formation used
at a time when spectroscopic methods were not available for establishing the
functional groups. It is of course true that information about the functional
groups in a carotenoid should in the first instance be sought by spectrometric
methods. Ideally, the molecular formula established by mass spectrometry
reveals the number of oxygen functions, while the fragmentation pattern
frequently suggests the type of functional groups present. The infrared, proton
magnetic resonance and, to a lesser extent, the visible light absorption spectrum
give further information on the nature of these groups. However, the informa-
tion obtained from the infrared spectrum is only qualitative and frequently
only indicative, and the proton magnetic resonance spectrum indicates the
functional groups in an indirect, although quantitative way.
Generally it is desirable to confirm the number and type of functional
groups suggested from spectroscopic evidence by chemical reactions. In cases
where limited quantities prevent the recording of the desired spectrometric
data, preparation of derivatives may indicate the number and character of
functional groups present. Whenever possible all spectroscopic data of a
derivative should be sought in order to confirm that the expected chemical
change indeed has occurred, and the chemical and spectroscopic data evaluated
together. With the modern micro techniques discussed in Sections A.3-A.7
chemical reactions in the carotenoid series may, if necessary, be carried out
on a microgram scale.
The purpose of the present section is to review the chemical reactions
available in the structural elucidation of carotenoids with particular emphasis
on the establishment of functional groups. Information about the location of
74 SYNN0VE LIAAEN-JENSEN

the functional groups relative to each other and to the main chromophore is
frequently deduced from the result of such reactions.

2. Carbon-carbon multiple bonds

In addition to the chemical reactions mentioned below, the biological
vitamin A activity test for carotenoids containing unsubstituted f:J-rings should
be remembered [2, 126].

a) Addition reactions
Miscellaneous. Theoretically information about the number of carbon-
carbon double bonds in a molecule can be obtained by quantitative measure-
ments of the addition of halogens (bromine [127]) or oxygen [128]. Such
methods, introduced in classical carotenoid chemistry and discussed else-
where [2], are no longer in use.
Hydrogenation. The most useful addition reaction is catalytic hydrogena-
tion, which generally results in reduction of all double bonds present, including
benzene rings [129], carbonyl groups and epoxy groups. However, the latter
two types react more slowly. The solvents and catalysts employed have been
reviewed [2]. The catalysts commonly used include palladium oxide, colloidal
platinum and platinum oxide on various adsorbents, and must be used in large
excess. Acetic acid and ethyl acetate are suitable solvents. Intermediates in the
catalytic hydrogenation of lycopene have been described [130], but generally
only the perhydro products have been examined. The micro hydrogenation
method of Kuhn and Moller [131] was previously a routine tool, the hydrogen
consumption revealing the number of double bonds present and the elementary
analysis of the perhydro product the number of rings present. However,
unequivocal differentiation between e. g. twelve and thirteen double bonds
requires high purity and ac~uracy. Moreover, carbonyl groups may be reduced
[132, 133] and hydrogenolysis of allylic hydroxy groups can occur. Today
consideration of the chromophore, revealed by the visible light absorption
spectrum, together with the proton magnetic resonance spectrum and the
mass spectrum is a safer method of establishing the number of carbon-carbon
double bonds present.
As to hydrogen consumption, a carbon-carbon triple bond of course is
equivalent to two double bonds. Acetylenic bonds are best detected by infrared
(normally weak absorption) and proton magnetic resonance spectra. In
principle, however, selective reduction by means of Lindlar catalyst [134] to
the corresponding cis polyene, as frequently employed in the synthetic field
[135, 136], is possible.
Cumulative double bonds are best detected by infrared spectroscopy.

b) Oxidative degradation
Ozonolysis. The classical use of ozonolysis for oxidative degradation of
carotenoids is discussed by Karrer and Jucker [2]. By this method oxidative
 III. Isolation, Reactions 75

cleavage of carbon-carbon double bonds was accomplished, and from the

carboxylic acids produced conclusions were drawn concerning the end groups
of carotenoids. Thus ozonization of P-carotene (3) gave geronic acid (1), while
IX-carotene (5) provided geronic acid (1) as well as isogeronic acid (2) because it
has dissimilar end groups [137] (Scheme 2). By modern identification methods,
using paper chromatography [102, 138] or gas chromatography of suitable
derivatives, the sample requirements are greatly reduced.

O~C02H o,
+-----

(2)
Scheme2

In quantitative isopropylidene determination [139] the acetone formed on

ozonization followed by permanganate oxidation is determined iodometrically.
One isopropylidene group gives ca. 0.8 moles of acetone, and it should be
noted that IX- and P-end groups give ca. 0.1 mole [140], and -C(CH 3 h0H and
-C(CH 3 h0CH 3 residues ca. 0.3 moles [141]. Although isopropylidene groups
today are normally easily revealed by proton magnetic resonance and mass
spectrometry, the method is still useful in certain cases [142].
Chromic acid and permanganate oxidation. These reagents represent alter-
natives for oxidative attack on double bonds. Stepwise degradation with
alkaline permanganate or chromic acid, extensively used in the classical period,
allowed the isolation of large degradation products like apo-carotenals and
ketonic products, which permitted conclusions to be drawn concerning the
structure of the natural carotenoid [2]. The methods are still in use [129,
143-146] when sufficient sample is available.
Chromic acid oxidation was developed into an analytical procedure by
Kuhn and Roth [147] for the determination of methyl groups attached to
the polyene chain as acetic acid. Today proton magnetic resonance spectro-
scopy is a superior tool for this purpose.

c) Rearrangements
Isomerization. Cis-trans isomerism is fully treated in Chapter V and is not
dealt with here. Alkali-catalysed isomerization of isolated double bonds into
conjugation is not readily effected in the carotenoid series. Isomerization of
zeaxanthin (67) to lutein (73) in low yield by treatment with sodium methoxide
has been reported [148]. o-Carotene (11) gave y-carotene (8) by the same
method [149]. Terminal methylene groups also appear to represent stable
configurations [85, 150, 151].
Trisubstituted allenes of a type present in neoxanthin (122) and para-
centrone (246) are resistant towards alkali [152]. The well-known alkali-
lability offucoxanthin (190) [153] is due to other structural elements.
76 SYNN0VE LIAAEN-JENSEN

Acid-catalysed isomerization of a carotenoid allene, neoxanthin (122), into

the acetylenic diadinochrome (3), as depicted in Scheme 3, has recently been
claimed [154]. An analogous conversion has been claimed for deepoxyneo-
xanthin (86) to diatoxanthin (66) [155]. Further evidence in support of these
transformations is required.

"y'-·~ ~ ··yb.~ - .~.,---0

H~OH ~OH ~H
(3)
Scheme 3

Cyclization. In vitro cyclization to P- or IX-end groups at the C 40 -level has

only recently been reported [156]. Carotenoids like y-carotene (8), rubixanthin
(45) esters and P-zeacarotene (9) with lycopene-type end groups were reported
to cyclize in the presence of TiC1 4 to the IX-ring isomer according to Scheme 4.

(( TiCL, [ Cl,Tiu@I RJ Cle-

ecR
,#

Scheme 4

3. Hydroxy groups
The partition data and chromatographic behaviour of a carotenoid afford
useful indications to the number of hydroxyl functions present (Sections A.4
and A.5). Spectrometric methods provide further information. Thus the presence
and character of any hydroxy groups present in simple carotenoids are indicated
by the infrared spectrum. Primary hydroxy groups are revealed by the methylene
signals and tertiary ones indicated by the signal position of any adjacent methyl
groups in the proton magnetic resonance spectrum. Losses of water, acetone
etc. in the mass spectrum are further indications of the presence of hydroxy
groups (see Chapter IV). However, establishment of the number and nature of
hydroxy groups in a carotenoid of fairly complex structure generally demands,
in addition, the use of chemical methods.

a) Grignard reaction
Zerewitinoff determination of active hydrogen by the Roth [157] procedure
is based on a Grignard reaction (Scheme 5).
ROH + CH 3 Mgl ------+ ROMgl + CH4
Scheme 5

The methane produced is determined manometrically. If more than three

hydroxy groups are present, a quantitative reaction is not obtained [2] and
 III. Isolation, Reactions 77

enolizable keto groups and carboxyl groups give rise to methane production
thus simulating hydroxy groups [158]. Considering the sample requirement
of this reaction, spectrometry and micro scale preparation of derivatives, as
discussed below, are preferable.

b) Esterification
Primary and secondary hydroxy groups are readily acetylated at room
temperature by acetic anhydride in dry pyridine [105, 142] while tertiary
hydroxy groups do not react under these conditions [105, 106]. When the
course of acetylation is followed by periodic chromatographic inspection, the
number of intermediary acetates formed gives information about the number
of hydroxy groups accessible for acetylation [46, 105, 150]. Thus while a
monool can only give one acetate, a diol can form one intermediary acetate if
the molecule is symmetrical, and two if not, in addition to the diacetate. A trial
can give a maximum of three monoacetates, three diacetates and one triacetate
etc. The method is only reliable when three or less hydroxy groups are reacting,
since with more, inseparable mixtures are formed. In such cases mass or proton
magnetic resonance spectroscopy may reveal the number of acetate groups
present in the fully acetylated product [46, 150].
Higher fatty acid esters of secondary or primary carotenols are generally
prepared by means of the corresponding acid anhydride or acid chloride [2].
Acid chlorides, to some extent, also react with tertiary hydroxy groups [106,
159]. Iodoacetates are difficult to obtain in the crystalline state suitable for
X-ray analysis [160, 161].

c) Silylation
The formation of trimethylsilyl ethers of primary and secondary as well as
tertiary carotenols proceeds smoothly at room temperature [162] or lower
[163]. The reactivity of even tertiary carbinols is probably a result of the
greater-length ofthe 0-Si bond compared to the 0-c bond (cf. acetates above).
The only hydroxy groups not accessible to silylation are those in 6,6'-positions
in azafrin (261)-like end groups [162], where the steric hindrance is great.
The number of tertiary hydroxy groups present in a carotenoid can be deter-
mined by submitting the fully acetylated product to silylation. Since the
silylation reaction is very fast, the reaction course should be studied at low
temperature [16, 85, 163]. In the author's laboratory mass spectrometry of
tertiary trimethylsilyl ethers is used extensively [16, 85, 162, 164].
Trimethylsilyl ethers are hydrolysed in alkaline media, tertiary ethers
considerably more slowly than secondary or primary ones [162]. This allows
an easy differentiation between secondary/primary and tertiary hydroxy groups.

d) Methylation
Methylation of primary, secondary and tertiary, non-allylic and allylic
hydroxy groups can be effected by various methods.
78 SYNN0VE LIAAEN-JENSEN

Rather surprisingly enolic hydroxy groups such as those in astacene (198)

or in the phenolic carotenoids 3-hydroxyisorenieratene (52) [144] and 3,3'-
dihydroxyisorenieratene (79) [144] are not methylated with diazomethane
[165, 166].
Methylation of non-allylic secondary hydroxy groups, e. g. in zeaxanthin
(67), may be achieved by treatment with potassium t-amyloxide and methyl
iodide [167] or better by the Kuhn procedure [168, 169] with methyl iodide
and silver oxide or barium oxide in dimethylformamide [106]. By the latter
method tertiary non-allylic hydroxy groups may also be methylated, as in the
case of bacterioruberin (231) [85, 106].
Secondary hydroxy groups in allyl position to the polyene chain, as in
isozeaxanthin (71) or in (x-rings as in lutein (73), may be methylated selectively
by hydrogen chloride in methanol [170-172] under conditions where non-
allylic hydroxy groups do not react [171, 172]. Primary allylic hydroxy groups
of the type present in decaprenoxanthin (226) are not methylated by this
procedure [142].
It has recently been reported [173] that lutein (73) bis(trichloroacetate)
undergoes selective methanolysis yielding two allylic methyl ethers, namely
lutein 3'-(methyl ether) and its allylic rearrangement product with a tertiary
methoxy group in 5'-position (see Scheme 66).

e) Oxidation
Oppenauer oxidation of the non-allylic hydroxy groups of carotenoids
with five-membered rings to the corresponding cyclopentanones have been
reported [174-176]. Under similar conditions the non-allylic hydroxy groups
of zeaxanthin (67) were not attacked [177]. a-Glycols ofthe azafrin (261)-type,
more specifically 5,6-dihydro-p,p-carotene-5,6-diol *,may be oxidized to semi-
P-carotenone (213) by means of lead tetraacetate or chromic acid [178, 179].
Oxidation of allylic secondary and primary carotenols to a,p-unsaturated
ketones and aldehydes, respectively, may be effected by various other reagents
besides that of Oppenauer [136]. Distinction between primary and secondary
allylic hydroxy groups is generally not made from their reactivity, but is
revealed by spectrometric analysis of the product. From the infrared (conju-
gated carbonyl), proton magnetic resonance (aldehyde) and visible light ab-
sorption spectrum (change in chromophore) the character of the original
hydroxy· group and its position relative to the main chromophore are inferred,
cf. Chapter IV. In particular allylic position to the polyene chain is easily
checked on the microgram scale. The reagents used for allylic oxidation in the
carotenoid field are discussed below.
p-Chloranil is useful for oxidizing hydroxy groups in allylic position to the
polyene chain [180] (e.g. of isozeaxanthin (71) to canthaxanthin (193) [181]).
Yields are improved if the reaction is carried out in light in the presence of
* When no formulae number is referred to, the new IUPAC nomenclature is used (see Appen-
dix of this book).
 III. Isolation, Reactions 79

iodine [181]. However, IX-glycol arrangements may prevent the oxidation.

Thus, neither the triol corresponding to flexixanthin (171) nor crustaxanthin (93)
are oxidized by this reagent [105], whereas plectaniaxanthin (76) is oxidized
[207]. Hydroxy groups allylic to an isolated double bond are not attacked [181].
2,3-Dichloro-5,6-dicyano-p-benzoquinone, a somewhat stronger oxidizing
reagent, is capable of oxidizing hydroxy groups allylic to the polyene chain,
including IX-glycols such as crustaxanthin (93) [182]. However, many caro-
tenoids are destroyed by this reagent. Lack of reactivity towards p-chloranil and
reaction with dichlorodicyano-p-benzoquinone may be a useful indication of
IX-glycol arrangement in the 3,4-position of P-rings. The characteristic reaction
behaviour of IX-ketols is discussed in Section B.4. IX-Glycols are otherwise not
easily characterized, as for example by acetonide formation. Oxidation with
sodium periodate or lead tetraacetate, leading to cleavage of the molecule, is
not frequently used [183].
Careful oxidation with air in the presence of iodine in light [181] may be
an efficient method when other reagents fail, and has been used successfully for
rhodopinol (80) [184] and violerythrol (158, Scheme 83) [41].
Active manganese dioxide [185] has been used for allylic oxidation [182,
186], but is generally too active in micro-scale experiments.
Silver oxide has also been employed for the oxidation of hydroxy groups
allylic to the polyene chain [187].
Nickel peroxide [188] may effect oxidation of hydroxy groups in allylic
position to the polyene chain as well as to isolated double bonds (e.g. in lutein
(73) [172] and decaprenoxanthin (226) [142]). Again many carotenoids are
destroyed by this reagent.
Silver carbonate on celite, which has proved so successful for allylic oxida-
tion of various terpenes [189], has been tested on lycoxanthin (62) with less
success [190] than nickel peroxide.

f) Dehydration
Primary, secondary or tertiary allylic hydroxy groups may be eliminated
as water on treatment with chloroform acidified with hydrochloric acid by a
reaction first introduced by Karrer and Leumann [191] for eschscholtzxanthin
(84) (Scheme 6a) and later standardized [192]. Allylic ether groups [170, 193,
194], including glycosides [46], react in an analogous manner. The reaction
results in decreased polarity and extension of the chromophoric system of the
products, which may readily be identified. The elimination is frequently
accompanied by allylic rearrangement (Scheme 6b). Intermediate (tertiary
carbonium ion) and product control appear to be important. Formation of less
sterically hindered retro products and long polyene chains appears to be
favoured. Several successive eliminations may occur, as shown in Scheme 6c
for reduced spheroidenone (4). Enolic intermediates do not appear to be
favoured. The result [191, 194,217, 259-261] and assumed mechanism of some
simple reactions are exemplified in Scheme 6a-c. If no hydrogen atom is
available for water elimination (e.g. 2,2'-dihydroxyspirilloxanthin (5) and
80 SYNN0VE LIAAEN-JENSEN

(84)

j- 2H 2 0
-2Hm

(35)
Scheme 6a

(40) ) -H 2 0

"<:::::: "<:::::: "<:::::: "<:::::: "<:::::: "<:::::: "<:::::: "<::::::

(!>

p
-::? -::? -::? -::? -::? -::? -::? -::? );:)
1-Hm

-::? -::? -::? -::? -::? -::? -::? -::?

(36)
Scheme 6b
 III. Isolation, Reactions 81

(4)
l-H 20

H,

~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~

(18)
Scheme 6c

capsorubol (6)) no elimination occurs in spite of allylic hydroxy groups being

present (Scheme 6d).
Concerning the steric requirements for elimination, it has been argued that
crustaxanthin (93), because of a trans cx-glycol arrangement not permitting trans
dehydration, does not react with acidified chloroform, whereas borohydride-
reduced astaxanthin, assumed to be a cis cx-glycol, does [ 40]. The reaction,

Carotenoids 6
82 SYNNOVE LIAAEN-JENSEN

OCH 3 OH

"":::: "":::: "":::: "":::: "":::: "":::: "":::: "":::: "":::: "":::: "":::: "":::: ""::::

t
OH CH 3 0
(5)

OH H®

OH
Scheme 6d

STARTING MATERIAL DEHYDRATION PRODUCT

( ) =Minor product
1,1'-Dihydroxy-1,2,1',2'- HO,I I
tetrahydrolycopene (81) ~R --~R
[85, 159]
Rhodopin (56) [196]
OH-Chlorobactene (53) [197]
OH-Spheroidene (107) [198]
OH-Spheroidenone (185)
[194]

1,1' -Dihydroxy-1,2,1',2'-
tetrahydrolycopene (81) [85]

Saproxanthin (75)
3-monoacetate [195] ~R
3,4,3',4'-Tetradehydro-
1,2,1',2'-tetrahydro-t/J,t/J-
carotene-1,1'-diol [127]
3,4-Didehydrorhodopin (55)
\
[199] ~R

5,6-Dihydro-P,P-caroten-5-ol
[200]
~·-~

Scheme 7
\(~)
 III. Isolation, Reactions 83

normally carried out in 0.03 N chloroform-hydrogen chloride, is very fast,

often being completed in 5 minutes at room temperature. The pigment recovery
is usually satisfactory and the reaction is extensively used. Activation by the
polyene chain presumably facilitates the eliminations, since the reaction has
no counterpart in other classes of compounds.
Dehydration of tertiary non-allylic carotenols may be effected by treatment
with phosphorus oxychloride in pyridine [159]. Primary and secondary
hydroxy groups react differently, forming hypophasic phosphate esters, and
must be protected, for example by acetylation [195]. Dehydration of rhodopin
(56) or its dihydroxy analogue (81) yields nearly exclusively lycopene (19);
this is in agreement with Saytzeff's rule. However, as seen from the results
compiled in Scheme 7, not all dehydrations proceed according to this rule. The
· reaction is most useful in the structural elucidation of natural tertiary carotenols
and has been extensively used [85, 159, 196-198, 200].

g) Character and vicinity

By means of the reactions and reagents discussed above it is, in most cases,
possible to establish the total number of hydroxy groups and their character
(primary, secondary, tertiary). Enolic hydroxy groups are generally identified
by their spectrometric and acidic properties (partition, chromatography).
Concerning the immediate surroundings of the various hydroxy groups,
those in allylic position to the polyene chain or to isolated double bonds are
easily identified. In the former case the bathochromic shift observed on allylic
oxidation may further reveal location in the aliphatic chain or an alicyclic ring.
Allylic in-chain hydroxy substituents are readily established by mass spectro-
metry [201-203], see Chapter IV. However, the location ofnon-allylic hydroxy
groups is not revealed by these methods. Oxidative degradation by means of
ozone or permanganate (cf. Section B.2) followed by analysis of the carboxylic
acids produced was introduced in the classical period for this purpose, see
Scheme 8. On oxidation of an unhydroxylated P-ring geronic (1), oc,oc-dimethyl-

0~C02H H02C
~
C0 2H
-+H02C~
C0 2H H0 2C
X C0 2H
(1) (7) (8) (9)
Scheme 8
84 SYNN0VE LIAAEN-JENSEN

glutaric (7), IX,IX-dimethylsuccinic (8) and dimethylmalonic (9) acids are obtained,
whereas in a hydroxylated P-ring the non-formation of (1) and (7) indicates
3-substitution [2]. By modem paper- or gas-chromatographic analysis of the
products the sample requirements are considerably reduced.
Examination of the influence on the chemical shift of neighbouring protons
on modifying the hydroxy substituent [204] (see Chapter IV) represents a
welcome alternative to oxidative degradation.

4. Carotenol esters
a) Hydrolysis
Hydrolysis of carotenol esters proceeds smoothly within a few hours in
weakly (1-5 %) alkaline solution at room temperature. In principle it is possible
to follow the course of hydrolysis chromatographically in order to determine
the number of ester groups. On hydrolysis of crystalline carotenol esters the
carboxylic acid liberated may be determined quantitatively [39, 205].

a)
CH 3- r- 3 ~
-o-C-R'
~· ~
CH 3- i - o - C - R 1
H- - 0 6
--+
H-~-0-~-R'
+ R 2 C02H

l
-r-
H-?-oe
CH 3
3
06 CH 3
~· 6
-T-o
H-T-o-~-R 1
R + R 1C02H R 0

b) rn,-f;-L' CH 3
~·
-T-o'c/o
Ho-T-0/
6

'R•
R R

l
~·
rn~' CH 3-~-o6
Ho- -O-C-R3
II
R + R 3 CO,H R 0

Scheme 9
 III. Isolation, Reactions 85

It should be mentioned that natural tertiary carotenol esters, not expected

to hydrolyse readily, do so provided they have a hydroxy, acyloxy or keto
group at the adjacent carbon atom [205, 207]. This has been explained by acyl
migration (Scheme 9). In the case of the hydrolysis of a tert. sec. oc-diester
(example a) a monool intermediate was not present to any appreciable extent.

b) Hydrogenolysis
When carotenol esters are subjected to hydrogenolysis with lithium
aluminium hydride the free carotenols are readily obtained. Ester groups are
generally considered not to react with sodium borohydride [209, 210].
However, exceptions are also known for carotenol esters [41].

5. Carbonyl functions
In addition to carotenol esters treated above, the carbonyl functions
encountered in natural carotenoids comprise keto, aldehyde, carboxylic acid,
carboxylic acid methyl ester and lactone. With few exceptions [111, 211] the
carbonyl functions encountered are conjugated with the polyene chain.

a) Reduction
Reduction products are usually readily distinguished from the parent
compound by their increased polarity and the hypsochromic shift.
For the reduction of the carbonyl function metal hydrides are commonly
used, either lithium aluminium hydride in ether [105] or tetrahydrofuran or
sodium borohydride in ethanol or ethanol-benzene [41, 182]. With the former
reagent the reaction is complete in a few minutes at room temperature. Reduc-
tion with sodium borohydride proceeds more slowly, several hours being
required. A practical difficulty with lithium aluminium hydride reduction is the
adsorption of strongly polar xanthophylls on inorganic precipitates in the
aqueous hypophase during the subsequent isolation of the product. Lithium
aluminium hydride reduces ketones and aldehydes as well as carotenoic acids
and their esters [105, 184, 212]. Sodium borohydride also reduces aldehyde
and keto groups, but free carboxylic acids and their methyl esters are not
reduced by this reagent [209, 212]. This difference in reactivity may be of
diagnostic value. Furthermore, aldehydes and ketones may be differentiated
according to their rate of reduction by borohydride under standardized
conditions [32]. It has been generally found [209, 210] that the steric course of
the reduction of keto groups varies somewhat with the reagent used. Thus
reduction of astaxanthin (203) diester with sodium borohydride is reported
to give the cis di-oc-glycol [213]. Favoured formation of the cis glycol has
also been assumed for the analogous reduction of actinioerythrin (272),
whereas lithium aluminium hydride appeared to favour formation of the trans
glycol [ 41].
The Meerwein-Ponndorfreduction previously used [214] offers no advan-
tage over metal hydride reduction.
86 SYNN0VE LIAAEN-JENSEN

The zinc-acetic acid-pyridine reduction of conjugated w,w' -diketones,

giving the unconjugated diketones, is a very useful reaction for identification
of this structural group [117, 179, 190, 215-217]. Thus rhodoxanthin (209)
gives 4,4'-dihydrorhodoxanthin (p,p-carotene-3,3'-dione), which is readily
dehydrogenated by atmospheric oxygen back to rhodoxanthin [179].

b) Oxidation
Oxidation of aldehydes to carboxylic acids, for example with silver oxide
[218], is not easily effected in the carotenoid series. Bixindial (10) has been
converted to the corresponding dicarboxylic acid (13) via the oxime acetate (11),
which on alkali treatment gave the dinitrile (12) subsequently saponified to
the dicarboxylic acid (13) [219], according to Scheme 10.

l
AcO-N=C '<:::::: '<:::::: '<:::::: '<:::::: '<:::::: '<:::::: '<:::::: '<:::::: '<:::::: C N--DAc

(11)

J
NC '<:::::: '<:::::: '<:::::: '<:::::: '<:::::: '<:::::: '<:::::: '<:::::: '<:::::: CN
(12)

l
Ho.c -<:::::: -<:::::: -<:::::: -<:::::: -<:::::: -<:::::: -<:::::: -<:::::: -<:::::: co.H
(13)
Scheme 10
 III. Isolation, Reactions 87

Neither the dial of decaprenoxanthin (226) [142], nor rhodopinal (144) or

{J-apo-2'-carotenal (233) [184] gave the corresponding nitrites by this procedure.
It is recognized that only anti oximes (or their acetates) undergo dehydration
to nitrites [220]. It is therefore possible that only the syn isomer was formed
in these reactions.
Attempted Cannizzaro disproportionation of carotenals has not been
successful [184].

c) Condensation
RNH 2 carbonyl-reagents do not always react with carbonyl groups in
carotenoids.
Oximes are the derivatives most frequently prepared. Thus ketones like
echinenone (148) and canthaxanthin (193) form a monoxime [221] and dioxime
[222] respectively, and rhodoxanthin (209) gives a dioxime [158]. However,
astacene (198) and {J-carotenone (216) form dioximes [223, 224] only, and
capsanthin (170), capsorubin (205) and fucoxanthin (190) give no oxime
[2, 160]. In the aldehyde series oximes have been prepared from {J-citraurin
(249), tX- and {J-apo-8'-carotenal (248), {J-apo-12'-carotenal (262) and bixindial
(10) [2, 225]. Cross-conjugated aldehydes like rhodopinal (144) also provide
oximes [184].
Oximes are readily acetylated or methylated with diazomethane [142].
Semicarbazones are reported for various carotenals: {J-apo-8'-carotenal (248),
bixindial (1 0) and {J-citraurin (249) [2].
Hydrazones. The 2,4-dinitrophenylhydrazone ofrhodopinal (144) has been
prepared [184].
For the identification of micro-samples of carotenals and ketocarotenoids
the formation of deeply coloured derivatives with the 2-diphenylacetyl-1,3-
indandione-1-hydrazone reagent has been described [226].
Quinoxaline derivatives. tX-Diketones like astacene (198) and violerythrin
(154, Scheme 83) condense with a-phenylenediamine to a bisphenazine [223]
and a bisquinoxaline derivative [41] respectively. The five-ring tX-diketone
reacts considerably faster under analogous conditions [41].
Methyl carotenoates. Carotenoid carboxylic acids are methylated with
diazomethane [17, 224].

d) Addition
Acetals. The formation of apo-carotenal acetals is involved in the enol ether
synthesis frequently employed in the total synthesis of carotenoids [135].
Proton-catalysed acetal formation of cross-conjugated aldehydes like rhoda-
pinal (144) proceeds smoothly [184].

e) Alkali reactions
Titration of carboxyl groups. Carboxyl groups may be determined quanti-
tatively by titration with alkali, preferably using the hydrogenated compound
[227, 228].
88 SYNN0VE LIAAEN-JENSEN

Hydrolysis of methyl carotenoates. Stronger alkaline conditions are required

for the hydrolysis of methyl carotenoates than for carotenol esters, the former
not being hydrolysed by the saponification involved in the standard isolation
procedures (Section A.3). Treatment with 10% alcoholic KOH overnight is
normally sufficient [212].
Oxidation of a.-ketols. Bis-a.-ketols like astaxanthin (203) are readily
autoxidized by atmospheric oxygen in the presence of a base to the correspond-
ing a.-diketone [117] (Scheme 11). When only one a.-ketol group is present, as

HJ?C 0

KOH

Scheme 11

in flexixanthin (171), the reaction is slower [105]. Cyclopentenonol esters,

exemplified by actinioerythrin (272), show complicated behaviour on base
treatment [41], but the corresponding bis-a.-diketone may be isolated.

6. Alkoxy groups
Alkoxy groups other than methyl ethers have not been found in natural
carotenoids.
Methoxy groups may be determined quantitatively by the Zeisel method
[205, 229], but proton magnetic resonance spectroscopy is preferable. It
should be noted that ozonolysis of carotenoids containing a 1-methoxy-1,2-
dihydro end group may give a low, false isopropylidene value [106].
Demethylation of methoxy groups has not been reported. Provided the
methoxy groups occupy a position allylic to the polyene chain, or may be
brought into allylic position by other eliminations, methyl ethers may eliminate
methanol, with the introduction of a double bond, on treatment with acidified
chloroform (see Section B.3 f).

7. Epoxy groups
Reports of epoxy groups in natural carotenoids were until recently restricted
to those occupying the 5,6-position of cyclic carotenoids, as in epoxidized
/3-rings, but now epoxides in the 1,2-position, as in epoxidized isopropylidene
 III. Isolation, Reactions 89

groups, are also known [230]. The furanoid rearrangement products of the
5,6-epoxides are frequently encountered. They may be natural carotenoids,
although in many cases they presumably represent artifacts formed during
the isolation procedure (see below).

a) Colour tests
In general 5,6-epoxides and hydrofuranoid carotenoids produce a stable
blue colour on shaking with 20% aqueous hydrochloric acid and ether,
provided two such groups are present, the test being less reliable with only
one group. Some carotenals and polyols also give blue colours [231]. Various
modifications of the colour test [232, 233] include spectrometric ones based
on treatment with mercuric chloride [234] and on the epoxide-furanoid
rearrangement discussed below [235]. The presence of epoxy or hydrofuranoid
groupings should be checked by more exact methods.

b) Epoxide-furanoid rearrangement
5,6-Epoxides rearrange readily on treatment with dilute acids by ring
expansion to a hydrofuranoid system (see Scheme 12), small amounts of the
parent hydrocarbons being by-products in these acid rearrangements [2, 236].
The reaction is frequently carried out in chloroform containing hydrochloric
acid [2] but organic acids may also be used [237, 238]. This very fast reaction
causes a marked hypsochromic shift of the visible light absorption spectrum.
Furanoid oxides are not affected by treatment with dilute acids under con-

~- ~
H®
0

~- ~
HEll

Scheme 12

trolled conditions [237]. Epoxide-furanoid rearrangement may also occur on

illumination of solutions [239] and upon contact with alumina [240].
Isomerization by means of A1Cl 3 has recently been discussed [241].

c) Hydrogenolytic cleavage
Reductive opening of 5,6- and 5,8-oxides on treatment with lithium
aluminium hydride to yield the parent olefins was first observed by Hungarian
workers [242, 243]. It was initially thought that infrared light was necessary
90 SYNN0VE LIAAEN-JENSEN

Scheme 13
 III. Isolation, Reactions 91

for this reaction [242, 243], but this could not be confirmed, and olefin
production was later stated to occur only when a large excess of hydride was
employed [200]. Hydrogenolysis of 5,6- and 5,8-oxides by lithium aluminium
hydride under relatively mild conditions results in the formation of the
5-ol [200, 244, 245]. The 5-ol is not an intermediate in the olefin formation
[245]. Possible mechanisms for the reaction have been discussed [200, 245, 246].
It may be commented that the claimed cis relationship between the 3-hydroxy
group and the epoxide in natural neoxanthin and 'antheraxanthin' [245] is
opposite to the conclusion arrived at for natural violaxanthin (135) based
on o.r.d. studies [247]. The correct structure of neoxanthin (122) and the
later identification of 'antheraxanthin' with diadinoxanthin (118) [248] is
irrelevant in this context.
The salient features of the hydrogenolytic opening of 5,6-epoxides and
furanoid oxides are summarized in Scheme 13, together with mechanisms
suggested for the hydrogenolytic opening of 5,8-oxides. Formation of the
olefin may be explained by hydride attack either at C-5 or C-8 followed by
1,4-elimination of water from the resulting 5- or 8-hydroxy derivative [246],
whereas formation of the 5-ol may be due to hydride attack at C-6 and
electron displacement [200]. In the same way formation of the olefin from
the 5,6-epoxide could be explained by hydride attack at C-5, followed by
allylic dehydration, whereas hydride attack at C-6 would lead to the 5-ol
(Scheme 13).

d) De-epoxidation
Fully satisfactory methods for de-epoxidation of carotenoid epoxides are
not available. In addition to the methods above (Schemes 12 and 13) a method
involving treatment with propylmagnesium bromide and ferric chloride
allowed transformation of P-carotene diepoxide (133) to P-carotene (3) and
of violaxanthin (135) to antheraxanthin (119) and zeaxanthin (67) [249]. By
the xanthate method [250] de-epoxidation of 5,6: 5',6'-diepoxides to the
parent carotene was achieved in low yield, whereas monoepoxides and
furanoid oxides were not de-epoxidized [251]. Sulphuric acid treatment gave
unsatisfactory results [251].

8. Glycosides
Secondary [46, 150] as well as tertiary [212, 252] glycosides of C40 -
carotenoids are encountered. Carotenoid glycosides are best identified as
such by mass spectrometry of the acetates [46, 150,212, 253], which also
provides information about the type of sugar involved, these being hexose
[212], deoxyhexose [46, 150], methylated deoxyhexose [253] etc. Supporting
evidence may be sought from infrared and proton magnetic resonance spectra
of the natural glycoside and the peracetate. To establish the stereochemistry
of the glycosidic linkage enzymatic hydrolysis [252, 254] and synthetic [252]
and spectrometric [150, 255] methods have been attempted. Proton-catalysed
92 SYNN0VE LIAAEN-JENSEN

hydrolysis liberates the sugar moiety but also leads to the destruction of the
aglycone. However, reductive cleavage or allylic elimination preserves the
structure of the carotenoid moiety to such a degree that conclusions may be
drawn about the original aglycone.

a) Hydrolysis
Enzymatic hydrolysis of crystalline carotenoid glucosides with o:- and
p-glucosidase [254] has not been successful; this is due to solubility problems
[252]. Impure carotenoid glucosides are said to respond to enzymatic
hydrolysis [256, 257].
Glycosides may be hydrolysed chemically by means of 0.15 N HCl in
methanol overnight, followed by hydrolysis of the resulting methyl glycoside
with aqueous polystyrenesulphonic acid at reflux temperature [ 46, 258]. The
sugar may be identified by paper chromatography in various systems [46].
o-Glucose may be subjected to enzymatic treatment with D-glucose oxidase
for its unequivocal identification [252, 254, 262].

b) Hydrogenolysis
Cleavage of allylic glycosides may be effected with lithium aluminium
hydride [ 46, 150, 253]. The end-group modifications introduced into the
carotenoid moiety by this procedure in the case of myxoxanthophyll (90)
and oscillaxanthin (95) are depicted in Scheme 14 [150] (cf. also Scheme 13).

+ Glycosyi-0 9

Scheme 14

c) Elimination
Like alkoxy ether groups, allylic glycosides may, on treatment with acidified
chloroform, eliminate the sugar moiety with introduction of a double bond
[46, 252] (cf. Section B.3t). Thus phleixanthophyll (77) gave 3,4-didehydro-
torulene (48, Scheme 28) [252], and analogous treatment of myxoxanthophyll
(90) is believed to result in aromatization to 3',4'-didehydrochlorobactene
(52, Scheme 29) by a sequence of successive eliminations [46].
 III. Isolation, Reactions 93

C. Other Partial Syntheses

1. Introduction
In the carotenoid field chemical conversions of carotenoids are conceived
as partial syntheses. Thus partial syntheses involve reactions of compounds
with a pre-formed carbon skeleton and usually imply no skeletal modifications
other than cyclization. However, oxidations leading to loss of carbon atoms
from the skeleton are considered as partial syntheses. In the present treatise
reactions resulting in an extension of the carbon skeleton are considered as
total syntheses (Chapter VI).
From these considerations the group reactions discussed in the preceding
section are considered as partial syntheses. In this section further reactions
classified as partial syntheses will be treated. They mainly involve introduction
rather than modification of oxygen functions, and other reactions not dealt
with before.

2. Dehydrogenation
Only reactions leading to the introduction of an additional carbon-carbon
double bond by formal removal of hydrogen atoms are discussed here.
N-Bromosuccinimide is the dehydrogenation reagent most extensively
used. Early work has been reviewed elsewhere [263]. Presumably allylic
bromides are formed as intermediates in this reaction. In an inert solvent the
bromides may eliminate hydrogen bromide either spontaneously or in the
presence of a suitable base, resulting in a new olefinic bond. At one time the
stepwise in vitro dehydrogenation [264, 265] of phytoene (32) to lycopene (19)
was of considerable interest for the structure determination of phytofluene
(30), (-carotene (26) and neurosporene (22) [263]. Dehydrolycopene (18) and
bisdehydrolycopene (17) have also been prepared by this method [266]
(Scheme 15).
fJ-Carotene (3) furnished five crystalline dehydrocarotenes, namely 3,4-
didehydro-fJ-carotene (2), 3,4,3',4'-tetradehydro-fJ-carotene (1), retro-dehydro-
carotene (36), retro-bisdehydro-fJ-carotene (14) and anhydroeschscholtz-
xanthin (35), three of which exhibited retro structures [263, 267, 268].
a-Carotene (5) also affords a mixture of hydrocarbons on N-bromosuccinimide
dehydrogenation, the carotenes (1), (35), (36) and 3,4-didehydro-a-carotene
(15) [263]. a-Zeacarotene (12) is reported to give b-carotene (11), and fJ-
zeacarotene (9) provides y-carotene (8) [269] by the same reaction (see
Scheme 16).
Treatment of canthaxanthin (193) with N-bromosuccinimide has been
reported to give the diacetylenic derivative (16) (Scheme 17) [270]. In a
reinvestigation, including full spectral characterization, the product is shown
to be 15,15'-didehydrocanthaxanthin (17) [271]. The formation of acetylenic
derivatives from an w,w'-diketone is remarkable.
 f II II II II II II
\
r );- );- );- );- );- >
f f
>
r );- );- f f f
f
f
f
f
z
> f f f f
el
z
f 00 f 00 f f 00 f 00 f 00 f 00 f
z z
~
~
z
~
z
~
z
~
z
~
~"' r- r- f r- r- f- f- f .....,.,
~ e
<!)
<!)
..=
..:I
~
f f f f f f f f ()
Cll
~
~ j f f f II II
~~
~
Cll
f f f f f
N'
~ I ~ f ~ f ~ f ~
~
f ~ f;
f -I( f f f f f
\
--< f f
"<t
f
\ --< ~ 00
~ _) _) _) f
\!_I I
0\
\~
 III. Isolation, Reactions 95

The formation of dehydrocarotenes on cleavage of the dark blue boron

trifluoride-carotenoid complexes with ammonia has been discussed [263].
Hydrolysis of the complex leading to oxygenated derivatives is, however, a
more important reaction.
More recently the application of other dehydrogenating agents has been
studied. Treatment of cryptoxanthin (39) and cryptoxanthin palmitate with
N,N'-dichlorourea gave the corresponding 3',4'-didehydro products [272].
y-Carotene (8) afforded the 3',4'-didehydro and 3,4,3',4'-tetradehydro deriv-
atives and rubixanthin (45) palmitate the 3',4'-didehydro derivative [273].
15-Carotene (11) gave the 3',4'-didehydro product [274], and lycoxanthin (62)
palmitate was stated to give a tridecaene product [275].
Treatment of titanium tetrachloride-carotenoid complexes with polar
solvents is reported to provide dehydro carotenoids [276]. Thus P-carotene
(3) yielded retro-dehydrocarotene (36), zeaxanthin (67) gave anhydroeschscholtz-
xanthin (35), and physalien (68) gave eschscholtzxanthin (84) dipalmitate in
good yield; a mechanism has been suggested [276]. Likewise retro-dehydro-
dodecaprenocarotene was obtained from the synthetic C 50-model compound
dodecapreno-P-carotene [277].
Treatment of cryptoxanthin (39) with lead tetraacetate is stated to give
retro-dehydrocryptoxanthin among other products. A radical mechanism
has been suggested [278, 279].
Finally it should be mentioned that vigorous oxidation of zeaxanthin (67)
with manganese dioxide gives rhodoxanthin (209) with retro structure and
an additional double bond [288].

3. Introduction of oxygen functions

a) Epoxidation
Carotenoids containing P-rings with or without substituents in the 3,3'-
positions are readily epoxidized by monoperphthalic acid under controlled
conditions [2, 231]. From inspection of models it has been assumed that in
the 3-acetoxy-p-ring epoxidation cis to the C-3 oxygen function is favoured
[146,247,280,281].
There is also a report on the formation of a 5,8-epoxide on treatment
of P-carotene (3) with lead tetraacetate [282].
Treatment of lycopene (19) with monoperphthalic acid is reported to
give the 5,6-epoxide in low yield [283].
Recently it has been reported that canthaxanthin (193) on treatment with
perbenzoic; acid provides the corresponding 9,10-epoxy- (17 a) and 13,14-
epoxy- (17 b) derivative (Scheme 17 a) [545]. The epoxide ring could be
opened hydrogenolytically with LiAIH 4 , but not with NaBH 4 • The allylic
alcohols (17 c and 17 d) obtained were readily dehydrated with acidified chloro-
form. On treatment of the two epoxides (17 a and 17 b) with hydrogen chloride
in methanol they reverted to canthaxanthin (193).
Further reactions of the epoxides have been dealt with in Section B.7.
96 SYNN0VE LIAAEN-JENSEN

(3)

(36)

(14)

NBS

(5)

(12)

(9)

Scheme 16
 III. Isolation, Reactions

NBS

(2)

(1)

NBS

(11)

NBS

(8)

Carotenmds 7
 Whereas no chemical method is available for the direct introduction of
non-allylic oxygen functions in a carotenoid, allylic hydroxy, keto, alkoxy
or acyloxy groups are conveniently introduced by the modified N-bromo-
succinimide procedure [170, 192] (cf. Section C.2). The presence of alcoholic
solvents or acetic acid results in the formation of oxygenated derivatives,
e.g. (18) and (19), explained by substitution of the allylic bromides first formed
(Scheme 18). Keto products are considered to be the result of hydrolysis of
intermediary ketals. This elegant reaction is used in the technical synthesis
of canthaxanthin (193) from P-carotene (3) via isozeaxanthin (71), as presently
carried out by Hoffmann-La Roche. The reaction has also been used in other
total syntheses of academic interest [284]. Of historical interest is the fact
that the structures of echinenone (148) and canthaxanthin (193) were established
on direct comparison of the natural carotenoids with p,p-caroten-4-one and
p,p-carotene-4,4'-dione produced in this reaction [170, 285].
Formation of boron trifluoride-carotenoid complexes in an inert solvent
under controlled conditions followed by hydrolysis or alcoholysis of the
complex may similarly lead to the introduction of allylic hydroxy or alkoxy
functions [263]. If a P-ring is present in the molecule this is the preferred
substitution site. However, lycopene derivatives oxygenated in 5,6(5',6')-
 III. Isolation, Reactions 99

OH

OH (17d)

j1
(17 b)

H"' Perbenzoic
acid

(193)
H"'l JP~rbenzoic
acid

0 (17 a)

Scheme 17a
100 SYNN0VE LIAAEN-JENSEN

(3)
~s y(O Br
C2H 5 0H

l2NBS

yc:Q 2C,H,OH ~H, C,H,OH

Br OC 2 H 5 (18)

NBSl

Q;:Q' C,H,OH
~;Q"'
Br OC 2 H 5 C2H 5 0 OC2Hs

l
NBS Q=;QH,
0

(3)
NBS
AcOH
yc:Q
OAc (19)
-KOH yc:O
OH (71)

Scheme 18
 III. Isolation, Reactions 101

positions have been prepared by this procedure, 5,6-dihydro-1/J,Ijl-carotene-

5,6-diol being a major product [283]. A mechanism involving a zwitter ion
intermediate, formed by addition of BF39 near the centre of the polyene
chain, has been suggested for the formation and cleavage of the tX-carotene
(5)- BF3 complex, giving p,e-caroten-4-ol [263].
Other oxygenations have also been reported. Thus treatment of crypto-
xanthin (39) with lead tetraacetate was stated to give p,fJ-carotene-3,4'-diol
[278, 279], and similar treatment of {3-carotene (3) gave a product considered
to be 5',8'-epoxy-5',8'-dihydro-{3,{3-caroten-4-ol [282]. A compound identified
as 3' -hydroxy-/J,/J-caroten-4-one (154) was obtained on treatment of crypto-
xanthin (39) palmitate with N,N' -dichlorourea and subsequent hydrolysis [272].

c) Diosphenols
Oxidation of canthaxanthin (193) to phoeniconone (195) and astacene (198)
may be effected in high yield with oxygen in the presence of a strong base
(potassium t-butoxide in t-butanol) [286]. The reaction has been formulated
as autoxidation of the enolate and decomposition of the resulting hydro-
peroxide [177] (Scheme 19). Further autoxidation of astacene (198) is reported
to give a compound believed to be (20) [177]. No details are published (cf.
oxidation of astacene below).

Cf R'0 6

~
02

HO
H
\w~ ~
- 0
.....-
~

HO

OH

HO
"""::: """::: """::: """::: """::: """::: """::: """:::

(20)

Scheme 19

4. Ring contraction
Oxidation of astacene (198) or its 15, 15'-didehydro derivative with man-
ganese dioxide gave the corresponding bis-cyclopentenediones, presumably
by initial formation of the 2,3,4-triones, followed by benzilic acid rearrange-
ment to the hydroxy acids, decarboxylation and further oxidation [287]
(Scheme 20).

(<YR
g
O~R
HO~R 1) -C0 2
Mn02
H02cJ----t__
2) Oxid.
HOY
o::r o::r
0 Scheme 20
102 8YNN0VE LIAAEN-JENSEN

D. Reactions and Structures of Natural Carotenoids

1. Introduction
In the foregoing Sections B and C there was provided for new carotenoids
information on the chemical methods available for establishing the functional
groups present and for elucidating the complete structure.
The present section gives a historical review of the chemical behaviour
of the naturally occurring carotenoids to which structures have been assigned,
providing at the same time the chemical evidence upon which the structures
compiled in Chapter XII are based. Whereas Karrer and Jucker's mono-
graph [2] of 1948 was exhaustive at that time, space limitations must make
the present treatise selective rather than complete. In addition to chemical
reactions, spectrometric data (Chapter IV) and total syntheses (Chapter VI)
are major factors in contemporary structure determination. However, evalua-
tion of the evidence on which a structure is based is made in the present section.
Stereochemical considerations are fully treated in Chapter V and are not
included here. The sources of the various carotenoids are not mentioned, for
which the reader is referred to Chapter II and the recent monograph by
Goodwin [289].
The carotenoids listed in Chapter XII are here divided into four main
groups: 1) carotenes (C 40 ), 2) C40 -xanthophylls, 3) apo-carotenoids and
nor-carotenoids with less than forty carbon atoms in the skeleton, and 4)
carotenoids with more than forty skeletal carbon atoms. Within the groups 2)
and 3) the carotenoids with the rarer structural features will be treated first.
These characteristic structural elements are often responsible for the specific
reactions of th_e compound.

2. Carotenes (C 4 o)
The naturally occuring C40 -carotene hydrocarbons comprise the first
thirty-eight compounds of Chapter XII. Eighteen of these are aliphatic, fifteen
are alicyclic, including five monocyclic and ten bicyclic representatives, and
five with either one or two trimethylphenyl end groups.

a) Aliphatic and alicyclic carotenes

The aliphatic carotenes do not readily form derivatives. Generally only
catalytic hydrogenation, dehydrogenation by means of N-bromosuccinimide
and oxidative degradation involving cleavage of double bonds have been
accomplished; cf. Section B.2.
Lycopene (19) is one of the classical, well established carotenoids [2],
several total syntheses of which have been performed (Chapter VI). Lycopersene
(34) has also been synthesized, but may not be a natural carotenoid (Chapters
II and VII).
The structures of phytoene (32), phytofluene (30), (-carotene (26) and
neurosporene (22) were of considerable interest from a biosynthetic point of
 III. Isolation, Reactions 103

view (Chapter VII). The colourless phytoene is an oil. Its structure elucidation
was based on oxidative degradation [126] and subsequent confirmation on
proton magnetic resonance spectroscopy and total synthesis [290]. Structure
(30) was assigned to phytofluene on the basis of oxidative degradation [291],
and the structure was subsequently proved by proton magnetic resonance
spectroscopy and synthesis [290]. Degradation studies on (-carotene supported
structure (26) for this carotene [291]. Confirmation by total synthesis was
later obtained [290]. The natural occurrence of the isomeric non-symmetrical
compound (25) was foreseen [5], and the isolation and structure determination,
based on spectroscopic [292] and synthetic [290] evidence, have recently
been reported [292]. Neurosporene [293] (22) was for a while considered to
be identical with 5,6,5',6'-tetrahydro-1/f,l/l-carotene prepared by total syn-
theses [294]. However, the dehydrogenation of (-carotene (26) to neurosporene
by N-bromosuccinimide was inconsistent with this formula. The structure (22)
for neurosporene was finally solved by proton magnetic resonance data and
total synthesis [290].
3,4-Didehydrolycopene (18) and 3,4,3',4' -tetradehydrolycopene (17) were
obtained by dehydrogenation oflycopene (19) with N-bromosuccinimide [266].
Total synthesis of (17) has since been reported [159].
While, as a general rule, the skeletal methyl groups of the carotenoids
have been tertiary, exceptions were found recently in 1,2-dihydroneurosporene
(23), 1,2-dihydrolycopene (21) and 3,4-didehydro-1,2-dihydrolycopene (20),
the structures of which were deduced from proton magnetic resonance and
mass spectrometric data [295] and that of 1,2-dihydrolycopene (21) confirmed
by total synthesis [538].
Generally the reactions of alicyclic carotenes resemble those of the
aliphatic ones but /3-rings may readily be epoxidized and allylic oxygen
substituents may be introduced into the 4,4'-positions (Section C.3). The
cyclic double bond is also more susceptible to oxidation (Section B.2). (X-Rings
represent a stable configuration, and isomerization into /3-rings can be achieved
only under forcing conditions.
The monocyclic carotenes may be considered as derivatives of y-carotene
(8). The relatively saturated representative /3-zeacarotene (probably 9) char-
acterized as an oil [269], has been dehydrogenated to y-carotene (8) (see
Scheme 16). The isomeric (X-zeacarotene (probably 12), lacking vitamin A
activity, has likewise be,en converted to c5-carotene (11) (Scheme 16) [269].
Total synthesis of solid 7',8'-dihydro-/3,1/f-carotene (9) has been achieved [296].
More recently also 7',8'-dihydro-e,l/f-carotene (12) has been synthesized [297].
y-Carotene [298] (8) is another of the classical carotenoids [2] whose structure
is proved beyond doubt by total synthesis (Chapter VI). The structure of
c5-carotene (11) was based on extensive degradation and is in agreement with
the optical activity and lack of vitamin A activity [149]. Alkali isomerization
under forcing conditions gave y-carotene (8) [149]. Total synthesis of racemic
c5-carotene by three routes has been reported [299]. The structure of torulene,
first described in 1933 [300], was solved by the total synthesis of 3',4'-di-
104 SYNNiiiVE LIAAEN-JENSEN

dehydro-P,t/J-carotene (7) [296], when the two carotenes were found to be

identical.
The bicyclic carotenes may be considered as derivatives of P-carotene (3).
Chemical or other conclusive evidence is lacking for the most saturated
representative 11-carotene, thought to be a cyclic analogue (4) of '-carotene
[301]. The structures of P-carotene (3) and ex-carotene (5) are fully established.
The evidence is reviewed in the earlier literature [2] and discussed elsewhere
in this book (Chapters IV, V and VI). e-Carotene [302] (10) is a more recent
representative, subsequently identified [303] with 'e1-carotene' prepared by
total synthesis [304, 305] (for absolute stereochemistry see Chapter V). Very
recently the natural occurrence of two non-reactive hydrocarbons, chromato-
graphically identical with synthetic [268, 306] 3,4-didehydro-p,p-carotene (2)
and 3,4,3',4'-tetradehydro-p,p-carotene (1), has been claimed [307], and a
C40 H 56 carotene, provisionally called P 444 [206], was shown by full spectro-
scopic characterization to be 'p, y-carotene' (3a) with a terminal methylene
group [206a].

(3a)

b) Aromatic carotenes
The structure determination of the aryl carotenes is mainly due to Japanese
work [129, 143, 308-311]. The three first members of the series were reported
in 1957 [308]. From physical data including elementary analysis and from
oxidative degradation with chromic acid or potassium permanganate out-
lined in Scheme 21, the structure (16) was ascribed to renierapurpurin, the
structure (14) to renieratene and the structure (13) to isorenieratene. Among
the products identified were isorenieral (21) and renieral (22).
These assignments were subsequently confirmed by total synthesis carried
out by the same worker [310, 311] and later also by others [312, 313].
Leprotene, isolated already in the late thirties [314], has been shown to be
identical with isorenieratene (13) [108, 315, 316].
Later the structure (15) was assigned to the monocyclic aryl carotene
chlorobactene [197] on the basis of spectral evidence (proton magnetic
resonance in particular), and subsequently confirmed by total synthesis [317].
The bicyclic monoaryl carotene P-isorenieratene (6), first made by chemical
synthesis [313], was later found to occur naturally [318].
The aryl carotenes are beautifully crystalline, rather insoluble, chemically
non-reactive carotenoids with characteristic spectral properties (Chapter IV).
Derivatives, other than oxidative degradation products (Scheme 21), perhydro
and dehydro derivatives, have not been prepared. Catalytic hydrogenation
with platinum oxide also reduces the benzene rings [129]. Dehydrogenation
of chlorobactene (15) with N-bromosuccinimide is reported to give the
3',4' -didehydro derivative [ 46].
 III. Isolation, Reactions 105

:::?' '-"::::: '-"::::: '-"::::: '-"::::: '-"::::: '-"::::: '-"::::: '-"::::: '-":::::

~
(16)

X:r"""
CHO
'-"::::: '-"::::: '-"::::: '-"::::: '-"::::: '-"::::: '-"::::: '-":::::
I
l
(22)

:::?' '-"::::: '-"::::: '-"::::: '-"::::: '-":::::

Cr0 3

'-"::::: '-"::::: '-"::::: '-":::::

/
~
(14)

Cr0 3
l Cr0 3
~
OHC '-"::::: '-"::::: '-"::::: '-"::::: '-"::::: '-"::::: '-":::::
CHO
eCHO
I

l
(267)

Cr0 3
/
HO,CCCO,H
:::?'

~
'-"::::: '-"::::: '-"::::: '-"::::: '-"::::: '-"::::: '-"::::: '-"::::: '-"::::: KMn0 4
~I

l
(13)

l Cr0 3 KMn0 4

'-"::::: '-"::::: '-"::::: '-"::::: '-"::::: '-"::::: '-"::::: '-":::::

CHO HO,C'(:r:CO,H
COzH

(21) ~ C0 2 H
Scheme 21

3. C40 -xanthophylls
The majority of the natural carotenoids belong to this class. Even with
the increasing number of new structural variations that have come to light
in recent years, this picture is not expected to change.
106 SYNN0VE LIAAEN-JENSEN

a) Acetylenes and allenes

Structures have been assigned to nine acetylenic and ten allenic C 40 -
xanthophylls. No single carotenoid has yet incorporated both these structural
elements.
The triple bonds are always located in 7,8(7',8')-positions, and the allenic
bond is always terminating the chromophore adjacent to a cyclic end group
of the fucoxanthin (190) or neoxanthin (122) type. The acetylenic and allenic
carotenoids exhibit characteristic spectral properties (Chapter IV), readily
disclosing these structural features. The acetylenic grouping exhibits few
characteristic reactions. Reactions specific for the allenic carotenoids have
already been mentioned (Scheme 3) and will be further discussed here. A more
thorough survey of the acetylenic and allenic carotenoids has recently been
presented [319].
The acetylenic xanthophylls with C40 -skeletons known at present comprise
nine carotenoids. The structural element (23) (Scheme 22) is common to all
of them.
~ R
(<Y'~

R'Y R2 (23) R 1 =HorOH,R 2 =H 2 or0

(24) R 1 =H or Ac
Scheme 22

The structure determination of the acetylenic carotenoids has taken place

since 1966, largely based on the spectroscopy of the native carotenoids or
their acetates. Thus alloxanthin (65) was identified as a diacetylenic derivative
of zeaxanthin (67), and monadoxanthin (72) and crocoxanthin (41) were shown
to be monoacetylenic derivatives of lutein (73) and zeinoxanthin (42) respec-
tively [320, 321]. The previously described pectenoxanthin and cynthiaxanthin
[322] were later shown to be identical with alloxanthin (65) [323]. A stereo-
isomer of alloxan thin (65) and of crocoxanthin (41) have since been synthesized
[319]. Diatoxanthin [324] has been formulated as a monoacetylenic lutein (73)
[321] on the basis of spectral evidence. Diadinoxanthin [324], which under-
goes the characteristic epoxide-furanoid oxide rearrangement (Section B.7),
was more recently ascribed the acetylenic structure (118), again mainly on the
basis of spectral data [248]. The latest representative, heteroxanthin, has been
ascribed the structure (94), on the basis of spectral properties and polarity
data [325]. In another recent report [547] the structure P-carotene-3,8,3'-triol
 III. Isolation, Reactions 107

5,6-epoxide (187) has been proposed for heteroxanthin on speculative and

contradictionary grounds. In support of structure (94) heteroxanthin forms
a diacetate which gives a mono(trimethylsilyl ether) and does not react with
acidified ether [325, 535, 547, 548].

~ ~ ~ ~ ~ ~ ~
~
JY"
HO (94)

OH

~ ~ ~ ~ ~ ~ ~ ~

OH
HO (1S7)

The first acetylenic ketone, pectenolone, was ascribed the structure (166)
on the basis of spectral evidence [323]. Asterinic acid [121], known for a
long time, was recently shown to be a mixture of the di- and mono-acetylenic
derivatives of astaxanthin (203), namely (200) and (202) [124, 326]. These
acetylenic astaxanthin derivatives undergo transformation to the correspond-
ing diosphenols (cf. Section B.5) on treatment with alkali, and are reduced
with complex metal hydride to the corresponding tetrols [124].
The allenic xanthophylls with a C 40 -skeleton, comprising fucoxanthin (190),
fucoxanthinol (189), isofucoxanthin (179), isofucoxanthinol (178), neoxanthin
(122) ( = trollixanthin ?), neochrome (131) ( = trollichrome ?), deepoxyneo-
xanthin (86) and vaucheriaxanthin (124), all have the structural element (24)
(Scheme 22) in common. The stability of the allenic function in these compounds
is presumably due to the adjacent cyclic end group.
Fucoxanthin (190), first isolated in the crystalline state in 1914 [327], is
the longest known and chemically best studied representative of the class.
Work leading to its structure determination has been compiled [39, 146].
Although modern spectroscopy was a necessary tool in the structure deter-
mination of fucoxanthin, this is an example where chemical degradation was
required. The main reactions of fucoxanthin (190) are summarized m
Schemes 23 a -c.
R 2 0:r:y0Ac

.~
-7

R 10
(190) R 1 = R 2 = H
(25) R 1 =Ac, R 2 =H
(26) R 1 =Ac, R 2 = Si(CH 3 h
108 SYNN0VE LIAAEN-JENSEN

Chemical evidence for the nature of the six oxygen atoms of fucoxanthin,
C42 H 58 0 6 , has been presented. The presence of two of these as an acetoxy
group is confirmed by quantitative determination of acetic acid after saponifica-
tion. One constitutes a hydroxy group in a position accessible for acetylation,
as demonstrated by the formation of a monoacetate (25). The fourth oxygen
atom is present as a tertiary hydroxy group, since fucoxanthin acetate (25)
forms a mono(trimethylsilyl ether) (26). The fifth is a conjugated carbonyl
group, demonstrated by hydride reduction to the semifucoxanthols (27) and
two epimeric fucoxanthols (28), all with shorter (octaene) chromophore
(Scheme 23 a). The final oxygen function constitutes an epoxy group in structural
surroundings permitting rearrangement to isofucoxanthin (179) and iso-
fucoxanthinol (178) under weakly alkaline conditions, and furthermore of
conversion of the fucoxanthols (28) to the fucochromes (30) on treatment
with acidified chloroform. The latter reaction presumably occurs by allylic
dehydration and subsequent epoxide-furanoid rearrangement.

HO:trOR

.:9'
(><Yy~~
Al 20 3
(190)
HO~OHI)
(178) R=H
(179) R=Ac

HO~OH

~X/~
-~
(27) R=Ac HCI-cHCI 3
(28) R=H
HO~
(30)

HO:trOH
1 LiAIH4

.:9' (67)

~~
HO
(189) X=

Scheme 23a
 III. Isolation, Reactions 109

The usual bicyclic carbon skeleton is demonstrated by the conversion of

fucochrome (30), on treatment with lithium aluminium hydride, to zeaxanthin
(67). The hydrogenolytic opening of the furanoid end group by such treatment
has been discussed in Section B.7, and the conversion of the allenic end group
parallels that of neoxanthin (122), discussed below. X-ray analysis [280] of
a derivative of the allenic methyl ketone (31) (Scheme 23c), obtained on
ozonolysis of fucoxanthin (190), provides strong support for the allenic end
group. Stereochemical aspects are discussed in Chapter V. The other reactions
quoted in Schemes 23 a -c are in full support of the structure (190) accepted
for fucoxanthin. Thus oxidation of the fucoxanthols (28) with dichlorodicyano-
p-benzoquinone (DDQ) furnishes fucoxanthinol (189); the tertiary hydroxy
group may be removed by dehydration with phosphorus oxychloride to
(25) (179)

j POCI 3
lA~<ym~~hOA<
~X/-;::;-
-~
AcO~OH()
(32) (34)

1 POCI 3

(36)

(33)
Scheme 23b
 110 SYNN121VE LIAAEN-JENSEN

products with unchanged chromophore (25 -+ 32 and 34-+ 35), and the anhydro-
acetate (32) may be reduced to the anhydrofucoxanthols (36), which in turn
may be converted to the furanoid product (33). Moreover, further oxidative
degradation products obtained with zinc permanganate include the aldehydes
(37}-(42), all in accordance with structure (190) for fucoxanthin.

0~ ~yx;
H:vOAc
(31) (37)

(190)

HO (39)

HO (40)

~ ~ ~ CHO OHC~CHO
(41) (42)
Scheme 23c

Isofucoxanthin (179) is presumably formed from fucoxanthin (190) by

nucleophilic attack at the C-7 methylene [39, 146] (Scheme 24). Since isofuco-
Be
;J
xU/R
HOu~~ -+
HO
W R

(190) (179)
Scheme 24
 III. Isolation, Reactions 111

xanthin (179) has been isolated from egg yolk [328] it must be considered to be
a natural carotenoid. Pentaxanthin [322, 329] has been shown to be identical
with the semisynthetic isofucoxanthinol (178) (Scheme 23a) [330].
Another common carotenoid with allenic structure is neoxanthin (122) [1].
Foliaxanthin [331] is now known to be identical with neoxanthin [332].
Neoxanthin, C40 H 56 0 4 [1, 246], yields a diacetate (43) [246, 333]. The di-
acetate contains a tertiary hydroxy group, which may be dehydrated to (44)
(Scheme 25) with phosphorus oxychloride in pyridine.

(122) R=H
(43) R=Ac

Neoxanthin undergoes the characteristic epoxide-furanoid oxide re-

arrangement [24, 334, 335] to the neochromes (131). Before the allenic
arrangement in neoxanthin was recognized by spectroscopic methods [336],
a partly correct structure was suggested for neoxanthin [333]. Neoxanthin
was later assigned structure (122) [336], confirmed by other workers [338, 339].
The neochromes (131), a mixture of epimeric furanoid oxides, may be
converted to zeaxanthin (67) on hydride reduction [243, 246]. The reduction
of the furanoid oxide is analogous to the reaction discussed in Section B.7c.
The reaction of the allenic end group has been explained by reduction of the
oc-hydroxy allene group with subsequent 1,2-elimination from the resulting
6',7'-dihydro derivative (45) [246] or by a concerted mech~nism [340]
(Scheme 25). The conversion of neoxanthin (122) to diadinochrome (3) in the
presence of acid [154] has already been commented on (Section B.7). The
stereochemistry of neoxanthin (122) is discussed in Chapter V. Neochrome
(131) may be a natural carotenoid [28, 30, 341] or an artifact produced in
the presence of plant acids during the isolation of neoxanthin.
Trollixanthin, C40 H 56 0 4 [360, 361], considered to have structure (123)
at a time when proton magnetic resonance and mass spectrometry were not
yet available, undergoes the characteristic furanoid rearrangement to trolli-
chrome (132) [22], supporting its epoxide character [342]. Support for the
presence of a tertiary hydroxy group in trollixanthin was derived from the
facts that trollichrome gave only a diacetate on acetylation and that per-
hydrotrollichrome acetate on saponification gave the dial anhydroperhydro-
trollichrome (C40 H 740 3 ). However, it has been argued that the bisallylic
tertiary alcohol structure is not likely to be preserved under the conditions
required to effect the furanoid rearrangement [319], and it has been suggested
that trollixanthin may be a stereoisomer of the allenic neoxanthin (122) [146].
 112 SYNNOVE LIAAEN-JENSEN

OAc

"""" """" """" """" """" """" """"

(43)
POCI 3
i:X~
AcO .· ,r•
(44)

OH

"""" """" """" """" """" """" """"

i:C~
HO (3)

(122)
~ OH

"""" """" """" """" """" """" """"

~ 0~
1
HO ,r:H (131)
LiAIH 4

Hl~~R
HO ~ "'~ (45)
H OH

l OH

"""" """" """" """" """" """" """" """"

HO (67)

Scheme 25

While the recent report of a direct comparison between trollixanthin and

neoxanthin supports this assumption [534], further criteria of identity are
desirable.
A partly characterized carotenoid has recently been considered to be
deepoxyneoxanthin (86) on the basis of the formation of a monoacetate, a
diacetate, a diacetate mono(trimethylsilyl ether), an allylic methyl ether with
acidic methanol, and conversion to diatoxanthin (66) on treatment with
acidified chloroform (cf. Section B.2c) [155]. Further characterization of each
compound would be desirable. Trollein [24] is thought to be identical with
deepoxyneoxanthin (86) [155].
 III. Isolation, Reactions 113
Vaucheriaxanthin [535] was recently assigned structure (124) [536] on
the basis of spectral characterization [537] (except proton magnetic resonance
spectrum) and chemical reactions on the micro-scale [535, 537]. Vaucheria-
xanthin, a natural monoester, gave a diacetate on acetylation and the diacetate
a mono(trimethylsilyl ether) on silylation. After saponification vaucheria-
xanthin furthermore underwent epoxide-furanoid rearrangement and gave
a dimethyl ether on treatment with acidified methanol. One of the allylic
methoxy groups was tertiary. Vaucheriaxanthin diacetate was stated to re-
arrange to the corresponding furanoid acetylene (cf. Section B.2 c) on treatment
with acidified chloroform. Experimental details are not given.

b) Epoxides and furanoid oxides

The discovery of this class of carotenoids is due to Karrer's school. The
fundamental studies carried out in the 1945-1950 period have been reviewed
in detail [2, 231] and further work up to 1967 ably summarized [240]. The
discovery of the acid-catalysed rearrangement of the epoxides (Section B.7)
merely by the use of aged chloroform has been expertly described [343]. The
recognition of the structural implications of this reaction and of the relation-
ship between the naturally occuring epoxides and those accessible by partial
synthesis (Section C.3a) was a major reason for this success.
The extensive use of trivial names in this series, often with no structural
implication, can be rather confusing. In both the epoxide and furanoid series
geometrical isomerism at C-5, C-6 or C-8 is encountered; for a full treatment
of the stereochemistry see Chapter V. Since in many cases chromatographic
separation of the isomeric epoxides and furanoid oxides may be accomplished,
this serves to complicate the picture. For convenience the trivial names of
several naturally occurring epoxides and furanoid oxides and their systematic
relationship to the parent carotenoid with P-type end group are given in
Table 1. In addition to the twenty-one derivatives listed there, the epoxidic/
furanoid C40 -carotenoids also include the acetylenic diadinoxanthin (118) and
the allenic fucoxanthin (190), fucoxanthinol (189), neoxanthin (122), trolli-
xanthin, vaucheriaxanthin (124), neochrome (131) and trollichrome, dealt with
in the previous section, as well as phytoene 1,2-epoxide (116) discussed below.
Attention should be given to the fact that it is difficult to prove beyond doubt
the natural occurrence of furanoid oxides, since these are so readily formed
by rearrangement of the corresponding epoxides during the isolation procedure.
In modern work proton magnetic resonance and mass spectrometry are
indispensable methods for establishing the structure of the epoxidic and
furanoid carotenoids (Chapter IV). Their reactions are treated here.
The P-carotene derivatives (Table 1) may be prepared by treatment of
P-carotene (3) with monoperphthalic acid [239, 344] giving the monoepoxide
(114), the diepoxide (133) and the partly rearranged furanoid derivative luteo-
chrome (136). When kept in acidified chloroform the monoepoxide (114)
yields its furanoid derivative mutatochrome (125), and the diepoxide (133)
gives luteochrome (136), aurochrome (139) and, most interestingly, also small

Carotenmds 8
114 SYNN0VE LIAAEN-JENSEN

Table 1. Epoxides and furanoid oxides derived from carotenoids with P-type end groups

Parent carotenoid Structural feature Trivial name

5,6-epoxide 5,8-furanoid
oxide

P-Carotene (3) + P-Carotene monoepoxide (114)

++ P-Carotene diepoxide (133)
+ Mutatochrome (125), citroxanthin,
flavacin
+ + Luteochrome (136)
++ Aurochrome (139)
Cryptoxanthin (39) + Cryptoxanthin monoepoxide (117)
+ Cryptoflavin (127)
++ Cryptoxanthin diepoxide (134)
++ Cryptochrome (140)
Rubixanthin (45) + Rubixanthin epoxide
+ Rubichrome (128)
Zeaxanthin (67) + Antheraxanthin (119)
+ Mutatoxanthin (129)
++ Violaxanthin (135), violeoxanthin
+ + Luteoxanthin (138)
++ Auroxanthin (141)
ex-Carotene (5) + ex-Carotene monoepoxide (115)
+ Flavochrome (126)
Lutein (73) + Taraxanthin (120)
{ Flavoxanthin (130)
+ Chrysanthemaxanthin (130)
Capsanthin (170) + Capsanthin monoepoxide (188)

+ present once; + + present twice.

amounts of P-carotene (3) (Scheme 26). Both citroxanthin [345] and flavacin
[222] have been identified with mutatochrome (125) [200, 346]. Hydride
reduction of mutatochrome results in P-carotene (3) and 5-hydroxy-5,6-di-
hydro-P-carotene (46); the latter is reported to give mainly a-carotene (5) on
dehydration with phosphorus oxychloride in pyridine [200]. Hydride reduction
of P-carotene monoepoxide (114) yields 5-hydroxy-5,6-dihydro-P-carotene (46)
[244] and P-carotene (3) [243], and similar treatment of P-carotene diepoxide
(133) gives P-carotene (3) in good yield [243].
The cryptoxanthin oxides (Table 1) may be prepared from cryptoxanthin
(39) acetate by perphthalic acid oxidation [236], but have been less extensively
studied chemically [34, 347, 348]. As already mentioned, treatment of crypto-
xanthin (39) with lead tetraacetate is reported to give cryptoflavin (127) among
other products [279].
Rubixanthin 5,6-epoxide and the furanoid derivative rubichrome (128)
have been described [349].
f f f
f f f f
f f f f
.
f j f f
~

0
0 f
0..
----+ ----+
f f f f
f f f f
til 0:::
G'
f N
:::- f ~ f ~ f :::-
~

f f f

\
:I:
. ;
. """'.,
~
(;)
.,
8
....l ~ :I: ..d
<.)
r/J

f f 0 f

f f f f
f f f f
..

---
:I:
f ~ f f (;) f
::l :I:
----+
f §: f §: f f
f f f f
;;;-
f §: f ....
~
:::-
f :::-
~
f ~
:::-
f f f f
f fo

a;'
(;) <
\ :I:....l I
116 SYNN0VE LIAAEN-JENSEN

The oxide derivatives of zeaxanthin (Table 1) have all been isolated from
natural sources and have been prepared by partial synthesis from zeaxanthin
(67) diacetate by perphthalic acid oxidation [350]; for the stereochemistry
see Chapter V. They undergo the same type of reactions as discussed for the
p-carotene oxides in Scheme 26. Thus violaxanthin (135), on treatment with
acidified chloroform, furnishes auroxanthin (141), mutatoxanthin (129) and
some zeaxanthin (67) [351], and on hydride reduction affords zeaxanthin
(67) [243].
oc-Carotene monoepoxide (115) has also been prepared by partial synthesis
from oc-carotene (5) [352], and may be converted back to oc-carotene by treat-
ment with lithium aluminium hydride [243].
Lutein epoxide (120) (Scheme 27) has been partially synthesized by the
general method [350]. A discussion of its natural occurrence appears in
Chapter II. Taraxanthin, characterized as a hydroxylated lutein epoxide
C40 H 56 0 4 [353], has recently been claimed to be identical with lutein epoxide
[354] without sufficient criteria of identity. Other reports [246, 355] suggest
that taraxanthin may have been a mixture, with lutein epoxide (120) as a
major component. Taraxien has been described as a natural fatty acid ester
of taraxanthin [356]. The furanoid derivatives of lutein epoxide, flavoxanthin
(130) [357, 358] and chrysanthemaxanthin (130) [350, 359], are presumably
epimeric at C-8.
OH

HO

(116)
Scheme 27

Capsanthin monoepoxide (188) [33] is not a fully characterized carotenoid.

The more recently described epoxide is of a new type and belongs to the
aliphatic series. Phytoene 1,2-epoxide (116) (Scheme 27), C40 H 640, cannot be
acetylated, saponified or reduced with sodium borohydride. The structural
assignment is based on mass spectrometric data and is in agreement with the
triene chromophore [230].

c) Glycosides
The first sugar derivative of a carotenoid studied, crocin (271), is not a
true glycoside, but a carbohydrate diester of a C 20 -dicarboxylic carotenoid
acid (see Section D.4e). A long time elapsed after this pioneering study by
Karrer's school [362] before the glycosidic carotenoids were recognized.
 III. Isolation, Reactions 117

Structures have been assigned to fourteen glycosides of C 40-xanthophylls, two

glycosidic apo-carotenoids (Section D.4d) and two glycosidic C 50-carotenoids
(Section D.Sc) during the last four years. Several more strongly polar caro-
tenoids are expected to be glycosidic in nature.
The structure elucidation has partly been based on proton magnetic
resonance and mass spectrometry of the acetylated glycosides (Chapter IV).
In addition to hydrolysis of the glycoside followed by paper-chromatographic
identification of the sugar moiety, elimination of allylic glycosides by treatment
with acidified chloroform or hydrogenolytic cleavage with lithium aluminium
hydride (Section B.8) have permitted the isolation of the aglycone in a structur-
ally modified form. The aglycone does not survive the conditions required to
effect acid-catalysed hydrolysis of the sugar residue. Because of their low
solubility in water enzymatic hydrolysis of the pure carotenoid glycosides has
not been accomplished. The glycosidic carotenoids are relatively polar and
are best purified in the form of acetates. The stereochemistry of the sugar
moiety and of the glycosidic linkage has been studied by enzymatic methods
and proton magnetic resonance spectroscopy. Since the glycosides have all been
studied in recent years, most of them are fully characterized spectroscopically.
In spite ofthe fact that tertiary glycosides in general are rare in Nature [363],
many of the carotenoid glycosides have proved to be tertiary: phleixanthophyll
(77), 4-ketophleixanthophyll (172), myxobactin (47), myxobactone (159), OH-
chlorobactene glucoside (54), rhodopin glucoside (57), rhodopin-20-al o-gluco-
side (145) as well as two apo-carotenoid glycosides, (253) and (255). The others
are secondary glycosides, namely myxoxanthophyll (90), aphanizophyll (94a),
4-ketomyxol 2'-(5-C-methylpentoside) (176), myxol 2'-(0-methyl-5-C-methyl-
pentoside) (91) and glucoside (92), oscillaxanthin (95) and oscillol 2,2'-bis-(0-
methyl-5-C-methylpentoside) (96), the last two being 2,2'-diglycosides. Two
primary glycosides have been reported in the C 50-series (SectionD.5c). Varia-
tions in the sugar moiety occur also, glucosides, 0-acylglucosides [413, 546], a
mannoside, rhamnosides and 0-methyl-5-C-methylpentosides having been
reported. In the case of some of the glycosides a large number of derivatives
have been prepared. Their more characteristic reactions are discussed below.
The glycosides first studied [252], phleixanthophyll (77) and 4-ketophlei-
xanthophyll (172), could be separated only as peracetates. The peracetates
furnished no trimethylsilyl ethers. On allylic oxidation with p-chloranil,
phleixanthophyll (77) provided 2'-dehydrophleixanthophyll (47), which formed
no elimination products on treatment with acidified chloroform. Phlei-
xanthophyll (77) itself, however, is smoothly converted to 3,4-didehydro-
torulene (48) on treatment with acidified chloroform (Scheme 28). Acid hydro-
lysis of (77) gave o-glucose. A partial synthesis of 2'-dehydrophleixanthophyll
(47) peracetate was achieved in low yield by a Koenigs-Knorr reaction of the
synthetically available aglycone.
Allylic oxidation of 4-ketophleixanthophyll (172) gives the correspond-
ing w,w' -diketone, which is selectively reduced by treatment with zinc-
acetic acid (Section B.5a). Treatment of 4-ketophleixanthophyll (172) and 4-
118 SYNN0VE LIAAEN-JENSEN

o-Glucosyl
?
p-Chloranil ~
H

(77)
?
HCI-CHCI 3 j o-Glucosyl

HCI-CHCI 3 1

HCI-CHCI, ~

I
o-Glucosyl
(49)

?
1
(172) D-Glucosyl

HCI-CHCI 3

(50)
Scheme 28
 III. Isolation, Reactions 119

hydroxyphleixanthophyll (49) with acidified chloroform gave 4-ketotorulene

(50) and 3,4-didehydrotorulene (48), respectively, in slow reactions. These
reactions differ in principle from the elimination reaction of phleixanthophyll
(77) giving 3,4-didehydrotorulene (48). Other reactions of (77) and (172) have
been described and discussed [252].
Myxoxanthophyll, first isolated and partly characterized in the classical
period [221, 364], has recently been shown to be a mixed carotenoid glycoside
in which the rhamnoside (90) (Scheme 29) is the main component [ 46]. By
spectrometric methods the presence of the hexoside (92) has also been de-
monstrated [46]. Both glycosides have the aglycone (51), called myxol [253],
in common. Many derivatives of myxoxanthophyll have been prepared [ 46].

HO

'<::::::: '<::::::: '<::::::: '<::::::: '<::::::: '<::::::: '<::::::: '<::::::: '<::::::: '<::::::: '<:::::::

(52)

HCI-CHCI, r ?

'<::::::: '<::::::: '<::::::: '<::::::: '<::::::: '<::::::: '<::::::: '<::::::: '<::::::: '<:::::::

HO (90)

LiAIH4 1 LiAIH4

HO
(44)

OH
(75)
Scheme 29
120 SYNN0VE LIAAEN-JENSEN

(90) gives a tetraacetate and the tetraacetate forms a mono(trimethylsilyl ether).

Treatment of myxoxanthophyll with acidified chloroform results in extensive
rearrangement to an elimination product thought to be identical with 3',4'-
didehydrochlorobactene (52) and a carbohydrate derivative with properties
similar to rhamnitol. Treatment of (90) with lithium aluminium hydride gives
3-hydroxytorulene (44) and sapro xanthin (75) (see Section B.8 b). Acid hydroly-
sis affords rhamnose. Myxol (51) further occurs as an 0-methyl-5-C-methyl-
pentoside (91) with a reaction behaviour analogous to that of myxoxantho-
phyll [253]. A minor carotenoid has been ascribed the related 4-ketomyxol
2'-(5-C-methylpentoside) structure (176) mainly on the basis of spectrometric
evidence and hydride reduction to the corresponding allylic alcohol.

OH OH

OH OH
(53)

OH Rhamnosyl-0

(}-Rhamnosyl OH
(95) 1) Acetylation
2) LiAlH 4

OH

(55)

OH

~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~

(54) OH
POC1 3

(57)
Scheme 30
 III. Isolation, Reactions 121

Oscillaxanthin [364], was recently shown to be a dirhamnoside (95) of a

tetrol (53) [127], called oscillol [253]. Oscillaxanthin provides a hexaacetate,
which on silylation gives a mono- and a bis-(trimethylsilyl ether). Hydride
reduction of the peracetate under forcing conditions yields the diol (54) and
the tetradecaene monool (55). Dehydration of the former with phosphorus
oxychloride gives (56) and (57) with tridecaene chromophore (Scheme 30).
Glycoside hydrolysis of oscillaxanthin gives rhamnose.
More recently an oscillaxanthin-like carotenoid has been shown to be an
oscillol 2,2'-bis(O-methyl-5-C-methylpentoside) (96) mainly on spectral evi-
dence and by acetylation and silylation reactions [253].
The structure of rhodopin glucoside (57) is based on spectral evidence for
the tetraacetate and identification of the sugar as glucose after hydrolysis [365].
The related rhodopin-20-al o-glucoside (145) has been spectroscopically charac-
terized. o-Glucose was identified after hydrolysis, and (145) furthermore under-
went the reactions characteristic of rhodopin-20-al (144) discussed below. The
ease with which (145) undergoes aldol condensation with acetone to the C 43
methyl ketone is remarkable [365].

d) Methyl ethers
The naturally occurring carotenoid methyl ethers, of which there are at
present twenty-nine, are all tertiary ethers with the methoxy group(s) in 1(1')-
position. Other structural features encountered in these methyl ethers are iso-
propylidene, trimethylaryl, tertiary hydroxy groups in !'-position, conjugated
keto groups in 2,2'- or 4'-positions and a cross-conjugated aldehyde group in
20-position. With the single exception of okenone (181) with one cyclic end
group the carotenoid methyl ethers are all aliphatic with a conjugated polyene
chain of 7-13 double bonds. They are structurally and biosynthetically (Chap-
ter II) closely related.
The carotenoids of this series have all been studied in recent years. The
majority are fully characterized spectroscopically (Chapter IV), and several
have been prepared by total synthesis (Chapter VI). They have no asymmetric
centre so the absolute stereochemistry poses no problems.
Type reactions of the methoxy groups have been commented on in Sec-
tion B.6; such characteristic reactions of the individual carotenoids, when
reported, are dealt with here. They will be treated according to the biosynthetic
series to which they belong. The four series will be regarded in historical order
of characterization.
Spirilloxanthin series. The longest-known methoxylated carotenoid, spi-
rilloxanthin (rhodoviolascin), was characterized in the middle of the thirties by
two schools [229, 366-370]. On permanganate oxidation two products,
identified as bixindial (10) and a C 30 -dial (58), were obtained [368, 369]
(Scheme 31). In a reinvestigation much later [371], using proton magnetic
resonance spectroscopy, the formation of the C 30 -dial (58) was confirmed. In
addition crocetindial (267) could be isolated together with larger amounts of
a C 25 -dial (59), but no bixindial (1 0). The formation of both bixindial (1 0) and
122 SYNN0VE LIAAEN-JENSEN

the C 25 -dial (59) are in accordance with the known structure of spirilloxanthin
(108). The number of methoxy groups was determined in the early work [229]
by means of the Zeisel reaction. Their position was later assigned from spectro-
metric data and the observed stability of spirilloxanthin towards acids [372,
373]. Methoxy groups in such structural surroundings have been shown to

~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~

(108)

JKMn0 4

CHO
OHC ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~

(58)
+

CHO
OHC ~ ~ ~ ~ ~ ~ ~ ~ ~

(59)
+

CHO
OHC ~ ~ ~ ~ ~ ~ ~ ~ ~

(10)
+

CHO
OHC ~ ~ ~ ~ ~ ~ ~

(267)
Scheme 31
 III. Isolation, Reactions 123

cause a low, false isopropylidene value [106, 372]. The structure of spirillo-
xanthin was subsequently proved by total synthesis [159].
The structure of the so-called OH-spirilloxanthin [374] or monodemethyl-
ated spirilloxanthin [375] (105) (Scheme 31), suggested from infrared evidence
and resistance towards acetylation [106, 198], has been confirmed by total
synthesis [376]. For a time this compound was thought to be identical with
a methylation product of bacterioruberin [377] (231) [85, 163] (cf. Sec-
tion D.Sc).
The 3,4,3',4'-tetrahydro derivative (110) of spirilloxanthin (108) (Scheme 31),
was assigned its structure on the basis of spectral evidence. The structure was
proved by total synthesis [378].
OH-P 481 [379], now considered to be identical with rhodovibrin [106,
229, 336], was first given a speculative structure (60) [373] (Scheme 32). The
later structure (106) was assigned to it on the basis of Zeisel determination of
methoxyl, isopropylidene value, resistance towards acetylation, the lack of
allylic elimination and spectral evidence [106, 380], and was confirmed by
total synthesis [337].

Scheme 32

The structure (97) of P 481 [369], now designated anhydrorhodovibrin

[106], was proved by total synthesis [337]. From spectral data (infrared and
visible light), isopropylidene and Zeisel determination and biosynthetic
considerations [106, 381, 372] structure (97) had been favoured, although
symmetrical location of the dodecaene chromophore could not be excluded
[106, 373].
A series of minor mono- and di-methoxy carotenoids have recently been
isolated under particular growth conditions [544]. The structures proposed:
'methoxyphytoene' (104), 'methoxyphytofluene' (103), 3,4,11',12'-tetrahydro-
spheroidene (102), 11',12'-dihydrospheroidene (101), 3,4-dihydrospheroidene
(100), 1-methoxy-1,2-dihydrolycopene (98), 3,4,3',4', 7',8',11',12'-octahydro-
124 SYNN0VE LIAAEN-JENSEN

spirilloxanthin (113), 3,4,3',4',7',8'-hexahydrospirilloxanthin (112), 3',4',7',8'-

tetrahydrospirilloxanthin (111) and 3,4-dihydrospirilloxanthin (109), are based
on visible light and mass spectrometry.
Spheroidenejspheroidenone series. The spheroidene-type compounds treated
below do not form many derivatives. Pigment Y(ellow) [382], later renamed
spheroidene [383], was partly characterized [193, 384]. The full structure (99)
followed from the proton magnetic resonance spectrum [385] and was con-
firmed by total synthesis [386].
Spectral (visible light and infrared) data and resistance towards acetylation
of the so-called OH-Y [198] or OH-spheroidene [387] were consistent with
structure (107). Proton magnetic resonance data and dehydration with phos-
phorus oxychloride (Scheme 33) to spheroidene (99) provided further con-
firmation [217].

~ ~ ~ ~ ~ "<::::: ~ ~ ~

(99)

~ POCI3

"<::::: ~ ~ ~ ~ ~ ~ ~ ~ ~

(107)
Scheme 33

Two minor carotenoids, characterized by electronic spectra and mass

spectrometry, have recently been assigned the structures 11',12'-dihydro-
spheroidene (101) and 3,4,11',12' -tetrahydrospheroidene (102) [388].
The spheroidenone (182) derivatives contain conjugated keto group(s) in
2(2')-position and are chemically more reactive than the spheroidene (99)
derivatives above. Spheroidenone [193] is synonymous with Pigment R(ed)
[382]. Hydride reduction revealed the presence of a conjugated carbonyl group
and Zeisel determination one methoxy group. An isopropylidene value of 1.43
was obtained on ozonolysis [193]. Analytical data were also reported by
others [384]. The structure was solved by proton magnetic resonance spec-
troscopy [385] and confirmed by total synthesis [386]. The reduction product
(4) of spheroidenone is reported to give 3,4-didehydrolycopene (18) on treatment
with acidified chloroform [194] (Scheme 34).
OH-Spheroidenone [387] (185) was earlier referred to as OH-R or
hydroxy-Red [194, 375]. The structure was suggested on the basis of spectral
evidence (visible light and infrared), resistance towards acetylation, and bio-
synthetic considerations, and confirmed by dehydration to spheroidenone (182)
(Scheme 34) and by proton magnetic resonance data [217].
 III. Isolation, Reactions 125

l
POCI 3

""" """ """ """ """ """ """ """ """

(182)

j
LiAIH4

""" """(4) """ """ """ """ """ """

- H 20 j
HCI--cHCI 3

[ "·
~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~
""" "J
j
- CH 3 0H HCI-CHCI 3

""" """ """ """(18) """ """ """ """ """ """ """ """ """
Scheme 34

2,2'-Diketospirilloxanthin (208), synonym P 518 [260], was first considered

to be a monoketo derivative of spirilloxanthin (108) [194]. However, when
larger amounts became available, proton magnetic resonance data [217]
revealed the diketo structure (208), later proved by total synthesis [389]. The
lack of reaction on treatment of its reduction product (5) (Scheme 6d) with
acidified chloroform [194], cf. capsorubol (6) [261], and the behaviour of(208)
126 SYNNiiJVE LIAAEN-JENSEN

on treatment with zinc-acetic acid [217], giving the unstable dihydro product
(61) (Scheme 35), are explained by the diketo structure.

Zn-AcOH 11KOH, 0 2

Scheme 35

The minor carotenoid 2-keto-OH-spirilloxanthin (184) was considered to

be 2-ketorhodovibrin (62) (Scheme 36) [390], at a time when the monoketo
structure was still assumed for (208). Spectral similarity with 2'-dihydro-
phillipsiaxanthin (177) [208] revealed the chromophore, but further data in
support of structure (184) is required. P 512 [375] may have been (208) or (184).

Scheme 36

Okenone series. The structure elucidation of okenone (181) followed from

full spectral characterization, reduction to okenol (63) and allylic elimination
of the hydroxy group to give anhydrookenol (64) (Scheme 37) [133]. Total
synthesis of (181), (63) and (64) has been achieved [284].
Rhodopinal series. Three methoxylated members of this series are reported.
3,4,3',4'-Tetrahydrospirilloxanthin-20-al (147) has been characterized spectro-
scopically [184, 202] and by the formation of the allylic alcohol (65) (Scheme 38)
and of aldehyde derivatives such as the dinitrophenylhydrazone, oxime, oxime
acetate and oxime methyl ether [184]. The location of the cross-conjugated
aldehyde group follows from spectral data [184, 202].
Methoxylycopenal, first considered to be identical with lycopen-20-al (143)
(Scheme 40) [391], was recently assigned structure (146) on the basis of mass-
 III. Isolation, Reactions 127

~ ~ ~ ~ ~ ~ ~ ~ ~ ~

CH 3
(181)

l
LiAIH 4

~ ~ ~ ~ ~ ~ ~ ~ ~ ~

CH 3 0
(63)

- H 20 l
HCl-cHCl 3

~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~

CH 3 0
(64)
Scheme 37

~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~

(147) CH,O

l
LiAIH4

~ ~ ~ ~ ~ ~ ~ ~

(65) CH 3 0

~ ~ ~ ~ ~ ~ ~ ~ ~ ~

(146)
Scheme 38

spectral evidence [202]. A minor compound (145a) is considered to have an

additional double bond. The assignment is supported by mass-spectral evidence
and the result of hydride reduction [365].
128 SYNN0VE LIAAEN-JENSEN

e) Carboxylic acids and methyl esters

In the C 40 -series only one carboxylic acid and its methyl ester are known.
Torularhodin [300] was shown to be a carboxylic acid from its behaviour
towards alkali and the preparation of a methyl ester [224, 300, 392]. The
structure P-apo-2'-carotenoic acid (Cd was considered [392] until total
synthesis of the C 40 -carboxylic acid (211) showed it to be identical with

(211)

KOH 11CH N2 2

(66/50)

l p-Chloranil

Scheme 39

torularhodin [393]. Also the methyl ester (212) is naturally occurring [394],
it has been saponified to torularhodin (211) and reduced to the alcohol (66)
which in turn has been oxidized to torularhodinaldehyde (142) (see Scheme 39).

f) Aldehydes
The aldehydes of the C 40 -series comprise seven carotenoids where a lateral
methyl group, either in 16'- or 20-position, has formally been oxidized to an
aldehyde group.
 III. Isolation, Reactions 129

Torularhodinaldehyde (142) (Scheme 39) is naturally occurring [395, 396]

and has also been prepared by total synthesis [397].
The other six are cross-conjugated aldehydes, four of which (145, 145a, 146
and 147) have already been dealt with under glycosides or methyl ethers.
Rhodopinal (144) [184, 202] is chemically the best studied representative.
Dehydration of (144) (Scheme 40) affords lycopen-20-al (143) [184, 202], which
is the seventh naturally occurring aldehyde.
0

l POCI 3

J
R

(80) R=CH 2 0H (68) R=CH(OCH 3 h

(67) R=CH 2 0Ts (69) R=CH=CHR'
(56) R=CH 3
Scheme 40

The more important reactions [184] of rhodopinal (144), earlier called

warmingone [398], are given in Scheme 40. Hydride reduction affords
rhodopinol (80), the tosylate (67) of which, on hydride reduction, gives rhodopin
(56). Aldehyde derivatives such as the dimethyl acetal (68), oxime, oxime
acetate, oxime methyl ether and dinitrophenylhydrazone have been prepared.
Rhodopinal (144) readily undergoes aldol or Wittig condensation to products
of type (69). The exact location of the aldehyde group follows from proton
magnetic resonance [184] and particularly mass spectral [202] data. The
stereochemistry is dealt with in Chapter V.

g) Ketones
The C40 carotenoid ketones are a major class. In addition to the thirty-two
representatives treated below, fifteen members (pectenolone, mono- and di-
acetylenic asterinic acid, fucoxanthin, isofucoxanthin, fucoxanthinol, isofuco-
xanthinol, capsanthin monoepoxide, 4-ketophleixanthophyll, 4-ketomyxol
Carotenmds 9
130 SYNN0VE LIAAEN-JENSEN

2' -(5-C-methylpentoside), spheroidenone, OH -spheroidenone, 2,2' -diketo-

spirilloxanthin, 2-keto-OH-spirilloxanthin and okenone) have been dealt
with in earlier sections.
Aliphatic ketones. The structure of phillipsiaxanthin (207) has been estab-
lished [208] by comparison with (207) (Scheme 41) prepared by total synthesis
[399]. Phillipsiaxanthin occurs as a tertiary diester, the ease of hydrolysis of
which by alkali [208] is explained by acyl migration [207] (cf. Scheme 9).
Hydride reduction gives oscillol (53). 2'-Dihydrophillipsiaxanthin (177)
presumably occurs as a natural diester and triester [208] and has not been
fully characterized.
OH

"<::::, "<::::, "<::::, "<::::, "<::::, "<::::, "<::::, "<::::, "<::::, "<::::, "<::::, "<::::, "<::::,

0
(207)
OH

OH
l
"<::::, "<::::, "<::::, "<::::, "<::::, "<::::, "<::::, "<::::, "<::::, "<::::, "<::::,

OH
(53)

OH
"<::::, "<::::, "<::::, "<::::,

0
(177)
Scheme 41

P-Carotenone (216) (Scheme 42), a characteristic oxidative degradation

product of P-carotene (3) [ 400-402], was recently reported to occur naturally
[211]. Treatment of P-carotenone (216) with methanolic potassium hydroxide
provides dianhydro-{J-carotenone (70), which on w,w'-reduction gives dihydro-
dianhydro-{J-carotenone (71) [402, 403]. Dihydro-{J-carotenone (72) is likewise
formed on w,w' -reduction of P-carotenone (216) [ 404].
Monocyclic ketones with the keto group in the aliphatic part. The structure
of 2' -dehydroplectaniaxanthin (162) [207] was proved by direct comparison
with a specimen previously prepared by total synthesis [ 405]. 2'-Dehydro-
plectaniaxanthin occurs as a tertiary ester, which is readily saponified (cf.
Scheme 9) to the tertiary alcohol (162); the acid has been identified as linoleic
acid [452]. 2'-Dehydroplectaniaxanthin gives no acetate on acetylation but
a trimethylsilyl ether on silylation. Hydride reduction of(162) affords plectania-
xanthin (76) (Scheme 43), which on treatment with acidified chloroform gives
3,4-didehydrotorulene (48); cf. analogous results for myxoxanthophyll (90)
(Scheme 29).
 III. Isolation, Reactions 131

(3)

1Cr0 3

~ ~ ~ ~ ~· ~ ~ ~

(213)

1Cr0 3

~ ~ ~ ~ ~ ~ ~ ~ ~

KOH (216)

KOH, 02 il Zn-AcOH

0
~ ~ ~ ~ ~ ~ ~ ~

(72)

il
0
(70)
KOH, 0 2 Zn-AcOH

0
(71)
Scheme 42
132 SYNN0VE LIAAEN-JENSEN

~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~

OH
(162)

p-Chloranilll LiAIH4

~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~

OH
(76)

1HCI-CHCI3

~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~

(48)
Scheme 43

~ ~ ~ ~ ~ ~ ~ ~ ~ ~

l
(73)

LiAIH4

~ ~ ~ ~ ~ ~ ~ ~ ~ ~

(164)

lPOCI 3

~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~

(74)
Scheme 44
 III. Isolation, Reactions 133

Semi-P-carotenone (213) has lately been found in nature [211]. It was first
prepared by oxidative degradation [ 404] of P-carotene (3); for reactions, see
Scheme 42. Semi-a-carotenone (214) was more recently reported to be of
natural occurrence [ 406]. It had previously been prepared by chromic acid
oxidation of er:-carotene (5) [ 407, 408]; cf. Scheme 42. Triphasiaxanthin is
considered to be 3-hydroxy-semi-p-carotenone (215), the position of the
hydroxy group being, however, not yet fully established [ 406]. Rubixanthone,
thought to be 4' -ketorubixanthin (163), has not been completely characterized
[ 409]. Another monocyclic 4' -keto derivative is OH-okenone (164), the
suggested structure of which is supported by small-scale total synthesis [391].
Micro-scale conversion to OH-okenol (73) and to the anhydro product (74)
has been reported [391]; see Scheme 44.
Monocyclic ketones with the keto group in the ring. The four ketones to be
described are all derivatives ofy-carotene (8). 1'-Hydroxy-4-keto-1',2'-dihydro-
y-carotene (160) has been converted to 4-keto-y-carotene (151) (Scheme 45) [ 410].

OH

(160)

(151)

OH

l HCI-CHCI 3

(76)
Scheme 45
134 SYNN0VE LIAAEN-JENSEN

Both pigments are naturally occurring [410, 411] and have been prepared by
total synthesis [412]. 4-Hydroxy-y-carotene (75), obtained on hydride reduc-
tion of (151), gives a dehydration product, considered to be the carotene (76),
on treatment with acidified chloroform.
Deoxyflexixanthin (158) has one more double bond in the polyene chain
than (160). The structural assignment is based on spectral (visible light and
infrared) evidence together with acetylation, silylation, dehydration and
reduction data [105]. Through micro-scale conversion to 3,4-dehydrotorulene
(48) (Scheme 46), deoxyflexixanthin (158) has been connected with flexixanthin
(171), described below, and other structurally well-established carotenoids.

(158)

JPOCI3

(150)
jI) LiAIH4
2) HCI-CHCI3

(48)
Scheme46

The oc-ketol flexixanthin (171) has been characterized by spectral data

(visible light, infrared and proton magnetic resonance) of the corresponding
diosphenol (77) and by several reactions carried out on a micro-scale [105].
The structure (171) readily lends itself to the preparation of derivatives, and the
more important reactions are summarized in Scheme 47. The tautomeric
diketo formulation of dehydroflexixanthin (77) is supported by the formation
of a phenazine derivative (78) with o-phenylenediamine. The enol acetate (79)
has been dehydrated to (80), subsequently reduced to the diol (81) and converted
to 3,4-didehydrotorulene (48) on treatment with acidified chloroform. Hydride
reduction of (79) gave the triol (82).
 III. Isolation, Reactions 135

H
HO
(171)

lKOH, 0 2

CC I
H2

er qx~
2

OH
N

j
N

(77)
Acetylation (78)

~ ~
POCI 3
OH I
A cO A cO

j 0
(79) LiAIH4 j 0
(80) LiAIH4

HO y OH
HO «~ H

l
H
(82) (81)
HCI-CHCI 3

~
I
(48)

......'<:::::, "<:::::: "<:::::: "<:::::: "<:::::: "<:::::: "<:::::: "<:::::: "<:::::: "<:::::: •·•
X=

Scheme 47
 136 SYNNOVE LIAAEN-JENSEN

:I:
0 0

~
f f f f
~
f f f f
~
f f 0 f f
:I:
u ~

-
f f .L
u f f
:I: ~
f f f f
~
f f f f
r::- ~ s...
f ...
<:1'1 f G'
~
~ f § f .,..,
~
----
""'~ ~ ~
~
f f f f
~
f f
:I:
0

0 0
:I:

A
:I: ~

""
"'

~
.,
00
'<!"
;? i
j ~ .,E
..<::
(.)
en
0.
0

0
f
0 0
:I:
u f f f5
f f f f
f f f f
:I:

-
f f f f
- ~
:I: :I:
0 0
~ 0 0
~
~

f f f f
f f ~ f f IC
r::-
~
~
s f f
f 0\
'<!"
~
f r-
~
=
Iii'
~
f f f
u
f :I:
0

:I:
0

0 0
:I: :I:
 III. Isolation, Reactions 137

Bicyclic keto carotenoids with the keto group in the aliphatic central chain.
The structure elucidation of the paprika ketones capsanthin (170) and capsorubin
(205) started in the thirties with the work of Zechmeister and Cholnoky [413 a]
and was finished in the sixties by the Cholnoky, Karrer and Weedon schools,
using modern spectroscopic techniques. Thorough reviews on the early
[2, 415] and more recent work [5, 240] have been given elsewhere. Chemical
reactions of capsanthin (170) and capsorubin (205) are given in Scheme 48.
Treatment of capsanthin (170) with hot alkali affords p-citraurin (249) by a
retro aldol cleavage [225]. Oppenauer oxidation of capsanthin (170) gives
the five-ring ketone capsanthone (83) [132, 176], and ozonolysis of capsanthin
(170) acetate provides 1,2,2-trimethyl-4-hydroxycyclopentane-1-carboxylic acid
(84) [415a].
In the same manner capsorubin (205), with two identical end groups, gives
crocetindial (267) on alkali treatment, capsorubone (85) [176] on Oppenauer
oxidation and the cyclopentanecarboxylic acid (84) on ozonolysis of its di-
acetate. As would be expected from structure (205), the corresponding allylic
alcohol capsorubol (6) does not eliminate water on treatment with acidified
chloroform [261], the carbon atoms 5 and 5' being tertiary. Capsanthol (86),
the reduction product of capsanthin (170) on similar treatment, loses 2 moles
of water [261] giving probably (87). The stereochemistry of (84), capsanthin
(170) and capsorubin (205) is dealt with in Chapter V. A total synthesis of
racemic capsorubin has been achieved [416].

(88)

rOppenauer

"""' """' """' """' """' """' """' """'

(157)

!KOH
CHO
"""' """' """' """' """' """' """'
(248)
Scheme 49
138 SYNN0YE L!AAEN-JENSEN

OH
CH 2 0Acyl

o
1 """ """ """ """ """ """ """ """
HO
(175)

lKOH

OH
HOH 2C

""" """ """ """ """ """ """ """

/
HO

NaBH 4
(174)
j 11 Aoo< '" <ioo
2) NaBH 4

O Ac
CH 2 0Ac

OH
""" """ """ """ """ """ """ """
AcO

l
(89)

POCI 3

O Ac
2 0Ac

""" """ """ """ """ """ """ """ """

AcO
(90)

lKOH

OH
2 0H

""" """ """ """ """ """ """ """ """

HO
(88)

~ HCI-CHCI,

OH
CH 2 0H

OH
""" """ """ """ """ """ """ """
HO
(91)
Scheme 50
 III. Isolation, Reactions 139

Cryptocapsin was given structure (157) [175] on the basis of spectral evi-
dence, Oppenauer oxidation to cryptocapsone (88) and alkali degradation to
P-apo-8' -carotenal (248) (Scheme 49).
Siphonaxanthin was isolated [ 417, 418], characterized [ 44, 419-422] and
has been recently assigned the structure (174) from spectral evidence [44, 422]
and from micro-scale conversion to loroxanthin (88) (Scheme 50). The yield
in the dehydration step from siphonaxanthol acetate (89) to loroxanthin tri-
acetate (90) was not reported [421], but would be expected to be low for a
secondary alcohol. Allylic dehydration of siphonaxanthol (91) has also been
stated to give loroxanthin (88) [ 422].
In accordance with structure (174) siphonaxanthin is reported to form an
allylic dimethyl ether [421, 422]. Siphonein (175) (Scheme 50) is a naturally
occurring ester of siphonaxanthin. Since one of the hydroxy groups of si-
phonaxanthin is acetylated faster than the other two and since such a difference
does not exist between the hydroxy groups of siphonein, it has been argued
that siphonaxanthin has a primary hydroxy group which is esterified in
siphonein [ 420]. From visible light absorption data hydrogen bonding was
assumed for siphonaxanthin, but not for siphonein. The assumption of
siphonein being a laurate [ 420] is not supported by mass-spectrometric
results [ 422].
Bicyclic ketones with the keto group in the ring. These are keto derivatives
of P-carotene (3) or ct-carotene (5) with or without hydroxy substituents.
Echinenone [329] (148), identical [424] with myxoxanthin [221] and further
identical [ 425] with aphanin [ 429], is 4-keto-p-carotene. The structure (148),

'<::::: '<::::: '<::::: '<::::: '<::::: '<::::: '<::::: '<:::::

l
(148)

LiAIH4 Oppenauer

'<::::: '<::::: '<::::: '<::::: '<::::: '<::::: '<::::: '<:::::

(40)
1HCI-CHCI3

,:?' ,:?' ,:?' ,:?' ,:?' ,:?' ,:?' ,:?'

(36)
Scheme 51
140 SYNN0VE LIAAEN-JENSEN

arrived at from spectrometric (visible light) data [ 424], was confirmed by

Oppenauer oxidation of 4-hydroxy-P-carotene (40) to echinenone (148) [ 426].
The former may be obtained from P-carotene (3) by the modified N-bromo-
succinimide procedure [285]. The structure of echinenone has been confirmed
by total synthesis [430]. Hydride reduction of echinenone gives isocrypto-

(198)

r
KO-t-Bu, 0 2

(193)

LiAIH4 11 Oxidation

!l
(ISS)

LiAIH 4 Oxidation

"':::, "':::, "':::, "':::, "':::, "':::, "':::, "':::, "':::,

(71) R=H

1 HCI-CHCI3
~ (92) R=CH 3

"':::, "':::, "':::, "':::, "':::, "':::, "':::, "':::,

(93)
Scheme 52
 III. Isolation, Reactions 141

xanthin (40), which on allylic elimination gives retro-dehydrocarotene (36)

(Scheme 51) [259]; cf. Scheme 6 b.
Phoenicopterone, considered to be 4-keto-cx-carotene (149), gives on
hydride reduction a product with properties (visible light spectrum and RF
value) identical with semi-synthetic 4-hydroxy-cx-carotene [ 427].
Canthaxanthin [ 428], lately shown to be identical [ 425] with aphanicin
[429], was found to be identical with 4,4'-diketo-P-carotene (193), obtained by
partial synthesis from P-carotene (3) [285]. The first rational synthesis was
reported in 1958 [ 430]. Characteristic reactions of canthaxanthin are given in
Scheme 52. The base-catalysed autoxidation to astacene (198) has been dealt
with in Section C.3c. Hydride reduction gives isozeaxanthin (71), which
readily reverts to canthaxanthin (193) via (155) [181]. Treatment of isozea-
xanthin (71) with acidified methanol gives the dimethyl ether (92) [431],
whereas treatment with acidified chloroform gives a complex mixture of
compounds, one of which has been identified as 3',4'-didehydroechinenone (93)
[285].
Diketopirardixanthin, considered to be 5,6,5',6' -tetrahydrocanthaxanthin
(194) [ 432], has not been satisfactorily characterized either spectroscopically
or chemically.
3-Hydroxyechinenone (153) has been prepared by total synthesis [182].
Its natural occurrence [ 433] has been confirmed by co-chromatography tests
of the corresponding diosphenol (94) (Scheme 53) [434]. Euglenanone [ 435]

~ ~ ~ ~ ~ ~ ~ ~

HO
(153)

lKOH, 0 2

~ ~ ~ ~ ~ ~ ~ ~

l
HO
(94)

LiAIH4

~ ~ ~ ~ ~ ~ ~ ~

HO (95) R=H
OR (96) R=C 2 Hs
Scheme 53
142 SYNN0VE LIAAEN-JENSEN

is different from (94) [177]. Hydride reduction of the diosphenol (94) gives
3,4-dihydroxy-p-carotene (95), reported to give the allylic ether (96) when
reacted with acidified ethanol [ 433].
3'-Hydroxyechinenone (154) has been characterized by the conversions in
micro-scale to products compatible with structures (97) and (1) indicated in
Scheme 54 [ 436]. The same structure (154) has been suggested for asteroidenone
[437].
Crystalline 4' -hydroxyechinenone (155), obtained by ally lie oxidation of
isozeaxanthin (71), has been characterized [181]; for reactions, see Scheme 52.
Its natural occurrence has been reported [111, 120]; further proof of structure
by oxidation to canthaxanthin (193) is desirable.
The diosphenol (195) corresponding to 3-hydroxycanthaxanthin (196) has
been prepared by synthesis [ 438]. Direct chromatographic comparison of the
diosphenol (195) with phoeniconone, obtained by alkali treatment ofphoenico-
OH

~ ~ ~ ~ ~ ~ ~ ~ ~

I
(154)
0

LiAIH4

OH

~ ~ ~ ~ ~ ~ ~ ~ ~

(97)

l
OH

- H20 HCI-CHCI 3

~ ,:?' ,:?' ,:?' ,:?'

- H 20 l
,:?'

HCI-CHCI 3
,:?' ,:?' ,:?' OJ

(1)
Scheme 54
 III. Isolation, Reactions 143

xanthin [ 427], and also with dehydroadonirubin similarly obtained from

adonirubin [433, 439], supports the structure 3-hydroxycanthaxanthin (196)
for phoenicoxanthin and adonirubin. Adonirubin (196) is reported to give a
monoester on acetylation, a triol (98) on reduction with borohydride and a
dimethyl ether (99) on subsequent treatment with acidified methanol [ 427, 433]
(Scheme 55). In view of the facile autoxidation of (196) to (195), the natural
occurrence of phoeniconone is doubtful.

HO
(195)

~KOH, Oz

~ ~ ~ ~ ~ ~ ~ ~

HO
(196)

1NaBH 4

~ ~ ~ ~ ~ ~ ~ ~

(98/85) R=H ~ (99) R=CH 3

Scheme 55

The structure 3,3'-dihydroxyechinenone (167) has been ascribed to hydroxy-

asteroidenone [437], adonixanthin [433] and to a carotenoid isolated from
green algae [ 440], but further characterization is required. Micro-scale tests
support the formation ofthe derivatives (100), (101) and (102) (Scheme 56) from
adonixanthin (167) [433].
From spectral evidence the structure 4-ketolutein (169) has been inferred
for a natural carotenoid, characterized as the corresponding diosphenol,
named a-doradecin (103) (Scheme 57) [441].
Astaxanthin (203) has been closely studied. Its isolation and structure elu-
cidation have been reviewed elsewhere [2, 240], and its occurrence in protein
complexes is treated in Chapters II and VIII. The synthesis of racemic asta-
xanthin has been reported [182]. Some characteristic reactions are summarized
in Scheme 58. Hydride reductions will be discussed under crustaxanthin (93)
144 SYNN0VE LIAAEN-JENSEN

OH

"""' """' """' """' """' """' """' """'

HO
(100)
0
rKOH, 02
OH

"""' """' """' """' """' """' """' """' """'

HO (167)

l
0

NaBH 4

OH

"""' """' """' """' """' """' """' """' """'

HO
(101)
OH
lH<ll, C2H 5 0H

OH

"""' """' """' """' """' """' """' """' """'

HO (102)
OC 2H,
Scheme 56

OH

"""' """' """' """' """' """' """' """' """'

(169)

l
0

KOH, 0 2

OH

"""' """' """' """' """' """' """' """'

HO
(J03/168a)
0
Scheme 57
 :I:
~ 0
0~ 145

f f f
f f f
f f f
f f f
f 0
--+-
f f
~

f f f
f ~ f i f ~
0
::=, e ~
f f e
f

~
0 0

y
~ 0
:I:

.,..

b
~
00
r£
:s
= ...
6
~
~

z
~
-5
Vl
e...

:I:
0
:I:
0
Q-z ~
z=

f f f
f f f
f f f
f f f
f f f
f f f
f .-;- f \0 f ~
=
!::!-
0
::=, ::=,
f f f

z
:I:

z=b#
0

0
~ -
:I: 0
:I: ~
Carotenmds 10
146 SYNNOVE LIAAEN-JENSEN

(Section D.3 h). Astaxanthin gives a diacetate while alkali treatment in the
presence of oxygen results in astacene (198) the reaction presumably occurring
via the deep blue enolate (1 04) [117]. According to infrared data [153] astacene
exists in the enol form (198) in neutral solutions. The tautomeric bis-0(-diketone
(198a) reacts with a-phenylenediamine to provide the phenazine derivative
(105) [223]. Astacene (198) gives the enolic diacetate on acetylation, but no
methyl ether on treatment with diazomethane [165]; it forms a dioxime (106)
possessing two active hydrogens [223]. Taking into consideration the facile
autoxidation of astaxa~thin (203) to astacene (198) under weakly alkaline
conditions, the natural occurrence of astacene is doubtful.
Guaraxanthin is suspected to be 7,8-dihydroastaxanthin (204) from spectral
(visible light), chromatographic and partition data [442]. Alkali treatment
provides an acidic carotenoid and hydride reduction a product with ca. eight
spectroscopically efficient carbon-carbon double bonds in the chromophore.
The isolation and structure determination of the retro diketone rhodo-
xanthin (209) has been reviewed elsewhere [2, 240]. Rhodoxanthin has been
prepared by partial synthesis from zeaxanthin (67) [288] and recently by
total synthesis [ 444]. Characteristic reactions are given in Scheme 59. Rhodo-
xanthin (209) gives a dioxime [158]. Reduction of (209) with zinc-acetic
acid affords dihydrorhodoxanthin (107), which also gives a dioxime [158].
Meerwein-Ponndorf reduction of dihydrorhodoxanthin (107) gives zea-
xanthin (67) [445], which in turn may be converted to rhodoxanthin (209)
by treatment with manganese dioxide [288]. It has been suggested that
eschscholtzxanthin (84) is an intermediate in this reaction [288]. Lithium
aluminium hydride reduction of rhodoxanthin (209) gives eschscholtzxanthin
(84). By using sodium borohydride the intermediary ketone eschscholtz-
xanthone (192) may be isolated [ 446]. The latter may be oxidized to rhodo-
xanthin (209) by silver oxide [ 446]. Oxidation of eschscholtzxanthin (84) with
manganese dioxide [ 443] or p-chloranil [ 447] gives rhodoxanthin (209).
Eschscholtzxanthone (192) is a naturally occurring carotenoid [ 446], which
on dehydration with acidified chloroform afforded 3-keto-retro-bisdehydro-
carotene (108) [116, 187, 448].

h) Alcohols and esters thereof

The carotenols are by far the major class within the C 40 -series. In addition
to the thirty-four compounds, containing no other oxygen function than
hydroxy groups, which are dealt with in this section, seventy-three of the
carotenoids discussed earlier in Section D are naturally occurring carotenols.
As already mentioned some of these also occur in nature in esterified form.
The carotenols treated here are tertiary, secondary or primary with one
to four hydroxy groups. They undergo the type of reactions discussed in
Sections B.3 and B.4.
M onools-aliphatic. The structures of 1-hydroxy-1,2-dihydrophytoene (61)
and 1-hydroxy-1,2-dihydrophytofluene (60) have been assigned on the basis
of mass-spectrometric data for the corresponding trimethylsilyl ethers (109)
 ,yO
Zn-AcOH
~~~~~~~~~"/ (Y~~~~~v~~
KOH, 0 2
(209) 0' - (107)
Ag 2 0 11 NaBH 4 Meerwein-
l
LIAIH4 Ponndorf

OH ~ OH
~
~
P' P' P' P' P' P' P' P' P' P' u~~~~~y~y~

(192) HO (67)

1NaBH4
"'~Xo·
~-
Mn0 2 or ~ ,OH
'
p-chloraml
LL
P' P' P' P' P'y~y~x a~~~~~y~y
_,?
HO
(84) 0 (108)
Scheme 59

.....
~
148 SYNN0VE LIAAEN-JENSEN

and (110) (Scheme 60) and are in agreement with visible light absorption
spectra and the inertness of the hydroxy groups towards acetylating agents
[449].
Chloroxanthin [ 450] was given structure (58) on the basis of proton
magnetic resonance data [385] and total synthesis [451].
Rhodopin [366], for a while considered identical with lycoxanthin (62)
[375, 379], was ascribed structure (56) on the basis of isopropylidene deter-
mination [380] and failure to undergo acetylation [366, 380]. Proton magnetic
resonance data confirmed this structure [196]. Rhodopin was later obtained
by total synthesis [317]. Dehydration of rhodopin (56) with phosphorus
oxychloride gives lycopene (19). The other free tertiary alcohols mentioned
above would be expected to undergo the same dehydration reaction. 3,4-Di-
dehydrorhodopin (55) has been characterized only by spectral data (visible
light and proton magnetic resonance) [510]. ·
~i(CH 3 h

""':::: ""':::: ""'::::

(109)
~i(CH 3h
0

~ ~

(110)
OH

""':::: ""':::: ""':::: ""':::: ""':::: ""'::::

(56)

1 POCI 3

""':::: ~ ""':::: ""':::: ""':::: ""':::: ""':::: ""':::: ""':::: ""'::::

(19)
Scheme 60

Lycoxanthin, as characterized in the classical period [414], was considered

for a long time to be 3-hydroxylycopene (111), but on obtaining further spectral
data (proton magnetic resonance, mass and infrared) the structure was revised
to 16-hydroxylycopene (62) [453--455]. Lycoxanthin gives a monoacetate (112)
[414, 453] on acetylation, an aldehyde (113) on allylic oxidation with nickel
peroxide [414] and a tetrahydropyranyl ether (114) on treatment with di-
hydropyran [114]. A synthesis ofthe latter (114) has been reported [114]. The
tetrahydropyranyl ether could not be converted to lycoxanthin (62). Treat-
ment of (62) with hydrogen chloride in chloroform under drastic conditions
 III. Isolation, Reactions 149

was stated to give 3,4-didehydrolycopene (18) [ 453] by a reaction which

cannot readily be explained from the structure oflycoxanthin (62) (Scheme 61).
Before the revised structure (62) of lycoxanthin was published, treatment of
the latter, as palmitate, with N,N'-dichlorourea was reported to give 3-hydroxy-
3',4'-didehydrolycopene (115) [275]. The structure of this tridecaene de-
hydrogenation product could probably be established by an allylic hydroxyl
test. The total synthesis of lycoxanthin has recently been effected [ 455 a].

HOH 2C ~

1
HCJ-CHCI 3

~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~

(18)

(112) R = CH 2 0Ac

(113) R=CHO
(114) R=CH 2 0
D 0

OH

~· ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~

(111)
OH

~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~

(115)
Scheme 61

M onools-monocyclic. Rubixanthin, first studied some time ago [ 456, 457],

was given the structure (45) (3-hydroxy-y-carotene) on the basis of quantitative
hydrogenation, acetone formation on ozonolysis, Zerewitinoff determination
and absence of vitamin A activity. Support for the assumption that the hydroxy
group occupied the 3-position was later obtained from resistance towards
allylic dehydration [197] and from proton magnetic resonance data of rubi-
xanthin and its acetate [204]. Gazaniaxanthin [ 458], studied in more detail
in recent times [204, 459, 460], has been shown to be the (lower melting)
5'-cis isomer (46) [460] of rubixanthin; see Chapter V. To celaxanthin has
been ascribed the tentative structure 3-hydroxytorulene (44) [461] and
150 SYNN0VE LIAAEN-JENSEN

although partial syntheses of the latter have been reported [195, 462] (Scheme
62), no direct comparison with authentic celaxanthin appears to have been
carried out.

HO (45)

1) Acetylation
2) NBS
j or 1) Palmitoylation
2) N,N'-Dichlorourea
3) KOH 3) KOH

HO

OH
,-A-,

(116/49)

R ~ ~ ~ ~ ~ ~ ~ ~ ~ ~

(48) (53) OH

1
POCI 3

R ~ ~ ~ ~ ~ ~ ~ ~ ~ ~

(8) (15)

(48) and (8): R =

ex Scheme 62
(53)~d(15)' ~· R-

Aleuriaxanthin also has the y-carotene-type chromophore. It differs

slightly from rubixanthin (45) in polarity, forms an acetate but no normal
ally lie dehydration products. Although the structure 3' -hydroxy-y-carotene
has been considered [ 463], unpublished spectral data (nuclear magnetic
resonance, infrared and mass) sug~est that aleuriaxanthin has an unsubstituted
{3-ring, a terminal methylene group and carries the hydroxy group in 2'- or
 III. Isolation, Reactions 151

3' -position (116) (Scheme 62) [ 464]. Aleuriaxanthin was the first example of a
natural carotenoid with a terminal methylene group; later the structure of
'{3;y-carotene' (3a) has been elucidated [206a]. Aleuriaxanthin occurs as a
fatty acid ester [ 463].
Another monohydroxy derivative of y-carotene (8) is 1'-hydroxy-1',2'-di-
hydro-y-carotene (48) [410]. The latter does not form an acetate and is de-
hydrated to y-carotene (8) according to Scheme 62. A total synthesis of (48)
has been carried out [317].
OH-Chlorobactene (53) has the same hydroxylated end group as (48) and
affords chlorobactene (15) on dehydration with phosphorus oxychloride [197]
(Scheme 62). A total synthesis has been reported [317]. The occurrence of
OH-chlorobactene as a glucoside (54) [257] needs confirmation.
M onools-bicyclic. Zeinoxanthin has been formulated as 3-hydroxy-
oc-carotene (42) [465]. This formulation is supported by its non-identity with
synthetic 4-hydroxy-oc-carotene, the negative test for allylic hydroxyl and the
absence of vitamin A activity. However, the structure (42) is not unequivocally

HO (42)
OH

(43)

established. oc-Cryptoxanthin is considered to be 3'-hydroxy-oc-carotene (43)

[ 466]. It forms an acetate and is vitamin A active. The 3' -position of the
hydroxy group is in accordance with the formation of an allylic methyl ether
with acidified methanol. The same structure has been advanced for physo-
xanthin [ 467] assumed by others [ 466] to be a cis isomer of 3-hydroxy-
{3-carotene (39) (Scheme 63).
Cryptoxanthin [ 468] was assigned structure (39) (3-hydroxy-{3-carotene)
on the basis ofthe visible light absorption spectrum, quantitative hydrogenation
data, Zerewitinoff determination and acetylation data (monoacetate). The
structure has been confirmed by total synthesis [ 469]. Treatment of crypto-
xanthin (39) with N,N' -dichlorourea in ethanol-free chloroform is reported
to give retro-bisdehydrocarotene (14) in 20% yield, whereas cryptoxanthin (39)
palmitate was stated to give 3',4' -didehydrocryptoxanthin {117) after saponifica-
tion [272] (Scheme 63).
Isocryptoxanthin or 4-hydroxy-{3-carotene (40) is available by partial
synthesis from {3-carotene (3) [192] and has also been prepared by total
synthesis [186]. Its natural occurrence has been reported [120, 470]; allylic
oxidation to echinenone (148) (Scheme 51) would substantiate the evidence.
152 SYNN0VE LIAAEN-JENSEN

-;:::? -;:::? -;:::? -;:::? -;:::? -;:::? -;:::? -;:::?

l
(14)

N,N'-Dichlorourea

~ ~ ~ ~ ~ ~ ~ ~

j
HO
(39)
I) Polm,.oylotioo
2) N,N'-Dichlorourea
3) KOH

~ ~ ~ ~ ~ ~ ~ ~

HO
(117)
Scheme 63

Recently the first phenolic carotenoids were isolated and prepared by total
synthesis [144]. 3-Hydroxyisorenieratene (52) gives an acetate on acetylation
[144], but no methyl ether on treatment with diazomethane [166].

~ ~ ~ ~ ~

(82)

~ ~ ~ ~ ~ ~ ~ ~ ~ ~

(81)

l POCI 3

~ ~ ~ ~ ~ ~ ~ ~

(19)

~ ~ ~ ~ ~ ~ ~ ~ ~

(83)
Scheme 64
 III. Isolation, Reactions 153

Dials-aliphatic. The structure of the most saturated representative,

1,1'-dihydroxy-1,2,1',2'-tetrahydro-(-carotene (82) [423], was put forward on
a very tentative basis. The related 1,1'-dihydroxy-1,2,1',2'-tetrahydrolycopene
(81) has been prepared by total synthesis [159]. Its natural occurrence has
been reported [196]. This tertiary dial gives lycopene (19) on dehydration [159]
(Scheme 64).
The structure elucidation oflycophyll (83) parallels that oflycoxanthin (62).
The accepted structure (83) [ 453, 454] is based on full spectral characterization.
Also lycophyll dipalmitate [414] and diacetate [453] have been characterized.
Rhodopinol (80) [184, 202] is another naturally occurring dihydroxy derivative
of lycopene (19); for reactions, see Scheme 40.
Diols-monocyclic. Saproxanthin has been assigned structure (75) on the
basis of spectral evidence (visible light and infrared) and of the preparation
of derivatives on a micro-scale [195], see Scheme 65. It forms a monoacetate

""::::: ""::::: ""::::: ""::::: ""::::: ""::::: ""::::: ""::::: ""::::: "":::::

j
OH
HO (75)

I) Aoo<Yl><km
2) POCI3

""::::: ""::::: ""::::: ""::::: ""::::: ""::::: ""::::: ""::::: ""::::: ""::::: ""::::: "":::::

A cO (118)

KOH !I) Aulyl•tioo

2) NBS

""::::: ""::::: ""::::: ""::::: ""::::: ""::::: ""::::: ""::::: "":::::

HO (45)

(44)

OH
(76)
Scheme 65
 .OH .....
Vl
~
~~~~~ ............ I HO,C----><
CO,H HO,CxCO,H
~a.n•••....,.4

HO- ........- ' I (8) (9)

(73)

1Ni0 2

H'",CH,OH

~0 I ~ ,OH
'

I OHC/~~~~~~"""'
""""' """"' """"' """"' """"' """"' """"' """"' \ (119)
(120/1S6a)

1 CH3 I, BaO

~o.

""""' """"' D~~~~~y~"""'

""""' """"' """"' """"' """"' """"'
(121) H (126)

lLiAIH4 rCH,OH

.OH " .OCOCCI 3

.OCH, ~ OCH,

CH,I, BaO

Scheme 66
 III. Isolation, Reactions 155

which on dehydration with phosphorus oxychloride gives 3-acetoxytorulene

(118), identical with a specimen prepared by dehydrogenation of rubixanthin
(45) acetate.
Plectaniaxanthin (76) (Scheme 65) has been prepared by total synthesis [ 405]
and occurs in nature as a diester [207]. The facile saponification of the
tertiary ester has been explained by acyl migration (Scheme 9). Plectania-
xanthin gives a monoacetate which, in tum, yields a trimethylsilyl ether; for
oxidation and allylic elimination [207], see Scheme 43.
Diols-bicyclic. For the representative with shortest chromophore, tuna-
xanthin [471, 472], the structure 3,3'-dihydroxy-e-carotene (78) has been
suggested [471]. In accordance with this structure tunaxanthin forms an
allylic dimethyl ether with hydrochloric acid in methanol, but has not been
fully characterized.
Lutein, referred to as xanthophyll by Karrer, was first isolated in the
crystalline state in 1907 [ 473]. Further work [2] led to the formulation of
lutein as 3,3'-dihydroxy-oc-carotene (73) [ 474]. Lutein has not yet been prepared
by total synthesis. However, its degradation and other reactions (Scheme 66),
as well as its spectroscopic properties, are fully consistent with structure (73).
Several diesters of lutein (73) have been prepared [ 475, 476].
Permanganate oxidation of lutein (73) affords oc,oc-dimethylsuccinic (8) and
dimethylmalonic acid (9) [477, 478] as well as oc-citraurin (119) [479]. The
isomerization of lutein (73) to zeaxanthin (67) under strongly alkaline con-
ditions has already been mentioned [148]. Oxidation of lutein (73) with nickel
peroxide gives 3'-dehydrolutein (120) [172]. By this means the non-allylic
monomethyl ether (122) could be selectively prepared via the ketone (121) [172].
The allylic methyl ether (123) is available by treatment of lutein (73) with
acidified methanol [170-172] and has been shown to be identical with the
monomethyl ether obtained on treating lutein (73) with methyl iodide and
barium oxide [ 480], the allylic hydroxy group being the more reactive.
Methylation of lutein (73) by the Kuhn procedure [168] gives the dimethyl
ether (124) [480]. Recently it has been reported [173] that methanolysis of
lutein bis(trichloroacetate) (125) gives the allylic methyl ether (123), as well
as the tertiary methyl ether (126), formally resulting from allylic rearrangement
of (123). The preparation of lutein epoxide (120) by perphthalic acid oxidation
has been dealt with before (Section C.3 a). Helenien (74) is lutein dipalmitate
[475, 476,481, 482] and occurs naturally.
Zeaxanthin is 3,3'-dihydroxy-fJ-carotene (67). The isolation and structure
elucidation has been described in detail elsewhere [2, 240]. The structure has
been confirmed by the total synthesis of racemic zeaxanthin [ 483]; for the
stereochemistry see Chapter V. Characteristic reactions of zeaxanthin are
given in Scheme 67.
Oxidative degradation of zeaxanthin (67) with potassium permanganate
yields P-citraurin (249) [ 484] while dry distillation gave toluene, m-xylene
and 2,6-dimethylnaphthalene [485, 486] (cf. mass-spectrometric fragmentation
of carotenoids, Chapter IV). Catalytic hydrogenation gives perhydrozea-
156 SYNN0VE LIAAEN-JENSEN

OH

P" P" P" P" P" P" P" P"

!')
HO (84)

P>lmi<oyb<ioo
2) NBS
3) KOH

OH

~ ~ ~ ~ ~ ~ ~ ~

HO (67)

lKMn04

H,
CHO Pd
~ ~ ~ ~ ~ ~ ~

HO (249)

OH

Scheme 67

xanthin (127) [ 487]. The monomethyl ether and the dimethyl ether have been
prepared by methylation with methyl iodide and potassium t-amyloxide [167],
and several diesters have been characterized [487, 488]. The preparation of
its epoxides and furanoid oxides has been dealt with in Section C.3 a. On
treatment of zeaxanthin (67) dipalmitate with N-bromosuccinimide, followed
by alkaline hydrolysis, conversion to eschscholtzxanthin (84) has been achieved
[488a]. Physalien (68) is the naturally occurring dipalmitate of zeaxanthin
[489].
Isozeaxanthin or 4,4'-dihydroxy-P-carotene (71) has been prepared by
total synthesis [186], or by partial synthesis from P-carotene (3) [192]. Isozea-
xanthin is stated to be a naturally occurring diol [120, 470]. Proof that the
naturally occurring carotenol is (71) by allylic oxidation to canthaxanthin (193)
(see Scheme 52) is desirable. The reactions of isozeaxanthin have been dis-
cussed in connection with Scheme 52.
Eschscholtzxanthin [ 491] w~s assigned structure (84) (3,3'-dihydroxy-
retro-dehydrocarotene) mainly on the grounds of allylic dehydration to
 III. Isolation, Reactions 157

anhydroeschscholtzxanthin (35) [191] (Scheme 6a). Eschscholtzxanthin (84)

may be conveniently prepared by reduction of rhodoxanthin (209) (Scheme 59)
or by dehydrogenation of zeaxanthin (67) dipalmitate (Scheme 67).
The phenolic 3,3'-dihydroxyisorenieratene (79) has recently been found in
nature and also prepared by total synthesis [144]. It forms a diacetate but,
like the monohydroxy analogue (52), no methyl ether with diazomethane [166].
Oxidation with silver carbonate--celite [189] affords an unstable dark blue
product, presumably the corresponding quinone (128) [166] (Scheme 68).

HO (79)

JAgC0 3

(128)
Scheme 68

Triols. The three carotenoids to be discussed are all bicyclic. A compound,

thought to be identical with 3,4,4'-trihydroxy-fJ-carotene (85), has been isolated
but not satisfactorily characterized [470]. (85) may be obtained on hydride
reduction of (196) (Scheme 55).
5-Hydroxy-5,6-dihydrozeaxanthin (87) is stated to be a natural carotenoid
on the basis of spectral (visible light) evidence, formation of a diacetate and
dehydration of the latter with phosphorus oxychloride to a product identified
as zeaxanthin (67) [ 492].
Loroxanthin has been assigned the structure 19-hydroxylutein (88) based
on full spectral characterization and preparation of derivatives [201, 422]
(Scheme 69). It gives a triacetate on acetylation and a monomethyl ether (129)
and a dimethyl ether (130) on treatment with acidified methanol. Oxidation
with dichlorodicyano-p-benzoquinone yielded loroxanthal (131) [201]. Per-
manganate oxidation of loroxanthin (88) triacetate and subsequent reduction
and hydrolysis gave the three apo-carotenols (132), (133) and (134), the
structures of which were deduced from mass-spectrometric and methylation
evidence [ 422].
Tetrols. Crustaxanthin has been given the structure 3,4,3',4'-tetrahydroxy-
fJ-carotene (93) with bis-trans-e,e' conformation [ 40, 490]. Crustaxanthin
occurs naturally as a tetraester [ 40]. Spectral (visible light and infrared only)
and chemical data for crustaxanthin have been presented. Acetylation is
stated to give a diacetate (135) only, due to hydrogen bonding (Scheme 70).
 ......
Vl
00

_OH
0

~~y~~l /'
(131) CH,OH

~ _.,CH,OH
r ~ ~ ~ ~ ~
~DDQ
HO'. . . ~-.......
OH (132)
H,OH I) Acetylation
~ ~ ,OH
2) KMno.
~ ~ ~ ~ ~ ~ ~ ~ ;:OH
3) NaBH4
HOH 2C ~
4) KOH
(88)
~
(133) ~ ,OH
lHCI-CH 3 0H

l HOH,C"'~~~~

~ 'v"OCH, (134)
,OH

~y~
~
HO' "-./
(129)

lHCI-CH 3 0H

, ~ _.......OCH 3
H 2 0CH 3

~~~y~y~

(130)
Scheme 69
 .OH nO-Acyl

"""" """" """" """" """" """" """" """" """" """" """" """" """" """" """"
HO Acyl-0 (136)
(198)
0
v 1 NaBH4 1NaBH4

y~~
"""" """" """" """" """" """" """" """" """" """" """" """"
(93) (137)
1 Acetylation 1HCI-CHC13
!===
OH._
"'0
--
!"""" s·!Sf
~~~~~~~ I o"l F
"""" """" """" """" """" """" """" """" """" :;a
I HO' ~ ......
...
I»
(135) a
s·
1 =
"'

?'~~~~~~~~'/ ~J
1
,?I
,:7 ,:7 ,:7 ,:7 ,:7 ,:7 ,:7 ,:7 ,:7 NaBH. ,:7 ,:7 ,:7 ,:7 ,?y~y -, ~

--- VI
\0
-
(35) (108)
Scheme 70
160 SYNN0VE LIAAEN-JENSEN

Reduction of a natural astaxanthin diester (136) with sodium borohydride

gave a product different from crustaxanthin, particularly with respect to its
behaviour towards allylic elimination (Scheme 70). Whereas crustaxanthin is
stable towards treatment with acidic chloroform, explained by the trans-glycol
arrangement not permitting trans elimination, the semisynthetic tetrol (137),
presumed to be a di-cis oc-glycol, gave mainly a product identified as 3-keto-
retro-bisdehydrocarotene (1 08). On borohydride reduction the latter underwent
spontaneous dehydration to anhydroeschscholtzxanthin (35). Borohydride
reduction of astacene (198) gave a mixture of stereoisomers one of which was
identified as crustaxanthin.
The kinetics of the hydrolysis of the natural tetraester was taken as support
for the preferred configuration of crustaxanthin; for further stereochemical
discussion see Chapter V.

4. Apo-carotenoids and nor-carotenoids with less than forty carbon atoms in the
skeleton
a) Introduction
Carotenoids contammg less than forty carbon atoms in the skeleton,
formally arising by oxidative cleavage of a carbon-carbon double bond of a
traditional C40 -carotenoid, are referred to as apo-carotenoids. Interestingly
enough the structure of the first member of this class, crocetin (269) [506],
was elucidated before the structure of the common C40 -carotenoids were well
established. Other representatives have been studied in more recent years.
The twenty-six best characterized apo-carotenoids will be discussed in the

y
~ ~ ~ ~ ~ ~ ~ ~
...;:•
-:::-
x""

11
9

x""
...;:•-:::-
~
10
15~
14' 12~) ~ 1~y
•.
~13')
15' H-o ""'o

~
11 15 14' 12' 10'
y
~ ~
...;:•
-:::- 10 12
x""
Scheme70a
 III. Isolation, Reactions 161

present section and are arranged according to the functional groups as in

Section D.3. The apo-carotenoids are usually aldehydes, methyl ketones,
carboxylic acids or esters thereof.
Recently the structure of the first member of the nor-carotenoids, actinio-
erythrin (272), has been established [41]. In the nor series, carbon atoms are
formally removed by processes other than cleavage of carbon-carbon double
bonds. In the case of actinioerythrin this occurs presumably by ring con-
traction resulting from oxidation, benzilic acid rearrangement and de-
carboxylation [ 493]; cf. Scheme 20. In the case of peridinin (273), a novel nor-
carotenoid with butenolide structure and formally lacking one lateral methyl
group and two carbon atoms from the polyene chain, the biosynthetic forma-
tion may be rationalized as in Scheme 70a [549].

b) Acetylenes and allenes

The first acetylenic apo-carotenoids have recently been characterized
[494], among these hopkinsiaxanthin [495] and a carotenoid referred to
as F1 . Their exact chemical relationship is not clear, but both give the diosphe-
nol hopkinsianone (138) on alkali treatment (Scheme 71). F1 (or hopkin-
Hopkinsiaxanthin

!KOH, 02

(138)

lKOH, 02

(<Y~
uoy 0
(247)

~~
HOAA (252)
Scheme 71

Carotenoids 11
162 SYNN0VE LIAAEN-JENSEN

siaxanthin ?) is considered to be (247), an apo-carotenoid of asterinic acid

(200/202),judged by visible light, infrared and mass spectra ofhopkinsiaxanthin
and hopkinsianone (138) acetate and the corresponding spectra, excluding
mass spectrum, for F1 .
Structure (252) is in agreement with the observed properties of triopha-
xanthin, i.e. spectral (visible light, infrared and mass spectra) and acetylation
data [494].
The only allenic apo-carotenoid so far reported is paracentrone (246) [330],
which has been fully characterized spectroscopically. Its acetate (139) and the
allylic alcohol, obtained on hydride reduction, have been prepared. By means
of an Oppenauer oxidation an elegant partial synthesis with 9% yield of
paracentrone (246) from fucoxanthin (190) has been achieved [152] (Scheme 72).
The conversion is explained by oxidation of the 3-hydroxy group to the ketone,
thus causing activation of the methylene group at C-4 followed by electron
displacement to paracentrone acetate (139). The acetoxy group was hydrolysed
in the final step under mild alkaline conditions.

"<:::::: "<:::::: "<:::::: "<:::::: "<:::::: "<:::::: "<::::::

~·~ (246)
HO :::>'OH

K 2C0 3 11 Acetylation

"<:::::: "<:::::: "<:::::: "<:::::: "<:::::: "<:::::: "<::::::

~·~ (139)
AcO :::>'OH

r
"<:::::: "<:::::: "<:::::: "<:::::: "<:::::: "<:::::: "<::::::

~·~
AcO :::>'OH

~ Oppenauer

OH

"<:::::: "<:::::: "<:::::: "<:::::: "<:::::: "<:::::: "<::::::

)::(~
AcO :::>'OH (190)
Scheme 72
 III. Isolation, Reactions 163

Peridinin, C 39 H 50 0 7 , was recently ascribed structure (273) [549]. The

structure is based on detailed spectral analysis of peridinin itself and of various
acetylated, silylated and reduced derivatives. Ozonolysis provided the allenic
ketone (31), also obtained from fucoxanthin (190). Sodium borohydride
reduction gave a series of products with pentaene chromophore, one of which
exhibited properties as expected for (139a), Scheme 72a. The results of treat-
ment of peridinin with acid and base [324, 550] are also compatible with
structure (273).

OH

AcO
JX& :H
~ ~ ~ ~

(273)

l NaBH4

HO

AcO
JX& :H
~ ~ ~ ~

OH CH 2 0H

(139a)
Scheme 72a

c) Epoxides
The structure of apo-10'-violaxanthal (259) has been tentatively assigned
in accordance with borohydride reduction to a product with visible light
absorption corresponding to that predicted for compound (140) (Scheme 73),
and furanoid rearrangement to a product compatible with structure (141) [ 496].
The rate of borohydride reduction [ 497] was taken to support the presence
of an aldehyde group [ 496].
Analogous experimental data [ 498] support the structure of apo-12'-
violaxanthal (263).
164 SYNN0VE LIAAEN-JENSEN

CH20H
'<:::::, '<:::::, '<:::::, '<:::::, '<:::::, '<:::::,

HO (140)

~NaBH4
CHO
'<:::::, '<:::::, '<:::::, '<:::::, '<:::::, '<:::::,

HO (259)

!H®

'<:::::, '<:::::,
CHO
'<:::::, '<:::::,

HO

"<::::: "<::::: CHO

HO

Scheme 73
d) Glycosides
The first glycosidic apo-carotenoid has been given the structure methyl
1-hexosyloxy-3,4-didehydro-1,2-dihydroapo-8'-lycopenoate (255) [212] on the
basis of full spectral characterization and chemical reactions; the more
~annosyl

C02H
'<:::::, '<:::::, '<:::::, '<:::::, '<:::::, '<:::::, '<:::::, '<:::::, '<:::::,

(142)

KOHl! CH 2N 2
annosyl

C02CH 3
'<:::::, '<:::::, '<:::::, '<:::::, '<:::::, '<:::::, '<:::::, '<:::::, '<:::::, '<:::::,

(255)

lLiAI~
CH 20H
'<:::::, '<:::::, '<:::::, '<:::::, '<:::::, '<:::::, '<:::::, '<:::::, '<:::::,

(253)
Scheme 74
 III. Isolation, Reactions 165

important conversions are shown in Scheme 74. Acetylation gives a tetra-

acetate. Under strong alkaline hydrolysis the natural methyl ester (255) is
saponified to the free carboxylic acid (142), which on treatment with diazo-
methane reverts to the methyl ester (255). The natural ester cannot be reduced
with sodium borohydride, but reduction is readily effected with lithium
aluminium hydride. The allylic alcohol (253) is resistant to allylic dehydration.
Glycoside hydrolysis of (255) gives a sugar assumed to be mannose. The allylic
alcohol (253) also appears to occur naturally.

e) Carboxylic acids and esters

Apo-carotenoids with the greatest part of the normal carotenoid skeleton
intact are discussed first.
Neurosporaxanthin [539, 540] (234) (Scheme 75) is a Cwcarboxylic acid,
which is methylated with diazomethane to the methyl ester (235), reduced

C02R
~ ~ ~ ~ ~ ~ ~ ~ ~

(234) R=H (235) R=CH 3

C02R
~ ~ ~ ~ ~ ~ ~ ~ ~

(143) R=H (243) R=CH 3

l LiAIH4

CH 20H
~ ~ ~ ~ ~ ~ ~ ~ ~ ~

(144)

jp-Chloranil

CHO
~ ~ ~ ~ ~ ~ ~ ~ ~ ~

(242)

lHID, CH 3 0H

CH(OCH 3 ) 2
~ ~ ~ ~ ~ ~ ~ ~

Scheme 75
166 SYNN0VE LIAAEN-JENSEN

by lithium aluminium hydride to the primary allylic alcohol, which gives the
corresponding carotenal on oxidation with p-chloranil [17]. The methyl
ester (235) has been prepared by total synthesis [393] and occurs naturally
[541].
Methyl apo-6' -lycopenoate (243) has recently been isolated from a natural
source. The structure followed from spectral characterization, chemical
0
C0 2H

0
(150)

t1) H 2
12) Pb (0Ac) 4

(261) R=H--> (146) R=CH 3

1Cr0 3

0
(147)

11) SOCI 2
2) NH 3

0
(148)

0
(149)
Scheme 76
 III. Isolation, Reactions 167

degradation (Scheme 75) and synthesis [ 499]. The natural methyl ester is
saponified under forcing conditions to the free carboxylic acid (143), which
may be reconverted to the methyl ester with diazomethane. Reduction of
(243) with lithium aluminium hydride gave apo-6'-lycopenol (144), which
on oxidation with p-chloranil gave apo-6' -lycopenal (242). The latter readily
formed an acetal (145).
Azafrin (261) is a free carboxylic acid and has a second unique end group.
Its structure was established by Kuhn's school, extensive degradation studies
being used [2]. Some of the characteristic reactions of azafrin will be discussed
(Scheme 76). Methylation with diazomethane gave the methyl ester (146).
Chromic acid oxidation of azafrin (261) gave azafrinone (147) which, via the
corresponding acid chloride, gave azafrinone amide (148) [500]. Treatment of
the latter with alkali gave anhydroazafrinone amide (149) by intramolecular
aldol condensation. In accordance with the ditertiary a-glycol arrangement
azafrin forms no acetates, and perhydroazafrin on treatment with lead tetra-
acetate according to Criegee [501] gave the diketone perhydroazafrinone
(150) [227]. Azafrin (261) gives a mono(trimethylsilyl ether) only, on silylation
[162], which is explained by severe steric hindrance around C-6. The stereo-
chemistry of azafrin is discussed in Chapter V.
Bixin (265) is another apo-carotenoid, whose structure was established
early in the classical period [2]. Methylbixin (151) has been prepared by several
total syntheses [502, 503]; for stereochemistry, see Chapter V. Bixin has been
submitted to extensive oxidative degradation [2]. Conversion into per-
hydronorbixin (152) [504], subsequently prepared by total synthesis in the
early period [505], was of major importance in the structure elucidation of
bixin (265) (Scheme 77).

C02H
H0 2C
(152)

11)2) H2
KOH

C0 2CH 3
H0 2C """<:::::: """<:::::: """<:::::: """<:::::: """<:::::: """<:::::: """<:::::: """<:::::: """<::::::

(265)

lCH2N2

C0 2CH 3
CH 3 0 2C """<:::::: """<:::::: """<:::::: """<:::::: """<:::::: """<:::::: """<:::::: """<:::::: """<::::::

(151)
Scheme 77
168 SYNN0VE LIAAEN-JENSEN

Crocetin occurs naturally as the free dicarboxylic acid (269), as the

digentiobiosyl ester crocin (271) [362] and as the dimethyl ester (270) [542, 543].
More recently the natural occurrence of crocetinmonoaldehyde (268) has also
been well documented [507] (Scheme 78). The structure elucidation of crocin
and crocetin was mainly performed by the Karrer school [2], and the structure
of the dimethyl ester (270) has been confirmed by total synthesis [508]; for
stereochemistry, see Chapter V. In crocetin (269) the formal head-to-head
linkage of the isoprene units in the centre of the carotenoid molecule was
encountered for the first time. Crocetin has been extensively studied chemically
both by oxidation and hydrogenation reactions [2].

(269) R=H
C0 2 R
R0 2 C ~ ~ ~ ~ ~ ~ ~ (270) R=CH 3
(271) R = Gentiobiosyl

CHO
H0 2 C ~ ~ ~ ~ ~ ~ ~
(268)

Scheme 78

f) Aldehydes
The aliphatic and then the monocyclic apo-carotenals will be considered,
dealing first with the representatives in which the largest part of the carotenoid
skeleton is preserved.
The aliphatic carotenals are compiled in Scheme 79. Apo-6' -lycopenal
(242) has been obtained by chromic acid oxidation of lycopene (19) [509],
and its isolation from a natural source has also been reported [395]. The
naturally occurring [395] apo-8' -lycopenal (254) is readily available by total
synthesis [317]. Crocetindial (267) is an important intermediate in the total
synthesis of carotenoids. Its natural occurrence is well documented by spectro-
scopic data [507].

CHO
~ ~ ~ ~ ~ ~ ~ ~ ~

(242)

CHO
~ ~ ~ ~ ~ ~ ~ ~

(254)

CHO
~ ~ ~ ~ ~ ~ ~
OHC
(267)
Scheme 79
 III. Isolation, Reactions 169

The three naturally occurring [395, 511] aldehydes P-apo-2'-carotenal

(233), P-apo-8'-carotenal (248) and P-apo-10'-carotenal (256) have been
prepared by total synthesis [397]. The corresponding allylic alcohols and
their acetates have also been made [397, 512]. P-Apo-8'-carotenal (248) has
also been obtained by permanganate oxidation of P-carotene (3) [513, 514]
(Scheme 80); it forms a semicarbazone. P-Apo-10'-carotenal apparently also
R

~ ~ ~ ~ ~ ~ ~ ~

(3) R=H
(67) R=OH
lKMn04

CHO
~ ~ ~ ~ ~ ~ ~

R
(248) R=H
(249) R=OH

(258)
Scheme 80

occurs as a hydroxy derivative, for which the 3-hydroxy structure (257) has
been considered [511]. The analogous 3-hydroxy-fJ-apo-8'-carotenal (249) is
the long-known P-citraurin [515]. P-Citraurin is obtained by permanganate
oxidation of zeaxanthin (67) [ 484] (Scheme 80) and is naturally occurring
[498, 511, 515]. Finally, azafrinaldehyde (258) is stated to be of natural
occurrence [395].

g) Ketones
Several apo-carotenoids have in recent years been shown to be methyl
ketones. For the ill-characterized tangeraxanthin the hypothetic retro structure
(236) has been. considered [32]. Reticulataxanthin [32] has been assigned
structure (238) on the basis of full spectral data and retro aldol cleavage to
P-citraurin (249) [516, 517] (Scheme 81). By analogous criteria citranaxanthin
is reported to have the corresponding structure (237), lacking the 3-hydroxy
group [517, 518]. Again retro aldol cleavage yielded acetone and P-apo-
8'-carotenal (248), and condensation of the latter with acetone in the presence
170 SYNN0VE LIAAEN-JENSEN

of alkali gave citranaxanthin (237) in ca. 80% yield [518] (Scheme 81). The
structure 8'-hydroxy-7',8' -dihydrocitranaxanthin (239) has been claimed for
another naturally occurring carotenoid [519]. The assignment was based on
spectral evidence (except mass spectrum) and conversion to citranaxanthin
(237) on treatment with hydrochloric acid in chloroform. Aldol condensation
of P-apo-8'-carotenal (248) with acetone was reported to give 8'-hydroxy-
7',8'-dihydrocitranaxanthin (239) in ca. 15% yield (see Scheme 81). Under
these conditions spontaneous dehydration to citranaxanthin (237) might be
expected.
The lower analogue sintaxanthin (244) has been characterized by elementary
analysis and spectral data (except mass spectrum) [520]. Reduction with

0
,;::? ,;::? ,;::? ,;::? ,;::? ,;::? ,;::? ,;::? ,;::? 1?'

HO
(236)

-
0

""""' """"' """"' """"' """"' """"' """"' """"' (249)

HO
(238)

CHO + CH 3COCH 3
""""' """"' """"' """"' """"' """"' """"'
KOH
(248)

!KOH

(239)

l HCI-CHCI 3

(237)
Scheme 81
 III. Isolation, Reactions 171
borohydride gave sintaxanthol (153), also prepared by the action of methyl-
lithium on p-apo-8'-carotenal (248). Oxidation of sintaxanthol (153) with
manganese dioxide gave sintaxanthin (244) [520] (Scheme 82). The structure

""""' """"' """"' """"' """"' """"' """"'

!l
(244)

NaBH4 Mn0 2

""""' """"' """"' """"' """"' """"' """"' """"'

I
(153)

CH3Li

CHO
""""' """"' """"' """"' """"' """"' """"'
(248)
Scheme 82

3-hydroxysintaxanthin (245) has been tentatively assigned to a minor, naturally

occurring carotenoid [512] on the basis of polarity properties and its visible
light absorption data and that of the compound formed by hydride reduction.
In view of the facile aldol condensation of carotenals with acetone providing
methyl ketones [365], it should be checked that the methyl ketones are not
artifacts produced during the isolation.
In addition to peridinin (273) described above, a structure was earlier
assigned to the first member of the nor series, namely the five-ring bis-IX-ketol
actinioerythrin (272). Although actinioerythrin was partly oharacterized in the
thirties [521-523], the structure (272) recently given [41, 493] is based on
full spectral characterization, preparation of some thirty derivatives (cf.
Scheme 83) and chemical conversion of astacene (198) to the five-membered
ring tetraketone violerythrin (154) (cf. Scheme 20), also obtained by careful
alkali treatment of actinioerythrin [287].
Actinioerythrin (272) forms a dioxime, but no acetate or silyl ether. Treat-
ment of actini<;>erythrin (272) with sodium borohydride leads to stepwise
reduction of the two carbonyl groups and hydrogenolysis of the two ester
functions, giving ultimately the tetrol violerythrol (158). The functional groups
of each intermediate, (155), (156) and (157), were established by further chemical
reactions. Oxidation of violerythrol (158) with air in the presence of iodine
gave actinioerythrol (159). Careful alkali treatment of actinioerythrin (272)
172 SYNN0VE LIAAEN-JENSEN

~ >. >.
<>
<>
:>: ~ :>: < :>: <

R
0 6 0 6

0~
- ~
iO'l iO'l
z" z"

~~ t(;
~

~~
0 0 9 0
:>:
0
:>:
0
:>:
~ >.
< ~

~l
f

~1
>.
f
f
<>
<
6 ..,
~
f
-
:>: :>:

0~ -
0 0~ f ""00.,
.,s
f ..c:
f u
r/J

~~ ~~
f
f
f
f 0 0 0
:>:
0
:>:
:>:
f
f

K
f 0 ~
II
:i
0
f ~

~
f
f 0 0~
-
:i

-
~
0

~~
f ~
~

M'
r-
>Q~ zr; ~z

0
~
~
0 0 0 0
~
<

and of actinioerythrol (159) in the presence of oxygen in both cases gives the
blue tetraketone violerythrin {154). Violerythrin reacts very smoothly with
o-phenylenedia,mine providing the bisquinoxaline derivative (160). The two acyl
residues of actinioerythrin are fatty acids which together contain 12-17 carbon
atoms; capric (C 10), undecanoic (C 11 ) and lauric (C 12 ) acids were identified
by combined gas chromatography and mass spectrometry after hydrolysis
and methylation. Actinioerythrin (272) as well as violerythrin (154) show
 III. Isolation, Reactions 173

complex behaviour on treatment with alkali yielding yellow hypophasic

products. After neutralization these are immediately transformed to yellow
epiphasic products, which slowly revert to the blue violerythrin (154). The
structural implications of these reactions have been discussed [41].

5. Carotenoids with more than forty carbon atoms in the skeleton

a) Introduction
Naturally occurring carotenoids with more than forty carbon atoms in
the skeleton escaped detection until mass spectrometry became a routine tool
in the structure elucidation of carotenoid pigments. The first class discovered
comprises C 50- and C45 -carotenoids which are isoprenoid in nature and have
formally arisen by addition of isoprenoid units to 2,(2')-positions at the iso-
propylidene double bond of a traditional C40-carotenoid. Structures have been
advanced for four C45 -carotenoids and twelve C 50 -carotenoids of this type.
Aliphatic, monocyclic and bicyclic representatives are known, and the com-
pounds will be discussed in that order. The structures assigned are mainly
based on spectrometric evidence. Chemical reactions have generally only
served to establish the number and type of functional groups present.
During the past year carotenoids with C 43 -skeletons have been isolated
from photosynthetic bacteria, four having been characterized. These have
formally arisen by aldol condensation of a C40 -caroten-20-al with acetone and
are now shown to be artifacts, since they cannot be isolated when acetone is
omitted in the isolation procedure [365].

b) C 45 -carotenoids
Three C45 -carotenoids are presented in Scheme 84. 2-Isopentenyl-3,4-
didehydrorhodopin (219) is the best characterized representative [164].

~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~

(219)

HOH 2 ~ ~ ~ ~ ~ ~

(218)

12•

HOH 2 C ~ ~ ~ ~ ~ ~

(217)

Scheme 84
174 SYNNiilVE LIAAEN-JENSEN

The assigned structure was based on full spectral characterization, resistance

to acetylation and the formation of a mono(trimethylsilyl ether).
The structures of nonaprenoxanthin (218) and 11',12'-didehydrononapreno-
xanthin (217) have been advanced on the basis of polarity data and visible-
light and mass spectra [524].

c) C 50 -carotenoids
The four aliphatic members are compiled in Scheme 85. Bacterioruberin
[525], at one time considered to be a C 40 -diol [106, 377, 526], has been
ascribed the C 50 -tetrol structure (231) on the basis of full spectral charac-
terization and chemical evidence [85, 163]. The four oxygen functions were
shown to be tertiary hydroxy groups by their resistance towards acetylation
and from detailed studies of the silylation and dehydration (phosphorus
oxychloride) reactions. The silylation reaction was shown to consist of four
consecutive steps giving ultimately the tetraether. The extent of silylation
was verified by alkaline hydrolysis and further silylation of each intermediate.
The dehydration reaction was similarly shown to involve four consecutive
steps. Monoanhydrobacterioruberin (230) and dianhydrobacterioruberin (228)
thus produced were later found to be naturally occurring [16, 85]. The triol
(230) and diol (228) undergo the expected dehydration and silylation reactions
[16, 85]. The fact that the dehydrations ofbacterioruberin (231) and dianhydro-
bacterioruberin (228) give (161) and (162), and do not lead to products with
prolonged chromophore [85] has been commented on earlier, Scheme 7.
Dianhydro-3,4,3',4'-tetrahydrobacterioruberin (229) has been characterized
by spectral evidence (visible-light and mass spectra) of the natural diol and
the bis(trimethylsilyl ether) [16].
The only monocyclic C 50 -carotenoid is a diol (221), fully characterized
spectroscopically, which, in agreement with acetylation and silylation evidence,
possesses one primary allylic and one tertiary hydroxy group [16] (Scheme 86).
Among the bicyclic C 50-carotenoids is the first known member of the
C 50 -class, dehydrogenans-P 439 (226) [142, 527], now referred to as decapreno-
xanthin [524]. The structure is in accordance with detailed spectroscopic
data. The two hydroxy functions were shown to be primary and allylic by
the formation of a diacetate on acetylation and a dialdehyde (163) on selective
oxidation with nickel peroxide (Scheme 86). The dial (163) gave a dioxime
which failed to give the corresponding nitrile on treatment with glacial acetic
acid [142]. Corynexanthin [528] has recently been reported to be a mono-
glucoside (227) of decaprenoxanthin [529].
Structure (223) has been suggested for deshydroxydecaprenoxanthin solely
on the basis of polarity data, visible light absorption and mass spectra [524].
Sarcinaxanthin [530], for a while considered to be identical with decapreno-
xanthin (226) [531], is, according to a recent re-examination [532], isomeric
with decaprenoxanthin, but carries one of the allylic primary hydroxy groups
in 18- or 18'-position. Structure (224) [114] or (224a) is now suggested for
sarcinaxanthin (Scheme 87). Sarcinaxanthin forms, like decaprenoxanthin, a
 @ n < ;=0 /\____(@ @~@

# <
# # # # I
~
'N -;::::; 'N 'N 'N
N 0\ 0\ ~
::!E ,!:::! .:::: ~ -!::
# # - # # ~ #
# # # # #
<
# # # # # <
# ......
......
......
......
til
# # # # # "'0
<> g-
::r
8 .._ .._ .._ .._ s·
# "C # "C # "C # "C
# F
00
"'v. 0 0 0 ::tl
n
,;-
n
,;-
n
,;- p0 I"
# # # # # "'~
s·
f
# # # #
=
"'

# # # #
<
# # # # i -<
;j

# # #
#
-<
i i < -l
V'l
-
 176 SYNN0VE LIAAEN-JENSEN

"<:::::, "<:::::, "<:::::, "<:::::, "<:::::, "<:::::, "<:::::, "<:::::, "<:::::, "<:::::,

(221)

CH 20H
ROH2 C "<:::::, "<:::::, "<:::::, "<:::::, "<:::::, "<:::::, "<:::::, "<:::::, "<:::::,

(226) R=H (227) R=o-Glucosyl

JNi0 2

CHO
OHC "<:::::, "<:::::, "<:::::, "<:::::, "<:::::, "<:::::, "<:::::, "<:::::, "<:::::, "<:::::,

(163)

Scheme 86

HOH 2 C "<:::::,

(224) R=CH 2 0H, R'=CH 3

(224a) R=CH 3 , R'=CH 2 0H

(222)

2 0H

HOH 2 C "<:::::,

(220)
Scheme 87

diacetate on acetylation and an oc,p-unsaturated dial on selective oxidation

with nickel peroxide [531, 532]. Recently a o-glucoside of a C 50-diol has
been characterized by mass spectrometry and hydrolysis [262] and since
 III. Isolation, Reactions 177

it occurs together with sarcinaxanthin it may be a .sarcinaxanthin mono-

n-glucoside (225). By mass-spectrometric studies of a deuterated derivative
it has been demonstrated that the hydroxy groups of the aglucone are located
at different sides of the molecule [262]. With the necessary reservations it may
therefore be inferred that sarcinaxanthin is most likely (224a).
Attempts to reisolate sarcinene [530] in recent years have not been
successful. Sarcinene is tentatively considered to be the bicyclic C 50-hydro-
carbon (222).
The final C 50 -diol to be discussed has two substituted P-rings rather than
the a-rings characteristic of decaprenoxanthin (226) and sarcinaxanthin (224).
This diol (220) (Scheme 87) [16], referred to as C.p. 450, is spectrospopically
fully characterized. It gives a diacetate on acetylation. The further character
and positions of the hydroxy groups follow from spectrometric evidence.
Confirmation of the p-glycol arrangement was sought by reaction with
benzaldehyde [533], but no condensation product was obtained under con-
ditions where the carotenoid survived [16].

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 189

IV. Spectroscopic Methods

W. VETTER, G. ENGLERT, N. RIGASSI and U. SCHWIETER
F. Hoffmann-La Roche & Co. Ltd., Basle, Switzerland

A. Introduction . . . . . . . . . . . . . . . . . . . 190
B. Ultraviolet and Visible Light Absorption Spectroscopy . 192
C. Infrared Spectroscopy . . . . . . . . . . 202
D. Nuclear Magnetic Resonance Spectroscopy. 204
1. End groups . . . . . . 205
2. 'In-chain' methyl groups . . . . . . . . 209
3. Olefinic protons . . . . . . . . . . . 209
a) Vitamin A analogues and apo-carotenoids 213
b) Carotenoids . . . 235
4. Recent developments . . . . . . . . 239
E. Mass Spectrometry . . . . . . . . . . 243
1. Vaporization and thermal degradation . 244
2. Fragmentation . 246
a) Polyene chain 248
b) End groups 252
References . . . . . 265
190 W. VETTER, G. ENGLERT, N. RIGASSI and U. SCHWIETER

A. Introduction

Physical methods have greatly influenced carotenoid research. Prior to the

systematic application of these methods polyene chemistry was a rather trou-
blesome field in comparison with the situation today.
About twenty years ago a main tool, for instance, for the analysis of chro-
matographic fractions was the Carr- Price reaction: A polyene dissolved in
chloroform, when mixed with a saturated solution of SbC1 3 in alcohol-free
chloroform, exhibits a very pronounced bathochromic shift. This reaction is
still used for vitamin A. A transient deep blue colouration of the previously
colourless chloroform solution occurs, and the extinction at 620 nm serves as
a measure of the purity of the sample. Other colour reactions have also been used
in a similar fashion as diagnostic aids: the addition of sulphuric or trichloro-
acetic acid to a polyene causes colour changes very similar to the one of the
Carr- Price reaction. The exact nature of the coloured intermediate of all these
reactions is unknown. The mode of proton addition, however, to conjugated
polyenes has been discussed for vitamin A and anhydrovitamin A (Fig. 1) [1].

CH 2 0Ac

(1)

jj

-:7 -:7 -:7 7"

(2)

jf
""" """ """ """
-:7 -:7 -:7 (f)
+------+

607 nm (H 2 S0 4 -CH 3 0H)

Fig.l. Postulated intermediate carbonium ions of vitamin A acetate {I) and anhydrovitamin A (2)
on reaction with aqueous methanolic sulphuric acid
 IV. Spectroscopic Methods 191

Since the protonation of lycopene (19) has been formulated [2], one might
argue that, in analogy to this, it is possible to explain the colour reaction of the
carotenes as is shown in Fig. 2.

~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~

(19)

Jf
E!>
~ ~ ~ ~ ~ ~ ~ ~ ~ ~

l
E!> c::? c::? c::? c::? c::? c::? c::? c::? c::? c::?

900nm (CC1 3 C0 2 H-C 6H6)

Fig. 2. Postulated intermediate carbonium ions on reaction of lycopene (19) with trichloroacetic
acid in benzene

The approximate position of the absorption maximum observed in these

reactions can be calculated thus:
Amax = 3()().5 + 65.5 X n nm
where n represents the number of effective double bonds (i.e. 5 for vitamin A,
10 for P-carotene) [3]. This equation yields good values for the reactions in 80%
sulphuric acid. It also seems to give satisfactory results for the reaction in other
solvents.
Besides these reactions, C and H analysis was the only method readily
available for characterizing new structures. These methods were of course
often insufficient for this purpose, and this led to time-consuming and some-
times unsuccessful syntheses.
With the availability of new physical methods, the pace of progress began
to quicken. With the ever increasing number of synthetic and naturally occur-
ring materials available for comparison, the physical methods led in turn to
valuable data ap.d empirical rules. It was possible to develop new methods for
the synthesis of polyenes [ 4] and elucidate new structures of minor or compli-
cated carotenoids. In the following pages, the most important results of the
physical methods including absorption-, n.m.r.- and mass-spectroscopy rele-
vant to carotenoid research will be discussed. Optical rotatory dispersion (o. r.d.)
together with X-ray crystallography will be treated in Chapter V.
192 W. VETTER, G. ENGLERT, N. RIGASSI and U. SCHWIETER

B. Ultraviolet and Visible Light Absorption Spectroscopy

Absorption spectroscopy has been used for the characterization of carot-

enoids for some fifty years. Many different types of spectroscopes and photo-
meters were used during the period, and this sometimes resulted in conflicts
with regard to the positions of the absorption maxima and the values of ex-
tinction coefficients [5].
Fig. 3 illustrates the absorption spectrum of lycopene (19) using a Zeiss
grating spectroscope. In the upper half of the picture, a number of dark absorp-
tion bands is visible. These bands are not present in the spectrum of the solvent
below. When the instrument was used visually the position of the absorption
bands had to be measured by moving cross hairs over the spectrum. In Fig. 3
they are positioned at 503 nm. A quantitative analysis could be performed by
measuring the blackening of a photographic plate.
An 'autocatalytic' influence of physical methods on carotenoid research
began in the late forties with the introduction of manually operated photo-
electric spectrometers, which allowed rapid and accurate quantitative analysis.
Spectra of polyenes show, in addition to the main absorption band with its
vibrational fine structure, several minor peaks with or without fine structure.
Under standard conditions the shape of the spectrum is determined by the
number of double bonds, the substituents, cis-trans isomerism and by the
solvent. Table 1 summarizes these influences.

Table 1. Effects of substituents and solvents on the main absorption maximum of carotenoids

Double bond Chain +7 to +35nm

Double bond Ring +5to + 9nm
First carbonyl Chain +28nm
Second carbonyl Chain +lto+ 7nm
First carbonyl Ring + 7nm
Second carbonyl Ring +5to+ 9nm
First carbonyl Ring retro + 9nm
Second carbonyl Ring retro + 5nm
First and second 5,6-epoxide - 8nm
First and second 5,8-epoxide -20nm
Trans-+ cis - 4nm
Normal-+ retro -lOnm
Solvent effect: Ethanol, hexane, petrol. ether Small effect
Benzene, chloroform +15nm
Carbon disulphide +35nm

As the length of a chain of conjugated double bonds increases, additional

double bonds show diminishing effects on the position of the main absorption
band. The apo-carotenal series in Table 2 provides a good example. The batho-
chromic shift for one additional double bond is 23 nm between the first two
compounds in contrast to only 10 nm between the last two members of this
series.
 Fig. 3
Visible light absorption spectrum of lycopene (19) (in ethanol)
taken with a Zeiss grating spectroscope
 IV. Spectroscopic Methods 193

Table 2. Visible light absorption maxima of P-apo-carotenals and their 15,15'-didehydro analogues
(in petroleum ether)

Compound Amax (nm)

P-Apo- 15,15'-
carotenals Didehydro-
(~) P-apo-
"' carotenals
(~)
,.

c2s 414 400

c27 437 417

C3o 457 432

c32 473 447

c3s 485 459

c37 498 469

c40 508 481

The 15,15'-didehydro analogues show the same decreasing contribution of

each additional double bond. At the beginning of the series that difference is
17 nm and at the end 10 to 12 nm. A conjugated triple bond exerts a marked
hypsochromic shift depending in magnitude upon its position in the conju-
gated chain. The hypsochromic shift is 14 nm when the triple bond is located
at the y,~-position of the P-apo-12'-carotenal (C 25 ), but reaches 29 nm when it is
close to the cen-tre of the conjugated chain; see Table 2, C 37 .
Of the C 40 -carotenes, lycopene (19) has its absorption maxima at the longest
wavelengths. Fig. 4 indicates the changes of the absorption spectra for carotenes
with chromophores having the same length but one or two cyclic ends (P-end
groups). When such a ring is present at one end of the molecule (y-carotene (8)),
a less marked fine structure, a hypsochromic shift as well as a hypochromic

Caroteno1ds 13
194 W. VETTER, G. ENGLERT, N. RIGASSI and U. SCHWIETER

200 .----.-----.-----.-----.----~-----,,----.

75~---J----~----~~~--~----r-++--+---__,
A
~
~~
!\
,,,
\ n
' !,,

50----- ~ f----ti'f--!i-1+l--+-+-------1
1 fr, 1n1 I \,
. . . . . . . . -~ n,l\~\1 1
~ !

~ :+---+----+---+---+---F---1-~" "1 '1-~- - -\-V\

.
-+\---1
~I 1I
,\-!+:

1
,
75+----+----_,----~------~.~~+----++-+-~

II)
it \, \,.
JJ '\ \
50+----+-----M----~----~~----+---~~-+-~

-I )'

210 2 )() 300 3 )() 400 450 500 550rm

Fig. 4. Ultraviolet and visible light absorption spectra of P-carotene (-),')'-carotene(----) and
lycopene (····)(in petroleum ether)

effect are observed. This is even more pronounced upon formation of the second
ring (P-carotene (3)).
If conjugated double bonds are lost on cyclization to e-end groups (Fig. 5),
hypsochromic shifts of about 15 nm are observed for each conversion. Com-
pounds with e-end groups retain the pronounced fine structure of the main
absorption band.
Comparison of Figs. 4 and 5 shows higher extinction coefficients for the
~:-analogues. A hypsochromic shift of only 6 nm for each conversion of a P- to
an e-end group indicates that the double bond of the P-end group is not fully
conjugated to the chain.
Fig. 6 shows the effect of the introduction of a conjugated carbonyl group
in a carotene molecule. If one carbonyl group is introduced at the 4-position
of P-carotene (echinenone (148)), a bathochromic shift is observed with a com-
plete loss of fine structure. The addition of a second carbonyl function in the
4'-position (canthaxanthin (193)) causes only a small additional bathochromic
displacement of the broad absorption maximum.
The end groups can be hypothetically removed by oxidation at the 7,8(7',8')-
double bonds, giving alcohols, aldehydes or carboxylic acid esters. P-Caro-
 IV. Spectroscopic Methods 195
200.----,-----,-----,,-----.-----.-----.-----.

1\
175+---~----~----~------~----~~;~;--+---~
II ~~ \. r.

150 ~
(\ 1\ \ n 1\
f-----tl-tl-11-tH\+\+H+----1
I I I;
·············~ II I
I
I\
I.
125,,_---,-----,------.-----r---~~~~+1+-H-~~
I f !I I \ .
'( h! Jl .~ \ i
~100+---~-----4------~----r1~~AWHV.+.11_\HlJ~\~i*-+-~
w I ,u !\) \I 1
I Vi i 11 I
1 11 I
I !i u I

JF
75+---~-----4------~----hr,_r-+-~r-+-~~

50 I
I\
251\
/- Af\
;y_~r:
j,!· / \ I .

\ fiX\\ //;' \ ~\
~-U !..../ I \. \_~ - L::/·/ \ \.
.· · '1---- .
210 250 300 350 400 450 5C 0 550rm
Fig. 5. Ultraviolet and visible light absorption spectra of e-carotene (--- -), ~-carotene(-) and
lycopene (····)(in petroleum ether)

tene, for instance, would yield /3-apo-8' -carotenal (248) if one end group were
removed, or crocetindialdehyde (267) upon the loss of both end groups
(Fig. 7). Interestingly the first shortening induces a small bathochromic, where-
as the second causes a clear hypsochromic shift and a more pronounced fine
structure.
The ester analogues, i.e. ethyl /3-apo-8'-carotenoate and crocetin diethyl
ester, exhibit a similar overall picture as is shown in Fig. 8.
The corresponding alcohols, i.e. /3-apo-8'-carotenol and the C 20 -diol show
the expected trend: removal of the /3-end groups and of conjugated double
bonds lead to hypsochromic shifts (Fig. 9).
The change from the linear all-trans configuration to an angular cis isomer
results in the appearance of a subsidiary peak about 140 nm below the ab-
sorption maximum of the longest wavelength of the corresponding all-trans-
carotene [6]. In Fig. 10 this effect is demonstrated for /3-carotene (3) and its
15,15'-cis isomer, the latter clearly showing, besides a lower extinction coeffi-
cient, the additional cis peak. In most cases a small hypsochromic shift of the
main absorption band is also observed. The spectrum of /3-carotene (3) in
ethanol is also included in this figure, the solvent effect not being very pro-
196 175,-.--------------,----r-------.----,
-~
150 ----~
. . . . . . .-~ (1\

'i' A:\
125--t---,-----.-----,---------t------+--t+:-1
y·----'<;-.,---+-------1

100~---+-----+----~----~--rL~/~L__~_-n~~----4
t~
w : '\
75d----d-----4-----4r-----~~--+----++..----4
~' ! ~~~
1 ~/: 1\1\
/i ,,
I 1 I\
5Q I \\

1·, /l/~ ~
-=v-p·
25 II I\

\'-5
'\_ £_:X- \;;· ·.
-- "'__....,'".
1\I ~
\
210 2 0 300 350 400 450 500 550nm
Fig. 6. Ultraviolet and visible light absorption spectra of fJ-carotene (--), echinenone (----)and
canthaxanthin (· · · ·)(in petroleum ether)

175,,------------------------.-----.------.-----.

150 ------- ~HO

210 400 4 0
Fig. 7. Ultraviolet and visible light absorption spectra of fJ-carotene (--), fJ-apo-8'-carotenal
(----)and crocetindialdehyde (· · · ·)(in petroleum ether)
 175T------------------------.----~------~--~ 197

~I
150 ------ ~O,C,Hrs_ _ _-t-------1~----i
.1.
................, .. C;,H 5o,c~C;,Hs

"'
~X
w

210 250 3(0 350 400 4 0 500 550nm

Fig. 8. Ultraviolet and visible light absorption spectra of {J-carotene (---), ethyl {J-apo-8'-
carotenoate (----)and crocetin diethyl ester(· · · ·)(in petroleum ether)

175,----------------~

150 ~OH~--~-----+----~

125+-·~····~····~··,···~····~·····-H~o~H,c,~-~~--~~,-~--~~~-r--~f-MI\____-r-------1
f~y, \ (\
:· :i! \ I~"' \
100·+-----+ -----+--- -·---+-------+--+~'/-:+-c4-'--+-+-----i
"'
~X
w
75·+---·j-
,' Jf: v \ \
----1rl-----l

:\:· \ .!1r,
I'./; \,
-~--··

: ... I I
50•+---~-t-----+- ---r--

II
. .!lj/!
I
.....
: \\ I ... I

\ ,\,IL
25·~---~--~-~---+-~-+--+-~-~~---i~-~

~~~~
210 250 300 350 400 450 500 550nm
Fig. 9. Ultraviolet and visible light absorption spectra of {J-carotene (--), {J-apo-8' -carotenol
(----)and crocetindiol (· · · ·)(in petroleum ether)
198 175
=~
150 -----~
125
(1\

"'I 100
~
X

75

50
\ .

25
II /\ ;/ \\

210 250 3 0 3 0 4 0 4 )0 500 5 Orm

Fig. 10. Ultraviolet and visible light absorption spectra of P-carotene ( - - in hexane, · · · · in
ethanol) and 15,15' -cis-P-carotene (----)(in hexane)

175

50- ~CO.C:tts

25

00
{) ~~.

~X
w 75
fi \\ \
\

//
I
\
\
\
\
~EthanrJ. \

\\
~xane----t
50

/;
25

~ ~ ~
-
VI
'"'
\"-'' _
\
\
\

....
230250 300 350 400 450 500 550rm
Fig. 11. Ultraviolet and visible light absorption spectra of ethyl P-apo-8' -carotenoate in hexane
(--)and ethanol(----)
 IV. Spectroscopic Methods 199

Table 3. Constants for the calculation of the main absorption maximum of polyenes according to
Hirayama [9]

Compound Solvent A B

Polyenes Hexane 36.62 38.72

Ethanol 36.98 39.10
Chloroform 38.60 40.72
IX-Carbonyl polyenes Petroleum ether 39.78 39.33
IX,IX'-Dicarbonyl polyenes Petroleum ether 37.69 34.67

nounced. A similar solvent effect is clearly demonstrated in Fig. 11 for ethyl

{J-apo-8'-carotenoate.
Such effects on electronic spectra of carotenoids have been studied and also
discussed from a theoretical point of view [7]. In a binary system, the increase
in concentration of the solvent which has the stronger dispersive interaction
with the polyene causes a bathochromic shift of the absorption maxima as well
as a lower extinction. Recently it has been shown that the addition of about
50% of water to water-miscible solvents causes the appearance of an additional
absorption band at about 370 nm [8], often accompanied by a loss of fine
structure and a pronounced hypochromic effect.

Table 4. Increments (n) for substituents and conjugated double bonds for the calculation ofthe main
absorption maximum of the polyenes according to Hirayama [9]

Conjugated double bond +1.0

Alkyl or hydroxyalkyl +0.1
IX,P-Epoxyalkyl +0.2
Furanoid oxide 0.0
Carboxylic acid +0.3

(X)J:i
ex··.
-0.8

-0.6

c:x· I

ex··.
-1.0

-0.2

OJ:i. -0.9

ex· C0 2 CH 3
-0.4
200 W. VETTER, G. ENGLERT, N. RIGASSI and U. SCHWIETER

200

-~
r
175 -----~ I I
I I
., ., ., .n
I I
I

·-· - u:
X. J.. ..l J. .l.
I
I
I
I
I
\
I
150
I
"'
I 1.: I I i
·"·f I I ;

r
I
1 I I;
v
,.•./ l
1 I I\
\ I
I ii I, \

)M
125 I \, .
I
I :
' I
'? :

~ 100 \I

I
0

w
X
I
I II I :
I
' I :
I
I
I
, I I
75

I
:
I

\
:
; I
I
'I I
I I
: I
50
/ I

·~:
/~\
I I
I I I I I
I I I I I
I
\· ,/ I \
~ LX ~ //
I
25 l \ I

\_
. I I
. I
..- I
\
' ' '---
210 250 300 350 400 450 500 550 600nm
Fig. 12. Ultraviolet and visible light absorption spectra of P-carotene (-), decapreno-P-carotene
(----)and dodecapreno-P-carotene (· · · ·)(in petroleum ether)

The main absorption band of the spectra can be calculated according to the
general formula [9] :
(A.maxf =(A-B X 0.920N) X 104 nm 2 .
The values of A and B depend upon the solvent and the type of polyene studied
(Table 3). N is determined by the number of conjugated double bonds and their
substitution pattern: N = n1 + n2 + n3 •••• Some values of n; are given in Table 4.
One conjugated double bond is equivalent to one unit. Subtractions are made
for double bonds which are not fully effective, as for instance the bond in the
P-end group.
The formula does not, however, always give satisfactory results. It is inter-
esting that it does allow the calculation of the absorption maximum for infinite
length. In the case of polyene hydrocarbons, this band would be expected to be
at 608 nm (ethanol).
So far, only the main absorption band (A. 1 ) and the so-called cis peak (A. 2 )
have been discussed. Further peaks are observed, however, when the con-
jugated chain is extended beyond the normal polyene skeleton. Fig. 12 shows
the spectra of decapreno- and dodecapreno-/3-carotene with 2 and 4 additional
 IV. Spectroscopic Methods 201

isoprene units. A number of additional absorption bands appear (A. 3 , A. 4 etc.).

The A. 1 band is due to an electronic transition from the highest occupied to the
lowest unoccupied n-orbital. It has been argued that the minor bands, which
correspond to transitions to higher unoccupied orbitals, are overtones [10].
Their wavelengths can easily be calculated, because in an aliphatic polyene
with n conjugated double bonds, the maximum of the A.x band will lie very close
to the maximum of the A. 1 band of a polyene with nix double bonds. The A. 2
band, therefore, corresponds to nl2, A. 3 to nl3 etc.
The calculation, best carried out graphically, is shown in Fig. 13. The
wavelength maxima of the three peaks of the main absorption band are plotted
against the number of double bonds. The curve has been extended beyond the
one published [10] by using the data for decapreno- and dodecapreno-P-
carotene, taking into account the relatively small bathochromic influence of
the two P-end groups (i.e., 14 effective double bonds instead of 15, and 18
instead of 19, respectively). The A. 2 peak for dodecapreno-fJ-carotene is then at
nl2= 1812=9, A. 3 : nl3 =6, etc. The values calculated and observed are indicated
in the figure. The overtones of even order represent transitions which according
to Dale [10] should be forbidden in linear all-trans compounds but allowed in
angular molecules, such as cis isomers. This rule is apparently not valid for
longer carotenes (see Figs. 12 and 13).
The occasional observation of bands of even order. in all-trans polyenes has
been explained by isomerization in solution, but it might be argued that steric

n
A
I II I
18 I I
17
16 I ! \ I I II
15 I II II fl
14 A ) I I I
13 I I 1I
12 I I 1 1

11 I I 1 I
10
n{2='A 2_ _ _ _ _ _ _ _ _ _ I
9 I
8 I
7
n{a ='A a- _ _ _ _ _ _ _ ,I
6 I
5 n/4='A~;-----­ I
4 nf5='As------ I
3 ~"6:'As---- I
n,7-X7____ I
2 I
I iI I
600nm

Fig. 13. Graphic calculation of the overtone bands of dodecapreno-P-carotene (in petroleum ether)
according to Dale [10]
202 W. VETTER, G. ENGLERT, N. RIGASSI and U. SCHWIBTER

distortion of the molecule could be the cause of the appearance of these bands,
an argument similar to the one that has been used to explain the subsidiary
maxima of vitamin A 2 [10].

C. Infrared Spectroscopy

This technique has not in the past played a major role in carotenoid chemis-
try, mainly because the conjugated polyene system gives rise to only very weak
bands. It has, however, proved to be of value for detecting certain special
structural features such as acetylenic, allenic, hydroxy and unreactive keto
groups, e.g. like those in fucoxanthin (190) and capsanthin (170).
The structures of allenic and acetylenic carotenoids were recently solved
with the help of i.r. spectroscopy [11, 12]. The unusual peak at 1928 cm- 1 of
HO:J;TOAc

HO~,· WAvaENGTH

25 3.5 4 5 6 7 8 9 10 15 25~
100
%
_,- 1- ~ --,_
80
II
/
" ,-..

'
......... n IIJ' VIV
I )I
I\ I IV IJ J I
\ I .I I lA
\1 ! v \W v

0
4000 3000 2000 1800 1800 1400 1200 1000 800 800 400 em-•
FREQUENCY
Fig. 14. Infrared absorption spectrum of fucoxanthin (KBr pellet) (Courtesy of B. C. L. Weedon)

~?~
~? WAVELENGTH
02.5 3 4 6 8 1
%
J '-.,
V\ \
,
v
~ f"1

"" I l
I ill ,.J
40 IV ,t
I
20 u
n
v
0
4000 3000 2000 1800 1600 1400 1200 1000 800 cm-1
FREQUENCY
Fig.15. Infrared absorption spectrum of alloxanthin (in CHC1 3 ) (Courtesy of B.C.L. Weedon)
 IV. Spectroscopic Methods 203

~
1000 900
-.,---,-
1000 900
i
-.,---,-
1000 900
.......--
1000 900

Fig.16. Infrared absorption bands for retro apo-carotenoids (KBr pellet)

~HO WAVEI..ENGTH
2.5 35 4 5 6 7 6 9 10 15 25 II
-r --.
100

"'
....... (\, ~I
"
80 rv \ IV
·~
d
/I\
I
\
20

0
4000 3000 2000 1800 1600 1400 1200 1000 800 600 400cm-1
FREQUENCY

~-~_)-~-(~..(_HO
WAVELENGTH
2~ 35 ~ 4 5 6 7 8 9 10 15 25f1
1
xi-- 1--....._ ~ ...-M
y .( " v
V\ ~"'~T ~~ \
rv
'
80
\ I
\
111
80
(
40
II
\ l

0
4000 3000 2000 1800 1600 1400 1200 1000 800 800 400 cm- 1
FREQUENCY
Fig.17. Infrared absorption spectra of all-trans- and of 15,15'-cis-P-apo-8'-carotenal (>2600,
1300-1100 and <900cm- 1 in CS 2 , 2600-1300 and 1100-900cm- 1 in CHCI 3 )
204 W. VETTER, G. ENGLERT, N. RIGASSI and U. SCHWIETER

fucoxanthin (190) (Fig. 14) could be assigned to an allenic group. The weak band
at 2170 em - 1 in alloxanthin (65) (Fig. 15) is characteristic for an acetylenic
carotenoid.
Carotenoids with trans-disubstituted ethylene show a strong band near
965 em - 1 due to C-H out-of-plane deformation vibrations [13]. For all-trans-
carotenes this is a sharp singlet. Retro carotenoids as well as some carbonyl-
substituted or cis polyenes show a doublet in this region. This double peak seems
to be conclusive evidence for the retro system [14]. In retro apo-carotenoids
the appearance of this doublet is dependent on the chain length, as seen in
Fig. 16. The C 12 -compound exhibits only a single peak, while the higher
vinylogues show the characteristic pattern [15].
Splitting of a peak into a doublet is also often observed with central-cis
carotenoids. The C-H out-of-plane deformation vibrations of cis-disubstituted
double bonds give rise to a strong absorption at 780 em - 1 (Fig. 17). A trisub-
stituted cis double bond shows weak absorption near 830 em - 1 , and sometimes
a peak is also observed at 1380 em - 1 .

D. Nuclear Magnetic Resonance Spectroscopy

N.m.r. spectroscopy, almost solely proton magnetic resonance (p.m.r.), has

played a major role in identifying and elucidating molecular structures in
carotenoid chemistry. This spectroscopic technique, complementary to the ones
already discussed, was first used in carotenoid research by Jackman, Weedon
et al. in 1960 [16]. Since that time, its application has been reported in a rapidly
increasing number of papers, clearly demonstrating that p.m.r. is a very im-
portant and effective tool for the elucidation of carotenoid structures. The
chemical shifts (!5) of the signals, and the magnitude of the splittings (J) caused
by mutual spin-spin coupling of the protons, directly reflect the structural
properties of the different parts of the molecules. This means that chemical
shifts and coupling constants are generally, to a good approximation, dependent
only on the immediate surrounding of the different protons, and the inter-
pretation of the p.m.r. spectrum therefore yields detailed information on
structural subunits present in an unknown molecule.
This 'additivity' also considerably simplifies the present discussion of the
large amount of experimental p.m.r. data gathered during the last 10 years.
In what follows one is able to break up the survey by discussing separately the
partial spectra of the end groups and of the chains of conjugated double bonds,
thus achieving a considerable simplification. The main emphasis will be on
chemical shifts rather than on a discussion of coupling constants. The latter are
usually within the normal ranges expected of them, and minor differences
( < 1 Hz) may sometimes even arise due to the fact that the reported coupling
constants have been obtained by 'first-order' analysis from normal routine
spectra rather than by a more rigorous treatment of highly resolved signal
patterns. In order to minimize the' variation of the reported chemical shifts,
 IV. Spectroscopic Methods 205

all data in this chapter will refer, if not otherwise stated, to deuteriochloroform
as a solvent. Internal tetramethylsilane (TMS, !5 = 0.00 p. p.m.) is employed as
reference.

1. End groups
In recent years, information on molecular structures was mainly derived
from the highfield part of the spectrum between about 0.5 and 5.5 p.p.m.,
containing the methyl signals of the conjugated chain (briefly discussed later)
and the methyl, methylene and methine signals of the different end groups.
Since identification of the structure of the end groups is a first important step
in the elucidation of an unknown structure, the following discussion of their
partial spectra will be given in detail.
A very useful survey on p.m.r. data of carotenoids and relevant literature
has been published recently [17], and further data are e.g._presented in [18]
and [19]. From these data and from numerous unpublished results from our
own laboratories, relevant chemical shifts and coupling constants for some of
the cyclic and acyclic end groups have been collected in Table 5. These data
may be helpful in identifying these structural sub-units in unknown carotenoids.
It is assumed that the chemical shifts, which are mostly average values from
a number of compounds, will generally vary only slightly (±0.03 p.p.m.)
provided no significant structural changes occur in the proximity of these end
groups.
The signals appearing at highest magnetic fields are assigned to the strongly
shielded protons of the geminal methyl groups at C-1. The observation whether
only one (6 H's) or two different signals (3 H's each) appear in the spectrum from
a specific end group usually gives a first hint of possible structures. For example,
the /3-end group occurring in vitamin A compounds and /3-carotene always
gives rise to a common signal for the geminal methyl groups at about 1.03 p.p.m.
This is presumably partly due to a rapid equilibration of the two half-chair
conformers of the cyclohexene ring. With s-end groups, on the other hand, one
ofthe conformers may be more favoured energetically, and the protons of the
two methyl groups differently shielded. When using the criterion of equivalence
of protons it should, however, be kept in mind that any coincidence of signals
observed could be purely accidental and eventually be removed by specific
solvent interactions or by applying higher spectrometer frequencies.
Additional information is obtained from the shifts of the CH 2 - and CH-
proton signals. In particular, the appearance of CH signals between about 3
and 5.5 p.p.m. reveals the presence of an O-R-substituted end group_; the
presence of CH 2 multiplets at about 2 and 2.5 p. p.m. indicates a neighbouring
double bond Qr a carbonyl function. Although the CH 2 signals of the different
end groups generally give rise to complicated patterns, these may well be
helpful in the identification of the different end groups. As an example, the
characteristic niultiplets of the 2-, 3- and 4-CH 2 protons of a /3-end group are
frequently easy to detect in the 220 MHz spectrum (see Fig. 24 on p. 239) at
about 1.47, 1.60 and 2.02 p.p.m. ( ±0.03). This finding may give an even stronger
206 W. VETTER, G. ENGLERT, N. RIGASSI and U. SCHWIETER

indication of the presence of this group than does the observation of a singlet
of the methyl groups at C-1, since the latter is also valid for other groups, e.g. a
P-3,4-didehydro-end group. Besides the characteristic shift ranges or signal
shapes of methyl and methylene signals, further hints are derived from the shifts
of olefmic protons and/or CH protons of the end groups. The observation of a
broadened doublet ( -9.5 Hz) at about 2.2 p.p.m. indicates the presence of an
e-end group (H-6), and the occurrence of a broadened quasi-singlet (even at
220 MHz) for H-7 and H-8 of a P-5,8-epoxide-end group at about 5.16 p.p.m.
is very helpful. In CS 2 , the two shifts are slightly different (5.05 and 4.95 p.p.m.)
and a small coupling J 7, 8 1-2Hz is observed. These epoxides are usually
mixtures of epimers, and the shifts of the minor component were given-as far
as the corresponding signals could be identified-in brackets. Similarly,
synthetic carotenoids with retro-end groups are generally mixtures of presum-
ably 6-cis-trans isomers as indicated by additional peaks in the spectra. It is
assumed that the main component corresponds to the trans structure. As far
as the signals of the minor component could be assigned, differences are best
observed with the shifts of the methyl groups at C-1 and C-5.
A similar doubling of signals has been observed with retro-3-keto com-
pounds where variable ratios of the two isomers have been detected [20]. The
chemical shifts enclosed in brackets are attributed to the isomer present in
smaller amounts; the others, due to the major isomer, agree closely with those
of natural rhodoxanthin (209), which has been assumed to be 6-trans [20]. The
direction of the shift changes from trans to cis corresponds to that observed
with the retro compounds.
In some cases shown in Table 5, the chemical shift differences between
corresponding signals are rather small and therefore some assignments are
uncertain, as is indicated. This is true, for example, for some C-1- and C-5-
methyl signals.
A few remarks should be made concerning the two isomeric forms of the
P-5,6-dihydroxy-end group. Natural azafrin (261) and its methyl ester presum-
ably correspond to the trans-dihydroxy structure, since no infrared evidence
has been found for the existence of an intramolecular hydrogen bond [21] as
would be expected, because of geometrical reasons, for the cis structure.
Synthetic 5,6-dihydroxy compounds, on the other hand, give slightly different
chemical shifts and might therefore possess the cis-dihydroxy structure. In
agreement with this assumption there is the observation that, in the p.m.r.
spectrum of the synthetic ester, one of the hydroxyl proton signals was found
to be shifted downfield to about 2.6 p.p.m. (CDC1 3 ) and is hence easily distin-
guished there from all other signals. A deshielding of this kind is expected if a
weak hydrogen bond occurs. The two isomers can also be distinguished from
each other by a further observation: in the spectrum of the synthetic product,
a doublet presumably of H-7 or H -8 is observed at 5.94 p. p.m., whereas in the
natural product the same signal is shifted downfield to about 6.22 p. p.m.
In Table 5, only three acyclic end groups have been included, but more data
can be found under [17].
 IV. Spectroscopic Methods 2CY7
Table 5. Approximate characteristic chemical shifts (mean values; in p.p.m.) and coupling constants (in Hz) of
selected end groups. Solvent: CDC1 3

cc·····
l.0 3

~:"'
y-6 1-6.5
-147 1.85 (t)
/3- -1.60
/3-4-Keto-
2.51 (t)
1.72
-2.02
0

a
1.30

HOJX,:,:,
1.07 y--6.1
/3-2,3-Didehydro-
4 . 0 ) ( J C · .... 3-hydroxy-
/J-3-Hydroxy-
HO t 1.73
4-keto-
0
2.02

2.04/2.40
(1-16.5)

a l.r8
1.14/1.20 A.···· l.QC02/104
/-6.13
fJ-7 ,8-Didehydro- !X~ /3-4-Hydroxy- I
.. ··
3-hydroxy-
HO t 1.90
4.03
H
4

2.29
(d, J-7)

0.95/1 10*
if.60/-6.3
.. ·· /3-3-Hydroxy-
/3-5,6-Epoxide 0
5,6-epoxide-
1.14*

ar
b 0 84/Ll4* c 0.89/l.IO*
5 94*

/3-5,6-Dihydroxy- H
.···
/3-5,6-Dihydroxy- ~-.... (I x OH 2.6)
H

1.19*
t t
1.20*

£
w
d 5.16/5.16

w
(5.16/5.07) d
1.10/Ll5
1.21/1.30 ~516
(l.I0/1.16) I 79 (1.~4)

/3-3-Hydroxy-5,8-
/3-5,8-Epoxide
epoxide-
HO
1.42 1.58
(I 48) (1.58)

1.02/1.06
/3-3,4-
Didehydro- -1.68/-2.19 ~ ...
/3-3,4-Didehydro-
5,6-epoxide- 5.83 ~~27···
5.78

(Continued)
 208 W. VETTER, G. ENGLERT, N. RIGASSI and U. SCHWIETER

Table 5. (Continued)

083/091 0.85/1 00
_.......5 51

e- 12/15 ~"""···· e-3-Hydroxy-

~··
-20 # '-22(1-95)
1 58 HOAA1.63
5 40

1.02/1 10
Og:.74/0.92
/ 6 04
e-6-Hydroxy- 2 29/2 4 8 J : e · · · .
.·
OH e-4,5-Epoxide- ~206
3-keto- (J-!7) oP # 1 92

t
6

a
5.93
0
3.07

e e 2.18 (2.26)
r680(s) (-6.55,s)
-5 77 (5 63) retro- -5 95 (- 5 95)
retro-
-2 10 (?) 3-Keto- 0 (124)

t
-1 50
t
2.36
(2.30)

ft
(?)

a
o~.
l93 -64

5 75 -:·
3,4-Diketo-2,5,5-
retro-3-Hydroxy-
HO 1 /1 44
trimethylcyclo-
pent-1-enyl- oH..41
4 35

2-Acetyl-5,5- Ox2~- P-5,6-Diketo-

dimethylcyclo- 2 64 (t) 5,6-seco-
pent-1-enyl- 1.75 (t) 4.29~-

a 1.82
1.08/1.35 ):::· ..

Fucoxanthin-
-type-
2.02
Fucoxanthinol-
-type- HOJ:r:~: ·.

1 35

-2 12

1/J-1-Hydroxy-1,2-
1.68xJJx. dihydro-
5 10 5.95

-2 12

*:Assignment arbitrary (small shift differences). (s), (d), (t): Singlet, doublet, triplet.
a: Values from [17]. b: Natural azafrin and derivatives, presumably trans dihydroxy [21]. c: Main component
of synthetic product, presumably cis dihydroxy. d: Normally mixture of the two 8-epimers; values given for both
components (for minor component in brackets). e:,Normally mixture of 6-cis-trans isomers; formula shows trans
isomer. Values in brackets presumably 6-cis [20]. 1
 IV. Spectroscopic Methods 209

2. 'In-chain' methyl groups

The chemical shifts of the methyl groups of the conjugated chain (e.g. at
C-9, C-13, etc.) are, in general, less informative, since their signals usually
appear in a rather narrow range between about 1.9 and 2.05 p.p.m. They may
be shifted upfield if the position of the methyl group is near the end of the con-
jugated chain (up to "'1.65 p.p.m.). Methyl groups close to an acetylenic bond,
such as C-13- and C-13'-methyl groups in 15,15'-didehydrocarotenes, absorb
at a slightly lower field ("' 2.1 p.p.m.) than the normal 'in-chain' methyls.
Methyl groups cis to a carbonyl or carboxyl group may be considerably
shifted downfield, and this has helped in the identification of corresponding cis
isomers of vitamin A and analogues [19, 22].
Chemical shifts of a number of 'in-chain' methyl groups are contained in
Tables 6, 8 and 10 (see also [16-19, 22]).

3. Olefinic protons
The spectral range between 5.5 and 7 p. p.m. containing the signals of the
majority of the olefinic protons has proved of little informative value in the
past because of its complexity. Often only the number of protons contributing
to the signals in this range could be estimated from the integrated signal areas.
The difficulty arises from the near coincidence of most of the chemical shifts
resulting in strongly overlapping signals and hence in the appearance of a
complex jumble when measured at 60 or even 100 MHz. Only a few signals of
olefinic protons may be detectable separately outside this region, namely at
higher field, of protons on isolated double bonds, of protons at the end of a
conjugated double bond system and at lower field signals of protons in the
vicinity of a functional group with strong diamagnetic anisotropy, e.g. carbonyl
or carboxyl groups.
Recently, new types ofp.m.r. spectrometers using superconducting magnets
have been introduced, and the corresponding increase of the spectrometer
frequency may considerably alleviate the interpretation of the very complex
patterns in the olefinic spectral range. The reason is that the chemical shift
differences (in Hz) between different protons increase proportionally to the
frequency, whereas the spin-spin couplings remain constant. Therefore,
complicating second-order effects in the spectra will be reduced. The con-
siderable simplification especially of the olefinic part of the spectra of vitamin A
compounds and carotenoids effected by using a Varian 220 MHz spectro-
meter has been previously demonstrated [19, 22]. It has been found that the
increase of the chemical shift differences by a factor of 2.2 compared to
100 MHz instr.uments is actually, in many cases, sufficient to resolve the
olefinic portion of the spectrum into more or less separated singlets, doublets,
etc. Thus, ordinary 'first-order' interpretation can often be applied, and
the derived chetnical shifts and spin couplings can be used successfully
for the elucidation of unknown structures, especially in cases of cis-trans
isomerism.
Carotenoids 14
210 W. VETTER, G. ENGLERT, N. RIGASSI and U. SCHWIETER

Lycopene

38

8 7 4 3 2

"2,2"

Fig. 18. Proton magnetic resonance spectrum at 60 and 100 MHz in comparison to the olefinic
region at 220 MHz of all-trans-lycopene and all-trans-Iycopene-15,15'-D 2 (in CDCI 3 ).
[Continued on p. 211]

The drastic simplification of the spectra due to an increase of the spectro-

meter frequency to 220 MHz will be seen in the following examples [19].
In Fig. 18 the p.m.r. spectra of lycopene (19) in CDC1 3 solution are shown.
Because of the molecular symmetry the spectra are expected to be relatively
simple. A comparison of the 60 and 100 MHz spectra shows that the former
already reveals all spectral subtleties of the aliphatic part of the spectrum,
whereas in the olefinic part the signals of only 2 of a total of 18 protons (H-2
and H-2' at about 5.1 p.p.m.) can be identified. All other olefinic protons give
rise to the complex pattern between 5.8 and 6.8 p.p.m. The situation is com-
pletely different in the third spectrum showing the olefinic part measured
at 220 MHz. Here all the signals are identified as doublets, triplets or quartets
as well as the more complicated signal patterns of the central 4 protons,
i.e. of H-14,14' and H-15,15'. Approximate shifts for the latter protons are
 IV. Spectroscopic Methods 211

220MHz

H,a,ts' Ht4,w
-6.82 •8.24
,..-._
Hn.tt

H2,2'
5.11

7 6.5 6 5.5

220MHz
H14,t4'
6.24

Htt,tf Hto,tO' Hs.e•

6.17 5.114

7 6.5 6 5.5 5p.pm.

Fig.18. [Continued from p. 210]

deduced by considering the signal areas and by means of decoupling experi-

ments, since H -14 and H -14', forming one half of the corresponding AA'BB'-
type spectrum, are additionally coupled (Ja11y1) to the methyl protons at C-13
and C-13'. Another, more complicated way to make the assignment is revealed
in the last spectrum, showing the same spectral range for the 15,15'-D 2 com-
pound. Here the 15,15'-proton signals have disappeared, and the signals of
H-14 and H-14' appear, as expected, as a slightly broadened singlet.
It should b~ pointed out that, not only in this case but also with the other
spectra discussed later, a more or less complete assignment of the signals of
the individual protons has been achieved. These assignments, which might in
a few cases be only tentative, cannot be justified here in full detail. It need only
be mentioned that they are based on one or more of the following arguments
and techniques: comparison of chemical shifts and spin couplings in related
212 W. VETTER, G. ENGLERT, N. RIGASSI and U. SCHWIETER

1-CH3
1.29 lOOMHz
Anhydrovitam~n A

S-CH 3
1.94

11!5
A B
nf-,,1-.

4 3 2 p.p.m.

220MHz
H!o
6.44 Hr
) , 6.38

J., A
11!5
B
11!4
6.45 2 5.22 5,05
H8 H11 6.19 H4 ~ ~
8.78 6.81 5.78

Fig.l9. 100 MHz and olefinic part of the 220 MHz proton magnetic resonance spectra ofanhydro-
vitamin A ( in CDCI 3 )

molecules, starting with relatively simple ones; consideration of the signal

intensities and their changes upon variation of spectrometer frequency; study
of solvent effects; 100 MHz spin decoupling or tickling experiments. In some
cases, replacement of hydrogen by deuterium at specific positions was used for
the identification of some of the protons, as, for instance, in the case oflycopene.
In Fig. 19, as a further example, the 100 MHz spectrum of anhydrovitamin A
is shown in comparison with the olefinic region of the 220 MHz spectrum.
Although the molecule is of rather low molecular weight, the olefinic part of
the 100 MHz spectrum is much obscured by the overlap of most of the signals.
In the 220 MHz spectrum all individual doublets and quartets to be expected
from this molecule of known structure are easily recognized and assigned to the
different protons. The assignments were based here on the expected multiplet
structures and spin couplings. Som'e of the olefinic protons have a small allylic
 IV. Spectroscopic Methods 213

coupling of about 1.4 Hz with their neighbouring methyl protons. Although

this coupling is not resolved at 220 MHz, the resulting broadening of the lines
helps in their identification.

a) Vitamin A analogues and apo-carotenoids

220 MHz spectra of vitamin A compounds have been shown and discussed
in [19] and [22]. Table 6 summarizes some pertinent p.m.r. data for a number
of vitamin A derivatives and their analogues with different end groups including
some cis isomers. The chemical shifts of such end groups, which may be obtained
quite accurately from Table 5, have usually been omitted. The data of Table 6
have been found to be very helpful in the understanding of the more complicated
spectra of the carotenoids. Only a few examples with different end groups have
been included, since, as has already been mentioned, only the shifts of H-7 and
H-8 are expected to be possibly altered by replacement of the end group
and not the others of the residual 'in-chain' olefinic protons.
Although these data cannot be discussed here in full detail, a few points
should be emphasized. It is seen that the shift ofH-7 in comparable compounds
as, for example, with the first eight all-trans vitamin A analogues show major
scattering between 6.11 and 6.35 p.p.m., whereas those of the other protons-
as far as they are not directly influenced by the different end groups-are fairly
constant (e.g. H-8, H-10 and H-12). A possible explanation of this observation
may be that, in solution, small differences of the average angle of rotation
about the 6,7-bond may occur. In the solid state, this angle, measured as the
deviation from a planar s-cis structure, is of the order of 50°.
It is generally assumed that the steric repulsion between the ring methyl
groups at C-1 and C-5 and the hydrogen atoms at C-7 and C-8 forces the
molecule out of the planar s-trans and s-cis configurations. That the chemical
shifts of H-7 or H-8 are strongly dependent on this steric effect can be seen
from the shifts with the 1,1-didemethyl analogue (see Table 6) where one of
these signals is considerably shifted downfield to 6.85 p. p.m. It seems likely that
this compound may adopt a planar or nearly planar s-trans configuration,
explaining the deshielding of H-7 as being partly due to steric effects of the
C-5-methyl hydrogens. In the 1,1,5-tridemethyl analogue, the corresponding
signal is accordingly shifted upfield to its normal position around 6.3 p.p.m.
A downfield shift due to steric interactions is also observed for the signals of
H-11, H-15 and H-15' in all-trans vitamin A analogues and carotenoids. This
deshielding is also caused by an expectionally strong steric interaction, mainly
ofH-11 with the methyl group at C-9 and ofH-15,15' with those at C-13 and
C-13', respectively. This downfield shift due to dispersion forces, which has
been termed a .van der Waals' shift, has been shown to exist in numerous cases
(see e.g. [23]). It is known that appreciable downfield shifts for two hydrogens
only occur if their internuclear distance is smaller than the sum of their van der
Waals' radii. In principle, from the additional downfield shifts, a mean distance
of the interacting hydrogens can be calculated, if the rotation of the methyl
group between different (presumably eclipsed) conformational positions is
214 W. VETTER, G. ENGLERT, N. RIGASSI and U. SCHWIETER

Table 6. Chemical shift data (in p. p.m.) for some vitamin A compounds and analogues.

All-trans compounds R Olefinic protons

3 4 7

11 13
CH 2 0H 6.14
R

"""'· """'· """'· CH 2 0CH 3 6.14

CH 2 0COCH 3 6.19

CHO 6.35

COOH 6.29
COOCH 3 6.27

CH 3 -6.11

CONHCH 2 CH 3 6.21

~
R COOCH 3 6.39
I

~R COOH 6.34

9,13-Didemethyl

~
COOH 6.33

13-Demethyl

~
COOH -6.31

9-Demethyl
COOH 5.43 5.66

~
. """"'

CHO 5.94 5.90*

~
R
H
~ 4 5

~
COOH 6.85*
R

1,1-Didcmethyl
 IV. Spectroscopic Methods 215

Solvent: CDCI 3 . (For shifts of end groups see Table 5.)

Olefinic protons (continued) Others

8 9 10 11 12 13 14

6.09 6.08 6.60 6.27 5.67 15-CH 2 : 4.29; 9-CH 3 : 1.97;

13-CH 3 : 1.85
6.09 6.09 6.60 6.29 5.62 15-CH 2 : 4.08; OCH 3 : 3.37;
9-CH 3 : 1.95; 13-CH 3 : 1.85
6.10 6.10 6.65 6.28 5.61 15-CH 2 : 4.72; COCH 3 : 2.05;
9-CH 3 : 1.96; 13-CH 3 : 1.89
6.17 6.19 7.15 6.37 5.97 CHO: 10.09; 9-CH 3 : 2.05;
13-CH 3 : 2.32
6.14 6.15 7.03 6.31 5.79 9-CH 3 : 2.01; 13-CH 3 : 2.37
6.11 6.13 6.99 6.27 5.79 OCH 3 : 3.70; 9-CH 3 : 2.01;
13-CH 3 : 2.36
~6.11 6.08 6.47 6.27 5.59 9-CH 3 : 1.95; 13-CH 3 : 1.81;
14-CH 3 ; 1.76
6.12 6.12 6.91 6.23 5.65 NH:5.54;NCH 2 :3.36;N--<:-CH3 :
1.16; 9-CH 3 : 1.98; 13-CH 3 : 2.36

6.17* 6.62 6.27* 7.35 5.85 OCH 3 : 3.75

6.16 6.50 6.20* 6.65 6.32* 7.42 5.84 5-CH 3 : 1.74

6.14 6.14 6.99 6.36 7.48 5.85 9-CH 3 : 1.99

~6.16 6.50 n.a. 6.73 n. a. 7.42 5.79 13-CH 3 : 2.32

6.13 6.14 7.02 6.30 5.80 H-6: 2.22 (doublet, ~9.5 Hz);
9-CH 3 : 1.95; 13-CH 3 : 2.35

6.47* 6.36 7.08 6.39 5.97 2-CH 2 : 2.27/2.46; 5-CH 3 : 1.91;

CHO: 10.09; 9-CH 3 : 1.98;
13-CH 3 : 2.32

6.27* 6.21 7.04 6.28 5.79 5-CH 3 : 1.85; 9-CH 3 : 2.02;

13-CH 3 : 2.36

(Continued)
216 W. VETTER, G. ENGLERT, N. RIGASSI and U. ScuwmTER

Table 6.

All-trans compounds R Olefinic protons

3 4 7
COOH H-5: 6.37*

~
R 5.87

I, I ,5-Tridemethy I
CHO 5.72 5.86 -6.31

~ '
COOH
CH 2 0H
5.74
5.72
5.85
5.85
-6.29
6.17*

~
COOH 6.90*

COOH 6.37
"
"""' """' """' """'
9, 13-Didemethyl-9-ethyl

COOH 5.98*

~
R COOCH 3 5.97*
0
CHO 6.06*

CONHCH 2 CH(CH 3 h 5.96*

CH 2 0COCH 3 5.90*

CH 2 0CO-n-C 15 H 31 5.89*

~
CHO -5.83 -5.78 6.14*
R

COOH 6.64
"
"""' """' """' COOCH 2 CH 3 6.62

CHO 6.64

COOH 6.61
R
CH 2 0H 6.47*
"""'· """'· """' """' """' """'
CH 2 0COCH 3 6.51
 IV. Spectroscopic Methods 217
(Continued)

Olefinic protons (continued) Others

8 9 10 11 12 13 14

6.24* 6.20 7.04 6.30 5.80 1,4-CH 2 : ~2.19;2,3-CH 2 : ~ 1.67;

9-CH 3 : 1.99; 13-CH 3 : 2.35

~6.31 6.21 7.14 6.37 5.96 CHO: 10.09; 5-CH 3 : 1.87;

9-CH 3 : 2.05; 13-CH 3 : 2.32
~6.29 6.19 7.05 6.33 5.79 9-CH 3 : 1.93; 13-CH 3 : 2.37
6.26* 6.14 6.62 6.30 5.67 2-CH 2 : ~2.07; 15-CH 2 : 4.28;
5- and 13-CH 3 : 1.86; 9-CH 3 : 1.96
6.68* 6.34 7.06 6.37 5.83 9-CH 3 : 2.09; 13-CH 3 : 2.39

6.02 6.08 6.97

6.33* 6.19 7.01 6.33 5.82 9-CH 3 : 1.96; 13-CH 3 : 2.35

6.28* 6.16 6.94 6.28 5.76 COOCH 3 : 3.68; 9-CH 3 : 1.96;
13-CH 3 : 2.34
6.33* 6.22 7.11 6.39 5.98 CHO: 10.12; 9-CH 3 : 2.01;
13-CH 3 : 2.33
6.29* 6.15 6.87 6.25 5.71 NH: 5.71; NCH 2 : 3.13;
9-CH 3 : 1.95; 13-CH 3 : 2.35
6.29* 6.14 6.61 6.30 5.63 15-CH 2 : 4.72; COCH 3 : 2.05;
9-CH 3 : 1.92; 13-CH 3 : 1.88
6.28* 6.14 6.60 6.29 5.62 15-CH 2 : 4.79; COCH 2 : 2.16;
9-CH 3 : 1.92; 13-CH 3 : 1.87
~6.40* 6.26 7.12 6.40 5.99 CHO: 9.74; 9-CH 3 : 2.01;
13-CH 3 : 2.33

7.38 6.55 7.01 6.48 5.89 COCH 3 : 2.12; 1-CH 3 : 1.17;

9-CH 3 : 2.03; 13-CH 3 : 2.37
7.38 6.53 6.98 6.46 T 5.89 COOCH 2 : 4.17;
(OCH 2 )CH3 : 1.29

6.24 6.20 7.12 6.36 5.96 H-2: 5.10; H-6: 5.97;

9-CH 3 : 2.02; 13-CH 3 : 2.31
6.25 "6.19 7.04 6.31 5.79 H-2: 5.10; H-6: 5.97;
9-CH 3 : 2.01; 13-CH 3 : 2.36
6.27 6.11 6.57* 6.11 5.67 H-2: 5.09; H-6: 5.92;
9-CH 3 : 1.95; 13-CH 3 : 1.85
6.28 6.13 6.64 6.23 5.62 H-2: 5.11; H-6: 5.94; 9-CH 3 :
1.95; 13-CH 3 : 1.88; COCH 3 : 2.05

(Continued)
218 W. VETTER, G. ENGLERT, N. RIGASSI and U. SCHWIETER

Table6.

Compound R Olefinic protons

3 4 7
All-trans

COOH 7.25*
13

7.22*

5.73 6.14

5.76 6.36

b
5.78 6.38

COOH• -5.17
COOCH 3 • -5.17

9-cis

(('\
R'

R'=CH 3 COOH 6.39

R'=CH 2 CH 3 COOH 6.41

CHO" 6.31
COOH 6.31
II

""" 13

""" R

COOH 6.32

IJ

""" R
13-Demethyl
 IV. Spectroscopic Methods 219

(Continued)

Olefinic protons (continued) Others

8 9 10 11 12 13 14

6.77• 6.30 7.04 6.37 5.83 COCH 3 : 2.26; 9-CH 3 : 2.03;

13-CH 3 : 2.36
6.75* 6.29 7.00 6.35 5.81 COCH 3 : 2.24; COOCH 2 : 4.15;
(OCH 2 )CH 3 : 1.26

6.65 10-CH 2 : 3.23; 5-CH 3 : 1.86;

9-CH 3 : 1.96

6.72 6.30 6.45 6.04 14-CH 2 : 2.54; 15-CH 2 : 4.18;

COCH 3 : 2.04; 3-CH 2 : -2.10;
2-CH 2 : 1.50; 5-CH 3 : 1.91;
9-CH 3 : 1.93*; 13-CH 3 : 1.86

6.78 6.44 6.61 6.19 6.45 H 2 -15: 5.22/5.05; 5-CH 3 : 1.94;

9- and 13-CH 3 : 1.91

-5.17 6.22 6.90 6.28 5.79 9-CH 3 : 1.78; 13-CH 3 : 2.33

-5.17 6.21 6.86 6.25 5.76 9-CH 3 : 1.78; 13-CH 3 : 2.32;
COOCH 3 : 3.70

6.73 6.10 7.89 5.82 9-CH 3 : 2.04

6.60 6.10 7.89 5.83 9-CH 2 : 2.42; 9-(CH 2 )CH3 : 1.17

6.64 6.06 7.20 6.27 5.94 CHO: 10.07

6.67 6.09 7.15 6.27 5.82 9-CH 3 : 2.01; 13-CH 3 : 2.35

6.66 6.06 7.09 6.30 7.48 5.84 9-CH 3 : 2.00

(Continued)
220 W. VETTER, G. ENGLERT, N. RIGASSI and U. SCHWffiTER

Table6.

Compound R Olefinic protons

3 4 7
9-cis
COOH 6.35

It

"""'
"
9,13-Didemethyl- """'
9-ethyl R

COOH 6.31

5.75 5.88· 6.17

 IV. Spectroscopic Methods 221

(Continued)

Olefinic protons (continued) Others

8 9 10 11 12 13 14

6.55 6.07 7.09 6.32 7.46 5.85 9-CH 2 : 2.38; 9-(CH 2 )CH3 : 1.14

6.54 6.07 7.14 6.27 5.80 9-CH 2 : 2.38; 9-(CH 2 )CH3 : 1.15;
13-CH 3 : 2.35

6.80 6.04 6.72 6.24 5.69 15-CH 2 : 4.32; 9-CH 3 : 1.99;

13-CH 3 : 1.92

6.62 5.98 7.13 6.22 5.79 H-6: 2.26; 9-CH 3 : 1.94;

13-CH 3 : 2.36

6.14 6.54 6.69 5.92 6.07 CHO: 10.10

6.14 -6.52 -6.60 5.91 5.90 9-CH 3 : 1.98; 13-CH 3 : 2.36
6.12 6.56 6.39 5.87 5.63 15-CH 2 : 4.71; COCH 3 : 2.06;
9-CH 3 : 1.93; 13-CH 3 : 1.92

6.36 6.72 7.23 5.85 CHO: 10.17; 11-CH 3 : 2.12

6.18 6.24 7.04 7.30 5.85 CHO: 10.21; 9-CH 3 : 2.03;

l3-CH 3 : 2.15
6.17 6.27 7.03 7.77 5.69 9-CH 3 : 2.00; 13-CH 3 : 2.10
6.10 6.13 6.65 -6.60 5.55 15-CH 2 : 4.30; 9-CH 3 : -1.96;
13-CH 3 : 1.93
6.10 6.12 6.69 6.59 5.47 15-CH 2 : 4.74; COCH 3 : 2.05;
9- and 13-CH 3 : 1.95/1.96
(Continued)
222 W. VETTER, G. ENGLERT, N. RIGASSI and U. SCHWIETER

Table6.

Compound R Olefinic protons

3 4 7
13-cis

CH 2 0-p-(phenylazo)- 5.73 5.86 6.21*

benzoyl

COOH 6.28

9,13-Didemethyl-13-ethyl

COOH 5.41 5.64

COOCH 3 5.42 5.63

COOH 6.84*

1,1-Didemethyl

H-5:
COOH 5.88 6.37*
R

1,1,5-Tridemethyl

9,13-di-cis

CHO" 6.36
"""' COOH 6.27

11,13-di-cis
CHO" 6.28

~
CH 2 0H 6.16

"""' R CH 2 0COCH 3 6.19

CH 2 0H 5.71 5.83 ~6.19

*: Assignment uncertain.
•: Values from [22].
 IV. Spectroscopic Methods 223
(Continued)

Olefinic protons (continued) Others

8 9 10 11 12 13 14

6.28* 6.18 -6.74 -6.74 5.64 15-CH 2 : 5.04;

9- and 13-CH 3 : 2.00

-6.19 6.39 6.49 6.72 7.66 5.66 13-CH 2 : 2.43; 13-(CH 2 )CH3 : 1.17

6.13 6.23 6.98 7.73 5.65 H-6: 2.22; 9-CH 3 : 1.95;

13-CH 3 : 2.10
6.14 6.23 6.95 7.78 5.64 H-6: 2.21

6.38* 6.32 7.00 7.74 5.64 5-CH 3 : 1.84; 9-CH 3 : 2.01;

13-CH 3 : 2.09

6.27* 6.32 7.00 7.76 5.66 1,4-CH 2 : -2.17;2,3-CH 2 : -1.67;

9-CH 3 : 1.99; 13-CH 3 : 2.10

6.68 6.16 7.16 7.25 5.87 CHO: 10.27

6.64 6.18 7.11 7.68 5.66 9-CH 3 : 2.02; 13-CH 3 : 2.10

6.08 6.20 6.77 6.11 5.98 CH0:9.71

6.04 6.03 6.49 5.86 5.54 15-CH 2 : 4.04; 9-CH 3 : 1.93;
13-CH 3 : 1.88
6.08 6.09 6.49 5.87 5.49 15-CH 2 : 4.73; COCH 3 : 2.02

-6.19 6.08 6.46 5.88 5.56 15-CH 2 : 4.07; 9-CH 3 : 1.95;

13-CH 3 : 1.89

h: Values given presumably for 6-trans compound, shown in formula (see also Table 5):
•: Two 8-epimers possible; values given for main component (see also Table 5).
224 W. VETIER, G. ENGLERT, N. RIGASSI and U. SCHWIETER

Table 7. Significant chemical shift differences {!cos-{!trans {>0.05 p.p.m.) of olefinic protons due to
isomerism of double bonds. Solvent: CDC1 3

Compound R Shift differences [) ...-{!"... {p.p.m.)

7 8 10 11 12 14

9-cis

"':::: R'=CH 3 CHO +0.47 -0.13 -0.10

R'=CH 3 COOH +0.53 -0.06 +0.12
""':::: R'=H COOH +0.52 -0.08 +0.10 -0.06
R' ~ t4

+0.54 -0.10 +0.10 -0.06

""""' 14

COOH +0.49 -0.16 +0.11 -0.08

""""' 14

ll-cis

~··
CHO +0.35 -0.46 -0.45 +0.10
COOH +0.37 -0.43 -0.40 +0.11
CH 2 0COCH 3 +0.46 -0.26 -0.41
R

13-cis

CHO -0.11 +0.93 -0.12

COOH +0.12 + 1.46 -0.10
CH 2 0H +0.33 -0.12
CH 2 0COCH 3 +0.31 -0.14

COOH +0.09 + 1.43 -0.15

COOH +0.12 + 1.46 -0.14

I, I ,5-Tridemethyl

{Continued)
 IV. Spectroscopic Methods 225

Table 7. (Continued)

Compound R Shift differences bc,.-b.,••, (p.p.m.)

7 8 10 11 12 14

9,13-di-cis

CHO +0.51 +0.88 -0.10

'""" COOH +0.50 +0.08 +1.37 -0.13
""':::

~ R

11,13-di-cis

~
CHO -O.o7 -0.09 -0.38 -0.26
CH 2 0H -0.11 -0.41 -0.13
CH 2 0COCH 3 -0.16 -0.41 -0.12

~
CH 2 0H -0.06 -0.16 -0.42 -0.11
'"""
'<::: R

taken into account. Qualitatively, however, it already follows from the magni-
tude of the effect ( "'o.2-0.3p.p.m.) that the steric interaction must be very
severe. This is in agreement with the well-known·fact that the corresponding
steric repulsions are partly relieved in such compounds by a bending of the
planar zigzag-chain with the methyl groups on the convex side (see Chapter V).
The assumption that the shift of H-11 is mainly influenced by the methyl
group at C-9 is substantiated by the finding that the H-11 shift of all-trans-
vitamin A acid (7.03 p.p.m.) is almost the same with its 13-demethyl analogue
(6.99 p.p.m.). However, the H-11 signal is found to be shifted upfield in the
9-demethyl (6.73 p.p.m.) and in the 9,13-didemethyl analogues (6.65 p.p.m.).
It is interesting that replacement of the C-9-methyl by an ethyl group
apparently produces similar steric repulsion, as is seen from the H-11 shift
(6.97 p.p.m.) in the 9,13-didemethyl-9-ethyl analogue.
Further information contained in Table 6 is concerned with the different
all-trans and cis isomers. A comparison of the chemical shift data of selected
protons of the trans and the different cis isomers indicates that larger shift
changes only occur for such protons as are attached, or in close geometrical
proximity, to the corresponding cis bond. This fact considerably simplifies the
assignment of the signals of all the other protons in complicated spectra, since
the variation of their chemical shifts is generally rather small. On the other hand,
the magnitude of the shift differences of protons at or near to the cis bond is
characteristic of its position in the molecule.
These shift differences (in p.p.m.) between the different cis isomers and their
parent all-trans compounds are summarized in Table 7. Differences of 0.05
p.p.m. or less have been omitted, since they are thought not to be significant.

Carotenoids 15
226 W. VETTER, G. ENGLERT, N. RIGASSI and U. SCHWIETER

Positive numbers indicate that the corresponding proton signals of the cis
isomer appear at a lower magnetic field. Again, the shift differences depend
approximately only on the type of isomerism and not on the type of the end
group.
The different isomers can be distinguished from each other by the shift
changes of some of the proton signals compared with the all-trans compound.
For instance, a significant shift of the signal of H-8 is only observed for 9-cis
isomers. The signal of H-10 is shifted considerably downfield only in the case
of the 11-cis compound. Conversely, the signals of the two protons at the cis
bond (H-11 and H-12) are shifted upfield. These and additional experimental
rules derived from the data presented in Tables 6 and 7 have proved to be
extremely useful in the elucidation of the structures of several cis-isomeric
carotenoids [19].
Fig. 20 shows the 100 MHz spectrum and the olefinic range at 220 MHz of
an apo-carotenoid: /3-apo-12' -carotenal. The situation is similar to that in
Fig. 19: only the 220 MHz spectrum can be completely analysed, because all
signals are separately visible. Some characteristic features seen in this spectrum
and common to all other spectra of related all-trans molecules should be briefly
mentioned: H-7 and H-8 normally give rise, at about 6.1 to 6.3 p.p.m., to a
singlet or an AB-quartet with J 7 8 ~16Hz and relatively small chemical shift
differences. Therefore, the outer two lines may be weak in intensity. All com-
pounds with a /3-end group have a slightly broader H-7 signal, which is due to
the long-range coupling of H-7 with the 5-methyl- and 4-methylene protons,
as can easily be proved by 100 MHz decoupling experiments. The doublets
observed for H-10, H-12 and H-14 are situated at relatively high magnetic
fields, while the quartets of H-11, H-15 and H-15' with total splittings of
about 26Hz, as expected for the all-trans configuration, appear, as already
explained, at relatively low magnetic fields. Other proton signals may be
detected there if large downfield shifts occur as a result of strongly anisotropic
groups. In the spectrum shown in Fig. 20, this is true for H-14', which is in a cis
relationship to the carbonyl function.
The p.m.r. data derived from the 220 MHz spectra of a number of /3-
apo-carotenoids are collected in Table 8. The structures of some of the cis
isomers included in Table 8 were previously deduced from their p.m.r. spectra
[19]. The fact that it was possible to assign most of the signals of the trans
compounds proved of great assistance in this task. The observation of specific
changes of the chemical shifts of the olefinic protons with the cis isomers led
to the elucidation of their structure as will become evident in the following
discussion.
In Fig. 21 the olefinic part of the 220 MHz spectrum of /3-apo-12'-carotenal
(262) in CDC1 3 is repeated together with those of two cis isomers, which were
isolated from the reaction mixture on partial hydrogenation of the 15,15'-di-
dehydro compound. These compounds are of an orange and a yellow colour.
A comparison of the spectra made it easy to deduce the structures of these
isomers.
 IV. Spectroscopic Methods 227

~CH3 1-CHa
-CHO 2.()4 .00 ~03

13'-CHa 5 -ctta
9.45 1.88 1.72

5 4 3 2 p.p.m.

~CHO
l.Jl.,." . . . .,......
all-tntns-B-Apo-12~cerotenal Hrlla 220MHz
8.22 8.14

6 s.sp.p.m.
Fig. 20. 100 MHz and olefinic part of the 220 MHz proton magnetic resonance spectrum of all-
trans-{J-apo-12'-carotenal (in CDCI 3 )

The second spectrum of Fig. 21 was obtained from the orange isomer. Here,
the AB pattern of H-7 and H -8 as well as the doublet of H -10 are detected at
their expected positions around 6.2 p.p.m. The doublet (J =12Hz) at a very
low magnetic field must be attributed to a proton close to a cis carbonyl, i.e. to
H-14', shifted further downfield compared with the trans compound. Although
the chemical shift differences in the 100 MHz spectrum were very small, a spin
decoupling experiment could be used to localize the signal ofH-15'. Irradiation
of the H-14' doublet led to considerable changes around 6.4 p.p.m. In this
region the 220 MHz spectrum reveals a two-proton signal, consisting of a
doublet with a splitting of 15Hz and a quasi-triplet with a total splitting ofless
than 23Hz. It follows from this that this quasi-triplet must be assigned to H-15'
and that the compound must be the 15-cis isomer. On the basis of this assump-
tion it is possible to assign all other signals. It can then be seen that, compared to
 228 W. VETTER, G. ENGLERT, N. RIGASSI and U. SCHWIETER

Table 8. Chemical shifts (in p. p.m.)

Compound R Olefimc protons
10 10' 11 11'

~
COOCH 3 6.27 6.11 613 6.99

COOCH 3 6.24 612 6.14 6.83

CHO 622 6.14 616 679

CHO (15-ml 6.22 6.14 6.15 6.80
CHO (13-m) 625 617 6.16 6.78
CH 20COCH 3 6.18 6.12 6.13 6.64

COOH 6.22 6.15 6.15 6.75

COOCH3 6.21 6.14 6.16 6.74

CHO 6.20 6.14 6.15 6.74 -616

CHO 620 614 615 6.94 -6.70* 6.62

14' 12' 10"

"""" ..
R CHO (9-CIS) 6.19 -666* 605 6.93 6.81* 6.63*
COOCH 3 6.18 6.14 6.14 7.28 6.71 n.a
CH20COCH 3 -6.15 -615* 6.15* 6.17* 6.65* 6.46

CHO 6.25 6.13 6.13 682

COOCH 3 6.23 613 6.13 6.76 596

CHO 6.24 6.17 6.12 6.78 6.23

14" CHO 6.23 6.14 6.13 -6.91 n.a. n.a

COOC2H 5 6.23 614 614 -7.27 6.75 -657

COOC 2H, 6.22 6.13 -6.13 6.51 6.74 747

(9'-cis)

COOCH3 5.90* 630* 6.19 6.72 5.87

COOCH 3 5 93* 6.30* 6 16 6.73 596

 IV. Spectroscopic Methods 229

for apo-carotenoids. Solvent: CDCI 3

Olefmic protons (contmued) Methyl protons Others
12 12' 13' 14 14' 15 15' 9 9' 13 13' 14'

6.27 579 I 02 I 71 2.01 2.36 COOCH 3 ·370

634 620 7.69 588 1.02 1.72 1.96 207 COOC:H 3 • 3 76

------
636 629 6.94 703 666 1.03 I 73 2.00* 2.02* 187 CH0·945
6.41 6.68 7.40 6.81 642 1.04 I 73 201* 204* 189 CH0·9.54
691 6.22 696 720 6.61 104 1.73 201 2.06 1.88 CH0.9.46
632 -6.18 -618 660* 640* 1.03 172 1.97 197 182 COCH 3 :209

6.32* 5.81 6.23 7.03 6 37* I 03 172 199* 2.03* 236

6.31* 5.79 622 700 6 36* I 03 1.72 200* 2.03* 2.36 COOCH 3 : 3.72

635 715 6.27 n.a na n a 104 1.73 200* 2.03* 196* CHO· 9.57

635 673 626 6.43* 677* 663 I 02 I 73 199* 190 2.01* 2.01* CH0·944
628 672 624 643 676* 6.61* 104 175 -200 190 -200 -200 CH0·944
6.35 6.53 6.24 n a. n.a n a. I 03 I 73 -2.00 -200 -2.00 -2.00 COOCH 3 376
634 634 -624 -624 -6.62 -662 103 172 1.98 1.84 1.98 195 8'-CH 2 • 456
COCH 3 209

636 5.74 6.57 I 03 172 2.00 215 199 CHO 9.48

635 7 35 5.70 6.05 1.02 1.71 199 2.12 206 COOC:H 3 3 77

6.35 715 572 6.15 I 03 1.72 1.99 2.14 2.10 CHO 959

636 n a. 572 595 103 1.72 1.99 1.91 2.14 2.14 CH0:947
6.36 -663 5.73 5 87 102 172 2.00* 2.01* 2.11 211 COOC:H 2 • 4 23
I
CH 3 133
636 645 5.72 582 1.02 I 71 197* 200* 211 2.11 COOC:H 2 • 4 25
I
CH 3 ·134

094*
636 738 6.25 650 678 6.60 115* 195 1.92* 1.99* COOC:H 3 • 3 76
110*

094*
6.34 7.35 5.72 6.02 115* 1.95 2.10 2.06 COOC:H 3 . 3 77
110*

(Continued)
 230 W. VETTER, G. ENGLERT, N. RIGASSI and U. SCHWIETER

TableS.
Compound R Olefmic protons
10 10' 11 11'

COOCH 3 -5.18 -518 6.20 6.57 5 87

5.24• 507'

COOCH 3 -516 -5.16 617 6.64 596

5.22• 5.06•

COOCH 3 6.22* 6.40* 6.22 6 70" 5.88

COOCH 3 5.94* 6.48* 6.22 6.69* 5.87

•: Assignment uncertain.
n. a.: Not assigned.
•: Values for the minor component of the 8-epimeric mixture; only few signals have been identified.

the all-trans compound, the signal of H-14 is considerably shifted downfield,

as was the case for H-8 in 9-cis and for H-10 in 11-cis vitamin A analogues. At
the same time H-15 and H-15' where shifted upfield as was found e.g. for H-11
and H-12 in 11-cis and 11,13-di-cis vitamin A analogues (see Table 7).
The last spectrum in Fig. 21 was obtained from the yellow isomer. Three
quasi-triplets and quartets due to the protons H-11, H-15 and H-15' can be
seen. All these signals have total splittings of 26 to 28 Hz indicating an 11- and
15-trans configuration. The typical AB pattern of H-7 and H-8 is at its usual
position with J 7 , 8 of about 16Hz, proving that the 7,8- and 9,10-double bonds
must also be trans. Therefore, only the possibilities of a 13-cis, 13'-cis or a
13,13'-di-cis isomer remain. The last two possibilities are excluded by the
observation that the chemical shifts of the aldehyde proton (see Table 8) as well
as that ofH-14' are the same as in the all-trans compound. The assumption of
a 13-cis configuration is consistent with the predictable shift differences:
compared to the all-trans compound the signals ofH-12 and H-15 are shifted
downfield, whereas the H-14 signal is shifted slightly upfield, which is to be
expected (see Table 7).
A further convincing example for the capabilities of 220 MHz spectroscopy
is given in Fig. 22, showing the olefinic range of the spectra of all-trans-P-apo-
8' -carotenal (248) and of one of it!\ cis isomers. Although these molecules con-
tain 12 different olefinic protons atisorbing in a range of less than 1 p. p.m., the
 IV. Spectroscopic Methods 231

(Continued)
Olefimc protons (contmued) Methyl protons Others
12 12' 13' 14 14' 15 15' 9 9' 13 13' 14'

110
6.31 7 38 623 649 678 6.59 142 1.75 197 1.91 COOCH 3 : 3.76
1.15
110'
145' 181'
116'

110
6.30 7 35 5 70 6.03 141 175 2.08 2.05 COOCH,: 3.75
1.14
1.10'
145' 180'
116'

084*
6.36 738 6.27 6.50 6.79* 6.60
114*
1.19* 1.98* 2.00* 1.92* - COOCH, · 3.76

089*
6.36 738 6.27 6.50 6.79* 660 1.20* 195* 1.99* 1.92* - COOCH 3 · 3.76;
1.09*
OH:-26

multiplets of all these protons can be recognized and in most cases quite
defmitely assigned to the different protons. Independently of how reliable all
the assignments were, the following observation led to the deduction of the
unknown stereochemistry: a comparison of the two spectra indicates that the
typical AB pattern of H-7 and H-8 at about 6.2 p.p.m. has now disappeared.
Only the signal of one of these two protons (H-7) is found at 6.19 p.p.m. The
signal of H-8 is presumably located around 6.66 p.p.m., i.e. about 0.5 p.p.m.
downfield as compared to the all-trans compound. This shift is explained by
the assumption of a 9-cis configuration of the compound. In agreement with
this, the doublet observed at 6.05 p.p.m. is assigned to H-10, now shifted slightly
upfield, as is the case in 9-cis vitamin A analogues (see Table 7).
The spectra of ethyl all-trans-15,15'-didehydro-fJ-apo-8'-carotenoate and
of an unknown cis isomer are compared in Fig. 23. The spectrum of the latter
shows a quartet at a very low field. The signal of H-10', which occurs at 7.27
p.p.m. with the trans compound, is shifted upfield by as much as 0.76 p.p.m.
indicating a 9: -cis configuration. On the basis of this assumption a complete
assignment of·an·signals has been achieved, leading to reasonable values for
the shift differences between the spectrum of the all-trans and the 9'-cis com-
pound.
In the preceding examples, only the analysis ofthe olefinic part of the spectra
could be used for the elucidation of the stereochemistry, because the shift
232 W. VETTER, G. ENGLERT, N. RIGASSI and U. SCHWIETER

~~HO
~-····,.-.,.-

all-tnma-B-Apo-12'-carotenal 220MHz

6 5.5p.p.m.

~...
v._----~
IS- CHO 220MHz

H14'
7.40
rf..,

7.5 7 6. 6 S.Sppm.

220MHz

CHC~
7.28
H!s

7.5 7 6.5 6 S.Sppm.

Fig. 21. Olefinic part ofthe 220 MHz protontmagnetic resonance spectrum of all-trans-, 15-cis- and
13-cis-P-apo-12'-carotenal (in CDCI 3 )
 IV. Spectroscopic Methods 233

~. HO

aH-t,.,.-8-Apo-a:..carotenal 220MHz

CHC~
7.25

6 5.5 5p.pm.

220MHz

CHO

7 6.5 6 5.5 p.pm.

Fig. 22. Olefinic part of the 220 MHz proton magnetic resonance spectrum of all-trans- and 9-cis-
{J-apo-8' -carotenal (in CDCI 3 )

differences observed with the signals of the methyl protons were rather small
and the deductions were therefore less reliable. This need not, however, be true
in all cases (see Table 6). Information on the stereochemistry may also be
obtained from the chemical shifts of certain aldehydic protons. This is demon-
strated by the chemical shift data collected in Table 9 for the aldehydic proton
of conjugated aldehydes with a methyl group at the rx-carbon. A comparison
of literature data and results from our own unpublished work for cis-f3-C 14-
aldehyde and 9'-cis-15,15'-didehydro-fJ-apo-8'-carotenal indicates that the
signals of the aldehydic proton of the trans compounds appear at considerably
higher fields (9.2 to 9.7 p.p.m.) than those of the corresponding cis isomers
(10.0 to 10.3 p. p.m.). The chemical shift of the aldehyde proton of the dialdehyde
prepared from decaprenoxanthin [24], therefore, indicated the trans config-
uration of the chain [25].
234 W. VETTER, G. ENGLERT, N. RIGASSI and U. SCHWIETER

Table 9. Chemical shifts (in p. p.m.) of the aldehydic proton of IX-methyl-substituted conjugated
aldehydes (for further shifts see Tables 6 and 8)

Compound trans cis

lCHO Tiglaldehyde 9.32

(CC1 4 )

~CHO
Angelaldehyde 10.10
(CC14 )

~CHO
IX-Sinensal 9.65
(CC14 )

P-Sinensal 9.33

~CHO (CC1 4 )
9.30
(CDC1 3 )

2-trans- 9.26

~CHO 2,6-Dimethyl-
2,6-octadienal
(CC14 )

2-cis- 10.01
~0 2,6-Dimethyl- (CC1 4 )
2,6-octadienal

trans-P-C 14 - 9.32

~~0 Aldehyde (CC1 4 )

9.45
(CDC1 3)

~rno
trans- 9.43
3,4-Didehydro- (CDC1 3 )
P-C 14-aldehyde

~0
cis-P-C 14- 10.22
P' Aldehyde (CC14 )

9'-trans- 9.47
15,15'-Didehydro- (CDC1 3 )
P-apo-
8' -carotenal

9'-cis- 10.33
15,15'-Didehydro- (CDC1 3 )
P-apo-
8' -carotenal

Dialdehyde from 9.46

decaprenoxanthin (CDC1 3 )
 IV. Spectroscopic Methods 235

220MHz
Ethyl all-trans-15,15~didehydro-ll-apo-&!..carotenoate

CHC~

"'•' ...,.
-6.63.-6.57

"'• ~3 ~~4
"""
5.87 ~
5.73
6.38

14' 12' 10'

~·~
l.)l.. '8 'i> ·;; 'iO CO,C.H,
220MHz

9'-cls

Z5 7 6.5 6 5.5p.p.m.
Fig. 23. Olefinic part of the 220 MHz proton magnetic resonance spectrum of ethyl all-trans-
15,15'-didehydro-{J-apo-8'-carotenoate and its 9'-cis isomer (in CDC1 3 )

b) Carotenoids
Table 10 summarizes some chemical shifts of carotenes, in CDC1 3 solution.
It is seen that all protons have been identified, even in compounds with as mu.ch
as 18 olefinic protons. It is interesting that the shifts of the olefinic protons of
the unsymmetrical molecules can be obtained by taking the values from the
two corresponding symmetrical compounds. The same method was previously
used for the assignment of the methyl protons [18].
As an example, Fig. 24 shows the aliphatic and olefinic parts of the 220 MHz
spectrum of P-carotene. Here the multiplets of the methylene protons in posi-
tions 2 and 3 are clearly separated as has been found in all other compounds
with P-end groups. In the olefinic range the assignment was again simplified
by a comparison with the spectrum of P-carotene-15,15'-0 2 shown in the upper
part of the spectrum.
 N
w
a-

Table 10. Chemical shifts (in p. p.m.) of some carotenoids. Solvent: CDCI 3

Compound Mamly olefm1c protons

Formula' Name 2 2' 4 4' 6 6' 7 7' 8 8' 10 10' II II' 12 12' 14 14' 15 15'

A P-Carotene - - - -614 -614 614 6.65 6.34 -624 -663 ~

P-Carotene-15,15'-0 2 - - - -616 -616 6.16 665 636 624 -
15-cJS-P-Carotene - - - -615 -615 6.15 667 641 -665 -639

B Lycopene 511 - 594 648 624 6.17 662 634 -6.24 -662
j
Lycopene-15,15'-0 2 5.10 - 5.94 648 624 617 662 6.34 624 - p
c e-Carotene - 540 217 5 51 6.10 611 660 633 -623 -6.62
o __
2,2' -Ounethyl- - 537 215 5.58 610 613 662 6.34 -624 -663
e..carotene I
E Zeaxanthm - - - -611 -611 614 6.63 6 35 -6.24 -662
;z
F Canthaxanthm - - - 621* 6 35* 626 663 641 -626 -665

G 15,15'-0!dehydro- - - - 7.21* 677* 629 674 642 5.77 - i

d1anhydro- ~
P-carotenone Po
~
H P-Carotenone - - - 654 7 39 -6.51 na n.a -6.35 -6.70 Vl

I o-Carotene - - - 540 - 217 -6.14 552 -614 610 614 611 6.65 6.60 6.35* 634* -624 -6.24 -662 -6.62

J y-Carotene - 511 - - - 5 95 -615 6.49 -615 624 614 617 665 6.63 635 6.35 -624 -624 -660 -6.60

K a-Carotene - 510 540 - 217 5.94 5 51 648 609 6.24 610 617 660 6.62 6.32* 6.34* -6.24 -624 -6.60 -6.60

- - 641 -627 -663

!
L Echmenone - - - - -621* 614 634* -614 na -614 na n.a 6.36

259
M Fucoxanthm - - - - - - - - 605 716 612 656* 675* 671 6.33 640 625 -6.64
3.68
I -- ·- ---- -- ·------

(Continued)
 Table 10. (Continued)
Compound Methyl protons Methylene protons Others

Formula a Name I I' 5 5' 9 9' 13 13' 2 2' 3 3' 4 4'

A P-Carotene 103 1.72 198 198 1.45 -160 2.02 -

P-Carotene-15,15'-0 2 103 I 71 1.97 197 1.46 -1.59 202 -
15-crs-P-C..rotene 102 I 73 198 198 1.45 -1.60 201 -
163
B Lycopene 1.83 -1.97 -197 - -212 -2.12 -
170
162
Lycopene-15,15'-0 2 1.83 -197 -197 - -2.11 -2.12 -
1.70
082 1.18
c a-Carotene 158 191 195 -200 - -
090 1.45
:;::
D 2,2' -Drmethyl- 0.78 en
159 191 198 na na - 2-CH,: -o8o
a-carotene 0.81
147 2.04
E Zeaxanthm 107 174 197 197 H-3,3'. -4 00
-
l8
170 2.39 n
"'0
F Cantaxanthm 1.19 1.87 199* 200* 185(t) 251(t) - -
~-
G 15,15'-D•dehydro- 1.29 - 201 201 174(t) 264(t) - COCH 3 :226 3::
dJanhydro- ~
p-carotenone 0
=-
p.
H P-Carotenone 1.16 - 1.99 199 n.a na 2.40 COCH 3 ·2.10 "'
0.81 -1.18
I a-Carotene 1.02 172 1.58 197 1.91 1.97 1.97 145 1.60 -2.00 -2.00 - -
0.90 -142
1.61
J y-Carotene I 03 172 183 197 197 197 197 145 - -160 -211 202 211 -
1.69
082 162 -119
K ~-Carotene 1.58 183 1.92 197 197 197 - 200 -211 - 211 -
0.90 1.69 -1.45
L Echmenone 119 103 188 172 -198 -1.98 -198 -198 185 -146 251 -1.60 - na -
096 108 COCH 3 : 2.04;
M Fucoxanthm 1.22 1.39* 195 182 1.99 1.99 n.a - na
I 04* 135* H-3. 3.81; H-3'· 5.36
~-- - - - --- _L__ ---- ---- ---

(Continued) N
t..>
*: Assignment uncertain. n.a.: Not assigned. (t): Triplet. •: See page 238. -.I
 6
0
..l~
"(
~

J:
/)·
~:
~ I'! ~
g ...,
!='
II~
-~
75
0
"' ::s
g·
~
~ :I:
<~
(: .);
)· )~
"' "' <
F"-
~ j"- ~
<
!) '=
j
<
~
~\ :>::
tf ~ ..
~
X
"' 0 "'
'IIH~HIM.H:JS '[1 puu ISSVDI)I'N 'nl!l1:DN3 'D ''I!Hll!IA 'M 8£Z
 IV. Spectroscopic Methods 239

.......,
, .
13,13'-c ...

Fig. 24. Proton magnetic resonance spectrum (220 MHz, in CDC1 3 ) of all-trans-P-carotene and
olefinic part of the spectrum of P-carotene-15,15'-0 2 (upper left)

Concerning the signals ofH-14,14' and H-15,15' of 15,15'-cis-P-carotene it

should be noted that in Table 10 the previous assignments [19] have been
reversed, bringing the shift differences observed between all-trans- and 15-cis-
p-apo-12'-carotenal (see Table 8) and those of the two P-carotenes into close
agreement.
Fig. 25 presents the olefinic parts of the 220 MHz spectra of the symmetrical
(p-, e- and lycopene) and the unsymmetrical carotenes (ex-, b-andy-). A detailed
comparison of the spectra of the symmetrical compounds demonstrated the
additivity already mentioned. Part of the assignments have actually been
carried out by superimposing the experimental traces.

4. Recent developments
The examples discussed so far clearly demonstrate the advantage of p.m.r.
spectroscopy at high spectrometer frequencies. Fortunately, instruments using
frequencies of up to 300 MHz are now commercially available (Varian Asso-
ciates, Palo Alto, Calif.). It is believed that the loss of resolution generally
connected with instruments using superconducting magnets (line width "'1 Hz)
as opposed to 'classical' instruments ( "'0.2 Hz) is more than outweighed by
the increase of chemical shifts, making highfield spectroscopy especially suitable
for carotenoid research.
Besides p.m.r. spectroscopy, in recent years carbon-13 n.m.r. has become
increasingly important. Severe sensitivity problems connected with the low
natural abundance of 13 C ("' 1.1 %) and long spin-lattice relaxation times have
previously hampered the application of 13 C n.m.r. on a routine basis. These
difficulties have been partly overcome or at least reduced by the application of
the Fourier transform technique [26]. In contrast to conventional n.m.r.
spectroscopy where, at any instant, only one single point of the spectrum is
observed during the field- or frequency sweep, with the Fourier technique the
nuclei are simultaneously and repetitively irradiated with a broad frequency
240 W. VETTER, G. ENGLERT, N. RIGASSI and U. SCHWIETER

220MHz
B-c.otana

5.5 pp.m.

~
. - -... , -· ., ... .,....
yt...~ -~ 220MHz
~:-Carotene

CHC~ H7,7' ~·
5.51 &.40

7 6.5 6 5.5 5p.pm.

220MHz

---
~15'
.8.82

Hn,11'

CHCI 3
H2,2'
5.11

7 6.5 6 5.5
Fig. 25. Olefinic region of the proton magnetic resonance spectra (220 MHz) of the carotenes
(in CDCI 3 ). [Continued on p. 241]
 IV. Spectroscopic Methods 241

~.l..J...!ll.Jt J£~.!::1.-i)
(j(_'~~,~~"r.'"'r-"'r'~ X 220MHz
lts,15'
ex-Carotene ~682

HCb

4 9

7 6 5.5 5ppm.

~4
220MHz

6-Carotene

1115,15'
N680
~

1111 Ha
860 6.09
CHCI3 (), A

220MHz
y-Carotene

1114,14'
H12 ,-A...
1112' Ha'
6.35 6.24
nh

~·
CHCI 3 5.11

7 6.5 6 5.5

Fig. 25. [Continued from p. 240]

Carotenmds 16
242 W. VETTER, G. ENGLERT, N. RIGASSI and U. SCHWIETER

band produced by equally-spaced short radio frequency pulses. This leads to

a considerable reduction of the time (by a factor of about 100) necessary for the
attainment of a given signal-to-noise ratio. The resulting response signals of
the nuclear spin system (free induction decay) following each pulse are accu-
mulated coherently in a time-averaging computer. The absorption spectrum is
obtained from the accumulated response by a Fourier transformation carried
out usually in less than a minute by a small digital computer linked to the
spectrometer.
A further increase of sensitivity (or reduction of recording time) is achieved
if, by broad-band irradiation (noise decoupling), at the same time all the protons
are effectively decoupled from the 13 C nuclei. The signals of the latter therefore
collapse to singlets. In addition, an increase in signal intensity due to an intra-
molecular nuclear Overhauser effect [27] is achieved. This again may reduce
the observation time. The 13 C spectra obtained by this technique consist of a
number of singlets cor:responding to the number of carbon atoms in the mole-
cule possessing different chemical shifts.
An example is presented in Fig. 26, showing the 13 C n.m.r. spectra at
22.63 MHz of /3-carotene (3) and of its 15,15'-dideuterio analogue. The concen-
tration of the latter compound was 50 mg dissolved in about 2.5 ml chloroform.
Some tetramethylsilane (TMS) was added as an internal reference (0 p.p.m.) as
is usually done in p.m.r., and the field-frequency lock was provided by the 19 F
signal of internal hexafluorobenzene. The spectrum of the dideuterio compound
was obtained by accumulating more than 16,000 responses (pulse repetition
0.4 sec) corresponding to a total observation time ofless than 2 hours. It should
be pointed out that, with correspondingly higher concentrations, observation
times of less than a minute are possible.
Since the above-mentioned Overhauser enhancement is negligible for
tertiary carbon atoms, these signals may be considerably weaker in intensity.
Similarly, under these experimental conditions, the signals of C-15 and C-15'
in the dideuterio compound are expected to become undetectable in the back-
ground noise. Although application of broad-band decoupling causes an
advantageous increase in the signal-to-noise ratio of most of the signals, the
use of intensities as an aid in the assignment of signals is, in general, more
difficult than when using p.m.r. [27].
The 13 C n.m.r. spectra of vitamin A acetate, /3-carotene and 15,15'-dide-
hydro-/3-carotene obtained by conventional sweep techniques have recently
been discussed [28], and a tentative assignment of the signals to the different
carbon atoms has partly been achieved by a comparison with suitable reference
compounds and with the aid of special double resonance techniques. It is
obvious that the identification of individual carbon atoms in complex sub-
stances provides an important non-degradative technique for the investigation
of specific sites of chemical and biochemical structures. It is beyond the scope
of this review to discuss the assignment of the different signals of Fig. 26. It
need only be pointed out here that the number of observed signals indicates
that most of the 20 (by symmetry) different carbon atoms give rise to separate
 IV. Spectroscopic Methods 243

13-Carotene

1357 C-15.15" 19B Oppm

1410 340/ T29!l TMS
1413 \1330
1283 341 131
3> 100
15,15'·H2 409

113

0 ppm
198 TMS

1358 131
1415 \1340 1300 T283
1413
1407 40.9
1318
15,15'-02

Fig. 26. 13 C n.m.r. spectra at 22.63 MHz of fJ-carotene and of its 15,15'-D 2 analogue (in CHCI 3 ),
obtained by Fourier transform technique. Protons decoupled by irradiation at 90 MHz (noise
decoupling). *: Impurity.
(Courtesy of Mr. T. W. Keller of Bruker-Physik AG, Karlsruhe-Forchheim)

signals. While the signals of the saturated carbon atoms appear at high fields,
those of the unsaturated carbon atoms absorb at low fields. In this part, one of
the signals appearing 133.0 p.p.m. below TMS ( ~61 p.p.m. above CS 2 ) has
disappeared in the spectrum of the 15,15'-dideuterio compound and must
therefore be assigned to carbons 15 and 15'.
From this brief discussion it is evident that the application of 13 C n.m.r. will
no doubt be of great importance in carotenoid chemistry, despite the fact that
the price of the rather sophisticated instrumentation is considerably high.

E. Mass Spectrometry
Three different kinds of information useful for structural analysis can be
obtained from a mass spectrum [29- 31] :
1. The molecular weight and, if measured to an accuracy of a few p.p.m., the
elementary composition of a compound.
244 W. VETTER, G. ENGLERT, N. RIGASSI and U. SCHWIETER

2. Certain structural features may be deduced from the fragmentation pattern,

provided that spectra of a number of structurally related compounds are
available for comparison.
3. Proof of the identity of different samples. Since mass spectra usually contain
numerous peaks and are easily reproducible, they are generally considered
to be excellent for this purpose.
Mass spectra of organic compounds can only be obtained from vaporized
samples. This creates problems for the mass spectrometric investigation of
carotenoids, which are known to suffer degradation when vaporization is
attempted. The development oftechniques for introducing high boiling samples
into a mass spectrometer has reached the point where most carotenoids can
be handled successfully. Since, however, partial decomposition of the samples
during vaporization is unavoidable, the reproducibility of the mass spectra
of carotenoids is very poor.
This poor reproducibility does not normally interfere with the use of mass
spectroscopy for molecular weight determinations. It may however cause
serious difficulties in interpreting the spectra in terms of structures, and it can
render the identification of compounds impossible if proper precautions are
neglected. It therefore seems appropriate to give considerable attention to the
vaporization and the thermal degradation of the carotenoids.

1. Vaporization and thermal degradation

In a paper published in 1933, Kuhn and Winterstein made a summary of
the knowledge acquired by themselves and others, about the behaviour of
various carotenoids when heated to 200-300 °C in vacuo [32]. Distillation of
the compounds was impossible in all cases; the distillates were, at most, only
slightly coloured, and a major portion of the samples had decomposed into non-
volatile, probably polymeric, products. The list of their volatile degradation
products contains the following compounds:
1. Toluene and m-xylene (possibly including traces of o-xylene).
2. 2,6-Dimethylnaphthalene.
3. A trimethyloctatetraenedicarboxylic acid dimethyl ester from crocetin di-
methyl ester (270), believed to be formed by expulsion of toluene from the
polyene chain.
4. m-Toluic acid, from carotenoids containing carboxy groups.
5. Also from crocetin dimethyl ester, a colourless crystalline compound, iso-
meric with crocetin dimethyl ester, containing four double bonds instead
of seven in the original molecule, and therefore named 'tricyclo-crocetin'
dimethyl ester.
These findings were later confirmed by a number of investigators [33-36].
In addition it was subsequently shown that P-carotene gives rise to a number
of hydroaromatic products [33}, mainly ionene [36], which originate from
the ring structures at the ends of the polyene chain.
 IV. Spectroscopic Methods 245

Except for the polymeric residue, all other pyrolysis products are readily
volatile and will, therefore, contribute to the mass spectra. In fact, the molecular
ions of the thermal degradation products expected can often be detected in the
mass spectra of carotenoids. The extent to which the electron impact fragmen-
tation patterns are really those of thermally degraded and isomerized carot-
enoids has not yet been established. A sample oflycopene (19), treated thermally
until its colour had disappeared, gave a mass spectrum still exhibiting most of
the features, including the molecular ion at m/e 536, characteristic for the mass
spectrum obtained from lycopene (19) itself [19].
It seems remarkable that mass spectrometry is nevertheless a very useful
tool for the elucidation of structures in the carotenoid field. This is apparently
due to two features of the thermal degradation. First, a portion of the samples
is converted into compounds whose mass spectra are readily interpretable in
terms of the original structures: isomerized cyclic compounds, and molecules
which have lost well-defined portions of their polyene chain. Second, the reac-
tions have a relatively good reproducibility, under carefully controlled con-
ditions. If, however, different techniques for vaporization are used for the same
compound, the spectra, although qualitatively still similar, show considerable
quantitative differences. Another consequence of the thermal instability of
these compounds is that they yield spectra which gradually change while the
sample is kept in the evaporation zone of the mass spectrometer [19]. Indeed,
total disappearence of the spectrum is often observed after a few minutes,
because the thermal decomposition of the sample has proceeded to completion,
in the sense that all volatile degradation products have been vaporized and
only polymeric material is left.

100
% ,..-69 ~ (a)

536
I
;91 444
1431 f67
300 400 500 m/0

100
% /69
~ (b)

/91

/378 ,536

200 300 400 m.e

Fig. 27. Mass spectra oflycopene (19) obtained under different experimental conditions: (a) Sample
introduced on a piece of heating wire at the fringe of the electron beam; temperature approx.
200 °C; heating period approx. 1 minute. (b) Sample introduced on the tip of a ceramic rod at a
distance of approx. 10 mm to the electron beam, temperature approx. 200 oc; heating period
approx. 5 minutes. Both spectra: MS 9, 70 eV, 100 J.lA, 8 kV, same pressure
246 W. VETTER, G. ENGLERT, N. RIGASSI and U. SCHWIETER

The differences to be expected between mass spectra of a carotenoid

obtained under slightly different experimental conditions are illustrated for
lycopene (19) in Fig. 27. Both mass spectra were taken at the same ionizing
voltage and ion source pressure with the same mass spectrometer but using
different vaporization techniques. In (a) the sample was deposited on a piece
of heating wire and inserted into the ion chamber at the fringe of the electron
beam. For the spectrum (b) the sample was deposited on the tip of a ceramic
rod and introduced at a distance of about 10 mm to the electron beam, inside a
channel leading to the ionization chamber. The minimum heat to achieve
sufficient vapor pressure was applied in both cases.
The gentler conditions of vaporization of the first mentioned technique are
reflected clearly in the mass spectrum (a), where the molecular ion is several
times more intense with respect to the total ion current than it is in the second
spectrum (b). In addition considerable differences in the intensity of some other
peaks, e.g. mje 91, can be observed.

2. Fragmentation
Since the electron impact-induced fragmentation patterns of the carotenoids
and of their thermal decomposition products cannot usually be separated, the
whole pattern obtained will have to be considered as the 'mass spectrum of the
carotenoid'. Only in exceptional cases will electron impact-induced reactions
and thermal reactions be differentiated. For the discussion of specific fragmen-
tation reactions, the structure of the carotenoids may be conveniently divided
into a polyene chain and two end groups [37], whereby a clear definition of the
three elements is irrelevant. Correspondingly the fragmentation reactions are
summarized in two groups: those occurring in the polyene chain, on which the
end groups have rather little influence, and those occurring within the end
groups or in the outer portions of the polyene chain and induced by the end
groups.
This classification also serves a practical purpose: the reactions of the first
class are highly characteristic for carotenoids and are, therefore, a good means
of distinguishing them from other classes of compounds. Fragmentation of the
end groups, on the other hand, may serve to distinguish individual carotenoids
from one another.
While fragmentation of the polyene chain can always be observed to some
extent, fragments related to the end groups vary widely in their intensities. End
groups containing structural features favourable to fragmentation give rise
to mass spectra dominated by the corresponding peaks. On the other hand,
there are certain end groups which can hardly be detected by mass spectroscopy,
because of a lack of specific fragmentation. Examples of two extreme cases are
given in Figs. 28 and 29. Fig. 28 shows the spectrum of P-carotene (3), whose end
groups show an almost complete lack of characteristic fragments, while peak
m/e 73 in Fig. 29 represents an ion arising from the very easy cleavage of the
end groups in 2,2'-diketospirillox~nthin (208) [38].
 IV. Spectroscopic Methods 247
100
%

300
I
378

400
r/10/444

500 m,e

Fig. 28. Mass spectrum of {3-carotene (3). Experimental conditions: see Fig. 27(b)

1
%

91 518 624
IJ(!Q
I I I
100 200 300 400 500 600mle

Fig. 29. Mass spectrum of 2,2' -diketospirilloxanthin (208). Experimental conditions: see Fig. 27 (b)

For most carotenoids the picture is more like Fig. 28: the fragmentation
of the chain prevails in the spectrum, while only a few peaks can clearly be
attributed to the end groups. Fig. 28 also shows another feature typical of the
spectra of most carotenoids: the crowding of intense peaks in the low mass
region and the scarcity of peaks towards the high mass end. As a consequence
of this ion current distribution, particular peaks, even those of small intensity,
can clearly be recognized in the high mass region. On the other hand, most
peaks shown in the low mass region of Fig. 28 have little structural significance,
as they occur in similar intensity ratios in most carotenoid mass spectra. In
this region only exceptionally intense peaks may be considered significant.
With the increasing possibilities of recording complete mass spectra at high
resolution, this situation may change at least in the case of oxygen-containing
carotenoids. By arranging the ions according to their oxygen content [30],
structurally significant peaks even of moderate intensity could be detected
among the uncharacteristic hydrocarbon peaks.
The following discussion of fragmentation deals essentially with low
resolution data. It reflects the present situation in that peaks of very low
relative intensity in the high mass region will be discussed, whereas only intense
peaks of low mass ions will be considered.
100
%

ra 444
r30/
100 200 300 400 500 m/e

Fig. 30. Mass spectrum of {3-carotene (3). Experimental conditions: approx. 12 eV ionizing voltage;
otherwise see Fig. 27 (b)
248 W. VETTER, G. ENGLERT, N. RIGASSI and U. SCHWIETER

Most peaks in the low mass region arise from complicated multiple frag-
mentation reactions, as is revealed by corresponding metastable peaks. Their
appearance potentials are generally quite high in relation to those of the mole-
cular ions, so that a spectrum like the one shown in Fig. 30 of /3-carotene (3)
is easily obtained at low voltage. Due to the relatively high abundance of their
molecular ions, such low-voltage spectra are very useful for obtaining molecular
weights of carotenoids, particularly from impure samples. In many instances,
the characteristic fragments are also revealed more clearly in the low-voltage
spectra than at 70 eV [19, 39].

a) Polyene chain
Apart from insignificant, though abundant, ions of low mass caused by
cleavage of the polyene chain, a number of highly characteristic peaks also
attributable to fragmentation of the chain are observed. Most spectra of
carotenoids contain peaks at M-92, M-106 and M-158, coinciding with the
peaks to be expected from the molecular ions of thermal products previously
mentioned [18, 19, 40]. M-92 represents the loss of toluene, M-106 the loss
of xylene and M-158 the loss of C 12 H 20 (presumably the precursor of 2,6-
dimethylnaphthalene) from the molecular ions. The molecular ions of toluene,
xylene and dimethylnaphthalene can also be detected in the low mass region.
The fact that the peaks M-92, M-106 and M-158 are at least partly due to
fragments formed from ionized carotenoids (possibly isomerized on vaporiza-
tion) and not solely due to molecular ions of thermal degradation products
has been demonstrated by appropriate metastable peaks. They can be observed,
when the very sensitive method of searching the first field-free region of a
double focussing mass spectrometer is employed [19]. It is, however, note-
worthy that only the loss of toluene gives rise to a very intense metastable peak
in conventional mass spectra, whereas the other two are not detectable. An
explanation of this difference between the loss of toluene on one hand and the
loss of xylene and C 12 H 20 on the other has not yet been found.
While the peak at M-158 is always oflow intensity ( < 5 %) and often absent,
the other two peaks, M-92 and M-106, sometimes reach considerable inten-
sities and have therefore attracted much attention. The relative abundance of
these ions appears to be determined mainly by the length of the polyene chain
and by its structure and, to some extent, by the type of end groups (see Table 12).
The smallest chain length at which the losses of toluene and xylene occur is
not clearly defined. In apo-carotenoids containing 8 double bonds, the corre-
sponding peaks are usually still fairly intense, whereas with 7 double bonds only
a very small peak is observed for the loss of toluene and there is no peak for
xylene. On the other hand, in long chains such as are present in dodecapreno-
/3-carotene or in spirilloxanthin (108), consecutive losses of two molecules of
toluene and/or xylene are possible [19].
However, it must be emphasized that, particularly with carotenoids of
very low volatility, the thermal loss of xylene and toluene from the molecules
may strongly influence the pattern (lbtained in the mass spectrum.
 IV. Spectroscopic Methods 249

For the thermal expulsion of toluene and m-xylene, Edmunds and Johnstone
proposed a mechanism involving a 4-ring intermediate [33]. Fig. 31 shows this
mechanism for the formation of toluene, having been modified in the light of
recent investigations of the cyclization of polyolefins [ 41] and particularly of
the rules for the conservation of orbital symmetry [ 42].
Applications ofthis mechanism to other portions of the chain gives m-xylene,
and starting the cyclization with a 12-electron system instead of the one with
8 electrons outlined in Fig. 31leads to a dimethylcyclodecapentaene. A number
of cis-trans isomerizations, known to occur in polyenes under thermal con-
ditions [ 43, 44], has to precede the operation of the mechanism. Since the exact
configuration and conformation of the chain undergoing the cyclization is
not known, the arrangement ofthe chain drawn in Fig. 31 is, of course, arbitrary.

R'

R2
-- 92
+ ~
I
R'
# R2

Fig. 31. Mechanism for the thermal formation of toluene from an arbitrarily folded and cis-trans-
isomerized polyene chain of a carotenoid

The same sequence of steps may be used to describe the electron impact-
induced reaction, involving radical cations throughout the procedure until
neutral toluene is split off in the last step [19]. It can be expected that all steps,
including the preceding cis-trans isomerization are more facile in the electron-
deficient species than in the neutral molecules. Since the electronic state of the
fragmenting radical ions is unknown, the stereochemistry indicated in Fig. 31
for the intermediates of the thermal reaction is, of course, also unknown for the
ions produced upon electron impact.
This mechanism requires that cleavage occurs only between carbon atoms
connected by double bonds in the original polyene. A scheme can be constructed
on this basis for every carotenoid, showing all possible reaction paths. In
Fig. 32 such schemes are shown for the loss of toluene and xylene from f3-ca-
rotene.
Only the nuinber of possible ways for formation of the products is evident
from such a scheme. The actual relative yields obtained are determined supple-
mentarily by electronic and steric factors.
These schemes have been tested with differently substituted polyene chains.
All experiments are consistent with this mechanism. In particular, results
250 W. VETTER, G. ENGLERT, N. RIGASSI and U. SCHWIETER

: 106
I
I
I
I
"<:::::::
I I
IL _________ .JI

: 92
I
I
I
I

I I I I I
I I I I I
L--~---L--.J I
L--~---L--.J
L __ .J ______ .JI
L _________ .J

Fig. 32. Schematic representation of the possible ways of formation of xylene and toluene from
P-carotene (3)

obtained from P-carotene-15,15'-D2 and lycopene-7,7'-D 2 showed that, within

experimental error, only the expected ions are formed. Thus, for example,
toluene-D 2 , toluene-D 1 and xylene are expelled from P-carotene-15,15'-D 2 , but
labelled xylene and unlabelled toluene are not. In contrast only toluene-D 1
besides unlabelled toluene and xylene is expelled from lycopene-7,7'-D2
[19, 45].
Further support for the mechanism was derived from the study of a syn-
thetic carotenoid, having the two central methyl groups oxygenated and
shifted from their normal13,13'-positions to 14,14'. Again, only products to be
expected from schemes of the type shown in Fig. 32 were found [ 46].
With the validity of the mechanism established, consideration of steric and
electronic factors influencing the course of the reaction at various stages may
eventually lead to reliable estimates of product yields. These could then be used
to elucidate the structure of carotenoids with substituted or modified chains.
A first step in this direction has been made by Enzell and Liaaen-Jensen, who
studied the fragment yields of various substituted carotenoids in relation to
steric crowding at the 4-membered ring of the corresponding intermediates [ 46].
It was concluded that such steric strain appears to have a marked influence on
the course of the reaction; those fragments requiring a more highly substituted
4-membered ring or one carrying bulky groups as, for example, the trimethyl-
cyclohexene group of P-carotene (3) are obtained in considerably decreased
yields.
In the light of these results, the decrease of the ratio M-92/M-106 observed
with increasing length of the acyclic polyene chain can be interpreted as being
caused by the generally increased possibilities for the sterically unhindered
formation of xylene relative to tolvene [ 40, 46].
 IV. Spectroscopic Methods 251

Substituents at the methyl groups attached to the polyene chain cause no

fundamental change in the fragmentation, the corresponding substituted
toluenes and xylenes being lost [ 47]. Knowledge of the influence of func-
tional groups on the quantitative course of the fragmentation is still, however,
very limited.
By reasoning along such lines, Katz and co-workers concluded that a
hydroxyl in the recently discovered carotenoid loroxanthin (88) has to be
located on the methyl groups in either position 9 or 9' [ 48].
Modifications within the polyene system of the carotenoids have widely
varying effects on the fragmentation. The spectra of 15,15'-cis- and 15,15'-trans-
P-carotene are indistinguishable from each other.lt is very likely that all other
cis-trans isomers will show the same mass spectra.
A triple bond in 15,15'-position totally inhibits the loss of toluene and xylene.
However, a small peak at M-90 can be observed presumably due to loss of
methylbenzyne [ 49]. A triple bond near the end of the chain has no such effect:
loss of92 can be observed (diadinoxanthin (118), Table 12: 45 A 50). Changing
the normal structure of carotenes to the retro structure does not appear to
alter the fragmentation of the polyene chain [20] significantly (rhodoxanthin
(209), Table 12:41 C 41).
The presence of an allene in position 9,10, as encountered for example in
fucoxanthin (190) (Table 12: 48 A 54), inhibits the typical chain fragmentation
[50]. To predict the behaviour of allenes in general from this single example
would, however, be premature.
Finally a further comment on the problem ofthe thermal origin offragmen-
tation products is necessary. In connection with the mass spectrometric
studies on P-carotene-15,15'-D2 , the reaction products formed by thermal
treatment were investigated using mass spectrometry to determine the distribu-
tion of the deuterium [19].1t was found that the results obtained with toluene,
xylene and 2,6-dimethylnaphthalene corresponded exactly to those expected
from the scheme in Fig. 32. If it is assumed, for the sake of argument, that the
peaks at M-92, M-106 and M-158 in the mass spectrum are mainly due to
molecular ions of thermally formed reaction products, then the above discussion
is irrelevant with regard to the events taking place after electron impact, and
the true electron impact fragmentation remains unknown. The solution of this
problem appears to lie in the study of the relevant metastable ions of labelled
molecules. Until such information is available, the only workable assumption
appears to be the one made at the outset: the peaks in the mass spectra are
considered to be due to electron impact fragmentation.
Frequently (M-2) peaks are encountered in the mass spectra of carot-
enoids [37, 39, 51] (see Table 12). Their intensities do not bear any obvious
relationship to the structures of carotenoids, and therefore they are of no use
for structure elucidation. Whether their origin is thermal or electron impact-
induced is not known. Presumably they are due to the loss of hydrogen from
cyclized polyene-chains, which acquire aromatic character by this reaction.
Another peak of low intensity ( < 3 %) appears in the mass spectra of most
252 W. VETTER, G. ENGLERT, N. RIGASSI and U. SCHWIETER

carotenoids at M-79 [40]. Like M-92 etc., this peak is an indication of a

carotenoid structure. The origin of the ion has not been investigated.

b) End groups
As is commonly observed in mass spectrometric studies, the extent of frag-
mentation of a particular group is not only dependent on its own properties
but also on the remainder of the molecular structure [31]. The latter effect is
strongly displayed in carotenoids, where the behaviour of one end group is
influenced by the presence of the polyene chain and the other end group.
Qualitatively there is usually no change in the fragmentation, so that particular
reactions can be taken as characteristic for certain functional groups [18].
Typical for carotenoid spectra is the occurrence of multiple fragmentations,
in which one ofthe characteristic reactions ofthe chain, as previously discussed,
takes place before or after a fragmentation reaction of an end group. It seems
that, in general, end group reactions taking place in the molecular ions also occur
in the ions formed by loss of toluene and xylene. The yields in relation to the
respective parent ions are usually of the same order of magnitude. Such sec-
ondary reactions are therefore ignored in the following discussion, but the peaks
may be quite useful in all those cases where the abundance of molecular ions
is low compared to that of the ions M-92 or M-106.
Table 11lists the functional groups together with the mass numbers of their
most characteristic fragments [37]. Intensity data are deliberately omitted
from this table because of their strong dependence on the overall structure
of the molecules. In consulting the table for information, it must be kept in
mind that the intensity of the peaks does have significance. A peak at a certain
mass number can only be accepted as indicative for a structural element if its
intensity has a certain value relative to the total ion current. For example, a peak
at mje 69 is present in the spectra of all carotenoids. However, if this peak is of
exceptional intensity with respect to the other fragment peaks and also to the
molecular ion, as in the case of lycopene (19) (Fig. 27), it is strongly indicative
of end group 1 (Table 11).
In order to evaluate the intensities of peaks expected from certain functional
groups in different molecules, Table 12 has been compiled [37], listing the
intensity values of the peaks at selected mass numbers. Besides peaks that are
characteristic for end groups, the table also contains some of those peaks
originating from the fragmentation of the chain: mje 91, M-92, M-106, M-2.
These peaks may be used, in addition to the others, for the identification of a
compound, provided that proper attention is paid to possible intensity changes
due to introduction conditions.
As can be seen from Table 12, many end groups give rise to characteristic
peaks of only very low intensity in the high mass region. Great care has then to
be taken when the interpretation of spectra from samples of less than perfect
purity is attempted.
In the high mass region of some spectra, peaks of low intensity are observed
which can formally be derived by ,cleavage of a bond of the polyene chain,
 IV. Spectroscopic Methods 253

Table 11. Mass numbers of characteristic peaks of some carotenoid end groups
{adapted from [37])
End group No. m/e

~-­ M-69, 69

~-· 2 M-69, M-137,69

3 M-69, M-137, M-205

4 M-69, M-82, M-122, M-135, M-148

5 M-16, M-18, M-85

6 M-28, M-29, M-83

R=H 7 M-18, 59
R=CH 3 8 M-30, M-32, 73

R=H 9 M-18, M-58, 59

R=CH 3 10 M-32, M-73, 73

/ ""--../~ l
CH 30~
y~~""'-·
~ l 11 M-32, M-101, M-129, 73
0

12 M-16,M-18,M-59,M-87,M-88,59
13 M-30,M-32,M-60,M-73,M-101, 73

HO~ -
'Y~~~~-·
l ~ 1 R=H
R = Rhamnosyl
14
15
M-16, M-18. M-58, M-59
M-18, M-58, M-164, M-180, M-205,
OR 113,153,171,273

16 M-28, M-29

17 M-43, 43, 109

~- 0
18 M-155, 127,109,69,43

R=H 19 M-44,M-45,M-99
R=CH 3 20 M-31, M-59, M-113
R=C 2 H 5 21 M-45, M-73, M-127

22 M-28, M-58, M (,0

{Continued)
 254 W. VETTER, G. ENGLERT, N. RIGASSI and U. SCHWIETER

Table 11. (Continued)

End group No. m/e

HO~ 23 M-18, M-16-18

HO

~·· 24 M-18, M-155, 127, 109

OH

~·· 25 125,83

~··
~­
"l
27
M-133, 133

R
ftA·· R=H
R=OH
R=OAc
28
29
30
M-137
M-18, M-153
M-60, M-195

~--
ROU- -
R=H 31 M-18, M-56, M-138
R=CH 3 32 M-32, M-56, M-152
R=Ac 33 M-60, M-180

R=H 34 M-56, M-123

~··
R=CH 3 35 M-70, M-137
R=~ 36 M-124

R=HOH,~· 37 M-140

38 M-56, M-138, M-151, M-204

R=H 39 M-56, M-138, M-203

R=OH 40 M-16, M-154, M-167, M-233

41 M-56, M-137, M-150, M-163,

M-190, M-203

Continued)
 IV. Spectroscopic Methods 255

Table 11. (Continued)

End group No. m/e

42 M-18, M-98, M-127, M-155, 109

43 M-18

44 M-138, M-151

45 M-18

46 M-16,M-154,203, 152,137

R=H 47 M-18
R=Ac 48 M-60

if
R=H 49 M-80, 165, 205
R=OH 50 M-18, M-80, 181, 221
R=OAc 51 M-60, M-80

.M
R

R=H 52 M-80, 165, 205

R=OH 53 M-18, M-80, 181, 221

~-·
ROUO~ . R=H
R=Ac
54
55
M-18
M-60

56 M-18, M-170

frequently accompanied by the shift of a H atom. The true mechanism of the

formation of these ions, which is probably much more complex, remains at
present unknown. Since these peaks may nevertheless serve to identify par-
ticular end groups, they have been included in the table.
256 W. VETTER, G. ENGLERT, N. RIGASSI and U. SCHWIETER

Table 12. Intensities of characteristic peaks in the

Compound Relative intensity at m/e
43 59 69 73 83 91
Lycopene 19 1A 1 16 0 100 8 47
Lycoxanthin 62 1A5 30 0 82 1 8 100
Lycopen-16-al 1A6 81 3 100 10 40 69
Rhodopin 56 1A 7 1 6 100 3 30 30
Anhydrorhodovibrin 97 1 A 10 4 0 10 96 100
Chlorobactene 15 1 A26 10 0 50 4 38
Rubixanthin 45 1 A 29 16 1 65 1 9 34
Rubixanthin acetate 1 A30 10(?) 0 19 0 3 20

Spheroidenone 182 2 A 13 5 0 27 100 25

11',12'-Didehydrononaprenoxanthin 217 2 A37

Nonaprenoxanthin 218 2 B37

Lycopersene 34 3(CH 2 )z3 4 0 100 0 5 3

Bisdehydrolycopene 17 4A 4 2 0 21 1 4 100
Lycophyll 83 5A 5 100 35 1 8 81
'Dihydroxylycopene' 81 7A 7 27 20 77 1 9 100
Rhodovibrin 106 7A 10 4 2 5 100 50
2-Ketorhodovibrin 7A 13 6 3 1 100 85
3,4,3' ,4'-Tetrahydrospirilloxanthin 110 SA 8 3 3 53 100 9 59
Didemethylspirilloxanthin 9A 9 23 100 15 2 4 67
Monodemethylspirilloxanthin 105 9A 10 5 13 3 100 1 88
Spirilloxanthin 108 10A10 3 0 100 0 16
Okenone 181 11 A 27 2 4 51 35 7 80
2'-Dehydroplectaniaxanthin 162 12 A28 48 36 37 26 13 100
2,2' -Diketospirilloxanthin 208 13 A 13 4 0 2 100 0 30
Plectaniaxanthin 76 14 A 28 30 27 34 1 10 100
Myxoxanthophyll 90 15 A29
Crocetindialdehyde 267 16H 16 12 0 0 3 36
Bixin dial 16 I 16 11 0 5 0 4 100
retro- Diapo-3,3' -carotenedial 16 J 16 34 1 24 2 13 73
Renieral 16D27 11 0 2 1 1 20
{J-Apo-2' -carotenal 233 16 G28 17 1 31 12 8 100
retro- Diapo-5,5' -carotenedione 17 C17 100 0 2 0 6 31
{J-Carotenone 216 18 A 18 100 0 70 4 37

Neurosporaxanthin 234 19E 28 27 48 14 57

Azafrin 261 19 A42 65 32 42 6 35

Dimethyl retro-diapo-3,3'- 20 J 20 14 35 11 2 3 100

carotenedioate
Azafrin methyl ester 20A42 59 9 39 5 24

Methyl {J-apo-10' -carotenoate 20A49 100 28 43 2 10 61

5,6-epoxide
Torularhodin ethyl ester 21 F 28 70 2 86 3 24 84
Violerythrin 22 A22
 IV. Spectroscopic Methods 257

mass spectra of some carotenoids (adapted from [37])

(%of base peak) Ref.
109 133 M M-2 M-92 M-106 M-x
19 22 22 10 1.5 4 M-69: 4 [37]
24 29 7 4.5 l.O 4 M-16: 8, M-18: 13, M-69: 0,5, M-85: 2 [37]
25 31 70 4 8 23 M-29: 1, M-69: 5, M-83: 2.5 [37]
26 21 7 0.5 2 8 M-18: 11, M-69: 0.5 [37]
2 2 1 0.1 0.3 5 M-69: 0.1, M-73: 0.1 [37]
7 100 73 l.3 ll ll M-69: 4, M-133: 1 [37]
17 26 100 5 15 24 M-18: 1.5, M-69: 4, M-153: 1 [37]
5 lO 100 4 15 34 M-60: 10, M-69: 1, M-195: 1.5, [37]
M-69-60: 2.5
2 6 9 8 l.5 12 M-30: 0.5, M-32: 2.5, M-60: 0.2, M-73: [37]
0.4, M-101: 0.6, M-137: 2.6
100 15 0 M-18: 15, M-137: 8, M-140: 11, [54]
M-92-137: 8
100 36 0 M-18: 30, M-140: 40, M-205: 32, [54]
M-92-140: 90, M-92-205: 90
12 3 3 0 0 0 M-69: 1.5, M-137: 2, M-205: l.3 [37]
16 ll 13 0 3 7 M-69: 0.5, M-82: 0.3 [37]
19 6 35 5 8 25 M-16: 4.5, M-18: 1.5, M-85: 4 [37]
25 28 18 0.5 7 20 M-18: 16, M-18-18: 12 [37]
2 3 1.2 0 0.3 5.3 M-18: 0.8, M-32: 0.2, M-73: 0.1 [37]
4 6 6.5 0 1.2 23 M-18: 2.2, M-30: 0.3, M-32: 0.4, M-73: 0.3 [37]
22 23 2.0 0.7 0.7 2.8 M-30: 1.2, M-32: 1.7 [37]
6 14 14 0.8 l.3 25 M-18: 1.4, M-58: l.O [37]
2 5 4.5 0.5 0.5 20 M-18: 1.9, M-32: l.O, M-58: 0.3, M-73: 0.3 [37]
1 1 0.5 0 0.1 3.2 M-32: 0.1, M-73: 0.1 [37]
7 100 15 1.5 1 7.5 M-32: 22, M-101: 0.5 [37]
16 21 52 5 4 37 M-16: 2.5, M-18: 4, M-88: 5.5 [37]
1 2 0.6 0.1 0.1 1.9 M-30: 0.2, M-32: 0.3, M-73: 0.1, M-101: 0.3 [37]
15 25 17 2 2.5 39 M-16: 3.5, M-18: 8, M-58: 3.7, M-59: 2.7 [37]
1 0 0.2 1.4 M-58: 0.5, M-146: 1.3, M-164: 7 [59]
7 lO 100 1 0.5 0 M-28: 2, M-29: 4 [37]
5 17 45 16 1 ll M-28: 6, M-29: 5 [37]
18 23 100 5 5 lO M-28: 1, M-29: 1 [55c]
2 42 100 4 4 9 M-28: 1, M-29: 1, M-133: 2.5 [37]
12 24 60 24 4 30 M-28: 1.5, M-29: l.5 [37]
18 5 17 1 0 0 M-43: 1.8 [57c]
97 3 0.9 0 0 2.2 M-18: 0.4, M-155: 1.9, M-106-127: 2.8, [56c]
127: 50
17 31 100 9 17 10 M-44: 3, M-99: 1 [37]
100 18 24 1 0 M-18: 19, M-44: 6, M-98: 6, M-127: 3.5, [37]
M-155: 7
4 18 35 2 16 M-31: 1.3, M-59: 1 [57c]

100 19 62 2 0 M-18: 15, M-31: 3, M-32: 4, M-59: 2, [37]

M-98: 4.5, M-127: 6, M-155: 8
18 15 42 4 0 M-15: 9, M-80: 15, M-31: 3, M-59: 4 [55c]

30 43 60 5 9 17 M-45: l.l, M-137: 3, 119: 100 [58c]

100 33 10 ll M-28: 4, M-58: 4, M-60: 4 [53]
(Continued)
Carotenmds 17
258 W. VETTER, G. ENGLERT, N. RIGASSI and U. SCHWIETER

Table12.
Compound Relative intensity at m/e
43 59 69 73 83 91
Violerythrol 23 A23
Capsorubin 205 24 A24 46 2 28 3 51 40

Capsanthin 170 24 A29 23 2 15 1 37 100

Capsanthone 197 25 A29 38 2 37 0 100 60
P-Carotene 3 28 A28 30 0 100 0 23 45
IX-Carotene 5 28 A34 38 0 92 2 20 94
2' -Methyi-IX-carotene 28 A35 41 0 79 3 27 80
Cryptoxanthin 39 28 A29 51 3 92 1 27 78
Echinenone 148 28 A39 4 0 10 1 2 9
Mutatochrome 125 28 A 52 60 1 91 0 21 72
Zeaxanthin 67 29 A29 95 6 45 2 24 76
Lutein 73 29 A31 36 4 34 1 14 100

3'-0-Methyllutein 29 A32 100 6 27 6 20 38

3'-Dehydrolutein 156a 29 A38
Antheraxanthin 119 29 A 50 90 4 51 5 30 57

Lutein diacetate 30A33 100 32 1 9 82

Antheraxanthin diacetate 30A51 100 1 65 5 56 20
a-Carotene 10 34A34 33 0 45 1 9 70
Deshydroxydecaprenoxanthin 223 36 A37
Decaprenoxanthin 226 37 A37
Canthaxanthin 193 39 A39 26 1 15 1 100 19
Astaxanthin 203 40A40 52 2 37 2 21 100
Rhodoxanthin 209 41 C41 12 12 2 10 89

Isozeaxanthin 71 43 A43 53 2 38 3 13 100

P-Carotene-3,3'-dione 44 A44 38 1 33 0 32 80
Diadinoxanthin 118 45 A 50 78 10 52 100

Neoxanthin 122 47 A 50 6 77 5 55 100

Neochrome 131 47 A 53

Fucoxanthin 190 48 A 54 100 20 11 68

Violaxanthin 135 50 A 50 68 4 46 100

A urochrome 139 52 A 52 33 0 14 35

•: No intensity values published.

b: List number (Chapter XII).
c: Experimental conditions: see Fig. 27(b).
d: The numbers represent end groups from Table 11, the letters represent the following central
fragments:

·~. ·~ '~/
A B c
 IV. Spectroscopic Methods 259

(Continued)
(% of base peak) Ref.
109 133 M M-2 M-92 M-106 M-x
29 4 10 16 M-18: 30, M-18-16: 11 [53]
100 16 5 2 0 7 M-16: 5, M-18: 9, M-16-18: 6, M-155: 9, [39]
127:35
87 20 17 0 1 25 M-18: 5, M-155: 4 [39]
20 21 38 8 7 24 M-16: 9, M-18: 17, M-153: 2, 125: 100 [39]
22 34 44 3 15 4 M-137: 1.5, 119: 72 [19]
29 35 100 5 22 12 M-56: 2, M-123: 3, 119: 100 [19]
29 43 45 7 16 8 M-70: 2.6, M-137: 3, 119: 100 [19]
34 46 83 2 35 8 M-18: 0.6, M-137: 2.1, M-153: 2.3, 119:100 [58°]
4 9 100 9 13 3 M-56: 0.5, M-203: 2 [37]
31 37 20 8 5 2 M-80:24,205: 100,165:55,336:17 [56°]
37 53 100 7 34 3 M-18: 12, M-153: 3 [37]
28 38 23 2 8 5 M-18: 31, M-56: 0.3, M-153: 1.5, M-138: [37]
2.5, M-158-18: 29, M-106-138: 41
21 16 4.2 0.6 2.5 1 M-18:0.2, M-32: 2.5, M-56:0.5, M-152: 7 [37]
100 3 16 9 M-18: 7, M-138: 5, M-151: 2, M-153: 3 [37]
30 31 52 6 19 2 M-16: 22, M-18: 38, M-80: 40, 352:21, [39]
221: 100, 181: 25
30 51 3 0 0.5 0.3 M-60: 5.5, M-195: 0.1 [37]
25 16 45 2 7 2 M-60: 8, M-80: 15, 394: 9, 263: 13 [39]
25 48 72 0 9 8 M-56: 2.5, M-123: 3, 123: 100 [19]
a
100 8 0 M-18: 26, M-124: 6, M-140: 8 [54]
100 25 3 M-16: 5, M-140: 43, M-106-140: 46 [52]
8 14 41 2 7 2 M-56: 0.5, M-138: 0.7, M-204: 2.5 [37]
24 56 6 4 2.5 3 M-16: 8, M-16-16: 6, M-154: 1.5 [37]
15 28 100 15 24 43 M-56: 0.5, M-137: 5, M-150: 4, M-163: 6, [37]
M-190: 4, M-203: 9
18 42 0.6 0.6 2.5 1 M-18: 4.3, M-20: 3, M-18-18: 7 [57°]
24 63 91 0 15 5 M-138: 3, M-151: 2, 119: 100 [57j
41 40 22 7 12 0 M-16: 8, M-18: 10, M-16-18: 5, M-80: 10, [39]
352:21,221:80,181:47
51 62 24 1 5 0 M-18:2l,M-80:2,352: 13,221:92,181:61 [39]
22 1 8 1 M-16: 10, M-18: 19, M-80: 10, 352: 18, [39]
221: 92, 181: 61
17 14 5 0 2 M-16: 3, M-18: 8, M-18-18-60: 8, 221:40, [39]
197:76
43 43 35 3 2 1 M-18: 5, M-80: 7, 352: 13, 221: 80, 181: 52 [39]
24 14 23 8 1 0 M-80: 7, M-80-80: 14, 336: 9, 205: 100, [39]
165: 63

.··~.· ·~···
E

··~·· .. ·""" """ """

""::/

F G
(Continued)
260 W. VETTER, G. ENGLERT, N. RIGASSI and U. SCHWIETER

Table 12. (Continued)

~- H

I J

An interpretation of those peaks whose origin appears to be reasonably well

established is given in the following paragraphs.
As with most other classes of compounds, carotenoids containing hydroxy
groups or derivatives thereof1at saturated carbon atoms tend to lose water,
acetic acid etc., respectively, in varying amounts [17, 31, 50]. Multiple cleavages
of this type are also quite abundant. The temperature necessary to achieve
sufficient vapour pressure of these compounds suggests that some of these
eliminations are of thermal origin. Loss of water in small amounts is also
observed from aldehydes, ketones and cyclic ethers [17, 37]. From compounds
containing a 3-acetoxy-~:-end group, loss of ketene is observed [50]. A usual
fragmentation, apparently typical of some hydroxy groups occurring in carot-
enoids, is the loss of oxygen [17, 37]. To what extent this reaction is thermal or
electron impact-induced remains to be determined [39]. Glucosidic carotenoids
show a strong tendency to eliminate the sugar residue from the molecular
ions [59, 60]. In order to ensure vaporization without excessive thermal
degradation of the molecules, the hydroxy groups of the sugar may be con-
verted to acetoxy or trimethylsilyl ether groups. The fragmentation pattern
of the sugar is clearly discernible in the spectra superimposed on the fragmenta-
tion pattern of the carotenoid structure [60].
The loss of methyl groups appears to be surprisingly rare. Even in com-
pounds such as {3-carotene, having several geminal methyl groups in allylic
positions, the intensity of the M-15 peak is usually less than 1% of the base
peak [18].
End groups connected to the polyene chain by a doubly allylic bond
(Table 11, No. 1-3) show the expected cleavages of this bond [18, 61]:
I
I
I ! I
R~--··'
I
I

R+54 : M-(R+54)
_ _ _ I_ __

In the case of lycopene (19) (R=CH 3 ), both peaks, mje 69 and M-69 are
observed, with mje 69 being much more pronounced. If one of the terminal
methyl groups is oxidized (R=C~ 2 0H or CHO) (Table 11, No.5 and 6), this
can be detected by the corresponding mass difference between the high mass
 IV. Spectroscopic Methods 261

peak and the molecular weight. The intensity of the low mass fragment, at
m/e 85 and m/e 83 respectively, is, however, considerably reduced in com-
parison to m/e 69 of lycopene.
End groups containing several doubly allylic bonds (Table 11, No.2 and 3)
undergo cleavages at all these bonds. Thus, in the spectrum oflycopersene (34)
(Table 12, 3 (CH 2 h 3) for example, the peaks M-69, M-137 and M-205 are
due to such fragments. At the low mass side only mfe 69 is prominent, however.
A conjugated chain extending from one end of the molecules to the other,
as in bisdehydrolycopene (17), shows no characteristic fragmentation for the
chain ends [37].
Termination of a chain by 2-hydroxyisobutyl or 2-methoxyisobutyl groups
leads usually to strong peaks in the low mass region owing to the formation of
favourable ions (Fig. 29; Table 11, No. 7-13) [37, 38, 61, 62]:
I
I
RO...._I: I .
~-····
I
I
R+58:
___ I

If the hydroxy group is in P-position with respect to the polyene chain, the loss

r
of acetone is observed in considerable amounts (Table 11, No.9, 14 and
15) [37]:

~·· 0 ~··
~./ - )l +

R R
M-58
An analogous reaction is found when a methoxy group is in IX-position to a
ketone (Table 11, No. 13) [37].
®e

?~J-
H2C~ (o
H
CH,O + ~J OH
M-30

Aldehyde, acid and ester groups (Table 11, No.16, 19,20 and 21) attached
directly to the.polyene chain give rise to weak peaks only, loss of CO, C0 2 and
RO, respectively, being the main reactions [20, 37]. In the low mass region of
the mass spectra of esters, the alkoxycarbonyl ion (at m/e 59 for methyl ester)
is sometimes fairly prominent.
In the spectra of methyl ketones an intense peak at m/e 43 and a small
one at M -43 clearly indicate a cleavage reaction next to the keto group
262 W. VETTER, G. ENGLERT, N. RIGASSI and U. SCHWIETER

(Table 11, No.17): Ql

II! I
~-- ....
I
I
43 II__
___ M-43
_

In addition, a peak at 109 may be attributable to an ion produced in the

following reaction [63, 64]:

n 0

109

Other ketones (Table 11, No. 18, 24 and 25) also show peaks of considerable
intensities, which are due to cleavage reactions of bonds next to the carbonyl
group [49], for example:

Surprisingly, in such cases the charge remains preferentially on the tertiary

carbon atom and not on the conjugated carbonyl group, as is indicated by a
strong peak at m/e 127 and only a very weak one at M-127.
Phenyl nuclei (Table 11, No. 26 and 27) at the chain end give rise to ions
formally due to cleavage of the adjacent double bond accompanied by a shift
of one H atom to the fragment containing the aromatic ring. The mechanism
of the reaction is unknown [37] :
I

~
A) i . . .
I
---J....-H
I
133 II __
___ M-133
_

Six-membered rings with a double bond in the 5,6-position (Table 11,

No. 28, 43 and 44) show the same cleavage but with very small intensity and
no other significant fragmentation [37]. On the contrary, 4,5-cyclohexene end
groups (Table 11, No. 31 to 37) undergo a retro Diels-Alder reaction with the
 IV. Spectroscopic Methods 263

charge remaining at the polyene [18]:

~r~·~+
Rl~ 4
M-(R 1 +55)
By this reaction substituents in positions 2 and 3 can be differentiated [52]. The
reaction is noticeably characteristic for such L1 4 • 5 -structures, but the cor-
responding peaks are sometimes of very low intensity.
Fragmentation of cyclohexenone end groups (Table 11, No. 38 to 40) gives
rise to rather insignificant peaks only. The corresponding diosphenols (Table 11,
No. 46), however, exhibit a pronounced and characteristic fragmentation [49].
The first process, leading to m/e 152 and by subsequent loss of a methyl group
to m/e 137, is not fully understood: I
I
I~
I
I
HO I
I
I
I

+-u
I
152
I
I

137
For a second important reaction a mechanism has been proposed by Weedon

andco-wor~ ·.. l··~

®e

HO~H{'~· ..
0
HO~J 0

j ..
HO~
m H~J 0
203
264 W. VETTER, G. ENGLERT, N. RIGASSI and U. SCHWIETER

The spectra of 5,6- and 5,8-epoxides are very similar, so that mass spectro-
metry is not a reliable tool for differentiating between the two types of structures
[39, 65]. Of all functional groups encountered, the epoxides are among those
with the strongest fragmentation-directing properties (Table 11, No. 49-56).
One type of fragmentation gives rise to two intense peaks in the lower half of
the mass spectrum. The formation of these peaks may be adequately described
by the following mechanism [65]:

R
1l

R~J
j
)):'l.
164+R 204+R
A second characteristic reaction is the loss of 80 mass units (C 6 H 8 ) from the
molecular ions. A mechanism, postulating the loss of a portion of the cyclo-
hexane ring [65], has been disproved by a labelling experiment [39]. Although
no other mechanism has been proposed, it is likely that this reaction is similar
to the expulsion of toluene and xylene from the polyene chain [39].
Epoxides, containing an additional functional group in the chain close to
the end group, do not exhibit the typical fragmentations just described. Instead
they may undergo reactions involving both functional groups, for example [39]:

M-170
 IV. Spectroscopic Methods 265

A number of further fragments occurring in the spectra of carotenoid

epoxides are listed in Table 12, e.g. mje 352 in diadinoxanthin (118) {Table 12,
45 A 50) or mje 197 in fucoxanthin (190) (Table 12,48 A 54). These ions must be
formed by cleavage reactions within the polyene chain, but the mechanism of
their formation is presently not understood [29].
The authors are indepted to several of their colleagues of F. Hoffmann-La Roche & Co. Ltd.,
Basle, in particular to Drs. U. Gloor, U. Leuenberger, H.J. Mayer, R. Ruegg, G. Ryser and H.P.
Wagner, for providing numerous compounds from unpublished work, and to Dr. L.Chopard-dit-
J ean for infrared spectra.

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[34] G. G. McKeown, J. Ass. Offic. Agr. Chern. 48, 835 (1965).

[35] W.C. Day and J.G. Erdman, Science 141,808 (1963).
[36] I. Mader, Science 144, 533 (1964).
[37] C. R. Enzell, G. W. Francis and S. Liaaen-Jensen, Acta Chern. Scand. 23, 727 (1969).
[38] U. Schwieter, R. Riiegg and 0. Isler, Helv. Chirn. Acta 49, 992 (1966).
[39] H. Budzikiewicz, H. Brzezinka and B. Johannes, Monatsh. Chern. 101, 579 (1970).
[40] C.R. Enzell, G. W. Francis and S. Liaaen-Jensen, Acta. Chern. Scand. 22, 1054 (1968).
[41] R. Huisgen, A. Dahmen and H. Huber, J. Arner. Chern. Soc. 89, 7130 (1967).
[ 42] R. Hoffmann and R. B. Woodward, Accounts Chern. Res. 1, 17 (1968).
[43] K. W. Egger and T.L. James, Trans. Faraday Soc. 66, 410 (1970).
[44] L. Zechmeister, Fortschr. Chern. Org. Naturst. 18, 264 (1960).
[45] H. Kj~sen, S. Liaaen-Jensen and C.R. Enzell, Acta Chern. Scand. 25, 85 (1971).
[46] C.R. Enzell and S. Liaaen-Jensen, Acta Chern. Scand. 25,271 (1971).
[47] C.R. Enzell, Pure Appl. Chern. 20,497 (1969).
[48] K. Aitzetmiiller, H. H. Strain, W.A. Svec, M. Grandolfo and J.J. Katz, Phytochern. 8, 1761
(1969).
[49] J. Baldas, Q. N. Porter, A. P. Leftwick, R. Holzel, B.C.L. Weedon and J. Szabolcs, Chern.
Cornrnun. 1969, 415.
[SO] R. Bonnett, A. K. Mallams, A.A. Spark, J. L. Tee, B. C. L. Weedon and A. McCormick, J.
Chern. Soc. C 1969,429.
[51] N.A. S~rensen, S. Liaaen-Jensen, B. B~rdalen, A. Haug, C.R. Enzell and G. W. Francis,
Acta Chern. Scand. 22, 344 (1968).
[52] S. Liaaen-Jensen, S. Hertzberg, O.B. Weeks and U. Schwieter, Acta Chern. Scand. 22, 1171
(1968).
[53] S. Hertzberg, S. Liaaen-Jensen, C. R. Enzell and G. W. Francis, Acta Chern. Scand. 23, 3290
(1969).
[54] O.B.Weeks, A. G. Andrewes, B.O. Brown and B.C.L.Weedon, Nature 224,879 (1969).
[55] N. Rigassi [F. Hoffmann-La Roche & Co. Ltd., Basle], unpublished results.
[56] U. Schwieter [F. Hoffmann-La Roche & Co. Ltd., Basle], unpublished results.
[57] H. Mayer [F. Hoffmann-La Roche & Co. Ltd., Basle], unpublished results.
[58] F. Leuenberger [F. Hoffmann-La Roche & Co. Ltd., Basle], unpublished results.
[59] S. Hertzberg and S. Liaaen-Jensen, Phytochern. 8, 1259 (1969).
[60] S. Hertzberg and S. Liaaen-Jensen, Phytochern. 8, 1281 (1969).
[61] B. H. Davies, E. A. Holmes, D. E. Loeber, T.P. Toube and B.C.L.Weedon, J. Chern. Soc. C
1969, 1266.
[62] U. Schwieter, H. Gutmann, H. Lindlar, R. Marbet, N. Rigassi, R. Riiegg, S.F. Schaeren and
0. Isler, Helv. Chirn. Acta 49, 369 (1966).
[63] W. Benz, Massenspektrornetrie organischer Verbindungen (Akademische Verlagsgesellschaft,
Frankfurt a.M. 1969), p. 127.
[64] A. F. Thomas, B. Willhalm and R. Miiller, Org. Mass. Spectrorn. 2, 223 (1969).
[65] J. Baldas, Q. N. Porter, L. Cholnoky, J. Szabolcs and B.C.L. Weedon, Chern. Cornrnun. 1966,
852.
 267

V. Stereochemistry
B.C.L. WEEDON
Department of Chemistry, Queen Mary College, Mile End Road, London, E.l., England

A. Introduction . . . . . . . . . 268
B. Geometrical Isomerism . . . . 268
1. Occurrence of cis isomers 268
2. Stereomutation . . . . 270
3. Number of cis isomers . . 272
4. General properties . . . 273
5. Visible and ultraviolet light absorption. 273
6. Infrared light absorption . . 275
7. Proton magnetic resonance . 275
8. Biosynthesis of phytoene . 280
9. Synthesis of cis isomers . 281
C. Conformation . . . . . . . 284
1. Light absorption evidence 284
2. X-ray crystallography 285
3. P.m.r. evidence . . . . . . 287
D. Absolute Configuration . . . . 288
1. Methods of investigation . . 288
2. Capsanthin and capsorubin. 292
3. Fucoxanthin and zeaxanthin 295
4. Fucoxanthinol and paracentrone 300
5. Violaxanthin . . . . . . . . . 300
6. Neoxanthin . . . . . . . . . 302
7. The allenic ketone from grasshoppers 303
8. Abscisic acid . 304
9. Alloxanthin . . . . . . 305
10. oc-Carotene . . . . . . . 306
11. Monochiral xanthophylls. 307
12. Heterodichiral xanthophylls 308
13. Trisporic acids . . . . . . 312
E. Annex. Specification of Absolute Configuration. 312
References . . . . . . . . . . . . . . . . . . 319
268 B.C.L.WEEDON

A. Introduction

The stereochemistry of carotenoids can be conveniently discussed under

three main headings: (i) geometrical isomerism about the carbon-carbon double
bonds, (ii) conformation about the carbon-carbon single bonds, and (iii) the
absolute configuration of any chiral groups.
The literature on the first of these divisions is extensive. It dates back to
1923 when Herzig and Faltis [1] reported a single, unreproducible, experiment
in which they isolated from Orlean a bixin (265) with a higher melting point,
and a longer-wavelength light absorption maximum, than the usual product.
Six years later Karrer et al. [2] tentatively assumed that the two bixins were
geometrical isomers, and showed that the normal bixin was converted into the
higher melting form on exposure to iodine. A cis form of crocetin (269) was
reported by Kuhn and Winterstein [3] in 1933, but it was the pioneer obser-
vations of Gillam [4, 5] about 1935, and the subsequent extensive studies of
Zechmeister and his collaborators, that revealed the full extent and importance
of geometrical isomerism in the carotenoid field.
A detailed review of cis-trans isomerism with carotenoids and vitamin A
was published in 1962 [6]. In this chapter attention will therefore be confined
to important general aspects of the subject, and to advances made in the last
decade. A much fuller account will be given of the conformation and absolute
configuration of carotenoids since these represent comparatively recent devel-
opments and have not previously been reviewed.

B. Geometrical Isomerism*

1. Occurrence of cis isomers

Phytoene (32), generally regarded as the initial C 40-compound in the
biosynthesis of carotenoids, is believed to be formed by the stereospecific
linkage of two geranylgeranyl residues (Chapter VII). It is therefore of particular
interest that samples of phytoene from carrots, tomatoes and Chiarella vulgaris
have the 15-cis structure (32) [8-1 0]. Whether the same is true ofall carotenogenic
systems is not clear; phytoene from diphenylamine-inhibited cultures of Flavo-
bacterium dehydrogenans seems to consist principally of the all-trans isomer
[11], but the possibility cannot be excluded that stereomutation has occurred
during isolation; cis-phytoene is surprisingly stable, though stereomutation has
been reported [12].
The main phytofluene (30) present in tomatoes is probably also the 15-cis
isomer, but rapidly isomerizes to the all-trans form on isolation, particularly
* In this chapter stereoisomerism about a double bond is specified by means of the two
configurational descriptors cis and trans. With 1,2-disubstituted double bonds the meaning of
these terms is unambiguous. By convention they are also used in the carotenoid field to specify the
relative disposition of the two substituent!. forming part of the main chain of carbon atoms [7].
 V. Stereochemistry 269

if exposed to light [6, 13]. The equally labile cis isomer of (-carotene (26)
which has been observed in some Chiarella mutants [14] has been shown to
have the analogous central-cis structure [10]. It is therefore conceivable that
the cis bond in phytoene is preserved, at least to the (-carotene stage, during
the dehydrogenation sequence in the biosynthesis of carotenoids by some
organisms [10].
The only naturally occurring cis isomer of neurosporene (22) so far reported
is the so called proneurosporene present in some tomato selections [6] and in
the berries of Pyracantha angustifolia [15]. This differs markedly from the cis
hydrocarbons mentioned earlier in that it appears to have a poly-cis structure
with a trans double bond in the 15,15'-position [6].
Substantial amounts of a 'prolycopene' have been found in tangerine
tomatoes [16]. Its structure is unknown, but it is believed to be a poly-cis
isomer of lycopene (19) and gives the latter pigment on iodine-catalysed
stereomutation in the presence of light [6]. Six other 'poly-cis' lycopenes have
been isolated from the fruits of Pyracantha angustifolia [17], and related (or
identical) compounds have been reported in a Chiarella mutant cultivated in
the dark [18], in the fungus Fusarium aquaeductum when strongly ventilated
in light [19], and in other organisms [6]. A pro-y-carotene has also been
isolated from the ripe berries of P. angustifolia and yields y-carotene (8) on
treatment with iodine [20].
Most vegetable tissues contain no poly-cis carotenoids, though a number of
plants contain trace amounts [6]. Their significance is not known. They cannot
be formed from the all-trans carotenoids in vitro, and there is no evidence that
they are normal precursors in the biosynthesis of the common all-trans caro-
tenoids. It is interesting that no poly-cis oc- or P-carotene has been detected.
There are many reports of the isolation of mono-cis and di-cis carotenoids,
but it is by no means always clear whether these represent true natural products,
or are merely artifacts formed by stereomutation during isolation. Bixin (265)
and the cis isomer of crocetin (269), which have already been mentioned, are
undoubtedly authentic natural cis isomers since they are not formed by stereo-
mutation of the corresponding all-trans carotenoids [6]. The same is true of
the neo- P isomer of y-carotene (8) present in Pyracantha berries [17], and there
seems no doubt that the neo-B isomer of P-carotene (3) is a genuine constituent
of Osmanthusfragrans flowers [21]. Gazaniaxanthin (46), the principal pigment
in the flowers of Gazania rig ens, was recently shown to be the 5'-cis isomer of
rubixanthin (45) [22].
The main pigment in the pollen of various Lilium species appears to be a
mono-cis isomer of antheraxanthin (119) [23]. However, violeoxanthin from
pansies [24], now known to be a cis isomer of violaxanthin (135) [25], and
'tareoxanthin' from dandelions [26], which seems to consist mainly of the
neo-A and neo-B isomers of lutein epoxide (120) [25], could conceivably be
derived from the corresponding all-trans carotenoids by stereomutation.
Other isomers of antheraxanthin (119) and violaxanthin (135) that have been
reported [27, 28] may also be artifacts. Neoxanthin (122) is usually isolated
270 B. C. L. WEEDON

as a cis form, but again it seems likely that this is the result in many, if not all,
cases of stereomutation of the natural all-trans isomer [29].
To the list of naturally occuring cis carotenoids must now be added the
unique pigments from the photosynthetic bacterium Chromatium warmingii.
The key member of the series is rhodopinal (144), but a number of related
pigments, including rhodopinol (80), lycopenal (143) and the mono- (146) and
di-methoxy (147) derivatives, have also been isolated. In all these compounds
the C-20 methyl group is formally oxidized, and the molecule has the 13-cis
configuration [30, 31].
Interest in the geometrical isomerism of natural polyenes received a major
stimulus with the demonstration that the structurally 'hindered' 11-cis-retinals
(1) and (2), derived from vitamin A1 and vitamin A2 respectively, are the
prosthetic groups of the visual purple of vertebrates, insects and molluscs
(cf. Chapter X). It is the bleaching of visual purple in the retina of the eye that

0 0
(1) (2)

constitutes the fundamental chemical process involved in the recording of light

impulses. Though bleaching gives the all-trans-retinals, reformation of the
purple pigments requires an initial isomerization of the retinals to the 11-cis
isomers [6, 32, 33]. It is known that this isomerization is catalysed by a specific
enzyme.
Like vitamin A, the trisporic acids (see Chapter II, Section M.3), the prin-
cipal sexual hormones in the Mucorales, are derived from P-carotene (3). The
unsaturation in the side chain of the trisporic acids can be either 1-trans,9-trans
or 1-trans,9-cis [34, 35]. It should also be mentioned that the natural plant
growth regulator, abscisic acid (see Section D.8 and Chapter II, Section M.3),
has the 1-trans,9-cis configuration in the side chain (carotenoid numbering).
In diffuse sunlight abscisic acid is slowly transformed into an approximately 1 : 1
mixture with the 1-trans,9-trans isomer, and the same mixture is obtained
starting from the latter isomer. This makes it difficult to ascertain whether the
1-trans,9-trans isomer is physiologically significant. Certainly it has no effect
on growth if bioassayed in the dark, and there is no evidence of enzymes which
catalyse its isomerization, at least if light is excluded [36].

2. Stereomutation
Zechmeister and his collaborators [6] have shown that most, if not all,
carotenoids can be converted into mixtures of geometrical isomers under
appropriate conditions. The principal methods are indicated below.
 V. Stereochemistry 271

a) Thermal methods
Stereomutation of a carotenoid begins immediately on solution. The
process is usually slow at room temperature. Thus in benzene or light petroleum
solution, in diffuse daylight, only ca. 1-2% of()(-, {3- and y-carotene undergo
stereomutation in 24 hours. However, with all-trans-lycopene (19), under
similar conditions, the proportion is 10%, with spirilloxanthin (108) 23% and
with bacterioruberin (231) as much as 42% [6, 37].
Stereomutation is more rapid at elevated temperatures, and with all-trans
carotenoids in boiling benzene or hexane a quasi-equilibrium mixture of
geometrical isomers is produced within 10-60 minutes. Since many of these
mixtures contain several isomers, it is obvious that both heating and storage
of carotenoid solutions should be avoided as far as possible in all routine work
with these substances.
Most cis isomers of carotenoids exhibit marked thermal lability. However
when solutions of (cis-)methylbixin (266a) are boiled (in the absence of iodine)
a di-cis isomer is formed and not all-trans-methylbixin (266) as might have been
expected [38]. Some poly-cis carotenoids are as thermally stable as the corre-
sponding all-trans forms. On melting prolycopene crystals an isomer is formed
which is believed to contain an additional cis bond [6, 39].

b) Photochemical methods (without catalyst)

All carotenoids appear to undergo cis-trans isomerization on irradiation
in solution, light of wavelengths corresponding to the main absorption bands
being most effective. A similar process probably operates in nature in the inter-
conversion of geometrical isomers.

c) Irradiation in the presence of iodine

The most commonly used method of stereomutation is to expose a solution
of the carotenoid, containing catalytic amounts of iodine, to light. A quasi-
equilibrium mixture of isomers is rapidly formed. Each isomer when isolated
and submitted again to rearrangement yields a mixture of approximately the
same composition.
Qualitatively and quantitatively different mixtures, or 'sets', of isomers may
be formed using different methods of stereomutation. The most stable isomer
of a carotenoid is normally the all-trans form, and this predominates in mixtures
formed by iodine-catalysed stereomutation. Exceptions to this generalization
are provided by crocoxanthin (41), by alloxanthin (65) and, presumably, by
related acetylenic carotenoids. With crocoxanthin (41) the main constituent of
the equilibrium mixture is the 9-cis isomer, and with alloxanthin (65) the
9,9' -di-cis isomer. It has been suggested that, in contrast to their polyene
analogues, these cis isomers in the acetylene series are somewhat less sterically
hindered, and therefore more stable, than their all-trans forms [ 40].
Further exceptions to the generalization that the all-trans carotenoid
predominates in the equilibrium mixture of stereoisomers are provided by
272 B. C. L. WEEDON

compounds of the rhodopinal (144) series. Here cross-conjugation with the

side-chain aldehyde group favours the 13-cis structures, and it is only the
allylic alcohol rhodopinol (80) without this group that can be partly isomerized
to the all-trans form [31].

3. Number of cis isomers

Calculation of the numbers of geometrical isomers theoretically possible
for a given carotenoid leads to some rather daunting figures. For a conjugated
system with n non-cyclic double bonds, the number of stereoisomers N is given
by the expressions :
N = 2n for unsymmetrical systems.
N = 2<n-ll/2 • (2<n-ll/2 + 1) for symmetrical systems, n odd.
N = 2<n/l)-l. (2nfl + 1) for symmetrical systems, n even.
Thus lycopene (19), with a symmetrical carbon skeleton and eleven con-
jugated double bonds, should be capable of existing in 1056 different geo-
metrical modifications. Of these only about forty have so far been reported.
Fortunately the situation is not, in practice, as complex as that indicated
by calculations of the above type. As was first pointed out by Pauling [41], the
double bonds in the acyclic polyene chain of carotenoids are of two main
types: those for which the adoption of a cis configuration involves very little
steric hindrance (between two hydrogen atoms), and those for which a cis
configuration leads to a serious clash between a methyl and a hydrogen atom
(Fig. 1). The methyl-substituted double bonds come into the first category, as
do the 15,15'-double bonds present in most carotenoids. All other disubstituted
double bonds are adjacent to a methyl side-chain, and therefore fall into the
second category.

Fig. 1. Overlapping of hydrogen atoms in CH-CH=CH-CH, and of methyl and hydrogen in

C(CH 3 )-CH=CH-CH, with cis configurations (from Pauling [41])

The Pauling classification is useful in indicating the double bonds about

which stereomutation is most likely to take place. A further calculation reveals
that the number of sterically 'unhindered' isomers of lycopene (19) is only 72.
Furthermore, in practice, relatively few isomers constitute the bulk of the
mixtures produced by stereomutation, as would be expected on purely statis-
tical grounds r6l.
 V. Stereochemistry 273

It must not, of course, be concluded, as was widely done at one time, that
carotenoids with a cis double bond of the second category are incapable of
existence. A number of these sterically 'hindered' isomers have been synthesized
(Section B.9) and, as pointed out earlier, the sterically hindered 11-cis isomers
of the retinals play a vital role in the visual process.
The reason for the comparative stability of the 9-cis isomers in acetylenic
carotenoids will now be apparent. Because of the presence of the triple bond
in the 7,8-position, there is no hydrogen atom at C-8 to interfere with the
hydrogen at C-11 when the molecule adopts the 9-cis configuration.

4. General properties
The carotenoids so far studied conform to the general rule that the all-trans
compound is the isomer oflowest solubility and highest melting point. Because
of stereomutation on fusion, some cis isomers exhibit the phenomenon of a
double melting point.
The cis forms of an optically active carotenoid may exhibit different
rotation from that of the all-trans isomer [6]. Marked differences in optical
rotatory dispersion have been reported between all-trans-neoxanthin (122)
and a commonly encountered isomer believed to have the 9-cis configura-
tion [29].
The geometrical configuration of a carotenoid has a profound influence
on its adsorption affinity. This means that even complex mixtures of isomers
can usually be ·separated by chromatography. It is therefore interesting to
note that two groups have been unable to separate rubixanthin (45) from its
5' -cis isomer, gazaniaxanthin (46) [22, 42], though this separation apparently
presented no difficulty when gazaniaxanthin was originally isolated from
Gazania rig ens grown in Portugal [ 43].
Thin layer chromatography on basic magnesium carbonate has recently
been advocated for the separation of cis-trans-isomeric xanthophylls [28].

5. Visible and ultraviolet light absorption

Cis carotenoids exhibit light absorption of lower intensity than their all-
trans isomers, and, as a general rule, the principal light absorption maxima are
shifted to shorter wavelengths (ca. 2-5 nm for one and ca. 10 nm for two
sterically 'unhindered' cis double bonds) [6, 44]. One of the most noticeable
features in the spectra ofmono-('unhindered'-)cis carotenoids is the appearance
of a subsidiary peak in the near ultraviolet region (Fig. 2). The wavelength
difference between this 'cis peak' and the longest-wavelength maximum of the
all-trans compound is practically a constant (142 ± 2 nm for C40 -carotenoids
possessing 10 or 11 conjugated double bonds) [6], though the cross-conjugated
carotenoids of the rhodopinal (144) series are said to be exceptional [31]. As
would be expected on theoretical grounds, the central-cis isomer has less
intense absorption in the region of the principal maxima than the other
'unhindered' mono-cis forms, but the most intense cis peak [6, 45]. Since the

Carotenmds 18
274 B. C. L. WEEDON

16~~~~------------,------,------.------.------·
~-Carotene

~ 1-------t-----J'-\1-----+---------1
14 all-trans

12 9-cis

10

...I
$2

~·~ \
......
,...,\' I
,,
\.

6 15-cis
\
----

Fig. 2. Light absorption spectra of all-trans-, 9-cis- and 15-cis-{J-carotene

(from Jaffe and Orchin [ 45])

intensity of the cis peak seems to depend primarily on the overall shape of the
chromophore, it is scarcely surprising that neither the poly-cis, nor the authentic
di-cis, isomers exhibit significant absorption in this region; in both instances
the molecules approximate to the linear shape of the all-trans form [6].
It has long been known that the 13-cis isomers of vitamin A1 and vitamin A2
(Chapter II, Section M.3) have maximal absorption at wavelengths which are
2-3 nm longer than those ofthe all-trans forms [6]. In contrast to the cis isomers
discussed above, the distance between the ends of the chromophores is exactly
the same as that in the all-trans forms. Presumably a cis configuration of the
terminal double bond in these chromophores raises the energy of the excited
state slightly less than that of the ground state. Recently a similar structural
relationship has been proposed for gazaniaxanthin (46) and rubixanthin (45)
[22]. Gazaniaxanthin exhibits neither the usual shift in Amax to shorter wave-
lengths, nor a cis peak, but this is consistent with the proposed cis configuration
about the terminal double bond of the polyene chromophore [22]. In the past
 V. Stereochemistry 275

other 'terminal-cis' carotenoids may well have been mistaken for all-trans
compounds.
With phytoene (32), which has a simple triene chromophore, the common
15-cis isomer has light absorption maxima at the same wavelengths as the all-
trans isomer, but of lower intensity [9].
Carotenoids which contain a sterically 'hindered' cis double bond give
spectra with very little fine structure, and in which the main light absorption
maximum is at much lower wavelengths than that of the all-trans isomer.
Although the evidence is not conclusive, the mono-('hindered'-)cis isomers also
seem to exhibit a cis peak [6].

6. Irifrared light absorption

A prominent feature in the i.r. spectra of carotenoids is the strong absorption
near 965 em- 1 due to C-H out-of-plane deformation vibrations in the trans-
disubstituted double bonds [46]. Many all-trans carotenoids with a normal
chromophore exhibit a single sharp peak in this region. However, two, and
occasionally more, peaks have been reported for carotenoids in which the
polyene chain is conjugated with a ketone or an ester group, and for some
carotenoids with retro chromophores, e.g. eschscholtzxanthin (84) [6, 44, 46].
Splitting of the peak into a doublet is also observed with some 15-cis carotenoids.
In these latter isomers the C-H out-of-plane vibrations of the cis-disubstituted
double bond results in a strong absorption band near 780 em- 1 [6, 46]. A
similar band occurs in prolycopene, a poly-cis isomer of lycopene (19) [6], and
the presence of a band at 758 em - 1 in the i.r. spectrum of phytoene (32) provided
the original evidence for the 15-cis configuration of this key compound [9].
Many carotenoids with a cis configuration about a trisubstituted double
bond have an i.r. band near 1380 cm- 1, which has been attributed to the
deformation vibration of the methyl group on the cis double bond [ 46]. This
band is not always observed and is therefore of limited diagnostic value.
Moreover, all-trans-oc-carotene (5) has a similar absorption peak [6].

7. Proton magnetic resonance

Extensive use has been made of proton magnetic resonance (p.m.r.) in the
last ten years to elucidate the structures of carotenoids [ 44, 47]. However, the
characteristic methyl signals are often little affected by changes in stereochem-
istry, and the olefinic proton region, as recorded at 40, 60 and even 100 MHz,
normally reveals such a complex pattern due to spin-spin coupling and the
overlap of signals that little useful information can be deduced. Fortunately
various favourable structural features have enabled a number of important
problems to be'tackled successfully.

a) The hydrolycopenes
The p.m.r. spectrum of squalene (3) exhibits methyl bands at f> 1.58 and 1.65
in the proportions of 3: 1 [10, 48]. Since squalene is known from X-ray crystallo-
276 B. C. L. WEEDON

graphic studies on its urea clathrate to possess the all-trans configuration [ 49],
these p.m.r. bands are assigned to the olefinic methyls which are, respectively,
trans and cis to the neighbouring olefinic hydrogen atoms. From the relative
intensities of the corresponding bands in the p.m.r. spectra of phytoene (32),
phytofluene (30), (-carotene (26) and neurosporene (22) it was concluded that
all non-terminal unconjugated double bonds in these hydrolycopenes also have
the trans configuration [10].

~ ~

(3)

b) Vitamin A series ( P-meth yl bands)

Differences in stereochemistry about the IXP-double bond in polyene
aldehydes and esters containing a methyl substituent at the P-position are
readily detected by p.m.r. Thus the C-20 methyl in all-trans-vitamin A 1 acid
methyl ester (4), and in its A 2 analogue, give a P-methyl band at lower fields
(<5 2.16 and 2.42 respectively) than the corresponding band (1.88 and 2.14
respectively) in the 13-cis isomers, e.g. (5) [50, 51]. Such effects are attributable
to the greater deshielding of the P-methyl protons in the trans isomers owing

(4) (5)

~ C02H
(6)

to the proximity of the P-methyl and the magnetically anisotropic carbonyl

group. Smaller differences also exist between the C-20 methyl bands of vitamin
A1 and vitamin A 2 [50], and between the corresponding bands of cis and
trans isomers of other P-methyl allylic alcohols, e.g. geraniol, farnesol and
phytol [52, 53]. No significant differences in 'in-chain' or 'end-of-chain'
methyl bands have been reported in the p.m.r. spectra of geometrical isomers,
except for the 11,13-di-cis isomer of vitamin A acid (6) and some related
di-cis polyenes [54]. It may be significant that a similar deshielding has been
observed in two of the 'in-chain' methyls of prolycopene [55].

c) Rubixanthin and gazaniaxanthin

Though both rubixanthin (45) and gazaniaxanthin (46) have the same gross
structure, their properties show hone of the differences usually associated
 V. Stereochemistry 277

with cis-trans isomers, except for a difference in melting point (see Sections
B.4 and B.5). The clue to their unique stereochemical relationship was provided
by their p.m.r. spectra. In most respects these were identical, but small differences
were noted in the absorption near(> 2.10 attributable to the allylic methylene
group. With rubixanthin the band had the general appearance of a doublet,
rather like the corresponding absorption in lycopene (19), whereas with
gazaniaxanthin the band was noticeably more complex, suggesting more
effective spin-spin coupling ofthe 4'-methylene group with the olefinic hydrogen
at C-6'. These qualitative differences parallelled those in a number of simple
models of known stereochemistry, and it was therefore possible to assign the
all-trans configuration to rubixanthin (45) and the 5'-cis structure to gazania-
xanthin (46) [22].
The above conclusion raises the question whether other carotenoids
possess cis bonds at the end of the chromophore since these would not have
been revealed by conventional light absorption studies (Section B.5). Though
this point must be borne in mind, there is no doubt concerning the stereo-
chemistry of the double bonds at the end of the polyene chain in lycopene
(19), (-carotene (26) and neurosporene (22). These have been synthesized
by routes which determined the trans configuration of the double bonds
under consideration, and the products had the same melting points as the
natural pigments.

d) N eo xanthin
Two crystalline forms of neoxanthin (122) are known, differing by about
10° in melting point. The isomers may be converted into one another by
iodine-catalysed stereomutation, and there is reason to believe that the high
melting form is the true natural isomer. Their visible light absorption properties
indicate that the high m.p. form is the all-trans isomer, and that the other con-
tains an 'unhindered' cis bond located near one end of the polyene chain [29].
The methyl bands in the p.m.r. spectrum of the high m.p. form are in ex-
cellent agreement with those that would be expected for a carotenoid with one
end group identical with those in violaxanthin (135) and the other with the
allenic end of fucoxanthinol (189). However, with the low m.p. form, minor
differences were observed in the bands associated with the methyls at C-1' and
C-5' in the epoxide end of the molecule. This was taken to indicate that the
'unhindered' cis bond is located near the epoxide end group, and the 9'-cis
structure was proposed [29].
Other isomers of neoxanthin have been reported, but they have not as yet
been fully characterized [56, 57].

e) Decaprenoxanthin
The chemical shift of the aldehydic proton in conjugated aldehydes with a
methyl substituent in the a-position has been found to vary with the stereo-
chemistry about the ap-double bond. The signals of the aldehydic protons of
278 B.C.L.WEEDON

the trans compounds appear at considerably higher fields (<5 9.26 to 9.70) than
those of the corresponding cis isomers (<5 10.01 to 10.33). This correlation has
been used to assign a trans configuration to the double bonds in the C-2 and
C-2' side chains of the dialdehyde from decaprenoxanthin, and hence to those
in decaprenoxanthin (226) [58].

f) Bixin and the methylbixins*

Bixin (265) was the first cis polyene to be recognized in nature, and it is
scarcely surprising that the stereochemistry of this readily available pigment
from Orlean should have attracted a good deal of attention.
Early attempts to solve the problem by purely chemical means led to the
proposal that bixin (265) and the derived diester, methylbixin (266a), have
11-cis structures [59]. Later it was recognized that such 'hindered' cis structures
were incompatible with the visible light absorption properties, and 9-cis
structures, which had not been definitely excluded by the chemical evidence,
were favoured [60, 61]. Subsequent infrared light absorption studies were held
to support a 7-cis formulation for methylbixin [46].
Though the signals due to most of the olefinic protons cannot be resolved
(at 40 or 60 MHz) in the p.m.r. spectrum of methylbixin, shielding by the ester
groups makes it possible to study both 1)(- and {J-protons [62]. In all-trans-
methylbixin (266) these 1)(- and {J-protons constitute two identical AB systems
(Fig. 3). Of the resulting two doublets (each equivalent to two protons) that at
higher field (<5 5.90) may be assigned to the two 1)(-protons, and that at lower
fields (<5 7.44) to the two {J-protons, by analogy with the results obtained earlier
with the muconic acids and other simple models [63]. Moreover the observed
coupling constant (J 15.5 Hz) confirmed the trans configuration of the two
terminal carbon-carbon double bonds. However the p.m.r. spectrum of cis-
methylbixin (266a), derived from natural bixin, showed that though the two
I)(- protons are virtually identical and give rise to doublets (<5 5.93 and 5.91;

J 15.5 Hz) closely resembling that observed with the all-trans isomer, the two
{J-protons experience different fields. One gave a doublet (<5 7.43; J 15.5 Hz)
again closely resembling that from the all-trans isomer, but of half the intensity.
The other gave rise to a new doublet (<5 8.02; J 15.5 Hz), also equivalent to one
proton. These differences could not be attributed to a cis configuration about
one of the Q(P-double bonds since the coupling constant was unchanged. The
result was, however, readily explicable in terms of a cis configuration about
one of the y<5-double bonds since this would bring the adjacent {J-proton into
a region where it is deshielded by the third double bond from the end of the
polyene chain. The correctness of this interpretation was confirmed by analysis
of the p.m.r. spectra of simple triene models, and computation of their spectra.

* In this chapter the numbering of the double bonds in bixin, crocetin and their derivatives
refers to the corresponding positions in the carbon skeleton of lycopene. This follows the current
carotenoid convention. The numbering therefore differs from that in the papers cited in which
C0 2 H=C-1.
 V. Stereochemistry 279

8 7 6 5p.p.m.
Fig. 3. Olefinic proton region in the p.m.r. spectra (60 MHz, CDC1 3 ) of all-trans-methylbixin (266)
(top curve), and of the dimethyl ester (266a) derived from natural (9'-cis) bixin (bottom curve)

This left no doubt that the methylbixin derived from natural bixin has the
9-cis configuration [62], but final proof has recently been obtained by the
stereospecific synthesis of 9-cis-methylbixin (Section B.9).
Examination of the cis apo-aldehydic methyl ester (7), prepared by oxidative
cleavage of the 7,8-double bond in bixin, led to a surprising result. Its p.m.r.
spectrum exhibited the bands characteristic of the cis end of the diester from
natural bixin. This showed that, contrary to all previous suggestions, bixin has
the 9'-cis configuration (265) [62].

(7)

By thermal and photochemical stereomutation of all-trans- and cis-methyl-

bixin, Zechmeister and Escue [60] obtained two further crystalline isomers,
neo A and neo C respectively, as the principal products. Light absorption
280 B. C. L. WEEDON

studies revealed that neo A has a mono-'unhindered'-cis structure, and a

strong 'cis peak' indicated that the cis bond is near the centre of the molecule.
Since neo A differs both from 'natural' cis-methylbixin and from the synthetic
15-cis isomer [ 46], it must have the 13-cis structure [62]. The visible light
absorption properties of neo-C-methylbixin are those of an isomer with two
'unhindered' cis bonds, and its i.r. spectrum indicates the absence of a central-
cis bond [ 46]. In the p.m.r. spectrum the band pattern due to the rx- and {3-
protons is very similar to that observed with 'natural' cis-methylbixin. This
confirms that the original cis bond is preserved during stereomutation of
cis-methylbixin, and indicates that neo C must be either the 9,13-di-cis or the
9,13'-di-cis isomer. Of these two possibilities the latter seems more likely since
neo C is formed under conditions which convert all-trans-methylbixin into the
neo A isomer [62].

g) 220 MHz p.m.r.

A major advance in p.m.r. spectroscopy comes from the recent introduction
of 220 MHz spectrometers. The increase in chemical shifts provided by these
instruments means that, with many polyenes, it is possible to resolve the ole-
finic portion of the spectrum into more or less separate singlets or multiplets
(Chapter IV). The possibility, even in rather complex cases, of being able to
assign all the olefinic protons has obvious and important implications for
studies on the geometrical isomerism of carotenoids.
The 220 MHz spectra of many geometrical isomers of vitamin A and its
derivatives such as the retinals and the corresponding acids have now been
run and analysed [64, 65]. A comparison of the chemical shifts of selected
protons of the trans and of the different cis isomers indicates that large changes
in chemical shift only occur for protons attached to, or in close geometrical
proximity with, the cis bond. The experimental rules derived from these pilot
studies have already proved extremely useful for the elucidation of the stereo-
chemistry of synthetic {3-apo-carotenals [65]. The extension of this work to
the study of natural cis carotenoids, including the intriguing poly-cis com-
pounds, holds considerable promise.

8. Biosynthesis of phytoene
Though no direct evidence was available from the original p.m.r. studies
concerning the stereochemistry of the 13- and 13'-double bonds in phytoene
(32), it seemed probably that these were trans like the four non-terminal
unconjugated double bonds. Support for this view was provided recently when
it was shown that squalene (3) and phytoene (32) synthesized from (4R)-meva-
lonic-2-14C-4-3H acid in isolated carrot root slices have the same 3Hjl 4C ratio.
This indicates that all trisubstituted double bonds in the C 15 - and C20 -inter-
mediates respectively in the biosynthesis of squalene and phytoene are formed
with the same configuration [66, 67]. Unless there is an inversion of geometrical
configuration either during, or after, the oxidative coupling process, the 13-
 V. Stereochemistry 281

and 13'-double bonds in phytoene must be trans, and (32) represents the
complete stereochemistry of this key compound in carotenoid biosynthesis.

9. Synthesis of cis isomers

Considerable success has been achieved in the synthesis of specific cis
isomers of vitamin A and related compounds. These include examples of the
sterically 'unhindered' 9-cis and 13-cis isomers, and of the 9,13-di-cis isomers.
Sterically 'hindered' forms have also been prepared, namely representatives
of the 11-cis and 11,13-di-cis series (for reviews see Chapter VI and references
[6, 54, 68, 69]).
With carotenoids, attention has hitherto been directed mainly to the
synthesis of the all-trans isomers. However, the routes developed often involve
the catalytic partial hydrogenation of an intermediate with an acetylenic group
at C-15. This gives initially the 15-cis isomer which may be isolated if appro-
priate precautions are taken. In this way the labile 'central-cis' isomers have
been prepared of a number of carotenoids including lycopene (19), P-carotene
(3), zeaxanthin (67), cryptoxanthin (39), canthaxanthin (193), echinenone (148),
methylbixin (266) and the dimethyl ester of crocetin (269). The sterically
'hindered' 11,11'-di-cis isomer of P-carotene has similarly been prepared from
the synthetic 11,11'-diacetylenic analogue and found to be surprisingly stable.
'Hindered' cis isomers oflycopene (19) have also been reported (see Chapter VI
and reference [6]).

a) Alloxanthin and crocoxanthin

As mentioned earlier (Section B.2c), the acetylenic carotenoids alloxanthin
(65) and crocoxanthin (41) are unusual in that iodine-catalysed stereomutation
gives mixtures consisting mainly of cis isomers. The geometrical configuration
assigned to the principal cis isomers has been confirmed by synthesis [ 40, 70].
The Wittig reagent (10) was synthesized from the ketone (8), derived from
isophorone [71], and the cis-methylpentenynol (9), an intermediate in the
manufacture of vitamin A [72], by the route outlined in Scheme 1. Reaction
at both ends of the trienedial (11) [73, 74] then gave 9,9'-di-cis-alloxanthin {12),
identical (apart from optical activity) with the main isomer from the stereo-
mutation of natural alloxanthin (65). A similar condensation of the Wittig
reagent (10) with oc-apo-12'-carotenal [75] gave 9-cis-crocoxanthin which was
identified with the main stereomutation product of natural crocoxanthin (41).

b) Crocetin
Both 9-cis [6] and 13-cis [76] structures have been proposed for the
unstable cis isomer of crocetin which was isolated as the dimethyl ester by
Kuhn and Winterstein [3]. It has been shown that reaction of the pentaenedial
(14) [77] with the phosphonate (13) derived from oc-bromopropionate gives a
mixture of the all-trans-dimethylcrocetin (270) and its 9-cis isomer {15) [70].
282 B. C. L. WEEDON

The latter has been isolated by chromatography and shown to have light
absorption and i.r. properties significantly different from those reported for
the natural isomer. It also proved to be comparatively stable.

c/J· (8) (9)

~~
1) He
2) LiAIH4
3) Ac 2 0 CH=P(C6 H,),

HO
(10)

_1__ ~ ~ ~CHO
(10) + OHC~ ~~I + {10)

(11)

j
~~
HO~

yyo"
~~
(12)

Scheme 1

From spectral evidence, natural cis-crocetin appears to be an isomer with

one, sterically 'unhindered', cis bond. Of the three conceivable structures,
15-cis [76] and 9-cis [70] have been excluded by synthesis. It therefore seems
probable that natural cis-crocetin has the 13-cis formulation favoured by
Kuhn et al. [76].
 V. Stereochemistry 283

CH 30 2C
_)___
PO(ORh +
~
OHC'~~~~~
.1. ~ ~ ~ ~CHO
(13) (14)

j
2 CH 3

~ ~ ~ ~ ~ ~ ~ CO,CH,
+ (270)
(15)

c) Methylbixin
Confirmation of the 9-cis structure assigned to the diester derived from
natural bixin (265) has recently been obtained by stereospecific synthesis
(Scheme 2) [78].

3 3
- - - tCH - - - tCH
HOr-----9 +

~
(R0) 2 0PCH 2 C0 2 CH 3
~C02H ~CHO
(16) (17) (18) (19)

(266a)

Scheme 2
284 B. C. L. WEEDON

Condensation of the lactol (16) with the phosphonate (17) gave the cis,trans
half ester (18) [54]. Reduction of its acid chloride with lithium tri-t-butoxy-
aluminohydride led to the corresponding aldehyde (19) with complete retention
of stereochemistry. Reaction of the aldehyde (19) with the Wittig reagent (20),
which was prepared from the trienedial (11 ), then yielded the 9-cis isomer
(266a), which proved to be identical in all respects with the methyl ester of
natural bixin [78].

C. Conformation

1. Light absorption evidence

The visible light absorption properties of acyclic all-trans carotenoids such
as lycopene (19) indicate that the polyene chromophore is essentially planar,
and that the conformation of the constituent single bonds iss-trans, or nearly so.
This arrangement allows good overlap of the n-orbitals and delocalization of
the n-electrons [6, 44]. The same conclusion can be drawn for the acyclic
portion of the chromophore in an all-trans cyclic carotenoid. Similar confor-
mations of the acyclic part of the chromophore would also be expected for
terminal-cis isomers, such as gazaniaxanthin (46), and for the favoured cis
isomers of the acetylenic carotenoids, e.g. 9,9' -di-cis-alloxanthin (12), since
these cis configurations do not involve steric interference of the types illustrated
in Fig. 1 that might offset the advantages of maximum orbital overlap.
'Unhindered' cis isomers usually exhibit light absorption maxima at
shorter wavelengths and of lower intensity than their all-trans analogues
(Section B.5). Though these changes might merely reflect the shorter distance
between the two ends of the chromophore, the light absorption evidence is
equally consistent with a small rotation about one of the single bonds adjacent
to the cis linkage occasioned by steric interference between two hydrogen
atoms (Fig. 1). With a sterically 'hindered' cis isomer the degraded spectrum,
with a large shift in Amax to shorter wavelengths and loss of fine structure,
clearly indicates that one at least of the single bonds has a skew geometry, and
that part of the acyclic polyene chain is far from co-planar with the rest.
As is well known, {3-carotene (3) exhibits light absorption maxima at
wavelengths which are ca. 24 nm shorter than those of lycopene (19) even
though both carotenoids contain a chromophore consisting of eleven conju-
gated double bonds [6, 44, 61]. The monocyclic isomer, y-carotene (8), again
with the same number of conjugated double bonds, has light absorption
maxima which are intermediate between those of lycopene and {3-carotene.
Such shifts in A-max with carotenoids containing a cyclohexene ring have long
been recognized as the result of a twisting of the molecule about the single
bond joining the ring to the polyene side chain [79]. This skew conformation
relieves the steric hindrance between the polyene chain and the ring methyls
that would exist in a planar arrangement, whether s-cis or s-trans, about the
 V. Stereochemistry 285

6,7-single bond. The resulting limitation in overlap between the n:-orbital of

the ring double bond, and those associated with the polyene side chain, explains
the observed shortening in wavelength of maximal absorption [6, 44].
A precisely similar situation exists in the aromatic carotenoids, such as
isorenieratene (13) [80] and chlorobactene (15) [81, 82], with methyl groups
at C-1, C-2 and C-5 on the aromatic ring (carotenoid numbering). Here steric
hindrance between the polyene chain and the two ortho methyl substituents
precludes conformations in which the aromatic ring and the polyene side chain
are co-planar. This limits the overlap of the n:-orbitals associated with the
aromatic ring and those of the polyene chain, and again there is a shift of Amax
to shorter wavelengths.
In contrast, a planar conformation is possible with the aromatic carotenoids
in which the ring methyl groups are at C-1, C-2 and C-3 (carotenoid numbering)
since there is only one substituent ortho to the polyene side chain. This is
reflected in the longer wavelength of maximal absorption of renierapurpurin
(16) compared with that of isorenieratene (13). As expected, renieratene (14) is
intermediate between the two [80, 83].
Contraction of the six-membered ring in P-carotene and its derivatives to a
five-membered ring would be expected to relieve the steric hindrance between
the ring methyls and the polyene chain. It is therefore interesting to note that
the cyclopentene ring and the polyene chain in actinioerythrin (272) and its
derivatives are nearly co-planar, judging from their visible light absorption
spectra [31, 84].

2. X-ray crystallography

Recent X-ray crystallographic studies have given much more precise

information on some of the conformational questions discussed above. Follow-
ing preliminary work on trans-P-ionylidenecrotonic acid (21) [85], its 9-cis
isomer (22) [86] and vitamin A acid (cf. (4)) [87], the determination of the
crystal and molecular structures of P-carotene (3) [88], canthaxanthin (193)
[89] and their acetylenic 15,15'-didehydro derivatives [90, 91] represent very
notable achievements. The following summary of the main findings concen-
trates on molecular features which are similar in all the structures determined

(21)
tr\ {22)
C02H

so far, and which accordingly appear to be dictated by intramolecular influ-

ences. For features which seem to be influenced mainly by the various packing
schemes, the reader must consult the original publications.
286 B. C. L. WEEDON

Contrary to earlier predictions of a rapid convergence of the bond lengths

of conjugated systems to a mean value on increasing chain length [92], an
alternation of shorter and longer bonds has been found (Fig. 4). A systematic
decrease in the alternative character of the single and double bonds towards
the centre of the molecule has only been observed for the two acetylenic
derivatives. In the latter the effect seems to be enhanced by the shortening of
the 14,15-single bond, which may well be due to the neighbouring triple bond.
The triple bond itself is not lengthened in spite of a presumed decrease in bond
order. Both 15,15'-didehydro compounds in the crystal phase have the trans
conformation about the 15,15'-triple bond [91], although spectroscopic studies
(admittedly inconclusive) seem to indicate the cis conformation as the stable
one in ether solution [93].

Fig. 4. Molecular structure of canthaxanthin (193); bond lengths and angles

(from Bart and MacGillavry [89])

The polyene chain is practically linear as expected from light absorption

studies. However, it is not a straight zigzag with valence angles of 125°. The
angles which are opposite methyl groups on the chain are significantly smaller
than 125°, thereby increasing the angles opposite the double bonds at C-9,
C-13, C-9' and C-13'. This means that the distances between C-7 and C-19,
between C-19 and C-11, between C-11 and C-20, and between C-20 and C-15
are about 3.0 A, instead of the 2.8 A which would be expected for a straight
zigzag chain. The resulting S-shaped bending of the polyene chain can be
clearly seen in Fig. 4. The effect appears to be due to the non-bonded inter-
actions between the side-chain methyl groups and the hydrogen atoms at C-11,
C-15, C-11' and C-15'. There is therefore little doubt that the same situation
 V. Stereochemistry 287

exists in 'free' molecules in solution and that it is not confined to molecules in

a crystal lattice. In addition to this well established in-plane bending of the
polyene chain away from the in-chain methyl groups, twisting of the planes of
the successive double bonds is a further, though probably less important,
factor in releasing steric hindrance in the polyene chain caused by the methyl
groups.
As anticipated from the light absorption studies, the cyclohexene ring
double bond is not co-planar with the polyene chain in any of the compounds
so far examined. In all-trans-P-ionylidenecrotonic acid (21) the deviation is
approximately 10° from the s-trans conformation about the 6,7-single bond.
In all other cases the observed arrangement is one in which there is a substantial
deviation from the s-cis conformation about the bond joining the ring and the
polyene chain. For triclinic crystals of canthaxanthin (193) the angle between
the plane of the cyclic double bond and that of the neighbouring double bond
in the polyene chain is slightly different for the two end groups (52° and 56°).
Both values are larger than (and in the opposite direction to) the 40° deviation
observed with P-carotene (3). Though the larger conjugated bond system in
canthaxanthin might have been expected to favour a planar conformation for
the molecule, the stiffening of the ring resulting from introduction of the keto
group apparently gives even more strain due to repulsion of non-bonded
groups. There is presumably a nice balance between conjugation, steric hin-
drance and the factors involved in crystal packing. The deviation from s-cis is
as much as 80° for 9-cis-P-ionylidenecrotonic acid (22) [91], possibly reflecting
the additional steric hindrance between the hydrogen atoms at C-8 and C-11.
A rotation about the 8,9-bond [86] can also be attributed to this cause.
Two further points concerning the conformation about the single bond
joining the ring and the polyene chain deserve mention. The angles between
the plane of the ring double bond and that of the adjacent double bond in the
polyene chain may be up to 10° greater than that between the ring double
bond and the overall 'best' plane for the polyene chain as a whole. Finally it
must be borne in mind that the conformations about the single bonds joining
the cyclohexene rings to the polyene chain are no doubt influenced by the
packing forces in the crystal lattice; these conformations may therefore be
significantly different for molecules in solution. Calculations based on the
extended Htickel method suggest that, in general, a non-planar s-cis confor-
mation should be the most stable one [94].

3. P.m.r. evidence
Little information is yet available regarding the conformation of the polyene
chain in cis ispmers. However, 220 MHz p.m.r. spectrometry allows some
interesting conclusions to be drawn concerning 11-cis-retinal (1), the sterically
'hindered' isomer involved in the formation of rhodopsin in the visual process,
and the related 11,13-di-cis compound [64].
Light absorption evidence clearly indicates that the severe steric crowding
between the 20-methyl (attached to C-13) and the hydrogen atom at C-10
288 B. C. L. WEEDON

results in a skew geometry for the polyene side chain in 11-cis-retinal {1). The
coupling constant 110 • 11 has nevertheless been found to be the same as that for
all-trans-retinal. Since the value of this coupling constant is a sensitive function
of the dihedral angle between the carbon-hydrogen bonds at C-10 and C-11,
the conformation about the 10,11-bond in 11-cis-retinal must be virtually the
same as in the all-trans isomer. This points to the 12,13-bond as the site of the
skew geometry in 11-cis-retinal.
If good overlap of n-orbitals in 11-cis-retinal extends only from C-7 to
C-12 because of a twist around the 12,13-bond, then the proton at C-12 should
resemble a proton at the end of a polyene chain and should appear at a higher
field than if it were in the middle of a polyene chain as in all-trans-retinal. This
is indeed the case, and an upfield shift of0.45 p.p.m. has been observed. A skew
geometry at the 12,13-bond should prevent tpe formation of a resonance
structure with a positive charge at C-11 and give rise to an upfield shift for the
proton at this position. This too has been observed (0.45 p.p.m.). It can thus be
concluded that steric crowding in 11-cis-retinal {1) is released mainly by a skew
geometry about the 12,13-bond. Consideration of the 220 MHz p.m.r. spectrum
of 11,13-di-cis-retinal leads to the conclusion that in this isomer also the
10,11-bond iss-trans, or nearly so, and that the 12,13-bond is twisted.

D. Absolute Configuration

1. Methods of investigation
Inspection of List A in Chapter XII shows that the question of absolute
configuration is relevant to about half the carotenoids that have been isolated
from nature. It is only recently, however, that much information on this
fundamentally important point has become available.

a) Optical activity
The intense visible light absorption of carotenoids renders optical meas-
urements difficult in the region of the principal absorption bands. It is therefore
scarcely surprising that there are a number of conflicting reports in the early
literature. Thus zeaxanthin (67) has been described as inactive [61], laevoro-
tatory [95], and dextrorotatory [96]. Capsanthin (170) and fucoxanthin (190)
have both been described as optically inactive [97, 98] and as optically active
[61, 99]. Some of these practical difficulties in classical polarimetry were
overcome by using the mercury C-line (656.3 nm) [100] or the cadmium line
(643.85 nm) [101, 102] instead of the sodium D-line (589 nm). In this way
optical activity was demonstrated in a-carotene (5) [102, 103], zeaxanthin (67)
[95], violaxanthin (135) [61], lutein (73) [96, 100], flavoxanthin (130) [104],
its epimer chrysanthemaxanthin (130) [104], capsanthin (170) [61], azafrin
(261) [105], taraxanthin (120) [61] and eschscholtzxanthin (84) [106]. How-
ever no optical activity could be 'detected with cryptoxanthin (39) [61] or
 V. Stereochemistry 289

celaxanthin (44) [107]. Again no activity was observed with capsorubin (205)
or its dipalmitate, though here rotations of the C-line were recorded with
cis isomers [97].
Despite the limitations mentioned, these classical studies were held to
support the view that, in all appropriate cases, natural carotenoids consist of
discrete optical isomers, and not of racemates or mixtures of isomers. This is,
of course, only to be expected from the probable stereospecificity of the enzy-
matic processes by which the carotenoids are believed to be formed. However,
it does not necessarily follow that a compound such as zeaxanthin (67) will
always have the same configuration regardless of the organism in which it is
produced. Neither does the detection of optical activity in a compound with
two or more sites for asymmetry show whether or not there is more than one
chiral element present.
In principle, the simplest way to avoid the complications associated with
the intense colour of carotenoids is to examine their perhydro derivatives. The
optical activity of zeaxanthin (67) [108, 109], cryptoxanthin (39) [110], lutein
(73) [100] and fucoxanthin (190) [111] has in fact been confirmed in this
way. However the method is by no means always as convenient as might be
supposed, and mention must be made of one report that perhydrozeaxanthin
is optically inactive [112]. Most carotenoids are only available in small amounts
and, quite apart from the desire to use non-destructive methods as far as
possible, the resulting perhydro derivatives are viscous oils and difficult to
handle on a small scale. Moreover it is now realized that it is not easy in
practice to achieve complete hydrogenation, particularly in carotenoids with
cyclic end groups, and that any contamination of the product with incompletely
reduced material might have a significant effect on the magnitude of the
observed rotation. Though this is not important if it is merely desired to deter-
mine whether or not a particular carotenoid is optically active, no comparisons
with other data are valid unless the products are first shown by mass spectro-
metry to be the required perhydro compounds.

b) O.r.d. and c.d. measurements

A more useful and general approach is provided by optical measurements
in the near ultraviolet region (200-400 nm) where carotenoids exhibit maxima
of comparatively low intensity (10- 3 e ca. 10-30) which have been attributed
to partial chromophores. In this range most carotenoids containing one or
more asymmetric carbon atoms exhibit optical rotatory dispersion (o.r.d.), with
characteristic Cotton effects [113, 114]. Some circular dichroism (c.d.) curves
[42, 114, 115] have also been determined, but this potentially very valuable
method has not as yet been fully exploited. Fig. 5 shows the relationship be-
tween the o.r.d., c.d. and the u.v.light absorption of zeaxanthin (67). As would be
expected, there is a good correlation between the position ofthe u.v. maximum,
the c.d. maximum and the mid-point between the extrema of the o.r.d. curve
(278, 286 and 284 nm respectively for the band oflonger wavelength). Very few
o.r.d. or c.d. measurements have been made with carotenoids in the neighbour-
Carotenotds 19
290 B. C. L. WEEDON

+5

+4 +20

+3

+2 , ... +10
I
I
I
+1 I
I
...
I
I
I }..(nm)
x
0

~
-1

-2 -10

-3

-4 +2 .I
.,.--.'\..\ -20
.\
...
___ ·" .;
. /
.\
b \ ..
'·...... __ _
+1
xw
}..(nm)

250 300 350

Fig. 5. O.r.d. (-), c.d. (----)and u.v. (-·-·-)curves for zeaxanthin (67) in dioxane
(from Bartlett et al. [114])

hood of the main absorption maxima, but the results reported for astaxanthin
(203) are of interest (Fig. 6) [116].
Samples of natural zeaxanthin (67) obtained from different plant sources,
Curcurbita pepo, Capsicum annuum flavum, Lycium halifolium and Physalis
alkekengi, from the brown alga Fucus vesiculosus [114], from blue green algae
and from bacteria [117], were found to have o.r.d. or c.d. curves in the u.v.
range of the same shape and magnitude (within experimental error), implying
the same absolute configuration at the asymmetric centres in all samples. The
same was true for samples of lutein (73) from alfalfa, pyrethrum (Chrysanthe-
mum cinerariifolium) and Chlorella vulgaris, for samples oflutein epoxide (120)
from Ranunculus acer, Impatiens' noli tangere and Taraxacum qfficinale, for
 V. Stereochemistry 291

X (nm)
400

.,..f, 1.25
I \
I \
I \
\
\
'
I
..!..
...
I
...
I I

52 52
X X
'S' w
L=o -0.1

0.25

26 22
v (cm-1 xl0-3)
Fig. 6. O.r.d. (----)and light absorption(-) curves for astaxanthin (203) in methylene chloride
(from Buchwald and Jencks [116])

samples of capsorubin (205) from Capsicum annuum and Encephalartos villosus,

for samples of (9-cis) neoxanthin (122) from spinach, maple leaves and the alga
Euglena gracilis, and for samples of alloxan thin (65), examined as the diacetate,
from the alga Cryptomonas ovata, the giant scallop Pecten maximus and from
the tunicate Halocynthia papillosa [114]. No instance has so far been found of
optical isomerism between samples of the same carotenoid isolated from
different sources.
Consideration of the o.r.d. properties of zeaxanthin (67) and of the related
carotenoid cryptoxanthin (39), which has one asymmetric end group struc-
turally identical with those in zeaxanthin, and one end group which is not
asymmetric, indicates that both end groups in zeaxanthin must contribute to
its optical properties [114]. This is confirmed by the molar rotations of the
hydro derivatives of zeaxanthin and cryptoxanthin prepared under comparable
conditions [109, 110]. A similar comparison of the o.r.d. properties of capso-
rubin (205) with those of cryptocapsin (157) again shows that both end groups
in the structurally symmetrical carotenoid are optically active [114]. Since
the structurally symmetrical carotenoids have a two-fold axis of symmetry
(C 2 ), this means that the two end groups must also be stereochemically iden-
tical. It is important to distinguish this situation from that in meso compounds
which have two identical end groups and a two-fold alternating axis of sym-
292 B. C. L. WEEDON

metry. No meso carotenoids have been observed in nature, though synthetic

zeaxanthin and synthetic capsorubin (see Chapter VI) presumably consist of
mixtures of DL and meso forms. Neither is there any evidence of a structurally
symmetrical carotenoid in nature which is optically active at one end and
inactive at the other.
It will be convenient at this point to classify optically active natural
carotenoids into three main stereochemical types: (i) homodichiral compounds
containing two identical chiral end groups, e.g. zeaxanthin (67) and capsorubin
(205), (ii) heterodichiral compounds containing two structurally non-identical
chiral end groups, e.g. capsanthin (170) and lutein (73), and (iii) monochiral
compounds with one chiral and one achiral end group, e.g. ex-carotene (5) and
rubixanthin (45) [114].

c) X-ray crystallography
The most direct way of determining the absolute configuration of any chiral
carotenoid would, of course, be by X-ray crystallography. No doubt this will
be possible before long using either a heavy atom derivative, or even the
carotenoid itself. However, as yet, this has not been achieved, and the only
successful X-ray crystallographic analyses so far reported with intact carote-
noids have been those of the achiral compounds mentioned in Section C.2
which have the advantage that the crystals are centrosymmetric. Meanwhile
considerable progress has been made using chemical and physical methods to
correlate the configurations of carotenoids with known absolute standards.
Another promising approach is to degrade the carotenoid to give, in effect, one
of the end groups which may then be more amenable to X-ray crystallographic
examination. Such methods will no doubt retain their value in the future, even
though the structure of some key carotenoids may be determined directly by
X-ray crystallography.
The evidence now available for the absolute configurations of various
carotenoids, and of a few structurally related compounds, is discussed in the
following sections. The configurations are shown by standard structural
formulae (- representing a bond projecting up from the plane of the paper,
and ..... one projecting below) and are specified by the RandS convention, which
is explained in Section E.

2. Capsanthin and capsorubin

The clue to the novel structure of capsanthin (170) came from the isolation of
the corresponding ketone, capsanthone (197), after Oppenauer oxidation and
its recognition as a cyclopentanone [118, 119]. To exclude the possibility that
the five-membered ring was formed during oxidation, capsanthone was reduced
with lithium aluminium hydride, and the resulting mixture of triols was
submitted to selective oxidation with chloranil. This gave, as expected, two
products, one of which was identical with capsanthin and the other was
formulated as the C-3' epimer [118]. Small differences in the carbonyl stretching
 V. Stereochemistry 293

frequencies in the i.r. spectra of the two compounds indicated intramolecular

hydrogen bonding in 3' -epi-capsanthin. It was therefore suggested that the
two oxygen substitutents on the five-membered ring in capsanthin (170) are
trans to one another (23) [118].

~H
;
-~~-
~. .~:b=O --o--
;
; 902"

OH OH
(23) (24) (25) (26)

The opposite conclusion was drawn by Faigle and Karrer [120, 121], who
isolated an optically active hydroxycamphonanic acid after ozonolysis of
capsanthin (170) acetate and capsorubin (205) acetate. Since the product
yielded an anhydride (25) when sublimed at 250°, it was assigned the cis con-
figuration (26) [121].
During studies on the structure of capsorubin, Cholnoky and Szabolcs [122]
noted that oxidative degradation yielded a mixture of acids which included
camphoronic acid (27). A similar degradation of capsanthin (170) was carried
out by Faigle and Karrer [121], who also showed that the camphoronic acid
formed had the same rotation as that reported earlier for a sample prepared
by degradation of (+)-camphor (28). Since the stereochemistry of the latter
was known [123], this observation by Faigle and Karrer established the
configuration of capsanthin at C-5', and of capsorubin at C-5 and C-5', assuming
that the degradation of the end group had proceeded normally.

~3
H02C--cH2-y--co2H
CH,-y--cH,
C02H
(27) (28)

Shortly afterwards Cooper et al. [124] reported the synthesis of the racemic
forms of both geometrical isomers (24) and (26) of the hydroxycamphonanic
acid. As expected, one lactonized very much more readily than the other, and
was therefore assigned the cis configuration (26). The two hydroxy acids were
then converted into the polyenes with the gross structure of capsorubin
(Chapter VI). The product from the trans hydroxy acid (24) had the same n.m.r.
and chromatographic properties as natural capsorubin (205), whereas the
isomer from the cis hydroxy acid (26) differed noticeably in both these respects
[124]. The end groups in capsorubin (205), and the corresponding end group
in capsanthin (170), were therefore assigned the trans configuration with the
absolute stereochemistry shown in (23) [124]. Later it was shown that, though
the hydroxycamphonanic acid derived from capsanthin yields an anhydride
when heated, this must involve an inversion of configuration at C-3; the (cis)
294 B. C. L. WEEDON

hydroxy acid formed on re-opening of the lactone was different from the
starting material and lactonized under much milder conditions. The n.m.r.
properties of these two hydroxy acids were in agreement with the revised
geometrical assignment for the natural end group [125].

-tJ- cro,H
_ <:02H 1) PC!, !

~ 2) C6 H,O.Na
3) Quinoline HO,AJ-
HO,c...~·
4) KOH

(28)
HBr/

/ ; c;:o,H

:JYJ-
' c;:o,H

H02~ +
H0 2
Br
/ Br

/ Na 2C0 3
Na 2C03 J

-er~=O
H0 2C'"'~ ........... 0
-11-
HO,C....
!

~
em
cro,H
uo;t)!"
; c;:o,H

(29) (30) (31)

1) CH2 N2
2) K2Cr20,-H2S04
3) H<tl

(28) ----+

H02~
;

OH
c;:o,H
----+
y· 0
(32) (33)

v
1
-v- -
,

(m
c;:o2H
H0 6
.
o~
........... ~=0
+

OH
(26) (25) (24)
Scheme 3
 V. Stereochemistry 295

Further confirmation of the stereochemistry of the cyclopentane end group

in capsanthin and capsorubin, and for that matter in cryptocapsin (157) (see
Section D.11), has been obtained by synthesis of the optically active forms of
the cis- (26) and trans- (24) hydroxycamphonanic acids from (+)-camphor (28)
[126, 127]. The route is summarized in Scheme 3. The preparation of the
hydroxy dicarboxylic acids (30) and (31) is based on the classical studies of
Bredt [128]. The subsequent oxidation to give (33) adopts the technique
developed recently by Marquet et al. [129] for the isomer (32) which was
derived from camphor by an alternative series of reactions. The optically
active trans hydroxy acid (24) synthesized from (+)-camphor had o.r.d.
properties in good agreement with those of a sample prepared by ozonolysis
of capsanthin [126, 127].
The configuration (23) for the cyclopentane end groups in carotenoids rests
on the previous assignment of stereochemistry to (+)-camphor. This formerly
depended on a lengthy, though apparently reliable, series of correlations, but
it has recently been verified by X-ray crystallographic studies on ( + )-3-bromo-
camphor [130]. The 3S,5R configuration (23) for the cyclopentane end group
in capsanthin and related compounds can therefore be regarded as firmly
established.
The characteristic end groups of capsanthin and capsorubin are believed
to be formed in nature from those of zeaxanthin (67) by reactions which do not
involve the oxygen substituent at C-3 (Chapter II; Scheme 8). Following the
elucidation of the stereochemistry of the cyclopentane ring it was therefore
suggested that the 'zeaxanthin' end group has the 3R configuration (52) [124].
It should be noted that, though C-3 is designated S in capsorubin and R in
zeaxanthin, this does not imply any inversion of configuration; the change is
merely the result of the application of the conventional sequence rule to specify
configuration (see Section E). This absolute configuration (67) for zeaxanthin
has recently been verified by the studies outlined below, and strong support
for the 3R,3' S,5' R configuration of capsanthin (170) has been obtained by o.r.d.
studies (Section 0.12).

3. Fucoxanthin and zeaxanthin

On reduction with lithium aluminium hydride, fucoxanthin (190) gives a
mixture of epimeric fucoxanthols [111, 131] whose gross structure (34) was
determined by Bonnettetal. [111]. That no reaction had occurred other than
reduction of the acetate and keto groups was established by selective oxidation
of the allylic secondary hydroxy group in the fucoxanthols with dichloro-
dicyanobenzoquinone to give fucoxanthinol (189); on acetylation under the
same conditio11s both fucoxanthin (190) and fucoxanthinol (189) gave the same
diacetate [111]. On treatment with traces of acid, the fucoxanthols (34) readily
lost the elements of water yielding a mixture of epimeric furanoid oxides, the
'fucochromes', which were shown [111] to have the same gross structure (131)
as that subsequently assigned to neochrome [29]. From their method of
preparation it seems safe to assume that these fucochromes preserve the
296 B. C. L. WEEDON

configuration of fucoxanthin at C-3 and C-3', and that the epimers differ in
configuration at C-8 and possibly at C-5.

~.yx;
HO
(34) HO~OH
Reduction of the mixture of fucochromes with lithium aluminium hydride,
under the conditions developed by Cholnoky et al. [132] for the conversion
of epoxides and furanoid oxides into their parent carotenoids, gave a product
identical in all respects, including o.r.d. properties, with natural zeaxanthin
(67) [111]. The reduction of the furanoid oxide end group, like that of a 5,6-
epoxide, presumably involves hydride ion attack to form a 5-hydroxy deriva-
tive which then undergoes elimination. The transformation at the other end
may be rationalized as involving reduction of the tX-hydroxy-allene group
(cf. reduction of tX-hydroxy acetylenes with the same reagent) to yield the
6',7'-dihydro derivative, followed by a 1,2-elimination of the substituent at
C-5'. Support for this interpretation comes from the observation [133] that
tX-hydroxy allenes (36) are intermediates in the conversion of 1,4-dihydroxy
acetylenes (35) into dienes (37) on treatment with lithium aluminium hydride

'\.
/y-c=c-ci\
OH
(35)
/

OH
- (36)
- '\.
/
C=CH-CH=C

(37)
/
'\.

[134]; with suitably substituted 1,4-dihydroxy acetylenes, the tX-hydroxy allenes

can be isolated in good yield [135]. The essential point for the present purposes
is that the C-3 and C-3' positions are not involved in the conversion of the
fucochromes into zeaxanthin. The C-3 and C-3' positions in fucoxanthin (190)
must therefore have the same configurations as the corresponding positions in
natural zeaxanthin (67) which, as pointed out previously, is homodichiral.

HO
(38)

o~·p
5 3
HO OAc

(39)
 V. Stereochemistry 297

Controlled partial oxidation of fucoxanthin (190) with zinc permanganate

furnishes a complex mixture of products which include the epoxy aldehyde (38)
and the allenic ketone (39) [111 ]. The relative configuration of the two oxygen
substituents at C-3 and C-5 in the allenic ketone (and hence at C-3' and C-5'
in fucoxanthin) was first determined by stereochemically controlled synthesis
of the racemic ketone (47) [136], from the readily available derivative (8) of
isophorone [71], by the method summarized in Scheme 4. Epoxidation of the
intermediate enyne diacetate (40) with monoperphthalic acid furnished, as
expected, a mixture of two epoxides (ca. 1 :2) which were separated by chro-
matography on silica gel. The minor, more strongly adsorbed isomer was
assigned the trans configuration (42) on the basis of experience with simple
models (see Section D.5). Reaction with lithium aluminium hydride gave the
epoxy diol (44) which, on more vigorous treatment with the same reagent,
yielded the allenic triol (45). Selective oxidation of the latter with manganese
dioxide then yielded the required allenic ketone (47). The cis acetoxy epoxide (41)
similarly gave the racemic ketone (46) with markedly different n.m.r. properties
[137]. Partial acetylation of the former racemate (47) gave the corresponding
acetate (48) identical in all respects (other than optical activity) with the degra-
dation product from fucoxanthin. It was therefore suggested that the oxygen
substituents at C-3' and C-5' in fucoxanthin are trans to one another [136].
It seems likely that the reduction of the acetylenic epoxides (43) and (44)
involves an organoaluminium intermediate of the type (50). If so, then in the
resulting allene the hydroxyl at C-5 and the substituent at C-8 would be ex-
pected to be trans to one another leading to the complete relative configurations
(46) and (47) for the two racemic ketones. Since, however, alternative reaction
pathways were conceivable for the key reductions, conclusive proof of the
stereochemistry of the racemate (47) was obtained by X-ray crystallographic
analysis of the p-bromobenzoate (49) [138].
In deuterioacetone the racemate (48) was found to exhibit n.m.r. bands which
were noticeably different from those observed with deuteriochloroform solu-
tions. The solvent shift for the allenic hydrogen signal was significantly larger
than those for the C-19 and other methyls. This is best interpreted as the result
of interaction of the C-5 hydroxyl with deuterioacetone with preferential
shielding of the neighbouring allenic hydrogen. The observation of the same
solvent shifts with the 'natural' allenic acetate from fucoxanthin showed that
this had the same geometrical configuration as that established by X-ray
crystallography for the synthetic racemate [138]. As the configuration at C-3'
in fucoxanthin was known to be the same as that at the corresponding positions
in zeaxanthin (67), and since these were believed on biogenetic grounds to be
the same as that established for C-3' in capsanthin (see Section D.2), the
absolute configuration (48) was proposed for the allenic end group in fuco-
xanthin (190) [138]. This assignment has recently been confirmed in the follow-
ing way.
Reduction of the 'natural' allenic acetate (39) from fucoxanthin (190) with
lithium aluminium hydride gave the allenic triol (45). Selective oxidation of
 298 B. C. L. WEEDON

1) Grignard
J:
+0~ 2) HEil

3) LiAIH 4
~u
HO --
OH

(8)

J:: ~D·~·
1) Ac 20
2) POC1 3

(40)
--
[OJ
o(
(41)
"'oAc
+

l l
•,

J: J:
~ (50)
OH
~D.,...
o...,

(43)
· '·ou
~~
;
(44)
..._OH

l l
·~·p
uo·~· ·ou
·~·n
uo •ou i
(45)

l l
Y·p Y·n HO
(46)
OH HO ! ''OR

R=H (47)
R=Ac (48)
Scheme 4 R = p- BrC6 H 4 CO (49)
 V. Stereochemistry 299

~ Br
Oo
0 c
Fig. 7. Molecular structure of the allenic p-bromobenzoate (49) prepared from natural fucoxanthin

the allylic hydroxyl group with manganese dioxide then furnished the allenic
ketone (47), which was reacted with p-bromobenzoyl chloride to give the
p-bromobenzoate (49). The structure of the latter (Fig. 7) was solved by X-ray
crystallography using the heavy-atom method [139]. The absolute configu-
ration was determined by the method of Bijvoet [140] using 15 pairs of reflec-
tions, all of which gave a consistent result. This X-ray study therefore provides
complete proof of the 3'S,5'R,6'R configuration (51) for the allenic end group
in fucoxanthin [139]. Furthermore, since the stereochemistry at C-3' in fuco-
xanthin is the same as that at C-3 and C-3' in zeaxanthin, the latter must have
the 3R,3'R configuration (67), with end groups of the type (52), as was proposed
earlier on biogenetic grounds [124].

.... ··~;·~-~
~......
HO~~'OAc HOA)l
(51) (52)

Regarding the epoxide end group of fucoxanthin (190), it will be recalled

that fucoxanthin has been converted into zeaxanthin (67) identical with the
natural homodichiral pigment. It follows that C-3 in fucoxanthin must have
the same configuration as C-3 and C-3' in zeaxanthin. This leaves the 5,6-
epoxide group, but unfortunately there is as yet no definite information con-
cerning its stereochemistry. Infrared light absorption studies with the degrada-
tion product (38) revealed no intramolecular hydrogen bonding between the
epoxide and the hydroxyl group at C-3 and a trans arrangement between these
two groups is therefore preferred [ 111 ], though the evidence cannot be regarded
as conclusive. Such a configuration would, however, be consistent with the
view that both ends of fucoxanthin, like those in violaxanthin (135), are formed
in nature from :?:eaxanthin by variations in a basically similar process [ 40, 138].
Since the configuration at C-5' in fucoxanthin is known unambiguously, and
the same configuration is favoured for C-5 and C-5' in violaxanthin (135)
(see Section D.5)", the corresponding configuration for C-5 in fucoxanthin, i.e.
5,6-epoxide trans to the hydroxyl at C-3, seems plausible. Though in principle
the point could be settled conclusively by X-ray crystallographic analysis of(38),
300 8. C. L. WEEDON

or one of its derivatives, all attempts to prepare a suitable crystal have so far
been unsuccessful. Subject to this single reservation concerning the stereo-
chemistry of the epoxide group, the absolute stereochemistry of fucoxanthin is
that shown in (53), and can be represented as 3S,5R,6S,3'S,5'R,6'R.

0
··''X)'
""' ..
~····"""<:::,•

HO
.
!
5' 3'
~..'OAc
(53)

4. Fucoxanthinol and paracentrone

Both fucoxanthinol (189) and paracentrone (246) from the sea urchin,
Paracentrotus lividus [141], have been identified by direct comparison with
samples prepared from fucoxanthin [141, 142]. The preparation of fucoxan-
thinol [111] has already been described. That of paracentrone involved
Oppenauer oxidation of fucoxanthin, spontaneous cleavage of the resulting
ketone (54) (Scheme 5) and hydrolysis of the paracentrone acetate thus pro-

Scheme 5

duced [142]. The agreement in the n.m.r. and chromatographic properties

between the natural and the semi-synthetic pigments shows that the relative
configuration of the asymmetric centres in the natural pigments is the same as
for the corresponding positions in fucoxanthin. Since fucoxanthinol and para-
centrone are almost certainly formed in the sea urchin from fucoxanthin [141],
their absolute stereochemistry can be assumed to parallel that of fucoxanthin.

5. Violaxanthin
Violaxanthin (135), on reduction with lithium aluminium hydride, gives
zeaxanthin which exhibits an o.r.d. curve in good agreement with that of
natural zeaxanthin (67). Violaxanthin must, therefore, have the same configu-
ration at C-3 and C-3' as zeaxanthin [114, 132].
It has long been known that treatment of zeaxanthin acetate with mono-
perphthalic acid, and subsequent hydrolysis, gives a product closely resembling
violaxanthin [143, 144]. Recently it was shown that the intermediate 'viola-
xanthin acetate' can be separated by chromatography into two isomers which
on separate hydrolysis gave 'semi-synthetic violaxanthin-A' and 'semi-syn-
thetic violaxanthin-B' [114]. N.m.r. studies indicated that the semi-synthetic
violaxanthin-A, like natural viola:!anthin, had two identical end groups, but
 V. Stereochemistry 301

+4 1\
I \
I \
I '
I '
I \
I '
II '\
+2 I \
I '
II '-... ... _
I
I
I
... I >..(nm)
I
I
0
X 250 300
~
I
I
I
I
-2 I
I
I
I
I
I
I
\ I
-4 ' I
' .;I
Fig. 8. O.r.d. curves for natural violaxanthin (135) (-)and semi-synthetic violaxanthin-A (--- -)
(from Bartlett et al. [114])

that the ring methyls in the semi-synthetic compound experience slightly

different shielding from those in the natural pigment. Semi-synthetic viola-
xanthin-B appeared from its n.m.r. spectrum to have one end group with the
same configuration as those in semi-synthetic violaxanthin-A, and the other
the same as those in natural violaxanthin.
The o.r.d. curve of semi-synthetic violaxanthin-A proved to be nearly
enantiomeric to that of natural violaxanthin (Fig. 8); since there was no
evidence of isomerism about any carbon-carbon double bond, this striking
difference was attributed to a difference in the stereochemistry of the epoxide
rings in the semi-synthetic violaxanthin-A and the natural carotenoid. As
expected, semi-synthetic violaxanthin-B, with one end group of each type, (55)
and (56), exhibited no detectable o.r.d. curve [114].

(56)
302
.
B. C. L. WEEDON

D
(57)
A cO
(58)
.0 +
D
AcO
(59)
0

Reaction of the simple model (57) under conditions similar to those used
to epoxidize zeaxanthin acetate, gave the trans (58) and the cis (59) epoxy
acetates in the proportion ca. 1: 2. The configurations of these two products
were assigned from their relative retention times on chromatography, and from
infrared light absorption studies on the derived epoxy alcohols; that which
exhibited intramolecular hydrogen bonding was assigned the cis configuration,
and that which did not the trans. A further model was provided by the enyne
diacetate (40) used in the synthesis of the allenic ketone (47) (Scheme 4). Here
two epoxides were formed (ca. 1: 2), and the trans configuration assigned to the
minor product was subsequently confirmed by X-ray crystallographic analysis
of the p-bromobenzoate (49) of the derived allenic ketone (47) [136, 138]. With
both models epoxidation with monoperphthalic acid gave more cis than trans
epoxy acetate, and therefore the end groups in natural violaxanthin (135) are
provisionally assigned the 3S,5R,6S configuration (55) in which the epoxide
ring is trans with respect to the C-3 hydroxyl group, and those in semi-syn-
thetic violaxanthin-A the 3S,5S,6R configuration (56) in which these two
functional groups have a cis relationship [114].

6. N eo xanthin
Neoxanthin (122) is remarkably labile to acids. On treatment with ca. 0.002%
ethereal hydrogen chloride it gives a mixture of neochromes (131). From their
mode of preparation these, like the fucochromes (Section D.3), can be assumed
to preserve the configuration of the starting material at C-3 and C-3', but to
differ in configuration at C-8' (and possibly about the 9',10'-double bond) [29].
Reduction of the neochromes with lithium aluminium hydride gave zeaxanthin
with o.r.d. properties in good agreement with those of the natural homodichiral
pigment (67) [29]. Neoxanthin, like fucoxanthin (190) and violaxanthin (135),
must therefore have the same configuration at C-3 and C-3' as zeaxanthin
[29, 114].
The n.m.r. properties of all-trans-neoxanthin are those that would be ex-
pected of a carotenoid with one end group identical with the allenic end of
fucoxanthinol (189) and the other with those in natural violaxanthin (135) [29].
This defines all chiral elements with respect to the known asymmetric centres
at C-3 and C-3'. Neoxanthin can therefore be represented by the 3S,5R,6R,
3'S,5'R,6'S configuration (60). The only uncertainty concerns the stereochemis-
HO......f'f"OH

~-~········~
 V. Stereochemistry 303

try of the 5',6'-epoxide, which depends on the correctness of the assignment to

the corresponding groups in violaxanthin (135). All other chiral elements are
derived from the X-ray crystallographic analysis of the p-bromobenzoate (49).

7. The allenic ketone from grasshoppers

The racemate (47) prepared by the route outlined in Scheme 4 had n.m.r.
properties in excellent agreement with those reported by Meinwald et al. for
the allenic ketone which they isolated from an ant-repellant secretion of the
large flightless grasshopper, Romalea microptera, and which they had shown to
have the same gross structure [145]. However, the possibility could not be
excluded that the grasshopper ketone has the (relative) stereochemistry (66),
tentatively favoured by Meinwald and Hendry [146], since it was not known
whether a difference in stereochemistry about the allene group would influence
the chromatographic and n.m.r. properties to any appreciable extent.

(61)

A possible mechanism for the biogenesis of the allenic end groups in

carotenoids was suggested by studies on the photosensitized oxidation of
simple derivatives (61) of P-ionone [147-149]. The products included small
amounts of allenic hydroperoxides which were subsequently reduced to the
corresponding alcohols (62) with end groups reminiscent of those in the allenic
carotenoids. These photochemical reactions were believed to occur by a con-
certed attack by singlet oxygen at the C-5 position and at the hydrogen attached
to C-7. The products were therefore assigned a (relative) stereochemistry in
which the resulting oxygen function at C-5 and the allenic hydrogen at C-8 are
trans to one another, i.e. the same as that tentatively proposed for the grass-
hopper ketone [146], but the opposite of that subsequently demonstrated for
the end group in the natural allenic carotenoids [139].
To resolve some ofthe issues raised above, the enyne diacetate (40), obtained
as an intermediate in the preparation of the two allenic racemates (46) and (47),
was converted into the diol (63), which on reduction with lithium aluminium
hydride furnished the (7-trans) diene diol (64) (Scheme 6) [150]. Photochemical
oxidation of the latter, in the presence of Rose Bengal as sensitizer, and reduc-
tion of the hydroperoxide thus produced, gave the expected allenic triol (65).
Selective oxidation of the allylic secondary alcohol function with manganese
dioxide yielded the corresponding allenic ketone, which was shown by X-ray
crystallographic analysis of its p-bromobenzoate (67) to be the racemate (66)
[151].
This result verifies the stereochemical course proposed for the in vitro
reactions leading to the simple allenes (62). It does not, of course, prove that
 304 B.C.L.WEEDON

J:~x ----- UOH

(63) (64)

--"0~·~
HO~OH
(65)
~~:Po. R=H (66)
R=p-BrC 6 H 4 CO (67)
Scheme 6

the photochemical oxidations necessarily proceed by means of a concerted

attack by singlet oxygen at C-5 and the hydrogen at C-7. Dioxetane or related
intermediates might well be involved [139, 152, 153]. Moreover nothing is yet
known regarding the configuration of the allene which is reported to be formed
on photochemical oxidation of P-carotene (3) using chlorophyll as the sensi-
tizer [154]. However, the n.m.r. properties of (66) were significantly different
from those reported for the grasshopper ketone. The latter must therefore have
the same geometry as that established for the racemate (47), from which it
differs only in melting point and presumably in optical activity [155]. This
strongly supports the view that the grasshopper ketone is formed in vivo by
degradation of the neoxanthin present in the leaf diet of the insects [ 40, 136].
Thus it is probable that (47) also represents the absolute configuration of the
natural ketone.

8. Abscisic acid
The plant growth regulator abscisic acid has a structure which resembles that
ofthe end groups in a number of carotenoids. The absolute stereochemistry (68)
has been assigned to natural ( + )-abscisic acid by Cornforth et al. [156] on the
basis of studies carried out with synthetic ( + )- and (- )-abscisic acid.
The methyl ester of synthetic ( + )-abscisic acid was reduced with sodium

,a·. ·~
borohydride, and the resulting mixture of cis (69) and trans (70) diol esters was

·~
'"<:::::,
OH
"<::::::

C02H
---+
,9
~~ "<::::::
OH
"<::::::

C0 2CH 3
+ a~
_...~ ,9
~·~ "<::::::
OH
"<::::::

C0 2CH 3
0 HO HO
(68) (69) (70)

HO~H•nr;(72)
 V. Stereochemistry 305

separated by chromatography. The cis diol ester (69) was identified by compari-
son with an authentic specimen of the racemate prepared by catalytic reduction
of the epidioxide (71 ), an intermediate in the synthesis of ( ± )-abscisic acid [157].
At the D-line, the cis diol ester (69) from ( + )-abscisic acid was more laevorota-
tory than the trans diol ester (70). According to Mills' empirical rule [158], for
which no exception seems to be known [156], the absolute configuration at C-3
(carotenoid numbering) of the cis and trans diol esters would appear to be (72)
and (73) respectively, from which it follows that the absolute stereochemistry
of the cis diol ester is (69), and that of natural ( + )-abscisic acid is (68). This
conclusion, which was also reached by parallel studies with synthetic (- )-
abscisic acid, depends on the reasonable assumption that Mills' rule is valid
in this series despite the presence of a chromophore capable of showing a
Cotton effect in the middle of the ultraviolet range.
It will be noted that the configuration assigned to ( + )-abscisic acid is the
opposite of that favoured for C-6 in violaxanthin (135) and neoxanthin (122)
(Sections D.5 and D.6). This obviously casts doubt on the suggestion that ( + )-
abscisic acid is a metabolite of the common carotenoid epoxides in plants,
unless the conversion is rather more subtle than that contemplated [142, 142a].
However, it is now known that, contrary to earlier indications, the laevorotatory
enantiomer of abscisic acid is, at least in one bioassay system, as potent a
growth inhibitor as the natural (+)-isomer [159]. Therefore it is not inconsistent
with the present stereochemical proposals for abscisic acid (68) and violaxanthin
(135) that growth inhibitory substances have been obtained in vitro by irradia-
tion of violaxanthin and neoxanthin [160, 161].
It is interesting to note that loliolide (digiprolactone), which is also possibly
a carotenoid degradation product, has been formulated as (74) [162], though
a different stereochemistry has also been proposed [163].

~0
HO~""'""'"[
(74) (75)

9. Alloxanthin
Insight into the absolute configuration of the acetylenic carotenoids was
obtained by perhydrogenation of alloxanthin (65) over a platinum catalyst.
The resulting saturated glycol exhibited optical rotation which agreed well
with that of a sample prepared by perhydrogenation of natural (3R,3' R)-zea-
xanthin (67) under identical conditions [164]. It was therefore concluded that
alloxanthin has the corresponding 3R,3'R configuration (65) with end groups
of the type (75) [114, 139].
Alloxanthin and zeaxanthin exhibit markedly different o.r.d. properties,
but this may be due, at least partially, to the different orientation of the (partial)
chromophores with respect to the chiral centres [114].

Carotenmds 20
306 B. c. L. WEEDON

The carotenoids pectenoxanthin and cynthiaxanthin (Chapter II, Section J)

were shown to have the same chromatographic and n.m.r. properties as allo-
xanthin. The similarity of the o.r.d. curves of the acetates of these diacetylenic
pigments indicated that the two carotenoids from marine animals have the
same absolute configuration as the algal pigment, i.e. that all three carotenoids
are completely identical [114, 164].

10. rx-Carotene
( ±)-rx-Ionone can be resolved via its menthylhydrazone [165]. The ( + )-
enantiomer has been converted into a product identical with naturalrx-carotene
(5), and the (- )-rx-ionone into the enantiomer ofnaturalrx-carotene, by reactions
which do not involve the asymmetric centre [114, 166]. Recently Eugster et al.
have shown that (- )-rx-ionone has the absolute stereochemistry (77) [167].
It follows that natural ( + )-rx-ionone, which occurs in Boronia megastigma [168],
Aplotaxis lappa [169], Acacia farnesiana [170, 171] and Rubus idaeus [172],
must be the enantiomer of (77), and that naturalrx-carotene (5) must have the

7/
end group (76) with the R configuration [167].

(76)

The main evidence (Scheme 7) for the S stereochemistry (77) for (- )-

rx-ionone was obtained by reduction to give the dihydro derivative (78), which
was converted via the corresponding rx-cyclogeranylacetic acid into the trans
<5-lactone (79), previously obtained by degradation of manool (80) [173].

x~·~o
..
~
I

(77) (78)

~,Do"
w~
(80)
wo l

(79)
Scheme 7

Although the optical activity of natural e-carotene (10) has not been re-
ported, it seems probable that the end groups of this carotenoid also have the
R configuration (76).
 V. Stereochemistry 307

11. M onochiral xanthophylls

A broad survey of the o.r.d. of carotenoids led to the hypothesis that the
perturbations by chiral centres at opposite ends of the molecule are, to a first
approximation, additive [114]. To test this hypothesis the o.r.d. curve for one
half of zeaxanthin was calculated by dividing the amplitudes in the experimental
curve for zeaxanthin (67) by two. The result was then compared with the
observed o.r.d. curves for cryptoxanthin (39), rubixanthin (45), gazaniaxanthin
(46), reticulataxanthin (238) and P-citraurin (249), all of which have one end
group of the zeaxanthin type and one end group with no chiral centre to
perturb the chromophore. As seen from Fig. 9 there is reasonable qualitative
agreement. It was therefore concluded that the single asymmetric centre in each
of the five monochiral compounds mentioned above has an absolute config-
uration identical to that ·in the zeaxanthin end group (52), namely R [114, 139].

."'"'·\
I
+2 ~

+1

...
I
!2 \'tQQ ;
X I (" i
~ I \ i
I
I I
/
I .I
I
I \..,/
-1 I
I
I
I
I
I

\
I
I
I
-2 I
I

Fig. 9. O.r.d. curves for! zeaxanthin(-), cryptoxanthin (39) (--- -), rubixanthin (4S) (- x- x- x ),
P-citraurin (249) (· · ·) and reticulataxanthin (238) (-·-·-) (from Bartlett et al. [114])
308 B. C. L. WEEDON

The o.r.d. curve for cryptocapsin (157), a monochiral carotenoid with an

inactive /3-end group, was similarly found to resemble the curve calculated for
'half-capsorubin'. Cryptocapsin was therefore assigned the same 3S,5R stereo-
chemistry (23) as capsorubin (205) [114].
Recently it has been shown that the diols formed on reduction of natural
semi-1)(-carotenone (214) with lithium aluminium hydride have c.d. properties
in the u. v. range which agree well with those of (6R,6' R)-~::-carotene (10). It
follows that semi-1)(-carotenone (214), like ()(-carotene (5) from which it is pre-
sumably derived, has R chirality [167 a].
Mention must also be made of another monochiral carotenoid, azafrin
(261) [105, 114]. No evidence of intramolecular hydrogen bonding has been
detected by infrared light absorption studies on azafrin methyl ester [174] or
on the monomethyl ether of the methyl ester [175]. It has therefore been con-
cluded that the two hydroxyl groups in azafrin have a trans diaxial arrangement.
However nothing is yet known concerning the absolute configuration of azafrin.

12. Heterodichiral xanthophylls

As a further check on the additivity hypothesis mentioned above the o.r.d.
curve for capsanthin (170) was compared with that calculated by addition of

+2

+1

...I A.(nm)
0 ~--~--~~----~~~r-----~---
x 1250 I
~ I I
I I
I \
I \
I \
I \ I
-1 I \ I
1 I \.//
I I
I I
1/
-2
Fig. 10. O.r.d. curves for capsanthi11 (170); experimental(---) and calculated(----)
(from Bartlett et al. [ 114])
 V. Stereochemistry 309

one half the molecular rotations of zeaxanthin (67) and of capsorubin (205)
[114]. The good agreement (Fig. 10) provided strong support for the configu-
ration assigned to the 'zeaxanthin' end group of capsanthin (170) on the basis
of the evidence summarized in Section D.2.
Zeinoxanthin (42) has end groups structurally identical with those in
zeaxanthin (67) and the chiral end of a-carotene (5) respectively. Its o.r.d. curve
(Fig. 11) serves to illustrate the general point that very approximate additivity
of half-carotenoid molecular rotations may still suffice to indicate the absolute
configuration. Included in Fig. 11 are the calculated curves for(! zeaxanthin +a-
carotene) and (! zeaxanthin+ the enantiomer of a-carotene). The remaining
two possibilities would, of course, be the mirror-images of these two calculated
curves, and it can be seen that the experimental curve resembles that calculated
for(! zeaxanthin+ a-carotene) and bears no resemblance to that calculated for
any other diastereoisomer. Zeinoxanthin can therefore be assigned the 3R,6' R
configuration (42) with some confidence [114, 139].
Diatoxanthin (66), with end groups of identical structure to those in zea-
xanthin (67) and alloxanthin (65), provides a similar situation. As shown in

+3

+1

}..(nm)

350

I
I
I
I
-2 I

I
I
I
I
I
I
-4 I
I
I

Fig.ll. O.r.d. curves for zeinoxanthin (42); experimental (-), calculated for (! zeaxanthin+
oc-carotene) (----)and calculated for(! zeaxanthin +enantiomer of oc-carotene) (-·-·-)
(from Bartlett et al. [114])
310 B. C. L. WEEDON

,.-.\
I i \
+2 i i \
i
i
i
i
I
i
+1 i
i
i
I
I I
I
... I i ;
I
I I A.(nm)
0
X i I 250 300 350
~ i I
i I /
; I ,I
j I I

i ; i. / /
-1 .I.; / I
I • I
.I \.../ I
\. I
I
I
I
I
I
I
I
-2 I
~

Fig. 12. O.r.d. curves for diatoxanthin (66); experimental (-), calculated for (! zeaxanthin+
t alloxanthin) (- - --) and calculated for (t zeaxanthin+ enantiomer oft alloxan thin) (- ·- ·-)
(from Bartlett et al. [114])

Fig.12 there is close agreement between the experimental curve for diatoxanthin
and that calculated by the addition of half the molecular rotations of zeaxanthin
and alloxan thin. Diatoxanthin is therefore assigned the 3R,3' R configuration
(66) [114, 139].
With crocoxanthin (41), which has end groups structurally identical with
those in alloxanthin (65) and the chiral end of a-carotene (5), the agreement
between the calculated and experimental curves was not sufficient to reveal
the complete stereochemistry of crocoxanthin. The experimental o.r.d. curve
had the same general shape as that calculated for either (a-carotene+t allo-
xanthin) or (a-carotene+the enantiomer of-} alloxanthin), but bore no resem-
 V. Stereochemistry 311

blance to either of the other calculated curves using the molecular rotations for
the enantiomer of a-carotene. The o.r.d. results therefore showed that the
'a-carotene end' of crocoxanthin has the same R configuration (76) as a-caro-
tene, but allowed no distinction between the two steric possibilities at the
alloxanthin end. However, since alloxanthin and crocoxanthin occur together
in nature it is highly probable on biogenetic grounds that both have the same
configuration at C-3. There therefore seems little doubt that crocoxanthin (41)
has the 3R,6'R configuration [40, 114].
An important example of a heterodichiral carotenoid is lutein (73). This
contains one end group structurally identical with those in zeaxanthin (67), but
authentic samples of monochiral or homodichiral carotenoids with the other
end group of lutein have not so far been available for o.r.d. measurements. The
approach used to assign stereochemistry to the three preceding heterodichiral
carotenoids is therefore not applicable. Some tentative conclusions can,
however, be drawn on the basis of the following arguments.
Experiments with etiolated maize seedlings and with Physalis alkekengi
have shown that it is the pro-R hydrogen from C-5 of mevalonic acid that is
lost in the introduction of the hydroxy functions in lutein (73) and in crypto-
xanthin (39) [176]. This suggests that the oxygen functions in lutein have the
same stereochemistry as that in cryptoxanthin, and, incidentally, that the
biological hydroxylation at C-3 in carotenoids, like many hydroxylations in
di- and tri-terpenes, involves replacement of a hydrogen atom without inversion
(see Chapter VII) [139]. It therefore seems likely that lutein (73) has a 3R,3'S
configuration corresponding to the 3R,3'R configuration of zeaxanthin (67).
It has therefore been possible to calculate the contribution of the 'hydroxy-a'
end to the molar rotation of lutein by subtracting the values for 'half-zeaxan-
thin' from the experimental values for lutein [114].
Reduction of natural lutein epoxide (120) with lithium aluminium hydride
gives lutein with similar o.r.d. properties to those of natural lutein [114],
indicating that the two natural carotenoids have the same configuration at
C-3, C-3' and C-6'. The acetate oflutein has been converted into semi-synthetic
lutein epoxide by the method used for the conversion of zeaxanthin (67) into
semi-synthetic violaxanthin-A (see Section D.5). As in the violaxanthin series,
semi-synthetic lutein epoxide had an o.r.d. curve opposite in sign to that of the
naturally occuring compound. The experimental o.r.d. curve for natural lutein
epoxide agreed reasonably well with that calculated for (t violaxanthin +
hydroxy-a), and the observed curve for semi-synthetic lutein epoxide was in
good agreement with that calculated for (t semi-synthetic violaxanthin-A +
hydroxy-a). These findings led to the conclusion that the epoxide end of lutein
epoxide has tqe same configuration as natural violaxanthin [114]. It therefore
seems likely that lutein epoxide has a 3S,5R,6S,3' S structure.
Recently it was shown that the product formed by elimination of the allylic
hydroxyl group from lutein (73) is identical with zeinoxanthin (42). The con-
figuration at C-6' in lutein and its epoxide (120) must therefore be the same as
that of the corresponding position in zeinoxanthin, namely R [176a].
312 B. C. L. WEEDON

Other carotenoids which are known from o.r.d. measurements to be

optically active include astaxanthin (203), crustaxanthin (93), monadoxanthin
(72) and decaprenoxanthin (226) [114], though there is as yet no information
on their absolute configurations. From a consideration of the polarity of
crustaxanthin (93), and its resistance to allylic dehydration, Nicoadi et al. [177]
have concluded that the two hydroxyls in each end group are mutually trans
to one another.

13. Trisporic acids

Ozonolysis of trisporic acid C (81), followed by lithium aluminium hydride
reduction, gave (- )-(R)-pentane-1,4-diol [178]. This confirmed an earlier
report [34] that ozonolysis of a hexahydro derivative of trisporic acid C gave
(R)-y-valerolactone [179]. To determine the stereochemistry at C-1, the circular

nnH ....H
OH OH

(81) (82)

dichroism of the tetrahydro derivative (82) was studied [35]. Three c.d. maxima
were recorded at 334, 245 and 217 nm. The two strong bands at 245 and 217 nm
were very similar to those observed with 3,20-dioxopregn-4-en-19-oic acid and
with nimbin [180] indicating the S configuration at C-1 in trisporic acid as
shown in (81). The implications of this result for the cyclization reaction leading
to the prochiral centres in the biosynthesis of /3-carotene (3) are discussed in
Chapter VII.

E. Annex. Specification of Absolute Configuration*

The absolute configurations of carotenoids discussed in this chapter have

been uniquely specified by means of the general R and S convention. The latter
was developed by Cahn, Ingold and Prelog [181-183], and the reader is
referred to a convenient introduction by Cahn [184]. The salient features for
the present purposes are summarized below.
In order to specify the configuration of an asymmetric carbon atom, the
four ligands attached to that atom are first assigned a descending order of
priority, a > b > c > d, on the basis of a sequence rule. If a model of the asym-
metric centre is then viewed from the side remote from the ligand of lowest
priority (d), a circle traced from a to b to c follows either a clockwise (I) or an
* The author is indebted to Dr. R. S. Cahn, Dr. G. P. Moss, Dr. J. W. Cornforth and Professor
C. H. Eugster for valuable comments on the 'preparation of this annex.
 V. Stereochemistry 313

anti-clockwise course (II). In the former case the asymmetric centre is designated
R, and in the latter S:
a a
A
~
- \:-c~
··'I
A
~
- "e-el
.ll
c·· I b' I
b R c s
(I) (II)

The sequence rule for assigning the priorities to the four ligands consists of
four sub-rules, only two of which are relevant here. The first of these (sub-rule 1),
'Higher atomic number precedes lower', suffices for all chiral centres in natural
carotenoids. It is applied to the four atoms by which the ligands are joined to
the asymmetric carbon atom. If the relative priorities of two (or three) groups
cannot be decided immediately in this way, the sub-rule is applied to a similar
comparison of the atomic numbers of the next atoms in the ligands, or if this
fails, of the next. The process is continued until a decision is obtained. If
branches are encountered in the ligands, the procedure is to consider the first
atoms in the prior branches (as determined again by the sequence rule), and
then, if necessary, the first atoms in the second branches, and finally the first
atoms in the third branches. This process is repeated working systematically
along the branches until a distinction is obtained (see Section 2.1 of reference
[183]; this differs in some respects from the procedure outlined in the earlier
paper [184]).
The treatment of multiple bonds in the application of the sequence rule
calls for special comment, and it is important to note that the convention which
was proposed in the third of the key papers [183] supercedes that adopted in
the first two [181, 182]. According to the current convention [183, 184], both
atoms of a double bond are considered to be duplicated, and then all atoms
other than hydrogen are complemented to quadriligancy by notionally adding
any necessary number of phantom atoms of atomic number zero. Thus the
carbonyl group C=O is treated as
y--?oo
( 0 )ooo
(C)ooo

and the carbon-carbon double bond C=C as

y-y
(C)ooo (C)ooo

where the duplicate atoms are shown in parentheses, and the phantom atoms
as o. Similarly the carbon-carbon triple bond C=C is treated as

(T)ooo (T)ooo
y-y
(C)ooo (C)ooo
314 B.C.L.WEEDON

Asymmetry resulting from the introduction of isotopic labels such as

deuterium is not, of course, covered by sub-rule 1. Such instances are dealt
with by sub-rule 2, which states' Higher mass number precedes lower'. However,
sub-rule 2 must only be invoked if no decision for a given chiral element is

'ill'
possible on the basis of sub-rule 1. x

C I iJ
I
y
(Ill) (IV)

In addition to chiral centres of the types discussed above, fucoxanthin (190),

neoxanthin (122) and a number of related carotenoids possess an allene group
which constitutes a chiral axis. This, notionally, is the result of taking a tetra-
hedron (III) with a chiral centre X and extending it to give the chiral axis
XY (IV). Although in all natural allenic terpenes known the four ligands a, b,
c, dare different, this is not a necessary condition for axial chirality (i.e. that the
mirror image is not superimposable on the original); it would suffice that
ligand a is different from b, and c from d, even though a= c and b =d. Therefore
the general application of the sequence rule to axial chirality requires an
additional rule: 'Near groups have priority over far groups'. The terms 'near'
and 'far' refer to the pairs a and b or c and d viewed from either end of the
axis XY. Fortunately it does not matter whether the models are viewed from
the X or the Y end of the axis; thus either way (IV) describes an R form.
The following examples illustrate the application of the conventions
summarized above.

Ex. 1. Zeaxanthin
The end groups of natural zeaxanthin (67) have the absolute configuration
shown in (V). Of the four ligands attached to the asymmetric centre at C-3, the
hydroxyl group clearly has highest priority (a) and the hydrogen atom the
lowest (d) (sub-rule 1). However, a distinction between the remaining two
ligands is not immediately apparent. It is therefore necessary to consider the

~...... """-
"<•"
...
HOAA
(V) (VI)
a
7 YH• H ?H H ~7
b 5--~i---!-Q)-1-- ~I c
I I I I I I
(C)ooo CH 3 H H H (C)ooo (C)ooo
d
(VIII)
 V. Stereochemistry 315

conventional partial expansion (VI), and the derived dissection (VIII) in which
the large numerals refer to different carbon atoms numbered according to the
normal carotenoid convention. Comparison of C-2 with C-4 provides no
distinction since both are of the type C(CHH), i.e. each consists of a carbon atom
to which the further linkages are to one other carbon and two hydrogens. The
prior branch from C-2 is clearly C-1 (and not one of the hydrogen atoms).
Similarly that from C-4 is C-5. However, comparison of C-1 with C-5 offers no
distinction since both are of the type C(CCC). Comparison of the second and
then the third branches from C-2 and C-4 is also fruitless since all consist of
hydrogen atoms. It is therefore necessary to consider the prior branches from
C-1 and C-5. The prior branch from C-1 is clearly C-6 since it is of the type
C(CCC), whereas both other branches (the methyl groups) are of this type
C(HHH). The prior branch from C-5 is also C-6, which is again seen to be of the
type C(CCC), whereas one of the other branches is of the type C(HHH) and the
other is of the type C(ooo). Since C-6 from either approach counts as C(CCC) it
is necessary to turn to the second branches from C-1 and C-5. These both
consist of a methyl group and are again equivalent. However, the remaining
branch at C-1 consists of a further methyl group whilst that at C-5 is the notional
duplicate of C-6. As the methyl is of the type C(HHH) whereas the duplicate
of C-6 counts as C(ooo), the former clearly has priority. It follows that, of the
two carbon ligands attached to C-3, that comprising C-2 and C-1 has preced-
ence over the C-4,C-5 ligand. Thus the order of precedence of the four
ligands is that indicated in (VII), from which the asymmetric centre is seen to
be R.

Ex. 2. a-Carotene
The chiral end group of natural a-carotene (5) has the absolute stereo-
chemistry indicated in (IX). Consider the conventional partial expansion (X)
and the derived dissection (XII) for the chiral centre at C-6. The hydrogen

(IX)
(X)

~
H--7--(C)ooo

b
v TH
3--2--!--@--5--4--3
3
I JH v
3
a
HI I 3
CH I I
(C)ooo I
(C)ooo
H
d
(XI) (XII)
316 B. C. L. WEEDON

atom clearly has lowest priority. Since C-7 is of the type C(CCH) it has lower
priority than either C-1 or C-5 both of which are of the type C(CCC). To
distinguish between the priorities of the ligands commencing with C-1 and C-5,
it is necessary to consider the next atoms along the prior branches, i.e. C-2
and C-4. These are of the types C(CHH) and C(CCH) respectively. The latter
has priority, and the order of precedence of the four ligands at C-6 is therefore
that indicated in (XI). From this the configuration is seen to be R. (Note
that the earlier convention [181, 182] for multiple bonds would lead to the
alternative conclusion.)

Ex. 3. Capsorubin
The two end groups of natural capsorubin (205) have the absolute confi-
guration (XIII). The appropriate partial expansion is (XIV) and leads to the
dissections (XV) and (XVI) for C-3 and C-5 respectively. From the former the

v
o I .·

OH
(XIII) (XIV)

a
f TH' v ?H v TH 3 (?)ooo
c 6--1--l--r--~--f--1--y---Doo b
CH 3 CH3 H H H I 7
d
(XV)

a
?00
(O)ooo-6--7
b
TH'
2--1--G)--4--3
I v c
&, I ~
CH,
d
(XVI)

hydroxyl clearly has the highest priority and the hydrogen the lowest. To
compare the ligand commencing at C-2 with that commencing at C-4 it is
necessary to proceed along the prior branches as far as C-5 and C-6 respectively.
The latter has priority since C(OOC) > C(CCC). The four ligands at C-3 can
therefore be ordered as shown in (XVII), which corresponds to S. The second
 V. Stereochemistry 317

dissection for C-5 indicates the labelling shown in (XVIII), and hence the R
configuration. The stereochemistry of the capsorubin end group is therefore
3S,5R.

v· ,. o~......

OH
(XVIII)

Ex. 4. Violaxanthin
The end groups of natural violaxanthin (135) have been provisionally
assigned the configuration (XIX). Application ofthe sequence rule in turn to the
chiral centres at C-3, C-5 and C-6 indicates that the ligands have the priorities
summarized in (XX), (XXI) and (XXII) respectively. The proposed configura-
tion (XIX) is therefore fully specified as 3S,5R,6S.

(XIX)

...... d
.... ··
. a ~

HO d 5R HO 6S
(XXI) (XXII)

Ex. 5. The allenic end group

The absolute configuration of the allenic end group in fucoxanthin (190),
neoxanthin (122) and related natural compounds is represented by (XXIII).
Considering first the two chiral centres at C-3 and C-5, application of the
sequence rule orders the ligands as shown in (XXIV) and (XXV) respectively.
The chiral centre at C-3 is therefore Sand that at C-5 is R.
318 B. C. L. WEEDON

The chiral axis of the allene group can be represented by the model (XXVI),
in which the numbers refer the carbon atoms in the carotenoid skeleton. When
viewed from the X end (cf. Newman projection XXVII) of the axis running
c

d
(XXVI) (XXVII)

through C-6, C-7 and C-8, the four ligands have the priorities indicated by
a, b, c and d. On inspection of the model from the side remote from d, a circle
traced from a to b to c follows a clockwise course, and the allene group therefore
has the R configuration. Thus the complete stereochemistry of the allenic end
group is specified as 3S,5R,6R.

Ex. 6. Abscisic acid

(+ )-Abscisic acid has been assigned the absolute configuration (XXVIII).
Consider the partial expansion (XXIX) and the dissection (XXXI). The
hydroxyl group obviously has first priority. Both C-5 and C-1 are of the type
C(CCC) and therefore have priority over C-7 which is of type C(CCH).

0
~~
AA'"O C:OlH

(XXVIII)

r
a

c
H fH·
3--!--1-@--~-4--3
fH· y b
&. I
~
H--r-- (C)ooo

(C)ooo
(t)ooo

d
(XXXI)
 V. Stereochemistry 319

Consider now the prior branches from C-5 and C-1, viz. C-4 and C-2. The
former, C-4, is of the type C(CCH) whereas C-2 is of the type C(CHH). The
C-5,C-4ligand therefore has priority over the C-1,C-2ligand. Hence the order
of the four ligands is that shown in (XXX) which corresponds to R. The original
specification [156] of structure (XXVIII) as S was on the basis of the 1956
convention [182], and illustrates an important difference between this and the
later 1966 convention [183].

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[179] W. v. E. Doering and R. W. Young, J. Amer. Chem. Soc. 74, 2997 (1952).
[180] G. Snatzke and K. Schaffner, Helv. Chim. Acta 51, 986 (1968).
[181] R.S. Cahn and C.K. Ingold, J. Chem. Soc./951, 612.
[182] R.S. Cahn, C.K. Ingold and V. Prelog, Experientia 12,81 (1956).
[183] R.S. Cahn, Sir Christopher Ingold and V. Prelog, Angew. Chem., Int. Ed. Engl. 5, 385
(1966).
[184] R.S. Cahn, J. Chem. Educ. 41, 116 (1964).
 325

VI. Total Syntheses

H. MAYER and 0. ISLER
Chemical Research Department, F. Hoffmann-La Roche & Co. Ltd., Basle, Switzerland

A. Introduction . 328
1. General considerations 328
2. Scope and limitation 328
B. Syntheses of Cyclic Components 331
1. C 9-Components 331
2. C 10 -Components 333
3. C 11 -Components 337
4. C 12 -Components 337
5. C 13 -Components 338
6. C 14-Components 344
7. C 15 -Components 347
a) C 9 +C 6 =C 15 347
b) C 10 +C 5 =C 15 351
c) Cll +C 4 =C 15 351
d) C 13 +C 2 =C 15 352
e) C 14 +C,=C 15 361
8. C 16 -Components 362
a) Cll +C 5 =C 16 362
b) C 13 +C 3 =C 11, 362
c) C 14 +C 2 =C 16 364
d) C 15 +C 1=C 16 366
9. C 17 -Components 366
a) C 13 +C 4 =C 17 366
b) C 15 +C 2 =C 17 366
10. C 18 -Components 366
a) C 9 +C 9 =C 18 368
b) C 14 +C 4 =C 18 368
c) C 15 +C 3=C 18 369
d) C 17 +C 1=C 18 369
11. C 19 -Components 370
a) C 19 -Aldehydes . 370
b) Other 3-oxygenated C 19 -components 372
12. C 20 -Components 373
a) C 10 +C 10 =C2o· 373
b) Cll +C 9 =C2o . 374
c) C 13 +C 7=C 20 . 375
d) C 14 +C 6 =C 20 . 379
e) C 15 +C 5 =C 20 . 379
f) C 16 +C 4 =C 20 . 385
g) C1s+C2=C2o · 386
h) C 19 +C 1=C 20 . 388
i) Retinyltriphenylphosphonium halides . 388
j) ,8-Retinylphosphonate. 389
k) 4-0xygenated vitamin A compounds 390
13. C 21 -Components 390
14. C 26 -Components 392
326 H. MAYER and 0. ISLER

C. Syntheses of Acyclic Components . 392

1. Cn C 3 - and C 4 -Components . 392
2. C 5 -Components 394
3. C 6 -Components 403
4. C 7 -Components 403
5. C8 -Components 408
6. C 9 -Components 408
7. C 10-Components 408
8. C 11 -Components 420
9. C 12 -Components 420
10. C 15 -Components 421
11. c,. -Components 424
12. C 17 -Components 425
13. C 20 -Components 426
D. Syntheses of Symmetrical Central Components . 428
1. C 4 -Components . 428
2. C8 -Components . 428
a) C 6 +C 2 =C 8 • . 428
b) C4 +C 4 =C 8 . . 430
c) C 3 +C 2 +C 3 =C 8 . . 430
d) C 1 +C 6 +C 1 =C 8 • . 430
3. C 10-Components . 431
a) C6 +C 4 =C 10 . • . 431
b) C 5 +C 5 =C 10 . • . 431
c) C 4 +C 2 +C 4 =C 10 . 433
d) C 3 +C 4 +C 3 =C 10 . 435
4. C12-Components . . . 436
a) C 6 +C 6 =C 12 • . . 436
b) C 5 +C 2 +C 5 =C 12 . 437
c) C 2 +C 8 +C 2 =C 12 . 438
d) C 1 +C 10 +C 1 =C 12 • . 438
5. C 14-Components . . . . 438
6. C18 -Components . . . . 439
a) C 4 +C 10 +C 4 =C 18 • . 439
b) C 3 +C 12 +C 3 =C 18 • . 440
E. Syntheses of Apo-carotenoids 440
1. tl-Apo-carotenals . . 440
a) tl-Apo-14'-carotenal . . 440
b) P-Apo-12'-carotenal . . 440
c) 3-Hydroxy-7,8-didehydro-Jl-apo-12' -carotenal 444
d) P-Apo-10'-carotenal . 444
e) P-Apo-8'-carotenal 445
I) P-Apo-6' -carotenal 448
g) P-Apo-4'-carotenal 448
h) P-Apo-2'-carotenal 450
2. IX-Apo-12'-carotenal . 450
3. Apo-carotenals with aromatic end groups 451
4. Apo-lycopenals . . . 453
a) Apo-15-lycopenals. 453
b) Apo-12'-lycopenals 454
c) Apo-8'-lycopenals . 455
d) Apo-4'-lycopenals . 455
5. P-Apo-carotenoic acid esters . 455
6. tl-Apo-carotenones . . . . 455
7. Apo-lycopenoic acid esters . 464
 VI. Total Syntheses 327

F. Syntheses of Diapo-carotenoids. 464

1. Diapo-carotenedials . . . . 466
2. Diapo-carotenediones . . . 466
3. Diapo-carotenedioic acid esters 469
4. Retro diapo-carotenoids . . 469
G. Syntheses of C 40 -Carotenoids . 474
1. Symmetrical building schemes 474
a) C2o+C2o=C4o . . . 474
b) Ct9+C2+C19=C4o · 480
c) C 18 +C 4 +C18 =C4o 482
d) C 16 +C 8 +C 16 =C 40 . 483
e) C 15 +C 10 +C 15 =C 40 . 489
f) Ct4 +C12+C14=C4o· 492
g) Ct3+Ct4+Ct3=C4o· 496
h) C10 + c2o + c.o = C4o 496
i) Cs+C3o+Cs=C4o · 501
2. Unsymmetrical building schemes . 504
a) C21 +C19=C4o 504
b) C2s+C1s=C4o 504
c) C3o+C10=C4o 504
d) C 35 +C 5=C 4o 516
e) C37+C3=C4o 517
H. Syntheses of Carotenoids with Less than or More than 40 Carbon Atoms . 517
1. C 20 -Carotenoids . . . . . 517
2. C 30 -Carbtenoids . . . . . 522
3. C 34- and C 37-Carotenoids. 522
4. C 50 -Carotenoids . . . . 522
a) C21+Ca+C2t=Cso· 522
b) C 20 +C 10 +C 20 =C 50 523
c) C 19 +C 12 +C 19 =C 50 526
5. C 60 -Carotenoids . . . . 526
a) C26+Ca+C26=C6o· 526
b) C2o+C2o+C2o=C6o 526
I. Syntheses of Related Compounds . 526
1. Analogues with altered carbon skeleton . 526
2. Abscisic acid . . . . . . . . 527
3. Grasshopper ketone . . . . . . . 535
J. Syntheses of Labelled Compounds . . 535
K. Annex. Synthetic Carotenoids (Tables) . 536
Table 46. Synthetic apo-carotenoids. . 536
Table 47. Synthetic diapo~carotenoids . 542
Table 48. Synthetic C 40 -carotenoids. . 545
Table 49. Synthetic carotenoids with more than 40 carbon atoms 564
References . . . . . . . . . . . . . . . . . . . . . . . . . 565
328 H. MAYER and 0. ISLER

A. Introduction

1. General considerations
Ever since the elucidation of the structure of fJ-carotene, lycopene, zea-
xanthin and vitamin A by the school of Karrer in 1930-1932 much effort has
been devoted to the total synthesis of carotenoids and related polyenes. In
1947 the first total synthesis of crystalline vitamin A was announced by Isler
et al. [1], and soon afterwards an industrial manufacturing procedure was
developed. The first total syntheses of fJ-carotene were reported by Karrer
and Eugster [2], Inhoffen et al. [3-5] and Milas et al. [6] in 1950. In the
succeeding decades the range of synthetic methods in the carotenoid field
has been greatly extended by teams led by Karrer, Inhoffen, Weedon, Isler,
Pommer, Surmatis and others, and large numbers of natural and unnatural
carotenoids as well as related and labelled compounds have been totally
synthesized. Based on these scientific achievements numerous syntheses have
been adapted to technical requirements so that today vitamin A, fJ-carotene,
canthaxanthin, /J-apo-8' -carotenal and ethyl /J-apo-8' -carotenoate are commer-
cially produced by total synthesis on a large scale [7-9].
Unambiguous total synthesis is being employed increasingly as an integral
part of structural and stereochemical studies so as to elucidate some molecular
feature for which other evidence is inconclusive or ambiguous. Moreover,
by making comparisons with synthetic materials, the unequivocal identification
of trace carotenoids has become possible. Total synthesis has also been
used successfully in many cases not only for the preparation of the familiar
all-trans isomer but also of specific cis isomers. With vitamin A, vitamin A2
and related compounds, the sterically 'unhindered' 9-cis, 13-cis and 9,13-di-
cis isomers as well as the hindered 11-cis and 11,13-di-cis isomers have been
prepared, thus establishing vitamin A and A2 (retinol and 3-dehydroretinol)
stereochemistry on a firm basis [10]. With carotenoids, partial hydrogenation
of synthetic carotenoid precursors possessing a central acetylenic group
afforded the labile 15-cis isomers of e.g. fJ-carotene, zeaxanthin, cryptoxanthin
and canthaxanthin [11]. In the last few years the stereochemically controlled
synthesis of carotenoids with methylated cis double bonds, e.g. of 9-cis-
dimethylcrocetin, 9,9' -di-cis-alloxanthin and 9-cis-crocoxanthin has received
considerable attention [12]. Successful synthesis also made many carotenoids
much more readily available for further investigations (including their use as
food colourants [13]) than would have been possible by the classical method
of solvent extraction of plant material.

2. Scope and limitation

Numerous reviews dealing with the tremendous amount of work devoted
to the total synthesis of carotenoids [7-29] and vitamin A [7-10, 29-37]
were published during the past two decades. The methods of polyene synthesis
have also been thoroughly reviewe'd, considering the relevant literature up to
 VI. Total Syntheses 329

the middle of 1961 [24, 38]. Strenuous efforts to develop economic processes for
the industrial production of vitamin A, P-carotene and other carotenoids have
rapidly increased the patent literature, which now covers a very large field.
It is the purpose of this chapter to give a summary up to the middle of 1970
of the main published methods for the synthesis of carotenoids and of the
necessary intermediates. To achieve this, general building schemes have been
developed which allow a systematic classification of the building units and of
the reaction types employed. Only well-established reaction sequences are
discussed in detail. Papers and patent claims without sufficient experimental
details are only briefly mentioned or even omitted. Partial syntheses, e.g. the
preparation of canthaxanthin from P-carotene, will not be dealt with in this
chapter (for a review see Chapter III).
Relatively few main reaction types have been employed for the synthesi~
of the carbon skeleton of carotenoids and intermediates. These are:
a) Wittig and Horner Condensations of carbonyl compounds with tri-
phenylphosphonium halides and dialkyl phosphonates, respectively, in the
presence of phenyllithium, n-butyllithium, alkali metal alkoxides, sodium
hydride, sodium amide, etc. (for an excellent review see [39]).
b) Grignard and Nef Reactions of carbonyl compounds with metal
acetylides.
c) Enol Ether Condensations of acetals with enol ethers in the presence
of zinc chloride or boron trifluoride.
d) Aldol Condensations, i.e. base-catalysed condensations of two carbonyl
compounds.
e) Reformatskii Reactions involving the reaction of oc- and y-halo-esters
and -nitriles with aldehydes or ketones in the presence of zinc, magnesium,
aluminium, etc.
f) Knoevenagel-Doebner Condensations of aldehydes with compounds
possessing activated hydrogens, e.g. malonic acid, in the presence of pyridine
and piperidine.
g) Robinson's Mannich Base Synthesis, i.e. the base-catalysed condensa-
tion of a polyene ketone and the methiodide of a Mannich base.
h) Reductive and Oxidative Dimerization Reactions.
i) Wurtz Reactions, i.e. the formation of hydrocarbons from alkyl halides
and alkali metals.
The present chapter has been subdivided into Sections B to J. In Section B
syntheses of cyclic components containing nine to twenty-six carbon atoms
are discussed. Starting with the Cwcomponents, the headings of the sub-
sections are given as follows: ex+ Cy = cz. The indices X and y indicate the
number of catbon atoms of the compounds used to synthesize the cyclic
component containing z carbon atoms, whereby z = x + y. Thus, the synthesis
of the P-Cwaldehyde (123) will be found in the subsection CwComponents,
and the syntheses of the P-C 1 s-aldehyde (180) are listed in the subsection
C1 s-Components under the headings C10 + C 5 = C 15 , Cu + C4 = C 1 s and
C13 + C 2 = C15 • Since vitamin A compounds are needed as important inter-
330 H. MAYER and 0. ISLER

mediates for cyclic e 20 -building units, their synthesis is described in the

subsection e 20 -eomponents.
Section e deals with the syntheses of the numerous acyclic components
with two to twenty carbon atoms which are mainly used for chain-lengthening
purposes and for the introduction of acyclic end groups into the carotenoid
skeleton.
In Section D syntheses of symmetrical central components, containing
four to eighteen carbon atoms, are described. These methods consist of com-
bining two molecules with different or identical carbon skeletons, or three
molecules, two of which have identical carbon skeletons, according to the
schemes ex+ ey = ez, ex+ ex= ez and ex+ ey +ex= ez, with increasing
size of the middle eY-component. In the case where three molecules are com-
bined in one step, the central component, eY, is always symmetrical corre-
sponding to the central part of the e 40 -carotenoid carbon skeleton.
Section E lists the methods leading to the various types of apo-carotenoids
including P- and oc-apo-carotenals and P-apo-carotenoic and apo-lycopenoic
acid esters. P- and oc-Apo-carotenals, apo-lycopenals, and apo-carotenals with
aromatic end groups are also employed as important building units in the
syntheses of e 40 -carotenoids. The headings of the subsections are given again
by the abbreviated building schemes explained above.
In Section F syntheses of diapo- and retro-diapo-carotenoids are discussed
comprising the vinylogous series of diapo-carotenedials, diapo-carotenedioic
acid esters and diapo-carotenediones. erocetindialdehyde and its central-
acetylenic analogue have been used extensively as important e 20 -building
units in numerous syntheses of symmetrical and unsymmetrical carotenoids.
The reaction schemes followed are very similar to those of Section D.
Section G deals systematically with the methods leading to the full e 40 -
carotenoid skeleton by combining either three or two components according
to the schemes ex+ey+ex=e40 and ex+ey=e40 , respectively. The head-
ings of the subsections are again represented as symmetrical and unsymmetrical
building schemes and progress with increasing size of the indices of the central
ey-component of the symmetrical scheme and of the ex-component of the
unsymmetrical scheme. Syntheses of different carotenoids following a given
building scheme are often recorded in the tables. To give an example, P-caro-
tene is commercially produced according to the scheme e 19 + e 2 + e 19 = e 40 ,
and its synthesis is discussed under this heading. Other carotenoids synthesized
in similar fashion are listed in Tables 24 and 25.
In Section H syntheses of 'carotenoids' with less than and more than
forty carbon atoms are discussed including the interesting e 50 - and e 60 -P-
carotene analogues decapreno-p-carotene and dodecapreno-P-carotene, re-
spectively. The building schemes used correspond to those of Section G.
In Section I compounds are collected which are in some way related to
the carotenoids; the list includes carotenoid analogues with altered carbon
skeletons, abscisic acid and 'grasshopper ketone'. The necessary building
units and the synthesized compounds are conveniently arranged in tables.
 VI. Total Syntheses 331
Section J lists the labelled compounds which have been prepared in the
laboratories of F. Hoffmann-La Roche & Co. Ltd., Basle.
For technical reasons it will not be possible to draw all the structural
formulae of the carotenoids discussed at the appropriate places in the text
and in the tables. To facilitate reference, the synthesized apo-, diapo-, C 40 -,
C 50 - and C 60 -carotenoids have therefore been assembled in the Tables 46-49
along with their structural formulae, appropriate building schemes, reaction
types employed and pertinent references. By the aid of the arabic number
assigned to each carotenoid the pertinent formula can be found readily.
Some typical transformations involving the carbon skeletons of carotenoids,
vitamin A and intermediates, such as dehydration, allylic rearrangement,
reduction, oxidation, etc., which have often been used as standard procedures,
are not dealt with in a separate section. They are, however, mentioned at the
appropriate places in the preceding sections.

B. Syntheses of Cyclic Components

1. C 9 -Components
The cyclic C 9 -building units 4,4-ethylenedioxy-2,6,6-trimethyl-2-cyclo-
hexen-1-one (5) and 4,4-ethylenedioxy-2,2,6-trimethylcyclohexanone (7) were
synthesized starting from commercially available isophorone (1). Reaction
with methylmagnesium bromide in the presence of 1 mole % of ferric chloride
yielded the p,y-unsaturated ketone p-phorone (2) [40], which on oxidation

)5, --- j i --- 0 ~CC ~ )5:0

-------
(J) (2) (3) (4)

cPO - (7)
}):0 cPO
~

(6)
~

(5)

&0 --- ((
(8) (9)
~

ex: ---
(JO)
(t
(11)
332 H. MAYER and 0. ISLER

with peracetic [41] or m-chloroperbenzoic acid [ 42] followed by treatment

with sodium hydroxide afforded the hydroxy ketone (3). Oxidation of the
latter with chromic acid in acetic acid [41] or with Jones' reagent [42] gave
the unsaturated diketone (4), which was transformed into the monoketal (5)
by treatment with ethylene glycol and p-toluenesulphonic acid [42-44].
Reduction of (4) with zinc in acetic acid [41] or rearrangement of (3) with
p-toluenesulphonic acid in benzene [42] led to the saturated diketone (6),
which was ketalized to the monoketal (7) in an analogous manner [41, 42].
2,6,6-Trimethyl-2-cyclohexen-1-one (11) was made from commercially
available 2-methylcyclohexanone (8). Methylation with methyl iodide-sodium
amide or methyl sulphate in anhydrous ether [45-47] yielded 2,2,6-trimethyl-
cyclohexanone (9), which was purified via the crystalline semicarbazone or
by fractionation [ 48]. Bromination with bromine in acetic acid followed by
dehydrobromination of the resulting bromo ketone (10) then gave the desired
product (11) [49, 50].
2-Chloro-1,3,3-trimethylcyclohexene (15) was prepared by a Wittig reaction
of 6-methyl-5-hepten-2-one (12) and (chloromethylene)triphenylphosphorane
(13) yielding a cis-trans mixture of chloro olefins (14) which were cyclized in
glacial acetic acid-cone. sulphuric acid at -20 °C. The 1: 1 mixture of chloro-
cyclohexenes (15 + 16) obtained could be readily separated by column chroma-
tography [51].

Ci (CoH5 hP=OICI
(13)
(Jca 0(+ ---+
cc
»0
(12) (14) (15) (16)

..J:i (17)
---+

(18)
---+
A):( (19)
---+
uo'cc (20)

The C 9 -dihydroxy ketone (20) was required for a synthesis of the 'grass-
hopper ketone' [52] (Chapter V). Lithium aluminium hydride reduction of
P-phorone (2) and acetylation of the product gave the acetate (17). Subsequent
epoxidation with m-chloroperbenzoic acid led to the epoxide (18), which was
transformed into the diol acetate (19) by treatment with aqueous perchloric
acid. Sarett oxidation of (19) followed by treatment of the product with sodium
carbonate in aqueous methanol then furnished the desired C 9 -component (20)
[52].
 VI. Total Syntheses 333

2. C 10-Components
P-Cyclocitral (24) has been prepared by the cyclization of citral (21). The
Schiff base of citral with aniline (22) was treated with concentrated sulphuric
acid giving a mixture of IX- (23) and P-cyclocitral (24) [53]. Pure P-cyclocitral
was obtained by treatment of the reaction mixture with potassium hydroxide in
ethanol [53, 54]. Alternatively, P-cyclogeranyltriphenylphosphonium bromide
(27) was treated with methanolic sodium methoxide in dimethylformamide
followed by reaction of the intermediate phosphorane with nitrosobenzene [55].

-- u~~.
--
l
(21) (22) (23) (24)

... - affi,lk-arn,ou
L________J

arn.~(<;H,),

(27) (26) (25)

P-Cyclogeraniol (25) was prepared by Ponndorf [56] or lithium aluminium

hydride reduction of P-cyclocitral. The direct cyclization of either geranyl
acetate or geraniol yielded IX-cyclogeraniol [57].
P-Cyclogeranyltriphenylphosphonium bromide (27) was prepared by
reaction of P-cyclogeraniol (25) with triphenylphosphonium bromide or
triphenylphosphine and hydrobromic acid in dimethylformamide, tetra-
hydrofuran or benzene [58, 59]. Alternatively, P-cyclogeraniol (25) was first
converted by reaction with phosphorus tribromide-pyridine into P-cyclo-
geranyl bromide (26), which then was quaternized with triphenylphosphine in
benzene [ 60, 61].
Since 3-oxygenated cyclic components had not been readily available
hitherto for the synthesis of higher polyenes, an improved synthesis of the
important intermediate keto-ester (30) has recently been studied by Surmatis
et al. [62]. The zinc chloride-catalysed condensation of mesityl oxide (28) with
ethyl acetoacetate (29) afforded a 4: 1 mixture of the two isomeric keto esters
(30) [63] and (31). The ester (31) could be selectively hydrolysed and decar-
boxylated to isophorone {1) which was removed by fractional distillation. (30)
was required for· the synthesis of additional 3-substituted cyclic building units,
such as 3,3-ethylenedioxy-fJ-ionone (112) and 3-ethoxy-3,4-didehydro-P-ionone
(115) [ 62] (Section B.5 f).
334 H. MAYER and 0. ISLER

(28)
+

(29)
-- 0~ Nco,c,H, +

(30)
c,H,o,c~

0~
(31)

I j

(34)
- HOAA
NCH,OH +

(33)
HOAA
NCH,OH

(32) (I)

1 1
ED
NCH,OH-- ~CH2 P(Cc;H,h Br 9

u(35)
~
(36)

Reduction of(30) with lithium aluminium hydride afforded the two isomeric
cyclohexene diols (32) and (33). Treatment of the latter with triphenylphos-
phonium bromide resulted in a dehydration yielding the C 10 - Wittig compound
(36) which was used for a synthesis of 3,4,3',4'-tetradehydro-P-carotene (1).
When (33) was stirred in methylene chloride containing a trace of hydrogen
chloride, the bicyclic ether (34) was formed which rearranged to the dienol (35).
Treatment of the latter with triphenylphosphonium bromide led to the same
phosphonium bromide (36). An Oppenauer oxidation of (33) led to the keto
alcohol (37), which was converted into the phosphonium salt (38) as mentioned.
The latter did not, however, form the corresponding phosphorane on treatment
with sodium methoxide but was cleaved instead to the dienone (39) and
triphenylphosphine.
 VI. Total Syntheses 335

[SJ
uc~
(40)
-- aro~,

(41)
I -- e:ro~,

(42)
--
0
;:x~·
OH

(43)

j
(Crn,OH --
(44)
I uo --- ceo
(45)
I
(46)

The synthesis of the C 10 -ester (43), representing the cyclic portion of

abscisic acid (855 b) has been reported [64, 65]. Air oxidation of P-cyclocitral
(24) gave P-cyclogeranic acid (40), which on bromination with bromine in the
presence of ultraviolet light, or with N-bromosuccinimide, followed by de-
hydrobromination with diethylaniline and esterification with diazomethane
furnished safranic acid methyl ester (41). Subsequent photosensitized oxidation
led to the epidioxide (42), which on basic aluminium oxide was rearranged to (43).
Lithium aluminium hydride reduction of(41) gave the allylic alcohol (44), which

[SJ
'(xrno --I
u~OH
I -- '(x • I
012 P(C6 H 5h Bre

(47) (48) (49)

XrCHO -- XYPH -- Xr •
I I I
012 P(C.H 5 h Br
e

(50) (51) (52)

RO
XxCHO Xr,OH -- Xx • -- THPO ~
I THPO
01 2 P(C.U 5h Br
e

(52 a) R=H (52 c) (52 d)

(52b)R=THP
336 H. MAYER and 0. ISLER

was oxidized with manganese dioxide to safranal (45). The latter compound was
also made from oc-cyclocitral (46) by successive bromination and dehydro-
bromination.
The C10-triphenylphosphonium bromides (49) and (52) were required for
the synthesis of isorenieratene (13) and renierapurpurin (16) [66]. Reduction
of 2,3,6- (47) and 2,3,4-trimethylbenzaldehyde (50) with lithium aluminium
hydride gave the alcohols (48) and (51), which were converted into the Wittig
reagents by bromination with phosphorus tribromide-pyridine followed by
reaction of the intermediate bromides with triphenylphosphine [66].
The C10-Wittig compound (52d) was used for the synthesis of 3,3'-iso-
renieratenediol (79) and 3-isorenieratenol (52) [67, 68]. 4-Hydroxy-2,3,6-
trimethylbenzaldehyde (52 a) was first converted into the tetrahydropyranyl
ether (52 b). Reduction with lithium aluminium hydride gave the benzyl
alcohol (52 c), from which the phosphonium bromide (52 d) was obtained by the
usual procedure [67, 68].

[SJ

o
Dm~~. ~ o~OH ~
o o::::>' #
Dm"' ~ .(fro~
o::::>' o:::?"
0

(53) (54) (55) (56)

J!~ -- y~ -- if· lm tt"

(56) (57) (58) (59)

J J

tf --
(62)
-tr -tf (63)
6u
(60)
v (61)

~·" __ ~
HON H~
(65) (66)
 VI. Total Syntheses 337

In the course of stereochemical and synthetic studies of capsanthin (170)

and capsorubin (205) Weedon's team synthesized a number of cyclopentane
derivatives which were used as valuable C10 -building units.
Autoxidation of the keto acid (55) obtained from the diketo ester (53) via
the keto alcohol (54) gave the diosphenol (56), which was converted into the
keto acid (57). Borohydride reduction yielded the cis- (58) and the trans-hydroxy
acids (59), which, on treatment with methyllithium, gave the cis- and trans-
hydroxy ketones (60) and (61), respectively [69].
Treatment of 1,2,2-trimethylcyclopentanecarboxylic (camphonanic) acid
(62) with methyllithium gave the methyl ketone (63) [70]. Reduction of cam-
phononic acid (64) with potassium borohydride in aqueous sodium hydroxide
gave the hydroxy acid (65), which was converted into the methyl ketone (66) in
an analogous way [71].

3. C 11 -Components
1-Ethynyl-2,2,6-trimethylcyclohexanol (67) and 2-ethynyl-1,3,3-trimethyl-
cyclohexene (69) were synthesized from 2,2,6-trimethylcyclohexanone (9).
Ethynylation was effected with sodium acetylide in liquid ammonia [47, 72]
yielding the ethynyl carbinol (67), which was dehydrated to (69) by treatment
with thionyl chloride in pyridine [47] or by distillation through a heated glass
tube containing a supported aluminium phosphate catalyst [ 47, 48]. Better
yields were obtained by pyrolysis of the corresponding acetate (68) [48] in
silicone oil in the presence of zinc oxide.

-+
?><K_~
UOH -+ uOAc
?><K_~
-+

(9) (67) (68) (69)

4. C 12 -Components
The C12 -keto ester (73) was prepared in analogy to the vinylogous C10 -ester
(43) by photosensitized oxidation of the triene ester (72) [65]. The latter was
obtained from rx-ionone (70) by halooxidation and subsequent esterification
giving the ester (71), which was converted into (72) by bromination with N-
bromosuccinimide followed by dehydrobromination of the product [65].
The synthe&is of the C12 -components (75) and (77) required for the prepa-
ration of some carotenoids with aromatic end groups was described in the
patent literature. Base-catalysed condensation of cinnamaldehyde with acetone
gave the aromatic ketone (74), which, on lithium aluminium hydride reduction
and treatment of the product with triphenylphosphine and methanolic hydro-
chloric acid, was converted into the triphenylphosphonium chloride (75) [73].

Carotenoids 22
338 H. MAYER and 0. ISLER

[g

~0
(70)
- ~co,cu,

(71)

1
0
~CO,CH,

(73)
H
-- ~co,cu,

(72)
I

~- (74)
~~(C.U,),
(75)
Cl"

CH 30
~co~
(76)
- CH 3 0
~CHO
(77)

A Reformatskii reaction of anisaldehyde and methyl y-bromo-{J-methyl-

crotonate (170) afforded the C 12 -acid (76) [74]. Lithium aluminium hydride
reduction of the corresponding ester followed by manganese dioxide oxidation
of the product then gave the aldehyde (77) [75].

5. C 13 -Components

a) {J-Ionone
Today all industrial syntheses of vitamin A, {J-carotene, canthaxanthin
and apo-carotenoids are based on {J-ionone (89). Four manufacturing proce-
dures have been developed for the total synthesis of this particularly impor-
tant key intermediate. Ethynylation of acetone with sodium acetylide in
liquid ammonia gave methylbutynol (78) [76-78], which was partially hydro-
genated over Lindlar catalyst yielding methylbutenol (79) [77, 78]. This was
reacted with diketene to form the corresponding acetoacetate (80), which on
pyrolysis was rearranged to methylheptenone (12) [77, 78]. Alternatively,
 VI. Total Syntheses 339

[SJ
-- (78)
--~ OH
-- cu.cocu2c~
(79) ~0)"'..

_1-"'- ~~
'c~u~o6'~
(82)
- ~OH ~
(81)
-~0
(12)

! !
(85) (84) (83)

!
~CHO ~CH(OAc) 2

(89) (86)
(88) -------

+
r
l.
CH 3 00__ /'..
-- '-./ '-./ 6.? ~ ~..:::,CHOAc
(91) (90) (87)

heating of methylbutenol (79) with ethyl acetoacetate at 170-190°C [79]

(Caroll reaction [80, 81]) or acid-catalysed reaction ofmethylbutenol (79) with
isopropenyl m~thyl ether [82] gave 6-methyl-5-hepten-2-one {12) directly.
Subsequent condensation with sodium acetylide in liquid ammonia led to
dehydrolinalool (81) [77, 83], which was transformed into pseudoionone (85)
by three different procedures: 1) treatment with diketene to give dehydrolinalyl
acetoacetate (82), which was pyrolysed to pseudoionone (85) [77, 78]; 2) acid-
catalysed reaction of dehydrolinalool (81) with isopropenyl methyl ether to
340 H. MAYER and 0. ISLER

give the allenic ketone (84), which was smoothly isomerized to (85) on treatment
with traces of alkali [84, 85]; 3) rearrangement of dehydrolinalyl acetate (83)
in the presence of silver catalysts to a mixture of citra} diacetate (86) and the
allenic acetate (87) which on hydrolysis gave citra} (88). The latter was converted
into pseudoionone (85) by base-catalysed condensation with acetone [86-88].
More conveniently, alkaline hydrolysis of the mixture of (86) and (87) in the
presence of acetone furnished pseudoionone directly [89]. Under controlled
acidic conditions pseudoionone was cyclized to fJ-ionone (89) [88, 90-92].
Citral (88) could also be obtained from 6-methyl-5-hepten-2-one (12) by
condensation with ethoxyacetylene followed by partial hydrogenation and
hydrolysis [93, 94].
In another procedure (81) was first converted into the methyl ether (90) by
acid-catalysed addition of methanol. This was reacted with ethyl acetoacetate
at 170-180 °C in the presence of aluminium isopropoxide yielding the pseudo-
ion one derivative (91) [95], which was then cyclized by treatment with sul-
phuric-acetic acid at 0 °C [96].
Lithium aluminium hydride reduction of /J-ionone yielded fJ-ionol (92),
which on treatment with p-toluenesulphonic acid was dehydrated to the triene
(96). Both compounds were required for the preparation of /J-ionyltriphenyl-
phosphonium halides and /J-ionylphosphonates.

b) fJ-Ionyltriphenylphosphonium halides and fJ-ionylphosphonates

P-Ionyltriphenylphosphonium bromide (94) was first prepared by reaction
of P-ionol (92) with phosphorus tribromide followed by treatment of the inter-
mediate bromide (93) with triphenylphosphine [61]. More conveniently, the
phosphonium bromide (94) and the corresponding chloride (95) could be made
from fJ-ionol by treatment with triphenylphosphine and hydrobromic acid in
benzene [58] or by treatment with triphenylphosphonium chloride in aceto-
nitrile [97]. It was shown that fJ-ionol could be substituted by the triene (96),
which, therefore, might be an intermediate in the above reaction sequence [98].

(92) (93) (94) X=Br

~
(95)X=Cl

(96) (97)
 VI. Total Syntheses 341

The synthesis of the C 13 -phosphonate (97) from the bromide (93) was found
not to be as clear-cut as that of the corresponding phosphonium halides in
that a ready formation of the triene (96) was observed on heating the bromide
with triethyl phosphite. The synthesis could, however, be achieved by heating
,8-ionol with a minimum excess of one mole of triethyl phosphite in the presence
of strong acids, e. g. hydrobromic acid [99, 100].

c) 3,4-Didehydro-,B-ionone
3,4-Didehydro-,B-ionone (99) has been prepared from ,8-ionone (89) by
bromination with N-bromosuccinimide in the presence of calcium oxide and
sodium carbonate in carbon tetrachloride and subsequent dehydrobromination
of the intermediate bromide (98) with diethyl- or dimethylaniline [101, 102].
,8-Ionone enol acetate (100) [103] and oc-ionone (101) [65, 104] have also been
used as starting materials. The bromination was effected by bromine in carbon
tetrachloride or chloroform at low temperature [103, 104] or by N-bromo-
succinimide [65].

[SJ

~0
(89)
-- ~ -- ~
Br
(98) (99)
I

~OM ~ ec~""o ··~

a¢
(100) (101) (102)

~OH hAo
(((9)
+
h~OH
?'
(103)
-- (104)
OH
-- ex~
(105)

d) oc-Ionone
(- )-(S)-oc-Ionone (102) and the enantiomer were required for the synthesis
of (- )-(S)- (901) and ( + )-(R)-oc-carotene (5) and of (- )-(6S,6'S)- (904) and ( + )-
(6R,6'R)-s-carotene (903). The enantiomeric oc-ionones were obtained by
resolution of the racemate (101) via the menthylhydrazones [105] and their
absolute configuration determined [106, 107].
342 H. MAYER and 0. ISLER

e) 7,8-Didehydro-P-ionone
The dehydration of carbinols possessing a P-ionol structure or ofvinylogues
thereof often leads to compounds with a retro system of conjugated double
bonds. This double-bond rearrangement could be prevented by the introduction
of an acetylene linkage at the 7,8- or 11,12-position. 7,8-Didehydro-P-ionone
(1 05) was synthesized by the condensation of 2,2,6-trimethylcyclohexanone (9)
with the lithium derivative of 3-butyn-2-ol (103) in liquid ammonia followed
by manganese dioxide oxidation and dehydration ofthe intermediate acetylenic
diol (104) [108].

f) Oxygenated P-ionones
The synthesis of 3-hydroxy-p-ionone (109) has recently been reported by
Weedon's team [109]. A Grignard reaction between the monoketal (7) and
3-butyn-2-ol (103), liberation of the protected keto group in the product, and
reduction with sodium borohydride or lithium aluminium hydride gave the
acetylenic triol (106) [110]. Treatment of the latter with acetic anhydride
yielded the enyne diacetate (107), which, on lithium aluminium hydride reduc-
tion, formed the diene glycol (108). Selective oxidation with manganese dioxide
then furnished 3-hydroxy-P-ionone (109).
3,3-Ethylenedioxy-P-ionone (112) and 3-ethoxy-3,4-didehydro-P-ionone
(115) were synthesized starting from the keto ester (30) (Section B.2) [62, 111].
Treatment with triethyl orthoformate and anhydrous ethanol in the presence
of sulphuric acid gave the enol ether (113). This was reacted with ethylene
glycol-p-toluenesulphonic acid yielding the ketal (110), which, on lithium
aluminium hydride reduction and subsequent manganese dioxide oxidation,
resulted in the aldehyde (111). Base-catalysed condensation of (111) with

~0~· (7) (103) (106)

l
~0
HO)__jl
(109)
- HO)__jl
~OH
- (fOB) (107)
 VI. Total Syntheses 343

[SJ

0
<(:o
~CO,CH,
-- ~CHO
<(:o
-- ~ <(:o
(110) (Ill) (112)

l
C2 H,O
nco~.

(113)
I -- C2 H50
DCHO

(114)
-- ~ C2 H50
(115)

acetone then afforded the substituted P-ionone (112). The substituted ionone
(115) was prepared from (113) following the sequence of reactions described for
(110).
3-Methoxy-P-ionone (116) was readily obtained when 3,4-didehydro-P-
ionone (99) was stirred in a solution of methanol and sulphuric acid at 5 °C for
24 hours [102].
6-Hydroxy-3-oxo-P-ionone (118) used for a synthesis of abscisic acid (855 b)
was obtained by photosensitized oxidation of 3,4-didehydro-P-ionone (99)
followed by base-catalysed rearrangement ofthe resulting epidioxide {117) [65].
The diketone (118) could also be obtained from P-ionone 5,6-epoxide by acid
treatment followed by chromic acid oxidation of the product [112].

[SJ

cu.o ~ (116)
~0 0

(117)
-- ~ 0
(118)
H

~
c 3

(119)
r (120)
-- r
(121)
344 H. MAYER and 0. ISLER

Photosensitized oxidation has also been applied to P- and oc-ionone yielding

4-hydroxy-P-ionone (120) [113]. Utilizing the intermediate 4-bromo-P-ionone
(98), 4-hydroxy- (120) and 4-methoxy-P-ionone (119) were made by treatment
with sodium formate and sodium carbonate, and methanol, respectively [101].
Chromic acid oxidation of (120) gave 4-oxo-P-ionone (121) [101].

6. C 14-Components
a) P-C14-Aldehyde
The P-C 14-aldehyde (123) represents a key intermediate in the technical
synthesis of vitamin A and carotenoids by F.Hoffmann-La Roche & Co. Ltd.
For the commercial preparation of this compound from P-ionone (89) the
Darzens glycidic ester synthesis has been developed into a very efficient
method of chain extension by one carbon atom by Isler et al. [1]. P-Ionone (89)
and ethyl chloroacetate reacted at -10 oc in the presence of sodium ethoxide
yielding the intermediate glycidic ester (122), which, on saponification and
decarboxylation, afforded the P-C 14-aldehyde (123) in 80% yield.
The Darzens reaction was originally applied to P-ionone by Ishikawa and
Matsuura [114], who assigned the isomeric structure (125) to the reaction
product. The same structure was also favoured by Milas et al. [115]. The
reaction was, however, reinvestigated by Heilbron et al. [116], who found that
the Japanese workers had in fact prepared the P-Cwaldehyde (123). Later it

~ ~
((+co~~
-- ~CHO
(89) (122) (123)

1
((+co~ ~ ~CHO
(124) (125)

~HOC~ ~CHO
(126) (127)
 VI. Total Syntheses 345

was shown that either of the aldehydes could be obtained in good yield via the
glycidic acid (124) [117].
Condensation of P-ionyltriphenylphosphonium bromide (94) with ethyl
formate under the conditions of the Wittig reaction gave P-C14-aldehyde enol
ether (126) [98].
The Darzens reaction was also applied to 0(-ionone (70) yielding the O(-C 14-
aldehyde (127) [116, 118].

bJ I so-, retro-dehydro-, dehydro-P- and 3-oxygenated C14-aldehydes

The synthesis of this series of C14-aldehydes as useful cyclic components
and intermediates was performed by Isler's team.
Condensation of 2,2,6-trimethylcyclohexanone (9) with ethoxyacetylene in
the presence of lithium amide in liquid ammonia gave the ethoxyethynyl
carbinol (128). Subsequent partial hydrogenation followed by acid-catalysed
hydrolysis furnished the iso-Cu-aldehyde (129), which was converted into the
iso-C 14-aldehyde (130) by propenyl ether condensation [119]. Alkaline hydro-
lysis of the enol acetate of (129) gave the P-C 11 -aldehyde (131) [119]. Alter-
natively, Wittig reaction of the Cu-triphenylphosphonium bromide (132) with
the keto acetal (133) led to the iso-C 14-aldehyde acetal (134), which was
hydrolysed to (130) [120].
Condensation of 2,6,6-trimethyl-2-cyclohexen-1-one (11) with ethoxyace-
tylene gave the retro-dehydro-C 11 -aldehyde (135), which was also obtained
from 1,5,5-trimethyl-6-methylenecyclohexene (137) by formylation with N-
methylformanilide and POC1 3 [121]. Further transformations of(135) into the
retro-dehydro-Cwaldehyde (136) and into the dehydro-P-C 11 -aldehyde (138)
were effected by propenyl ether condensation and by alkaline hydrolysis of the
enol acetate of (135); respectively [119]. The dehydro-P-C 14-aldehyde (139)
was obtained via the enol acetate of(136) [119].
The synthesis of the 3-oxo-iso-C14-aldehyde (142) was accomplished by the
condensation of the keto acetal (7) with the lithium derivative of the acetylenic
carbinol (140) to give the intermediate glycol (141). Subsequent lithium alu-
minium hydride reduction of the triple bond and acid hydrolysis then gave
(142) [41]. This was further transformed into the 3-acetoxy-iso-C14-aldehyde
(143) by the following sequence of reactions: acetalization of the aldehyde
grouping, lithium aluminium hydride reduction, acetylation and hydrolysis
of the acetal grouping [122]. Isomerization of (143) to the 3-acetoxy-/J-C14-
aldehyde (145) was readily achieved via the corresponding enol acetate (144),
which on hydrolysis with aqueous methanolic NaHC0 3 gave (145) [122].
3-0xo- [44, 122] and 3-acetoxy-retro-dehydro-C14-aldehyde [122] (148
and 149) were. synthesized starting from the monoketal (5). Addition of
lithium ethoxyacetylide gave the ethoxyethynyl carbinol (146), which was
converted into the 3-oxo-retro-dehydro-Cu-aldehyde (147) by successive
partial hydrogenation, treatment with POC1 3 -pyridine and acid hydrolysis.
Subsequent propenyl ether condensation then gave the 3-oxo-retro-dehydro-
C14-aldehyde (148), which could also be obtained by manganese dioxide
346 H. MAYER and 0. ISLER

a,.coc,u, ~

C( --
(9) (128)
OH
-- (CCHO(129)

~ l
c:Jcrno ~HO
(131) (130)

i
(Crn,P,c.u,,, +~~J -- ~0
Br6

(132) (133) (134)

ex-
(11)
(X'rno - ~0
(135) (136)

l / l
Cc (C'rno
(137) (138)
~0
(139)

oxidation of the keto alcohol (153). Wittig reaction of (147) with the phospho-
rane (150) led to the C14-keto ester (151), which, after ketalization, was reduced
with lithium aluminium hydride to the corresponding ketal alcohol (152).
Further transformation into the C 14-phosphonium bromide (155) was effected
by the following steps: acid-catalysed hydrolysis of the ketal grouping to give
 VI. Total Syntheses 347

~~CH 20CH3
7 OH
-
(7) (140) (141)

j
AcO
~0 ~0
(143) (142)

j
~CHO~~ ~CHO
AcO A cO
(144) (145)

the keto alcohol (153), reaction of this alcohol with PBr 3 and treatment of the
resulting bromide (154) with triphenylphosphine. The crystalline dark green
phosphorane (156) was obtained by the careful treatment of an aqueous
solution of the Wittig salt (155) with sodium hydroxide [44]. The synthesis
of the 3-acetoxy-retro-dehydro-C14 -aldehyde (149) was achieved similarly to
the reaction sequence (142)-+(143).
3-Methoxy-P-C14 -aldehyde (157), an intermediate for 3-methoxy-P-C19 -
aldehyde (296), was synthesized from 3-methoxy-p-ionone (116) by the Darzens
reaction [102].

7. C 15 -Components
a) C9+C6 =(~\s
The acetylenic triphenylphosphonium bromide (165), required for the
synthesis of several oxygenated carotenoids, e.g. zeaxanthin (1004), was syn-
thesized by the· following sequence of reactions [123]: condensation of the
monoketal (7) with the Grignard derivative of trans-3-methyl-2-penten-4-yn-
1-ol (158a) gave the acetylenic diol (159), which on refluxing with sodium
348 H. MAYER and 0. ISLER

-- oAA ~CHO

(5) (147)

j((;,;HshP=~~2C2Hs
(150)

~CHO ~C02C2Hs
0~ 0~
(148) (151)

J
~CHO ~CH,OH
A cO~
(149) (152)

~CH=P(Cc;H5) 3
0~
(156)
-- (153)X=OH
(154) X=Br
(Jl

(155) X= P(C 6 H 5 h Br 9

~CHO
CH.O
(157)
 VI. Total Syntheses 349

~CH 20H
l ~CH20R
'*
OH
'*
OH
HO 0
(162) (160) R=H
(161) R=Ac

l ~CH20R

RO~
~'*

(163) R=Ac
-- (165)
(164) R=H

(166a)

l
~
~ '*"' I CH=P(C H 6 5), - - ~ '*
~ zOAc

HO~ AcO~
(166) (166b)

u
~i

(167)
+
OHC I
~(CH 3 )z

(168)
-- (169)
350 H. MAYER and 0. ISLER

acetate in aqueous acetic acid yielded a mixture of the diol (160) and the acetate
(161). Treatment of this mixture with lithium aluminium hydride resulted in
the triol (162), which on refluxing in acetic anhydride-glacial acetic acid was
converted into the diacetate (163). Subsequent saponification with methanolic
sodium hydroxide led to the diol (164), which was transformed into the desired
Wittig compound (165) by treatment with triphenylphosphonium bromide.
The acetylenic C 15 -phosphorane (166), required for the synthesis of9,9'-di-
cis-alloxanthin (1002) was synthesized in an analogous way from the mono-
ketal (7) and cis-3-methyl-2-penten-4-yn-1-ol (158b) [27].
A novel synthetic principle for compounds of the vitamin A series is re-
presented by the condensation of (2,6,6-trimethyl-1-cyclohexen-1-yl)lithium
(167) with 5-dimethylamino-3-methyl-2,4-pentadienal (168) yielding, after hy-
drolysis with perchloric acid, the immonium perchlorate (169) of P-ionylidene-
acetaldehyde [51].

BrCH 2
~C02CH, u
+ NCHO +
BrCH 2
~CN
""'=:::

(170)

l
(24)

l (173)

cA
l (171)
l (174)

~NH,
(172b) (175) (172a)
OC2 H,

C:XCH(OC2 H,h

(176)
+ ~CHOC 2 H 5
- ~ffi(OC,H,),

(178)

~CHO
(180) (179)
 VI. Total Syntheses 351

b) C1o+Cs=C1s
The Reformatskii reaction of P-cyclocitral (24) and methyl y-bromo-P-
methylcrotonate (170) in the presence of zinc gave the C 15 -lactone (171), which
on treatment with sodium ethoxide in absolute alcohol yielded cis-fJ-ionylidene-
acetic acid (172 b) in quantitative yield [35].
cis-fJ- Ionylidenea~etic acid (172 b) could also be obtained by the application
of y-bromo-fJ-methylcrotonitrile (173) instead of methyl y-bromo-fJ-methyl-
crotonate (170) yielding the intermediate <5-imi<lolactone (174), which on
treatment with water gave the lactone (171) [35].
When the <5-imidolactone (174) was heated in vacuo, cis-fJ-ionylideneacet-
\lmide (175) was formed, which was smoothly transformed into trans-P-
ionylideneacetic acid (172 a) on treatment with alkali [35].
According to the same scheme P-ionylideneacetaldehyde (180) was obtained
by the condensation of P-cyclocitral diethyl acetal (176) and 1-ethoxy-3-methyl-
1,3-butadiene (177) in the presence of zinc chloride furnishing a mixture of the
ethoxy acetals (178) and (179) of P-ionylideneacetaldehyde and retinal, respec-
tively, followed by treatment of (178) with orthophosphoric acid in dioxane
[124].
c) C11 + C4= C1s
P-Ionylideneacetaldehyde (180) could also be obtained by the condensation
of the Grignard derivative of 2-ethynyl-1,3,3-trimethylcyclohexene (69) and
4,4-dimethoxy-2-butanone (181) affording the acetylenic C 15 -hydroxy acetal

(69) (181) (182)

~l
H
CH(OCH 3 ),

(183 a) (183 b)

~CHO
(180)
352 H. MAYER and 0. ISLER

(182), which was partially reduced to the trans (183 a) and the cis hydroxy acetal
(183 b), respectively. Subsequent acid hydrolysis of both trans and cis acetals
led to the same isomeric mixture of(180) [125].

d) C13+C2=C1s
ex) P-Ionylideneacetaldehyde. P-Ionylideneacetaldehyde (180) has been pre-
pared by numerous routes according to the scheme C13 + C 2 = C 15 . A synthesis
yielding both trans- (180a) and cis-P-ionylideneacetaldehydes (180b) was
reported by Huisman et al. [126]. The Reformatskii reaction of P-ionone (89)
with ethyl bromoacetate gave the P-C 15 -hydroxy ester (184), which on treatment
with ethanolic hydrochloric acid followed by hydrolysis yielded the retro-C 15 -
acid (185). This was rearranged to the normal P-ring system when the acid
chloride (186) was prepared with phosphorus trichloride in benzene. Hydrolysis
of (186) furnished a mixture of trans- (172 a) and cis-P-ionylideneacetic acid
(172 b) which could be separated into the pure isomers by fractional crystalliza-
tion. Subsequent lithium aluminium hydride reduction afforded the corre-
sponding alcohols (187 a) and (187 b) [127, 128], which were oxidized to the
aldehydes (180a) and (180b) by manganese dioxide [128].
It is a general observation that acid-catalysed dehydration of a molecule
with a P-ionol structure or a vinylogue thereof often leads to mixtures of com-
pounds possessing the so-called retro-ionyl, e.g. (185), and the normal P-ring
system, the former always predominating. This difficulty could, however, be
completely prevented by the conversion of the hydroxy ester (184) into the
corresponding acetate (188) followed by base-catalysed elimination of the
acetate moiety yielding ethyl trans-P-ionylideneacetate (189a) [35].
Ethyl P-ionylideneacetate (189) and P-ionylideneethanol (187) have also
been made by the Wittig reaction of P-ionyltriphenylphosphonium bromide
(94) with ethyl glyoxylate [98] and glycolaldehyde [98, 129], respectively.
The undesirable retro rearrangement could also be avoided by the applica-
tion, in the Reformatskii reaction, of bromoacetonitrile [35] or acetonitrile in
the presence of lithium amide in liquid ammonia [130], yielding the hydroxy
nitrile (190), which was readily dehydrated to P-ionylideneacetonitrile (191)
[35] under the usual acidic conditions, only traces of the retro compound
having been formed. With catalytic amounts of N-bromosuccinimide, N-iodo-
succinimide or N-iodophthalimide, however, the P-isomer (191) was formed
exclusively [35].
P-Ionylideneacetonitrile (191) was also synthesized by the following
procedures:
1) Knoevenagel condensation of P-ionone (89) with cyanoacetic acid or
methyl cyanoacetate [131-136]. The decarboxylation of the intermediate
P-ionylidenecyanoacetic acid (192) occurred during the condensation and could
be completed by either heating in pyridine [136] or with copper in toluene
[135]. The pure cis- and trans-P-ionylideneacetonitriles have been prepared
in a similar way [136]. Alternatively, a P-ionone Schiff-base was used in the
condensation with cyanoacetic add [137].
 VI. Total Syntheses 353

~0
~C02C2H5
Ul em
(89) (184)

l
~OO,H
(1~ (1~

~00~
(172 a)
+
~
(172 b)
C0 2H

l l
~CH,OH
(187a)
~ CH 20H

j
(187 b)

1
~CHO
(180a)
~
(18Gb)
CHO

~OO,C,H, ~ ~OO,C,H,
(188) (189a)

Carotenmds 23
354 H. MAYER and 0. ISLER

~CN
(89) (190) (191)

r
u
~0
(89)
+
NCCH.co.H
~CN
u fu.H
(192)

II
+ (C2H 50).PCH 2CN
(193)
0

--
~0
(89)
ED
+ (C6H 5 ).PCH 2CN
Cle
(194)
-- ~CN
(191)

+
C2HsO 0

c.H,
'\.II
/

(195)
PCH2CN
--
2) Horner reaction of P-ionone (89) with (diethylphosphono)acetonitrile
(193) [138], (cyanomethyl)triphenylphosphonium chloride (194) [139] or
(ethoxyphenylphosphinyl)acetonitrile (195) [140].
3) Treatment of ({J-ionylideneethyl)triphenylphosphonium chloride (204)
or sulphate (206) with amyl nitrite or N-methyl-N-nitroso-p-toluenesulphon-
amide in the presence of sodium methoxide or diethylamine, respectively [141].
The conversion of P-ionylideneacetonitrile (191) into P-ionylideneacet-
aldehyde (180) was readily achieved by reduction with diisobutylaluminium
hydride [135] or lithium aluminium hydride [134], or by catalytic reduction
over Raney nickel and hydrolysis of the intermediate aldimine [142].
The following additional methods have been devised for the synthesis of
P-ionylideneacetaldehyde (180) from P-ionone (89):
1) Horner reaction with ethyl (diethylphosphono)acetate (196a) in the
presence of sodium amide in tetrahydrofuran yielding ethyl trans-P-ionylidene-
acetate (189a), which on subsequent lithium aluminium hydride reduction and
manganese dioxide oxidation was converted into (180) [143].
 VI. Total Syntheses 355

0
II
(C 2 H,O),PCH 2 C0 2 C2 H,

(196a)
+
u
~0
+
0
II
(C 2 H,O),PCH 2 CH (OC 2 H,),
(199)
(89) or uc=cOC 2 H,

l j
~oo,c,H, ~CHO
(189a) (180)

r
~0 CH,CH~C,H,
(89) (197) (198)

2) Aldol condensation with N-ethylidenecyclohexylamine (197) in the

presence oflithium diisopropylamide followed by hydrolysis of the intermediate
carbinol (198) [144].
3) Horner reaction with diethyl (2,2-diethoxyethyl)phosphonate (199) in
the presence of sodium amide in tetrahydrofuran [145].
4) Condensation with the Grignard derivative of ethoxyacetylene followed
by partial hydrogenation and hydrolysis [93, 146].
5) Ethyl vinyl ether condensation with the diethyl acetal [147] or ethylene
acetal [148] of P-ionone.
6) Condensation with ethyl formate, followed by treatment of the enolate
of the reaction product with methyl sulphate, acetalization, a Grignard reaction
with methylmagnesium iodide, hydrolysis and dehydration [149].
7) Manganese dioxide oxidation of vinyl-P-ionol (201) under acidic con-
ditions [150].

p) Vinyl-P-ionol. Vinyl-P-ionol (201) was prepared from P-ionone (89) by

condensation with sodium [151], lithium [152-154] or calcium [152] acetylide
followed by partial hydrogenation [153-157] of the resulting ethynyl-P-ionol
(200), or by reaction with vinylmagnesium bromide [154] or chloride [150].
356 H. MAYER and 0. ISLER

(89) (200) (201)

y) (P-I onylideneethyl )triphenylphosphonium halides. Numerous methods

have been reported, mainly in the patent literature, for the preparation of
(/3-ionylideneethyl)triphenylphosphonium halides which are important cyclic
C 15 -components for the synthesis of /3-carotene, /3-apo-carotenoids and vita-
min A. Thus (/3-ionylideneethyl)triphenylphosphonium chloride (204) has been

~
(201) ~

~CH,OH ~ ~$
~ CH 2 P(C6 H 5 )a X
e

(I8ry / (204) X=Cl

(205) X=Br
(206) X=HS0 4

~CH,OAo

(202)

~CH~•
(203) (207)

(208) (209)
 VI. Total Syntheses 357

readily obtained from vinyl-P-ionol (201), P-ionylideneethanol (187) or the

corresponding acetate (202) by the action of triphenylphosphonium chloride
[158, 159] or triphenylphosphine and hydrochloric acid in various solvents,
e.g. tetrahydrofuran, methanol or dimethylformamide [153, 158, 160]. The
corresponding triphenylphosphonium bromide (205) [158-160], iodide [158],
sulphate (206) [160] and p-toluenesulphonate [160] have been prepared in an
analogous way. Alternatively, ({J-ionylideneethyl)triphenylphosphonium bro-
mide (205) was synthesized from (187) by reaction with phosphorus tribromide
and treatment of the intermediate bromide (203) with triphenylphosphine
[161, 162], or by treatment of the C 15 -hydrocarbon (207) with triphenyl-
phosphonium bromide in dimethylformamide [163]. Although the triphenyl-
phosphonium salts are crystalline compounds [158, 160], they were usually
not isolated but subjected immediately after formation to a Wittig reaction
with the appropriate carbonyl compounds. ,
The preparation of the C 15 -phosphonate (208) from (187) or (201) and
triethyl phosphite was found to be not as satisfactory as that of the correspond-
ing triphenylphosphonium halides in that a mixture of (208) and the tertiary
phosphonate (209) was formed [100].

[>) 3,4-Didehydro-fJ-ionylideneacetaldehyde. 3,4-Didehydro-P-ionylidene-

acetaldehyde (213)-an important intermediate in vitamin A2 syntheses [164]-
has been synthesized by two different routes:
1) trans-P-Ionylideneacetic acid (172 a) was transformed into the corre-
sponding methyl ester (210), which, on bromination with N-bromosuccinimide
and subsequent dehydrobromination, gave methyl 3,4-didehydro-p-ionylidene-
acetate (211). This was saponified and the mixture of cis-trans isomeric C15 -
acids (212a and 212b) separated by fractional crystallization and chromato-
graphy. The pure isomers were then converted into trans- (213 a) and cis-3,4-
didehydro-P-ionylideneacetaldehyde (213 b) by successive lithium aluminium
hydride reduction and manganese dioxide oxidation [164].
2) Darzens reaction of P-ionone (89) with chloroacetonitrile in the presence
of excess alcoholate (glycidimidic ester synthesis) gave the epoxy imido ester
(214), which could be readily converted into the retro IX-hydroxy ester (215) by
acid treatment. Dehydration with diethyl chlorophosphite in the presence of
tertiary bases, followed by saponification, yielded 3,4-didehydro-P-ionylidene-
acetic acid (212), which was transformed into the 3,4-didehydro-C 15 -aldehyde
(213) as mentioned [35].

c) (3,4-Didehydro-{J-ionylideneethyl)triphenylphosphonium bromide. The

Wittig compound (218) was prepared similarly to the CwWittig salt (205).
3,4-Didehydro~P-ionone (99) was condensed with lithium acetylide yielding
the ethynyl carbinol (216), which was partially hydrogenated over Lindlar
catalyst. Treatment of the resulting vinyl-3,4-didehydro-P-ionol (217) with
triphenylphosphonium bromide in methanol then afforded the crystalline
C 15 -component (218) [165].
358 H. MAYER and 0. IsLER

~co~,
(210)

1
~co.cH3
~
(211)

/
~C~H ~
~ to 2H

(212a) (212b)

1 1
~CHO
(213a)
~HO
(213b)

~0
(89)

1
~co.H
~
(212)
- ~co.c.H,
u
(215)
bH
 VI. Total Syntheses 359

~0
lvl
(99) (126)

1
CH 2 ~(CoHsh
Br 9
::::::,...

(218) (217)

G rx-Ionylideneacetaldehyde. A Reformatskii reaction of rx-ionone (101)

with ethyl bromoacetate gave the hydroxy ester (219), which on treatment
with p-toluehesulphonic acid in benzene was dehydrated to ethyl rx-ionylidene-

~CO,C,JI,
(101) (219)

1
~CHO ~CO,C,JI,
(221) (220)

acetate (220) [128]. The latter compound was more conveniently obtained
by a Horner condensation of rx-ionone (101) with ethyl (diethylphosphono)-
acetate (196a) iii the presence of sodium methoxide [166]. Reduction of (220)
with lithium aluminium hydride, followed by manganese dioxide oxidation
of the product, then gave rx-ionylideneacetaldehyde (221) [128, 166].
360 H. MAYER and 0. ISLER

17) ( rx-Ionylideneethyl)triphenylphosphonium halides. Partial hydrogenation

over Lindlar catalyst of ethynyl-rx-ionol (222), derived from rx-ionone (101)
and lithium or calcium acetylide [152, 153], furnished vinyl-rx-ionol (223)
[153, 166], which reacted with triphenylphosphonium bromide in methanol
to give the Cwtriphenylphosphonium bromide (224) [166]. Similar treatment
of (223) with triphenylphosphonium chloride produced the corresponding
chloride (225) [153]. The enantiomeric Wittig salt (224a) used for a synthesis
of ( + )-(R)-rx-carotene (5) has been prepared from ( + )-(R)-rx-ionone in an
analogous way [106].

~0
(101) (222)

~CH,~(c,H,), x•

(224) X=Br (223)

(225)X=Cl

(224a)

8) 3-0xygenated Cwcomponents. 3,3-Ethylenedioxy-{J-ionylideneacetalde-

hyde (226) and 3-ethoxy-3,4-didehydro-{J-ionylideneacetaldehyde (227) were
readily synthesized from the appropriate {J-ionones (112) and (115) (Section B.S f),
respectively, by the following steps: Horner reaction with ethyl (diethyl-
phosphono)acetate (196a), lithium aluminium hydride reduction and man-
ganese dioxide oxidation [167].
Methyl trans- (228) and cis-3-methoxy-{J-ionylideneacetate (229) have been
synthesized by the application of the Reformatskii reaction to 3-methoxy-
{J-ionone (116) [168].
The synthesis of [(3-hydroxy-{J-ionylidene)ethyl]triphenylphosphonium
bromide (231) has recently been •reported [109]. Reaction of 3-hydroxy-
 VI. Total Syntheses 361

~CHO ~CHO
A)l
\:-o C2 H50

(226) (227)

~CO,CH, ~
CH 3 0 CH.O
}__)l tolcH.
(228) (229)

~0 ~
HO)__)l
HO)__)l C>H
(109) (230)

j
~CH2~(C6Hs) 3 Br 8

HO)__)l
(231)

P-ionone (109) with vinylmagnesium bromide gave the vinyl carbinol (230),
which was treated with triphenylphosphonium bromide.

e) C14 +C1 =C 15
Russian workers [169] prepared the P-Cwaldehyde (180) from the P-Cw
aldehyde (123) utilizing the cyanohydrin synthesis with acetone cyanohydrin.
The resulting hydroxy nitrile (232) was dehydrated to P-ionylideneacetonitrile
(191), which was then reduced with diisobutylaluminium hydride.

~CN
~CHO u bH
(123) (232) (191)
362 H. MAYER and 0. ISLER

8. C 16 -Components
a) C11 +Cs=C16
A method was described for the synthesis of the P-C 16 -aldehyde (236)
by reaction of the Grignard derivative of 2-ethynyl-1,3,3-trimethylcyclohexene
(69) with the cyclopropyl ketone (233). The resulting acetylenic carbinol (234)
was reduced with lithium aluminium hydride to yield the carbinol (235),
which, on treatment with 2 N HCl, was converted into (236) [170].

Oc¢~~~
(69)
+ o~o-n-C.JI9
(233)
- (234)

~CHO
(236)
- ~~~
(235)

b) C13+C3=C16
The Reformatskii reaction of P-ionone (89) and propargyl bromide leading
to the c16-building units (237) and (238) has been intensively investigated.
Under the usual reaction conditions in the presence of zinc [35, 171-173]
the acetylenic C 16 -carbinol (237) was obtained in varying yields and purity,
always containing small amounts of an allenic by-product of structure (241).
The formation of (241) could, however, be suppressed when magnesium [174]
or aluminium [35, 175] in the presence of mercury salts were used instead
of zinc.
The acid-catalysed dehydration of (237) yielded, depending on the con-
ditions, a varying mixture of mainly two hydrocarbons, namely the liquid
P-C 16 -hydrocarbon (238) and its crystalline retro isomer (239). The desired
P-hydrocarbon (238) was obtained as the main product when the dehydration
was carried out with phosphorus oxychloride and pyridine [176]. Complete
prevention of the formation of the retro compound could be achieved by
first converting the acetylenic carbinol (237) into the corresponding tertiary
 VI. Total Syntheses 363

(89) (237) (238)

j +

~·~ ex+¢ c
~¢
-?'"

#
(241) (240) (239)

~CH& 0

ec---i~/
#
H
~¢
OH
#
(242) (243) (244a)

UCHO
I
(47)
---+
~ (245)
---+
~¢
(246}

XYCHO
I
(50)
---+
~ (245 a)
---+
~¢
(246a)

acetate (240) followed by elimination of the acetic acid moiety with potassium
t-butoxide [175].
A cis-trans isomeric mixture of the C 16 -hydrocarbon (238) has recently
been obtained by the Wittig reaction of P-ionyltriphenylphosphonium bromide
(94) with propargylaldehyde [177].
Besides propargyl bromide, the Grignard derivative of 1,3-dibromo-
2-propene was used [178] for the preparation of the acetylenic carbinol (237)
yielding the bromo carbinol (242), which on refluxing with sodium ethoxide
gave (237).
364 H. MAYER and 0. ISLER

The C16-carbinols (243) [179], (244a) and (244b, Table26) [107, 179a],
(246) [180, 181] and (246a) [182] have been similarly prepared by theRefor-
matskii reaction of the appropriate ketones with propargyl bromide. The
C13 -ketone (245) was readily accessible by the base-catalysed condensation of
2,3,6-trimethylbenzaldehyde with acetone [181-183].

c) C14 +C 2 =C 16
An alternative method has been reported for the synthesis of the C 16 -
hydrocarbon (238) [183]. Condensation of the P-C 14-aldehyde (123) with
sodium or lithium acetylide in liquid ammonia [115, 184, 185] yielded the
acetylenic C 16 -carbinol (247), which on treatment with phosphorus tribromide
in ether was converted into the rearranged secondary bromide (248). The
latter was readily dehydrobrominated by stirring a benzene solution with
1,5-diazabicyclo[4.3.0]-5-nonene (249) affording (238) in high yield.
In the technical manufacturing procedure of P-carotene by F. Hoffmann-
La Roche & Co. Ltd., the P-C 16 -aldehyde (236) was synthesized from the
P-Cwaldehyde (123) by the vinyl ether condensation. This reaction type
was shown by Isler et al. [41, 119, 186-188] to be a powerful tool for the
extension of polyene chains by two carbon atoms. The P-C 14-aldehyde (123)
was first transformed into the corresponding diethyl acetal (250) by treatment
with triethyl orthoformate and a trace of acid. The ocP-unsaturated acetal (250)
was condensed with ethyl vinyl ether (250a) in the presence of zinc chloride
or boron trifluoride etherate at low temperature in order to avoid further
condensation of the C 16 -acetal (251) formed. The latter compound was then
hydrolysed with e.g. moist acetic acid in the presence of sodium acetate [186].
(The related propenyl ether condensation is discussed in Section B.11 a y.)
The C 16 -aldehydes (252}-(259) [119, 122] and (260) [102] were prepared
from the appropriate Cwaldehydes in an analogous way, the aldehydes

~CHO
(123) (247)

(238) (249) (248)

 VI. Total Syntheses 365

~CH(OC,H,), ~CH(OC2H 5) 2
Ul 6c2u,
(250) (250a) (251)

j
~CHO ~CHO
(252) (236)

~CHO ~CHO
(253) (254)

~CHO ~CHO
0~ A cO
(255) (256)

~HO ~CHO
0~
AcO
(257) (258)

~CHO ~CHO
A cO~ CH 3 0
(259) (260)

(236) and (252}-(254) also by condensation of the C 14-aldehydes with ethoxy-

acetylene [119].
366 H. MAYER and 0. ISLER

d) C1s + C1 = c16
The enol ether (261) of the fJ-C 16 -aldehyde was obtained by reaction of
(fJ-ionylideneethyl)triphenylphosphonium chloride (204) or bromide (205)
with ethyl formate in the presence of sodium ethoxide [129, 159].

~uoc~,
(261)

9. C 17 -Components
a) C13+C4=C11
P-Ionylidenecrotonic acid (fJ-C 17 -acid) (266)-used as an intermediate
in several vitamin A syntheses-was obtained in low yields by the Reformatskii
reaction of fJ-ionone (89) and ethyl y-bromocrotonate (262) [189-191]. The
reaction was intensively studied by Eiter et al. [35], who showed that under
specified conditions about 70% of the desired hydroxy ester (264) and 15%
of a by-product (270) were formed. Dehydration of (264) with N-iodosuccin-
imide followed by saponification gave exclusively all-trans-fJ-ionylidene-
crotonic acid (266). No retro rearrangement of the double-bond system was
observed.
Similarly, y-bromocrotononitrile (263) in the presence of zinc led to the
hydroxy nitrile (265) besides small amounts of the by-product (271). In contrast
to (264), this hydroxy nitrile could be dehydrated with the usual dehydrating
agents yielding the nitrile (268), which was further hydrolysed to (266) [35].
P-Ionylidenecrotonic acid (266) and the corresponding methyl ester (267)
have also been obtained by hypochlorite oxidation of the fJ-C 18 -ketone (275)
followed by esterification [192].
Reduction of methyl P-ionylidenecrotonate (267) with diisobutylaluminium
hydride followed by manganese dioxide oxidation of the product furnished
P-ionylidenecrotonaldehyde (269) [192].

b) C15 +C 2 =C 17
Methyl all-trans- (272 a) and 9-cis-11-trans-3,4-didehydro-fJ-ionylidene-
crotonate (272 b) have recently been synthesized [193] by the Horner con-
densation of trans- (213 a) and cis-3,4-didehydro-fJ-ionylideneacetaldehyde
(213 b), respectively, with methyl (diethylphosphono)acetate (196b) (Tablel).

10. C 18 -Components
The P-C 18 -ketone (275) was used as a valuable intermediate for the
synthesis of fJ-carotene and a large number of vitamin A components. It could
be prepared by four different routes.
 VI. Total Syntheses 367

(89)

BrCH 2 ~co./cu. ~BrCH2 ~cN

(262) (263)

u
~C0 2C 2H 5
u~CN
6u 6u

(264) (265)

j j
~co~
(266)R=H
- ~CN
(268)
(267) R=CH 3

l
~CHO
(269) (270) R=C0 2 C 2 H 5
(271) R=CN

~co.cu.
~
(272a) (272b)
368 H. MAYER and 0. ISLER

a) C 9 + C 9 = C1 8
Condensation of 2,2,6-trimethylcyclohexanone (9) with the C 9 -acetylenic
carbinol (273) yielded the C 18 -acetylenic glycol (274), which was transformed
into (275) by successive lithium aluminium hydride reduction, manganese
dioxide oxidation and acid dehydration [ 48, 194].

(9)

j (273)

(274) (275)

bJ C14 + C4= c1s

Condensation of the P-C 14 -aldehyde (123) with the lithium derivative of
the C 4 -enol ether (276) gave the product (277), which on acid hydrolysis was
transformed into 11,12-didehydro-{J-C18 -ketone (278). Subsequent partial
hydrogenation then afforded (275) [195].

I c14+C4=C1s
I
~
~CHO+
(123)
?;- ~OC2Hs
(276)
-- ~"

(277)
H
oc,n,

1
"":::: "":::: ""=o
~,.Ao ""::::
-+----

(275) (278)
 VI. Total Syntheses 369

c) C1 5 +C3=C1s
all-trans- and 9-cis-P-C 18 -Ketone were also synthesized from the isomeric
P-C 15 -aldehydes (180a) and (180b) by the base-catalysed condensation with
acetone [126, 127, 196]. Photochemical isomerization of the all-trans ketone
gave the 11-cis isomer [196]. Alternatively, P-ionylideneacetaldehyde aldimine
was condensed with acetone to an amino ketone which, on treatment with
calcium chloride in ethanol, was deaminated to (275) [197]. A one-step
preparation of P-C 18 -ketone (275) by the Oppenauer oxidation of P-ionylidene-
ethanol (187) in the presence of acetone was reported [198].
The cx-C 18 -ketone (279) was synthesized in an analogous way [199].
The base-catalysed condensation of the isomeric 3,4-didehydro-P-C15 -
aldehydes (213 a) and (213 b) with acetone led to all-trans- and 9-cis-3,4-di-
dehydro-P-C18-ketone (280) [196, 200].

~CO,H~ (275)

(266)

(279) (280)

(281)
OH
- (282a)X=Br
(282b)X=CI

d) C11+C1=C1s
The P-Cwketone (275) was obtained from P-ionylidenecrotonic acid (266)
by reaction with methyllithium [189, 191, 201]. 9-cis-P-C18 -Ketone [202]
and 3,4-didehydro-P-C18 -ketone (280) [203] were synthesized in an analogous

Carotenoid& 24
370 H. MAYER and 0. ISLER

way. Starting from P-C 17 -acid chloride, the methyl group was introduced with
dimethylcadmium [190] or dimethylzinc [204].
The C18-triphenylphosphonium bromide (282 a) and the corresponding
chloride (282 b) were prepared by treatment of the P-Cwalcohol (281) with
triphenylphosphine and hydrogen bromide in dimethylformamide [36] or
hydrogen chloride in methanol [75], respectively.

11. CwComponents
a) CwAldehydes
The synthesis of these important building units has been achieved by
three different routes.
rx.) c14 + Cs = c19· The Wittig reaction of P-C14-aldehyde (123) with the
C 5 -phosphonium bromide (283 a) in the presence of sodium methoxide gave
the diethyl acetal of P-C19 -aldehyde (284), which was hydrolysed by dilute
phosphoric acid to a mixture of cis-trans isomers of the P-C19 -aldehyde (285)
[124].
Alternatively, the zinc chloride-catalysed condensation of P-Cwaldehyde
diethyl acetal (250) with 1-ethoxy-2-methyl-1,3-butadiene (286), followed by
treatment of the condensation product with dilute phosphoric acid, gave the
P-Cwaldehyde (285) [124].
Methyl y-bromo-rx.-methylcrotonate (444) was employed by Inhoffen's
team [118, 205] for the synthesis of the P- (285) and rx.-Cwaldehyde (287 c).
Reformatskii reaction with P- (123) and rx.-C14-aldehyde (127) produced the

(123)

(285) ~ (284)

~ffi(OC,H,), + ~CHoc.u,
(250) ~286)
 VI. Total Syntheses 371

~CHO ~CHO
{123) {127)

j + BrCH2~co2cH.
(444)
+
j

(286a) R=C0 2 CH 3 (287a) R=C0 2 CH 3

(286b) R=CH 2 0H (287b) R=CH 2 0H
(285) R=CHO (287c) R=CHO

C19 -esters (286a) and (287 a), which on lithium aluminium hydride reduction
gave the corresponding alcohols (286b) and (287 b). Subsequent manganese
dioxide oxidation then yielded the desired aldehydes (285) and (287 c).
Another synthesis of the {3-Cwaldehyde (285) was reported by Inhoffen
et al. [206] by chain lengthening of the {3-Cwaldehyde (123) with the acetylenic
carbinol (464) (Section C.2g).
/3) C15 + C4 = C19 • Two patents granted to the Badische Anilin- & Soda-
Fabrik AG [159, 207] describe the synthesis of the Cwenol ether (290) by
the Wittig reaction of ({3-ionylideneethyl)triphenylphosphonium chloride (204)
with the ethoxy aldehyde (289).

(289)

(290)
372 H. MAYER and 0. ISLER

y) C16 + C 3 = C19 • In exact analogy to the vinyl ether condensation des-

cribed in Section B.8 c the propenyl ether condensation was applied to the
technical preparation of the P-C 19 -aldehyde (285) from the P-C 16 -aldehyde
(236) by F. Hoffmann-La Roche & Co. Ltd. [186]. The chain lengthening
by three carbon atoms was achieved by the use of propenyl ether, e.g. ethyl
propenyl ether (415), instead of ethyl vinyl ether.
The C19 -aldehydes (291}-(295) [41, 119, 187, 208], (295a) [41] and (296)
[102] have been synthesized in an analogous way.

c16+C3=C19 \

-:::7 ?" ?" am

(291) (292)

-:::7 ?" ?" CHO

(293) (294)

?" ?" ?" CHO

CH 3 0
(295) (296)

A cO
(295a)

b) Other 3-oxygenated C 19 -components

C13 + C6 = C19 . A novel approach to the synthesis of 3-oxygenated caro-
tenoids by the use of the CwWittig compounds (300) and (302) has recently
been reported by Surmatis et al. [111, 167] (see also Section B.2). Condensation
of 3,3-ethylenedioxy-P-ionone (112) with the Grignard derivative of the
acetylenic carbinol (297), which was readily available by ethynylation of
methacrolein, yielded the diol (298). Acid-catalysed dehydration of the latter
gave the carbinol (299), which, on treatment with triphenylphosphonium
bromide in benzene, furnished the triphenylphosphonium bromide (300) [167].
 VI. Total Syntheses 373

~+
~~0
I
+ OH

0 (112) (297)

~~~~~0~~~
j
(298) / (299)

HO

"'
"""'~ I ®
O ,:::? CH2P(CoHsh Br 8

(301) (300)

(302)

The fully conjugated Wittig salt (302) resulted from an analogous treatment
of the carbinol (301), which was obtained by partial hydrogenation of (299)
over Lindlar catalyst [111].

12. C 20 -Components
a) C 10 +C10=Czo
Condensation of P-cyclogeranyltriphenylphosphonium bromide (27) with
the Cw-triene .aldehydic acid (303) or ester (304) in the presence of sodium
methoxide yielded all-trans-retinoic acid (305 a) and the corresponding ethyl
ester (306) [36, 209]. Although the Wittig reaction usually gives a cis-trans
mixture, the newly formed double bond is forced into the trans configuration.
Reduction of the ester with lithium aluminium hydride or sodium ethoxy-
ethylaluminohydride furnished vitamin A (307 a) [36].
374 H. MAYER and 0. ISLER

+ +

OH~CO,R OH~CH 20Ac

(303)R=H (308)
(304) R=C 2 H 5

j j
C02 R CH 2 0Ac
~ ~ ~ ~ ~ ~

(305a) R=H (309a)

(306) R=C 2 H 5

j
CH 2 0H
~ ~ ~

(307 a)

Vitamin A acetate (309a) was synthesized by an analogous reaction with

8-acetoxy-2,6-dimethyl-2,4,6-octatrienal (308) [36, 209].

b) C11 + C9= C2o

This reaction scheme was followed by Attenburrow et al. [ 48] for the
synthesis of compounds possessing the vitamin A skeleton. The Grignard
reaction of the ethynylcyclohexanol (67) with the ketone (310) gave the
acetylenic diol (311), which on acid treatment was rearranged to the diol (312).
Lithium aluminium hydride reduction of the triple bond resulted in the
formation of the tetraene diol (313). Dehydration of the corresponding acetate
(314) then produced a mixture of vitamin A acetate (309 a), anhydrovitamin A
(370) and starting material.
 VI. Total Syntheses 375

(67) (310)

h~
OH
~CH20H
~~ ~'*
UOH UOH
j
(311) (312)

(309a) (313)R=H
(314) R=Ac

(69) (315)

The same sequence of reactions was carried out with 2-ethynyl-1,3,3-tri-

methylcyclohexene (69) whereby 7,8-didehydrovitamin A (315) was obtained
[48].

c) C 13 +C7=C2o
The Wittig reaction of {J-ionyltriphenylphosphonium bromide (94) with
the aldehydic acid (316) or the aldehydic ester (317) gave a mixture of all-trans-
(305 a) and 9-cis-retinoic acid (305 b) and the corresponding ethyl esters (306)
and (318), resp!!ctively [36, 210]. In general, good yields were obtained but
the method gained little practical importance because of the presence of a
high percentage of 9-cis isomer which could not be isomerized to the all-trans
configuration.
Vitamin A acetate (309a) has been prepared accordingly from 6-acetoxy-
4-methyl-2,4-hexadienal (319 a) [36].
376 H. MAYER and 0. ISLER

~e
~ P(C H ).Br
e
6 5

{94) {316) R=H

j {317) R=C 2 H 5

(305a) R=H (305b) R=H

{306) R=C 2 H 5 (318) R=C 2 H 5

OH~CH 20Ac
(319a)

The Wittig reaction of P-ionone (89) (Section B.5a) with the C 7 -phos-
phonium bromide (320) is of no importance because of the poor reactivity
of P-ionone (yield < 3 %) [36]. The yield could, however, be substantially
improved when ethyl 6-(diethylphosphono)-3-methyl-2,4-hexadienoate (321)
was employed, but again the product was a mixture of all-trans- and 9-cis-
retinoic acid ethyl ester [211].
A novel approach to the synthesis of the vitamin A skeleton has been
reported by Jacobs et al. [212, 213]. Condensation of P-ionone (89) with the
lithium or sodium derivative of the acetylenic ether (322) yielded the acetylenic
C 20 -carbinol (323). Lithium aluminium hydride reduction or partial hydro-
genation of the triple bond produced the carbinol (324), which on acid hydro-
lysis was converted into vitamin A aldehyde (retinal, 325 a).
According to the C13 + C 7 = C 20 scheme the following compounds of the
vitamin A series have been synthesized: retinal (325 a) and vitamin A ethyl
ether by the condensation of P-ionone (89) with the acetylenic carbinols (326)
[214] and (327) [124], respectively; retinoic acid ethyl ester (306), 11,12-di-
dehydrovitamin A methyl ether and vitamin A methyl ether (366) by the
Reformatskii reaction of P-ionon'e (89) with the bromides (328) [215] and
 VI. Total Syntheses 377

I c13 +C7 =C2o

~~I CHOCH
3

(322)

~~~CHOCH, ~
~
OH
,:9' ,:9' ::7CHOCH 3

(323) (324)

l
CHO
~ ~ ~ ~

(325 a)

I
~.~CH(OC 2H 5) 2 ~~CH20C2Hs
OH
OH
(326) (327)

~C0 2 C 2 H 5 OHC~CH 2 0CH 3

BrCH 2 / *'
(328) (329)

~CH 2 0CH 3
BrCH 2/ *'
(328a) (330)

(328a) [216, 216a], respectively; vitamin A methyl ether (366) and 11,12-di-
dehydroretinal by the Wittig reaction of {3-ionyltriphenylphosphonium
bromide (94) with the aldehydic ether (329) [124] and the acetylenic aldehyde
(330) [177].
 w
-.l
I c,4+C6=Czo l 00

*~CH20H
0~CHzOH
*~H,OH r
*~ CH20H (158b) ~CHO (158 a) ~
H ~ H
~
(331 a) (123) (331 b)

1 1 p::
a:::
>
-::Py~ ~*~
u CH20H
~*~CH20H ~
I»
OH ~ _...CH20R s..
u~ 9
(332) R=H (334a) ......
(333) R=Ac (334 b) ~:=

1 1 1
~_...CH 2 0R ~~ ~~~

~ _.-CHzOH
~

R
(307a) R=H (307b) (307c) R=CH 2 0H
(309a) R=Ac (325e) R=CHO
all-trans 11,13-di-cis 11-cis
 VI. Total Syntheses 379

d) C14+C6=Czo

The technical synthesis of vitamin A developed by Isler et al. [1, 217]

in the laboratories of F. Hoffmann-La Roche & Co. Ltd., Basle, follows the
route C14 + C 6 = Czo.
The P-C 14-aldehyde (123) was condensed in a Grignard reaction with
cis-3-methyl-2-penten-4-yn-1-ol (158b) yielding the acetylenic diol (331 a).
Partial hydrogenation of the condensation product over Lindlar catalyst
afforded the glycol (332), which, after acetylation of the primary hydroxyl
group (333), was dehydrated and rearranged to all-trans-vitamin A acetate
(309a). Subsequent saponification then gave the corresponding vitamin A
alcohol (307 a). The rearrangement of the 11,12- and 13,14-cis double bonds
of (332) seems to occur during the dehydration step of the synthesis.
· By a variation of the procedure sterically hindered vitamin A isomers
have been prepared [218]: the Grignard reaction of P-Cwaldehyde (123)
with trans-3-methyl-2-penten-4-yn-1-ol (158a) led to the acetylenic diol (331 b).
Acetylation followed by dehydration furnished 11, 12-didehydroretinol (334 b),
which on partial hydrogenation was transformed into 11-cis-retinol (307 c).
Manganese dioxide oxidation of the latter gave the corresponding 11-cis
aldehyde (325 e) [219]. 11,13-Di-cis-retinol (307 b) was obtained by a similar
sequence of reactions starting from the acetylenic glycol (331 a) via 13-cis-11,12-
didehydroretinol (334a).
When the P-C 14-aldehyde (123) was replaced by the py-unsaturated aldehyde
(125), 9-cis-vitamin A acetate (336) was obtained instead of the all-trans
isomer. Dehydration of the intermediate (335) was achieved by successive
treatment with phosphorus tribromide and 1,5-diazabicyclo[4.3.0]-5-nonene
(249) [183, 220].

"'<:::::: "'<:::::: CH 2 0Ac

OH

(335) (336)
9-cis CH 20Ac

Starting from 3,4-didehydro-P-Cwaldehyde (139), all-trans- (337 a),

11,13-di-cis- (337 b) and 11-cis-vitamin A 2 (337 c) as well as the corresponding
aldehydes (338 a)-(338 c) have been synthesized in an analogous way [164, 221].

e) C1 5 +Cs=Czo
rx.) P-Ionylideneacetaldehyde routes. An interesting procedure that allows
the synthesis of four unhindered vitamin A isomers from trans- (180a) and
cis-P-Cwaldehyde (180b) was reported by Matsui et al. [222, 223]. Con-
 380 H. MAYER and 0. IsLER

~CHO

I \
(139)

l
""':::: R ""'::::
(337a) R=CH 2 0H (337b) R=CH 2 0H (337c) R=CH 2 0H R
(338a) R=CHO (338b) R=CHO (338c) R=CHO

all-trans 11,13-di-cis ll-cis

densation of trans-P-Cwaldehyde (180a) with ethyl senecioate (339) in the

presence of potassium amide in liquid ammonia or anhydrous ether yielded
all-trans-retinoic acid (305 a). Interestingly, when sodium or lithium amide
were employed, 13-cis-retinoic acid (305 c) was obtained. The 9-cis- (305 b)
and the 9,13-di-cis-retinoic acid (305 d) were prepared accordingly from
cis-P-Cwaldehyde (180b). Lithium aluminium hydride reduction of the
methyl esters then furnished the corresponding vitamin A isomers (307 a),
(307 d), (307 e) and (307 f). Manganese dioxide oxidation gave the corresponding
aldehydes (325 a), (325 b), (325 c) and (325 d) [219, 224, 225]. All-trans-vitamin A
aldehyde (325 a) has also been obtained from vitamin A by oxidation with
chloranil [226].
The same isomers had been prepared earlier by a very similar route through
the condensation of the isomeric P-C 15 -aldehydes with methyl P-methyl-
glutaconate [128].
13-cis-Retinoic acid (305 c) has also been obtained by the Reformatskii
reaction of the P-C 15 -aldehyde (180) with methyl y-bromo-P-methylcrotonate
(170) [35].
Starting from the isomeric 3,4-didehydro-P-Cwaldehydes (213 a) and
(213 b), the Matsui condensation was also applied successfully by Schwieter
et al. [164] to the synthesis of the vitamin A 2 acid isomers (340a)-(340d)
and of the corresponding alcohols (337 a) and (337 d)-(337f) and aldehydes
(338 a) and (338 d)-(338!).
IX-Vitamin A compounds have been prepared in an analogous way starting
from IX-ionylideneacetaldehyde (221) [166]. Condensation with ethyl senecioate
(339) in the presence of potassium amide gave IX-vitamin A acid (341). The
 VI. Total Syntheses 381

~CHO

KNH2 I (180a)

~C02C 2 H 5
(339) \ NaNH 2

C0 2 H

(305 a) (305c)

(307a) R=CH 2 0H (307d) R=CH 2 0H

(325a) R=CHO (325b) R=CHO
all-trans 13-cis

~
\N-,
CHO

(180b)

KNH,/ ~C0 2C2Hs

(339)

~ ~

~ ~

(305b) ~
C0 2H
(305d) ~

~ ~
C02H

~ ~

~ ~

R
~ ~

(307e) R=CH 2 0H R (307j) R=CH 2 0H

(325c) R=CHO (325d) R=CHO

9-cis 9,13-di-cis
382 H. MAYER and 0. IsLER

~CHO

I (213a) \

(340a) R=C0 2 H (340b) R=C0 2 H

(337 a) R = CH 2 0H (337d) R=CH 2 0H
(338a) R=CHO (338d) R=CHO

all-trans 13-cis

~HO
I (213b) \

"""" R
(340c) R=C0 2 H
""""R (340d) R=C0 2 H
(337e) R=CH 2 0H (337/) R=CH 2 0H
(338e) R=CHO (338!) R=CHO

9-cis 9,13-di-cis

corresponding methyl ester, on lithium aluminium hydride reduction, yielded

ex-vitamin A (342). Treatment of the latter with triphenylphosphonium chloride
gave the phosphonium salt (343), and oxidation with manganese dioxide
produced ex-retinal (344).
The Wittig and Horner reactions have been widely used for the synthesis
of vitamin A compounds according to the scheme outlined below. Thus
retinoic acid esters were obtained from the {J-Cwaldehyde (1 80) and the
phosphonium salt (345) [227] or the phosphonate (346) [36]. Vitamin A
acetate (309a) and vitamin A acid nitrile (361) (see Section B.l2g) were made
by the condensation of {J-Cwaldehyde (1 80) with the phosphonium salt
(348) [36] and the phosphonate (349) [140], respectively.
 VI. Total Syntheses 383

I C,s+Cs=C2o I

~CHO + ~C02C2Hs
(221) (339)

E9
(341) R=C0 2 H (343) R=CH 2 P(C 6 H 5 lJ Cl 9
(342) R=CH 2 0H (344)R=CHO

Vitamin A acid nitrile (361) has also been obtained from the P-C 18 -ketone
(275) and cyanoacetic acid (see Section B.12g) and by treatment of P-retinyl-
triphenylphosphonium chloride (367) with N-methyl-N-nitroso-p-toluene-
sulphonamide in the presence of sodium methoxide [141] (see also Section
B.7d).

~CHO ~CHO
c ..
lvl
(180) (213)

Br
9 Ell ~C02 CH 3
(C6 H,).PCH 2 ""<::::::
~ ~C0 2 R
(C 2H 5 0).PCH 2 ""<:::::: •
(345) (346) R=C 2 H 5
(347a) R=CH 3

Br
9 Ell ~CH 20Ac
(C6 H,).PCH 2 ""<::::::
~ ~CN
(C 2H,O).PCH 2 ""<::::::
(348) (349)

~C0 2CH 3 (C6 H 5 ) 3 P=CH

~CN
(C6 H,).P==CH ""<::::::
(350) (351)
384 H. MAYER and 0. ISLER

Wittig reaction of (180) with (4,4-diethoxy-2-methyl-2-butenyl)triphenyl-

phosphonium bromide (283 b) [124] produced retinal (325 a). The latter
compound has also been obtained by the enol ether condensation of P-C 15 -
aldehyde diethyl acetal with 1-ethoxy-3-methyl-1,3-butadiene (177) [124], by
the base-catalysed condensation of(180) with 4,4-dimethoxy-2-butanone (181)
followed by reaction with CH 3 Mgl and acid hydrolysis [228], and by
Oppenauer oxidation of a mixture of P-ionylideneethanol (187) and p,p-di-
methylallyl alcohol [229]. The Horner condensation of the isomeric 3,4-di-
dehydro-C15-aldehydes (213 a) and (213 b) with a mixture of the phosphonates
(347 a) and (347 b) (Table 3) produced all-trans- (340a) and 9-cis-vitamin A 2
acid (340c) [193]. Treatment of 3,4-didehydro-Cwaldehyde (213) with the
phosphoranes (350) and (351) in acetonitrile resulted in the formation of
vitamin A 2 acid methyl ester and vitamin A 2 acid nitrile, respectively [230].

PJ (P-Ionylideneethyl)triphenylphosphonium halide routes. The technical

manufacturing procedure of vitamin A developed by Pommer [36] in the
laboratories of the Badische Anilin- & Soda-Fabrik AG, Ludwigshafen, was
based on the Wittig reaction of a (P-ionylideneethyl)triphenylphosphonium
halide, e.g. the chloride (204) (see Section B.7dy), with y-acetoxytiglaldehyde
(352) in the presence of sodium methoxide [159, 163, 231]. The crude reaction
product was a mixture of all-trans- (309a) and 11-cis-vitamin A acetate (309b)
from which the crystalline all-trans isomer could be obtained by subsequent
isomerization and crystallization.
Vitamin A acid (305 a) and the corresponding ethyl ester (306) have been
prepared accordingly by employing P-formylcrotonic acid (353 a) and ethyl
P-formylcrotonate (354), respectively [159, 163, 231, 232]. The formation of
9-cis-vitamin A acid (305 b) was claimed when cis-P-ionylideneethanol (187 b)
was used for the preparation of the CwWittig salt [159].

OHC
~C0
"""
2R
(204) (352) (353a) R=H

j
(354) R=C 2 H 5

CH2 0Ac
"":::, "":::, "":::,
+

(309a) (309b) H 20Ac

 VI. Total Syntheses 385

(205)
+
·~ 0
(355)

j
+
"'<:::::: C0 2 H 0 2H

(305e) (305 c)
11,13-di-cis 13-cis

(356) (357)

Starting from (3,4-didehydro-{3-ionylideneethyl)triphenylphosphonium

bromide (218), an analogous reaction sequence was employed for the synthesis
of all-trans- (337 a) and 13-cis-vitamin A 2 (337 d) via the corresponding acids
(340a) and (340b), respectively [165].
Interestingly, the formation of 13-cis- (305 c) and of the hitherto unknown
11,13-di-cis-retinoic acid (305 e) has recently been achieved [233-235] b-y the
Wittig condensation of ({3-ionylideneethyl)triphenylphosphonium bromide
(205) with 4-hydroxy-3-methyl-2-buten-4-olide (355). The pure isomers were
obtained by fractional crystallization. By controlled stereomutation the di-cis
isomer could be isomerized to all-trans- (305 a) and 13-cis-retinoic acid (305 c),
respectively.
The preparation of vitamin A acid ethyl ester (306) and of vitamin A
(307 a) has been claimed by the condensation of ethyl {3-formylcrotonate (354)
with the Grignard derivatives ofthe acetylenic C 15 -compounds (356) and (357),
respectively [236, 237].
'
f) C16+ C4= C2o
The synthesis of vitamin A developed by the laboratories of Distillation
Products Ltd., Rochester, follows the scheme C 16 + C 4 = C 20 [174]. A Grignard
reaction of the Cwacetylenic carbinol (237) with 4,4-dimethoxy-2-butanone
(181) gave the C 20-diol acetal (358), which was partially hydrogenated over

Carotenoids 25
 386 H. MAYEll and 0. ISLER

ex+¢ ~CH(OCH 3 h
I
+
0

(237) (181)

l. .CH(OCH 3h
<9-'6~
H --

(358) (359)

~ ~ ~ CHO

(325a)

a palladium-on-charcoal catalyst to the diol acetal (359). Treatment of the

latter with quinoline hydrochloride in ethyl methyl ketone furnished retinal
(325 a) which could be purified via a crystalline hydroquinone adduct [238,
238a] and reduced to vitamin A with sodium borohydride [239].
Vitamin A aldehyde (325 a) has been synthesized in a similar way from
the acetylenic carbinol (237) and 4-methoxy-3-buten-2-one (417) (Section C.l)
[240] or from the P-C 16 -hydrocarbon (238) (Section B.8b) and 4,4-dimethoxy-
2-butanone (181) [175].

g) C1s+Cz=Czo
The P-C 18 -ketone (275) was the starting material of choice for the pre-
paration of many vitamin A compounds and intermediates according to this
scheme.
In a procedure of N.V. Philips' Gloeilampenfabrieken, the P-C 18 -ketone
(275) [241] or its Schiff base [137] was condensed with cyanoacetic acid
yielding vitamin A acid nitrile (361) via the cyanovitamin A acid (360) [136].
Reduction of the nitrile (361) with diisobutylaluminium hydride gave retinal
(325 a) after hydrolysis of the aldimine intermediate.
Vitamin A aldehyde has also been prepared from P-C 18 -ketone by the
following reaction sequence: condensation with ethyl formate, methylation
of the enolate, acetalization followed by a Grignard reaction with methyl-
magnesium chloride, hydrolysis and dehydration [149].
 VI. Total Syntheses 387

I C,s+C2=C2o I

"""" """"
""'o
-- CN
"""" """" """"C0 H
2

(275) (360)
1
CHO CN
"""" """" """" .._ """" """" """"
(325 a) (361)

All-trans- (325 a) and 9-cis-vitamin A aldehyde (325 c) have been obtained

from cis- and trans-P-C 18 -ketone and ethoxyacetylene [242, 243].
A large number of vitamin A compounds were synthesized from the
P-C 18 -ketone (275) by condensation with various C 2 -triphenylphosphonium
bromides listed in the following chart [36].. The yields reported were low
because of the poor reactivity of the P-Cwketone. 11-cis-Vitamin A acid
methyl ester has also been obtained recently by the Horner condensation of
11-cis-P-C18 -ketone and methyl (diethylphosphono)acetate [196].
The Reformatskii reaction with bromoacetic acid esters has been used
by several workers for the synthesis of vitamin A from the P-Cwketone (275)
[126, 127, 189,201,204, 244]. Dehydration of the resulting hydroxy ester (362)

(275)
R
Ell
Br 9 (C 6 H 5 hPCH 2 CH 2 0Ac ------+ CH 20Ac
Ell
Br 9 (C 6 H 5 hPCH 2CH 20H ------+ CH 2 0H
Ell

~}eN
Br 9 (C 6 H 5 hPCH 2 CN
0
II
(C 2H 5 0hPCH 2 CN
Ell
Br 9 (C 6 H 5 ) 3 PCH 2 C0 2 C 2 H 5 ------+ C0 2 C 2H 5
Ell
Br 9 (C 6 H 5 ) 3 PCH 2 CONH 2 ------+ CONH 2
Ell
Br 9 (C 6 H 5 hPCH 2CH(OC 2 H 5 ) 2 ------+ CH(OC 2 H 5 h
388 H. MAYER and 0. ISLER

co,c,H,

j
(362) (364)

co,c,H, co,c,H,
~ ~ ~ c::P' c::P' c::P'
+

(306) (363)

gave mainly retro-vitamin A acid ethyl ester (363). Hydrolysis to the corre-
sponding acid followed by treatment with PC1 3 furnished a mixture of isomeric
vitamin A acid chlorides which could be reduced directly with lithium
aluminium hydride to vitamin A or hydrolysed to vitamin A acid.
The C 20-acetylenic carbinol (364) required for a synthesis of P-carotene
was readily available from (275) and lithium acetylide [245].

h) C19+ C1 = C2o
Treatment of the P-C 19-aldehyde (285) with acetone cyanohydrin furnished
the C20 -cyanohydrin (365), which was dehydrated to vitamin A acid nitrile
(361) with phosphorus oxychloride and pyridine [245a]. The conversion of
this nitrile into retinal has already been discussed.

CN

OH

(365)

i) Retinyltriphenylphosphonium halides
P-Retinyltriphenylphosphonium halides are important C 20-building units
for the synthesis of P-carotene and P-apo-carotenoids. They could be prepared
by relatively simple procedures starting from vitamin A or vitamin A acetate.
Thus P-retinyltriphenylphosphonium chloride (367) was obtained by treat-
ment of vitamin A (307 a) [246, 247] with triphenylphosphonium chloride or
triphenylphosphine and ethanolic or methanolic HCl in ethanol, methanol
or dimethylformamide. Vitamin A methyl ether (366) [248], anhydro- (370)
[246] and 13-cis-vitamin A (307 d)' [249] have also been employed. Treatment
 VI. Total Syntheses 389

"""" CH2 0R

(307a)R=H (367) X=Cl

(309a)R=Ac (368) X=l
(366) R=CH 3 (369) X=HS0 4

(370) (371)

of vitamin A with triphenylphosphine, potassium iodide and ethanol-hydrogen

chloride afforded the corresponding iodide (368) [250]. When vitamin A
acetate (309a) was stirred in methanol with triphenylphosphine and concen-
trated sulphuric acid the sulphate (369) was formed [251, 252]. The p-toluene-
sulphonate and fluoroborate have been prepared similarly [251].
The retro-C 20 -triphenylphosphonium bromide (371) has also been used
as a C 20 -component for synthesizing fJ-carotene. It has been prepared from
the corresponding retro-retinyl bromide and triphenylphosphine [253].

j) fJ-Retinylphosphonate
The synthesis of fJ-retinylphosphonate (374) required for a synthesis of
fJ-carotene has recently been reported by Surmatis and Thommen [254, 255].
Polyene phosphonates were usually prepared by the Michaelis-Arbuzov

~CH 2 Br
~*
l ) l ~r
(372)

0
I II
~*~CH2 P(OC2 Hsh

u"""'
(373) (374)
"""" 0II
H 2 P(OCzHsh
 390 H. MAYER and 0. ISLER

reaction from appropriate halides and triethyl phosphite. As there is no

known procedure, however, for preparing a halide from vitamin A, the follow-
ing reaction sequence was devised. The C 20 -diol (331 a) (see Section B.l2d),
which is an intermediate of the industrial vitamin A synthesis of F. Hoffmann-
La Roche & Co. Ltd., was treated with phosphorus tribromide to yield the
rearranged dibromide (372). Treatment of this dibromide with triethyl phos-
phite gave the phosphonate (373), which was partially hydrogenated over
Lindlar catalyst to P-retinylphosphonate (374).

k) 4-0xygenated vitamin A compounds

The C 20 -components described in this section are readily accessible from
retinal or vitamin A acetate by partial synthesis. They were used for the
synthesis of 4-oxygenated carotenoids, e.g. echinenone (148), isozeaxanthin
(71) and canthaxanthin (193). Thus, bromination of retinal (325 a) with
N-bromosuccinimide in methylene chloride and acetic acid yielded 4-acetoxy-
retinal (375) [167], which on sodium borohydride reduction gave 4-acetoxy-
vitamin A (377) [256]. 4-Hydroxyretinal (376) was obtained when the retinal
enol acetate (378) was treated with selenium dioxide in tetrahydrofuran [257].
The Wittig compounds (379) and (380) were prepared by bromination of
P-retinyltriphenylphosphonium sulphate (369) with N-bromosuccinimide in
the presence of acetic acid [167] and by treatment of 4-acetoxyvitamin A (377)
with triphenylphosphine and methanolic hydrogen chloride [256], respectively.

~
CHO
~ ~ ~

OR OAc
(375) R=Ac (377)
(376)R=H

CHOAc
-:::? -:::? -:::? ,;?"

(378) (379) X=HS0 4

(380) X=Cl

13. C 21 -Components
The synthesis of a novel type of 3-oxygenated C 21 -building units has
been reported recently [111, 167]. Condensation of 3,3-ethylenedioxy-{1-ionone
(112) with the C 8 -acetal (381) by means of a Grignard reaction furnished the
 VI. Total Syntheses 391

(112) (381)

j
(~,'-~CH(OC,H,),
~o ~, 6c,H.
(382)

j
~"~uo
(383)

(384)

product (382), which was subsequently hydrolysed with sodium acetate, acetic
acid and water to yield the C 21 -keto aldehyde (383). Selective hydrogenation
then gave the fully conjugated C 2 rketo aldehyde (384).

C 1 s + C 3 = C21 ; C19 + C 2 = C 21 . The C 21 -acetylenic carbinols (385) [258],

(386) [259], (387) [118, 186] and (388) [260] were used for syntheses of P-ca-
rotene and unsymmetrical carotenoids. They were prepared from the appro-
priate C 18 -ketones and C 19-aldehydes by the Reformatskii and the Nef reac-
tions, respectively.
392 H. MAYER and 0. ISLER

C,s +C3=C21
I

"""'::: """'::: """':::

OH
"* """'::: """'::: """':::
OH
"*
(385) (386)

C 19 +Cz=Cz,
I

c:P c:P c:P "* c:P c:P c:P

1:--

OH OH

(387) (388)

I C23 +C3 =Cz6 I

"""'::: """'::: """'::: """':::

OH
"*
(389)

14. C 26 -Components
C 23 + C 3 = C 26 . The C 26 -acetylenic carbinol (389), required for a synthesis
of decapreno-/3-carotene (1035), was made by the Reformatskii reaction of
13',14'-dihydro-20'-nor-/3-apo-13'-caroten-14'-one (724, Table 15) with pro-
pargyl bromide [261].

C. Syntheses of Acyclic Components

1. Cz-, C 3 - and C 4 -Components
A large number of carotenoids and vitamin A derivatives as well as numer-
ous suitable intermediates have been synthesized by employing in Wittig and
Horner reactions the readily available C 2 - and Crphosphonium halides,
-phosphoranes and -phosphonates listed in Tables 1 and 2.
The phosphonium salts were prepared by reaction of the appropriate
halides with triphenylphosphine and tris(dimethylamino)phosphine, respec-
tively. On treatment with alkali they were converted into the corresponding
phosphoranes. The phosphonates were obtained from the appropriate halides
by heating with triethyl phosphite and diethoxyphenylphosphine, respectively
 VI. Total Syntheses 393

(Arbuzow reaction). In the case of the C 3 -phosphonates (406) and (408), the
methylation of the corresponding Crphosphonates (196a) and (193), respec-
tively, with methyl iodide in the presence of potassium t-butoxide was used as
an alternative procedure.
The C 4 -building units (181), (417), (419) and (289), mostly used in chain
lengthening by four carbon atoms, were prepared by simple procedures. The
base-catalysed condensation of acetone and methyl formate (414) followed by
treatment with methanolic hydrochloric acid gave 4,4-dimethoxy-2-butanone
(181) [278]. Subsequent heating in the presence of sodium methoxide resulted
in the enol ether (417) [279]. Both components have been applied to the syn-
thesis of retinal (325 a) [174, 240].
Condensation of ethyl propenyl ether (415) and triethyl orthoformate (416)
in the presence of boron trifluoride afforded methylmalonaldehyde bis(diethyl
acetal) (418). This was either treated with sulphuric acid followed by benzoyla-
tion of the product yielding the enol benzoate (419) [208], or converted into
the enol ether (289) by treatment with aqueous p-toluenesulphonic acid [280].

Table 1. C 2 -Components

Compound References
ED
Br 9 (C 6 H 5 ) 3 PCH 2 C0 2 CH 3 (390) (262]
(C 6 H 5 ) 3 P=CHC0 2 CH 3 (391) (262]
ED
Br 9 (C 6 H 5 hPCH 2 C0 2 C 2 H 5 (392) [262-265]
ED
Cl 9 (C 6 H 5 hPCH 2 C0 2 C 2 H 5 (393) [266]
(C 6 H 5 hP=CHC0 2 C 2 H 5 (394) (262, 264-266]
(C 6 H 5 hP=CHCHO (394a) [267]
ED
Cl 9 (C 6 H 5 hPCH 2 CH 2 0Ac (395) [264]
ED
Cl 9 (C 6 H 5 hPCH 2 CH 2 0H (396) [264]
ED
Cl 9 (C 6 H 5 hPCH 2 CN (194) [266,268]
ED
Br 9 (C 6 H 5hPCH 2 CN (397) [269]
(C 6 H 5 ) 3 P=CHCN (398) (266,268,269]
ED
Cl 9 (C 6 H 5 hPCH 2 CONH 2 (399) (266,268]
(C 6 H 5 hP=CHCONH 2 (400) [266, 268]
(C 2 H 5 0h0PCH 2 C0 2 C 2 H 5 (196a) (211, 270, 271]
(C 2 H 5 0hOPCH 2 C0 2 CH 3 (196b) [234]
(CH 3 0h0PCH 2 C0 2 CH 3 (196c) [27]
(C 2 H 5 0h0PCH 2 CN (193) [138, 270, 271]
(C~H 5 )(C 2 H 5 0)0PCH 2 CN (195) [140]
(C 2 H 5 0h0PCH 2 CH(OC 2 H 5 h (199) [271]
394 H. MAYER and 0. ISLER

Table 2. C 3 -Components

Compound References
CH 3
ml
Br 9 (C 6 H 5hPCHC0 2 CH 3 (401) [262]
CH 3
I
(C 0 H 5 hP=CC0 2 CH 3 (402) [262]
CH 3
ml
Br 9 (C 6 H 5 ) 3 PCHC0 2 C 2 H 5 (403) [262]
CH 3
I
(C 6 H 5 hP=CC0 2 C 2 H 5 (404) [262]
CH 3
ml
Br 9 [(CH 3 hN] 3 PCHC0 2 C 2 H 5 (405) [247]
CH 3
I
(C 2 H 5 0h0PCHC0 2 C 2 H 5 (406) [211, 272]
CH 3
I
(C 2 H 5 0h0PCHC0 2 CH 3 (407) [273]
CH 3
I
(C 2 H 5 0h0PCHCN (408) [138]
m
Cl 9 (C 6 H 5 hPCH 2 COCH 3 (409) [274]
(C,H 5 hP=CHCOCH 3 (410) [274]
(C 2 H 5 0h0PCH 2 COCH 3 (411) [271, 275]
m
Br 9 (C 6 H 5 hPCH 2 C=:CH (412) [276]
(C 2 H 5 0h0PCH 2 C=:CH (413) [277]

2. C 5 -Components
a) y-Acetoxytiglaldehyde
Several procedures have been devised for the synthesis of this important
C 5 -building unit.
1) Treatment of a methanolic solution of propargylaldehyde (420), made
from propargyl alcohol [21, 281], with boron trifluoride etherate and mercuric
oxide gave methylglyoxal dimethyl acetal (421 a) [21, 282]. Subsequent ethy-
nylation yielded the ethynyl carbinol (422), which on partial hydrogenation
was transformed into 1,1-dimethoxy-2-methyl-3-buten-2-ol (423 a) [283].
Reaction of the latter with phosgene or thionyl chloride [284, 285] afforded
y-chlorotiglaldehyde (424), which on treatment with sodium or potassium
acetate gave y-acetoxytiglaldehyde' (352) [286].
 VI. Total Syntheses 395

(414) (181) (417)

C,H,( + HC(OC 2 H 5 ), --+ (C 2 H 5 0),HC

~CH(OC H )z
2 5

(415) (416) (418)

/ 1
OHC~CHOC2H5 OHC~CHOCOC6H5
(289) (419)

Alternatively, y-acetoxytiglaldehyde (352) was obtained directly by heating

the tertiary acetate (423 b) in the presence of copper powder, Cu(l) or Cu(II)
compounds [287].
2) A convenient route investigated by Weedon's team [288] started from
methyl /3-methylcrotonate (425), which on bromination with N-bromosuccin-
imide gave methyl y-bromo-f3-methylcrotonate (170). A Krohnke reaction of
the latter gave methyl trans-/3-formylcrotonate (426) [289] without detectable
loss_of stereochemistry about the double bond. The methyl ester was converted
into the corresponding acetal (427 a), which was reduced with lithium alumini-
um hydride to the primary alcohol. Acetylation of the latter and subsequent
acid hydrolysis of the resulting acetate then gave the desired y-acetoxytigl-
aldehyde (352).
3) The base-catalysed condensation of propionaldehyde (429) with the
hemiacetal of ethyl glyoxylate (430) in the presence of di-n-butylamine gave
ethyl /3-formylcrotonate (354) [290]. This compound was transformed into
y-acetoxytiglaldehyde (352) following the lines already described with methyl
/3-formylcrotonate (426) by reduction of the corresponding acetal (428) with
LiAlH 4 , Na[A1H 2 (C 2 H 5 )(0C 2 H 5 )] or Na[AlH 2 (0C 2 H 5 h] [36, 291], sub-
sequent acetylation and acid hydrolysis [36].
4) An interesting approach to the synthesis of various C 5 -building units has
been reported by Makin and collaborators [124]. 1-Ethoxy-2-methyl-1,3-
butadiene (286) and 1-ethoxy-3-methyl-1,3-butadiene (177) were utilized as
key intermediates which were prepared by the condensation of 1,1-diethoxy-
ethane (431) and 2,2-diethoxypropane (433), respectively, with propenyl (415)
396 H. MAYER and 0. ISLER

~
oucc=cu ~C0 2CH3 OHj

(420) (425) (429)

l l HO
+

\.
(RO),HC~O BrCH 2
~C02CH 3 /
CHC0 2C2H5
C2HsO

(421 a) R= CH 3 (170) (430)

/
(42Ib) R=C 2 H 5

l l
(CH,0) 2HC~ :;::,. OHC
~C02R
OH "'<:

(422) (426) R=CH 3

(354) R=C 2 Hs

l l
(CH30),H~
OR

(423a) R=H (427a)R=CH 3

(423b) R=Ac (428) R=C 2 H 5

l ~~
(424) (352)
 VI. Total Syntheses 397

~
(C 2H 5 0},HCCH 3

(431)
+
~ OC2Hs
(415)
- 0CH(OC2Hsh
OC 2 H 5
(432)
- ~CHOC2H 5
(286)

*OCzHs
OC2Hs

(433)
+ ~OCzHs

(250a)
- ~CH(OCzHslz
OCzHs
(434)
- ~CHOC 2 H 5
(177)

(C2H 5 0) 2 H
~CH2Br
(435)
- (C 2 H 5 0},HC
~CH
"""
(436)
2 0CzHs

1 1
(CzHsOhH
~CH 2 0Ac OHC
~CH 2 0CzHs
(438) (437)

1
OH~CH 2 0Ac
(352)
- (CH 3 0},H
~CH 2 0Ac
(439)
- (CH 3 0),H
~CHO
(440)

and vinyl ether (250a) in the presence of boron trifluoride etherate. The initially
formed 1,1,3-triethoxyalkanes (432) and (434) were then pyrolysed to give the
desired butadienes (286) and {177), respectively.
y-Acetoxxtiglaldehyde (352) has been obtained from 1-ethoxy-2-methyl-
1,3-butadiene (286) by bromoethoxidation with N-bromosuccinimide in
anhydrous ethanol yielding the bromo acetal (435), which was transformed into
the acetate (438) by reaction with potassium acetate. Acid hydrolysis then led
to (352) [124].
5) y-Acetoxytiglaldehyde (352) could also be prepared by treatment of
isoprene with t-butyl hypochlorite in acetic acid [292] followed by Sommelet
reaction of the product [293].
398 H. MAYER and 0. ISLER

b) 4,4-Dimethoxy-3-methyl-2-butenal
Acetalization of y-acetoxytiglaldehyde (352) with trimethyl orthoformate
gave the corresponding acetal (439). Subsequent alkaline hydrolysis and man-
ganese dioxide oxidation of the product furnished the C 5 -building unit (440)
which was used for a synthesis of the C 10 -aldehydic ester (536) (Section C.7)
[294].

c) {3-Formylcrotonates
The synthesis of methyl {3-formylcrotonate (426) from methyl {3-methyl-
crotonate (425) and of ethyl {3-formylcrotonate (354) from propionaldehyde
(429) and the hemiacetal of ethyl glyoxylate (430) has already been mentioned
(Section C.2a). Similarly, n-butyl {3-formylcrotonate has been obtained by the
condensation of propionaldehyde with n-butyl glyoxylate [165]. The diethyl
acetal of ethyl {3-formylcrotonate (428) could also be prepared by the conden-
sation of methylglyoxal diethyl acetal (421 b) and ethyl (diethylphosphono)-
acetate (196 a) [211].

d) trans-{3-Formylcrotonic acid
trans-{3-Formylcrotonic acid (353 a) was prepared by treatment of 4-hy-
droxy-3-methyl-2-buten-4-olide (355) with sodium methoxide in methanol
followed by acidification with hydrochloric acid [234] or by alkaline hydrolysis
of the diethyl acetal of ethyl {3-formylcrotonate (428) and subsequent removal
of the acetal grouping [246].
The lactol (355) could be synthesized by three different procedures:
1) by acid hydrolysis of methyl {3-formylcrotonate (426) [234, 295];
2) by condensation of methylglyoxal diethyl acetal (421 b) with methyl
(diethylphosphono )acetate (196 b) yielding an isomeric mixture of acetals (427 a)
and (427 b) which on acid hydrolysis readily furnished the lactol (355) [234];
3) by bromination with N-bromosuccinimide of 3-methyl-2-buten-4-olide
(441) followed by hydrolysis of the initial bromide (442) [295].
The lactol (355) can also be regarded as the cyclic form of cis-{3-formyl-
crotonic acid (353 b) which, however, could not be obtained as such, but
cyclized spontaneously [295].

e) y-Ethoxytiglaldehyde
y-Ethoxytiglaldehyde (437) has been obtained from the bromo acetal (435)
by treatment with sodium ethoxide followed by acid hydrolysis of the resulting
acetal (436) [124].

f) Methyl trans-f31ormyl-rx-methylacrylate
This compound was prepared from methyl rx-methylcrotonate (443 a).
Bromination with N-bromosuccinimide gave methyl y-bromo-rx-methyl-
crotonate (444), which was transformed into the desired formyl ester (445) by
the Krohnke procedure [234].
 VI. Total Syntheses 399

- OHC
h C0 2H

(353 b)

~' C02 H
OHC~ (C2H 50),HC~
C0 2 R

(353 a) (427b) R=CH 3

0
II

(C2HsO),HC A 0
(C2Hs0) 2PCH 2C0 2R

r
(421 b) (196a) R=C 2 H 5
(196b) R= CH 3
(C2H 5 0),HC
~C0
""""
2R
(427 a) R = CH 3
(428) R=C 2 H 5

~0 0

(441) (442)

~C0 2 CH 3 ~C0 2 CH 3
~C0 2 R - BrCH2 I - OHC I
(443a) R=CH 3 (444) (445)
(443 b) R = C 2 H 5

g) C 5 -Phosphonium salts and -phosphonates

A number of C 5 -phosphonium salts and -phosphonates listed in Table 3
have been employed as convenient building units for the attachment of suitable
end groups and for chain lengthening by five carbon atoms.
The phosphonium salts were readily prepared by reaction of the appro-
priate halides with triphenylphosphine in suitable solvents. Conversion into
the corresponding phosphoranes was effected by treatment with alkali (sodium
hydroxide, triethylamine, etc.). The phosphonates were obtained by heating of
400 H. MAYER and 0. ISLER

Table 3. C 5 -Phosphonium salts and -phosphonates

I
Compound References

Br e (C6 H 5 hPCH
$ ~
2 "":::: C0 2CH 3 (345) [58, 227, 296]

(C,H,O),OPCH,~co,cH, (347 a)
[234, 296]
(C 2 H 5 0h0PCH~C02CH 3
(347b)

(C 2 H 5 0h0PCH 2 ~C0 2 C 2 H 5 (346) [211]

Br 9 (C 6 H 5 hPCH 2 ~CH(OC 2 H 5 h (446) [124]

(C 6 H 5 hP=CH~CH(ORh (446a) [297]

E9
Br 9 (C 6 H 5 hPCH 2 ~C0 2 CH 3 (447) [298, 299]

(C 2 H 5 0h0PCH 2 ~C0 2 CH 3 (448) [300]

E9
Cl 9 (C,H,),PCH,-"(CHO (449))
[301]
(C 6 H 5 hP=CH~CHO (450)

Cl 9 (C 6 H 5 h~CH 2 ~CH :::g~ [302]

(450a) R=CH 3 ; (450b) R=isopropyl

$
X 9 (C 6 H 5 hPCH 2 ~CH(OC2H 5 h [124]

(283a) X=Br; (283b) X=Cl

(C 2 H 5 0h0PCH 2 ~CH(OC 2 H 5 h (451) [124]

E9
Br 9 (C 6 H 5 hPCH 2 ~ (452) [303]
E9
Br 9 (C6 H 5hPCH 2 T (453) [296, 304]
EB -¢
Br 9 (C 6 H 5 hPCH 2 ~ (453a) [305]
e $~
Br (C 6 H 5 hP 11"" C02CH 3 (454))
[306, 307]
(C 6 H 5 hP~C0 2 CH 3 (455)

(C 6 H 5 hP=CHi< (456) [308,309]

OCH 3
 VI. Total Syntheses 401

the appropriate bromides with triethyl phosphite (Arbuzow reaction). The

bromide (345) could also be obtained from methyl y-hydroxy-fJ-methylcroto-
nate and triphenylphosphonium bromide [58]. When methyl y-bromo-fJ-
methylcrotonate (170) was subjected to the Arbuzow reaction, a cis-trans-
isomeric mixture ofphosphonates (347 a) and (347 b) was obtained which could
be separated into the pure isomers by fractional distillation and preparative
gas-liquid chromatography [234, 296]. The hemiacetals (450a) and (450b) were
made by treatment of the Wittig compound (449) with isopropanol or trimethyl
orthoformate, respectively, in the presence of p-toluenesulphonic acid [302].
The phosphorane (456) was synthesized from 1,1,1-trichloro-2-methyl-2-
propanol (457) in four steps. Treatment with aqueous potassium hydroxide and
methanol gave cx-methoxyisobutyric acid (458), which on reaction with oxalyl
chloride was transformed into cx-methoxyisobutyryl chloride (459). Refluxing
of the latter with the methylenephosphorane (460) followed by addition of
aqueous sodium hydroxide then furnished the phosphorane (456) [309].

- -
0
Cl~
f<oCH,
Cl3 C'-,C H0 2 C'-,C
I"OH I"OCH,
(457) (458) (459)

1 (C6 H 5 ),P==CH2
(460)

C2H 5COCI (C.H 5 ),P==CH ~ 3

~OCH
(456)

1 CH2==CH2

- (462)

(463)
- 0 + C H20CH 3
~ OH
(464)

Carotenoids 26
402 H. MAYER and 0. ISLER

h) Other C 5 -components
The C 5 -component 1-diethylamino-3-pentanone methiodide (462) was used
for a synthesis of echinenone (148) and canthaxanthin (193). It was synthesized
by reaction of propionyl chloride with ethylene in the presence of aluminium
chloride yielding 1-chloro-3-pentanone (461), which on treatment with diethyl-
amine and subsequent quatemization with methyl iodide gave the methiodide
(462) [310].
The synthesis of the acetylenic carbinol (464) was achieved by ethynylation
with lithium acetylide of 1-methoxy-2-propanone (463), which was obtained
by oxidation of 1-methoxy-2-propanol or by reaction of methoxyacetonitrile
with methylmagnesium bromide [206].

[;]
0~
(465)
- ~~
(466)
\~~,OH
~

(I 58 a)

~CH 2 0CH3
~
¢~ H 2 0H
(467) (158b)

- -
OH
OH~CHOC 2 H, ~CHOC2H, ~CH(OC 2 H 5 ) 2
~ ~

(468) (469) (470)

0
~CH 2Cl

(471)
- ~
~CH 20H

(472)
- ~
~CHO

(473)

l
~C0 2 H
~

(47'1)
 VI. Total Syntheses 403

3. C 6 -Components
The synthesis of 3-methyl-2-penten-4-yn-1-ol (158) was performed according
to the following scheme: the condensation of methyl vinyl ketone (465) with
sodium acetylide in liquid ammonia afforded 3-methyl-1-penten-4-yn-3-ol
(466). Subsequent rearrangement in the presence of sulphuric acid led to an
isomeric mixture of 3-methyl-2-penten-4-yn-1-ols (158a and 158b) [311], which
could be separated into the pure isomers by fractional distillation. The pre-
dominant lower boiling isomer possessing the cis configuration (158 b) [218,
312] represents the important C6 -building unit of Isler's industrial manu-
facturing procedure of vitamin A [1, 217].
1-Methoxy-3-methyl-2-penten-4-yne (467) was employed as a C6 -building
unit in the first total synthesis of vitamin A methyl ether (366) by Isler et al.
[313, 314]. It was synthesized by treatment of 3-methyl-1-penten-4-yn-3-ol
(466) with phosphorus tribromide [314] or hydrochloric acid [216] followed by
reaction of the intermediate primary halogenide with sodium methoxide.
Alternatively, (467) could be obtained directly from (466) by treatment with
methanolic sulphuric acid [313, 314].
The C6 -building unit (470), the so-called 'C 6 -acetal', was required for the
technical synthesis of P-apo-carotenals (Section E.1 b p). It was synthesized by
the condensation of sodium acetylide with methylmalonaldehyde enol ether
(468), yielding the acetylenic carbinol (469) [280], which was converted into
(470) by treatment with triethyl orthoformate in the presence of phosphoric
and p-toluenesulphonic acid [280].
The acetylenic C6 -alcohol (472), used by Ahmad and Weedon [315] in a
synthesis of all-trans-methylbixin (266), was prepared by the treatment of the
epichlorohydrin (471) with sodium acetylide [315]. Manganese dioxide oxida-
tion of (472) afforded the corresponding aldehyde (473), whereas with chromic
acid the acid (474) was formed.

4. C 7 -Components
The unsaturated C 7 -aldehydic ester (317 a) was synthesized starting from
ethyl P-formylcrotonate (354). Its diethyl acetal (428) was condensed with
ethyl vinyl ether (250a) in the presence of boron trifluoride etherate yielding the
triethoxy ester (475), which was hydrolysed with p-toluenesulphonic acid
[36, 316]. Alternatively, the cyclic acetal (476) was similarly reacted with ethyl
vinyl ether and the reaction mixture hydrolysed with 85 % phosphoric acid
[148]. The C 7 -acetoxy aldehyde (319a) was prepared by lithium aluminium
hydride reduction of the cyclic-acetal ester (477), followed by acetylation and
hydrolysis of the acetal grouping [36].
Alternatively, the C 7 -component (319a) was obtained by vinyl ether
condensation of the C 5 -acetal (438) giving the intermediate triethoxy compound
(479), which was hydrolysed with acetic acid [124].
The C 7 -methoxy aldehyde (329) was synthesized in a similar fashion
starting from the C 5 -acetal (478) via the tetraalkoxy compound (480) [124].
404 H. MAYER and 0. ISLER

~COzC 2 Hs ./'--_ l_~,COzCzH 5

(C 2 H 5 0),HC "'-'::::: (C2 H 5 0)zHC' ~

OCzHs
(428) (250a) (475)

l
[>w-L-co,c,u, OH~C02R
(476) (250a) (317a) R=C 2 H 5
(317b) R=H

1
OH~CH 2 0R Cd'-CH~COzCzHs
(319a) R=Ac
(319b) R=H (477)
(329) R=CH 3

I
./'--_~J__~,CH 0R2 ~OC 2 H,
(C 2 H,O),HC
~CH
"'-':::::
20R
(C 2 H 5 0)zHC'
OC 2 H 5
(479) R=Ac (250a) (438) R=Ac
(480) R=CH 3 (478) R=CH 3

~CH 2 0CH 3 ~CH 2 0CH 3

HOCH 2 / '* BrCH 2/ '*
(481) (328 a)

The C 7 -bromo ether (328a) was applied in a synthesis of 11,12-didehydro-

vitamin A methyl ether (Section B.12c) [216]. It was formed by the reaction of
the Grignard complex of the acetylenic ether (467) with formaldehyde, yielding
the intermediate hydroxy ether (481), which was transformed into (328a) by
treatment with phosphorus tribroinide in pyridine [216].
 VI. Total Syntheses 405

The synthesis of 1-methoxy-3-methyl-1,3-hexadien-5-yne (322), required for

a novel synthesis of retinal (325 a), has been reported by Jacobs et al. [212].
Condensation of 4,4-dimethoxy-2-butanone (181) with propynyllithium [212]
or propynylsodium [317] in liquid ammonia, or with propynylmagnesium
bromide [317] gave the acetylenic C 7 -carbinol (482). Addition of ethyl vinyl
ether in the presence of p-toluenesulphonic acid afforded the C 7 -diacetal (483),
which, on treatment with potassium amide in liquid ammonia, was converted
into the acetylenic ether (322). The product proved to be a mixture of the all-
trans and the 1-trans-3-cis isomers.
6-Ethoxy-4-methyl-4-hexen-1-yn-3-ol (327) used for a synthesis of vitamin A
ethyl ether (Section B.12c) was prepared by the condensation of y-ethoxy-
tiglaldehyde (437) with lithium acetylide [124].
The synthesis of the C 7 -component (330) has recently been reported [177].
Condensation of the Grignard complex of propargyl alcohol tetrahydro-
pyranyl ether (484) with methyl vinyl ketone (465) gave the intermediate (485),
which was hydrolysed to the acetylenic diol (486). Jones oxidation of the latter
then furnished the desired aldehyde (330).
The 4-cis-2-trans aldehydic ester (490a) used for a synthesis of natural
methylbixin (266a) has recently been synthesized by Weedon's team [27, 288].
Condensation of the lactol (355) with the phosphonate (196c) furnished the
2-cis-4-trans-3-methylmuconic acid monoester (487), which could also be
obtained by treating 3-nitro-p-cresol with concentrated sulphuric acid to give
the lactonic acid (488), followed by esterification and alkaline treatment of the
resulting ester (489). The half acid (487) was transformed into the corresponding
acid chloride, and subsequent reduction of the latter with lithium tri-t-butoxy-
aluminohydride yielded the aldehydic ester (490a) with complete retention of
stereochemistry. Iodine-catalysed isomerization of (490a) afforded the corre-
sponding all-trans isomer (490b), which was also obtained from (487) by the
imidazolide route.
The C 7 -dialdehyde monoacetal (492) was required for a synthesis of 11',12'-
didehydro-{:1-apo-10'-carotenal diethyl acetal (705). It was synthesized by con-
densation ofthe Grignard complex of trans-3-methyl-2-penten-4-yn-1-ol (158a)
with triethyl orthoformate yielding the hydroxy acetal (491) [247], which was
converted into (492) by manganese dioxide oxidation [249]. Partial hydrogena-
tion of (491) followed by acid hydrolysis of the product (493) furnished the
C 7 -hydroxy aldehyde (319b).
A patent granted to F. Hoffmann-La Roche & Co. Ltd. describes the syn-
thesis of the C 7 -phosphonium bromide (497) [298]. Condensation of methyl
4-oxo-2-pentenoate (494) with lithium acetylide in liquid ammonia followed by
partial hydrogenation of the product furnished the carbinol (495), which was
transformed into the primary bromide (496). Treatment of the latter with
triphenylphosphine then gave the Wittig compound (497). Alternatively, (497)
could be obtained by the Wittig reaction of y-acetoxytiglaldehyde (352) with
(carbomethoxymethylene)triphenylphosphorane (391) to give the acetoxy ester
(498), which was then treated with triphenylphosphonium bromide [288].
406 H. MAYER and 0. ISLER

CH(OCH3 h
~
/
(181) (482) R=H
(483) R=CHOC 2 H 5

j I
CH,

~ I
THPOCH,C=CH + 0~ ~OCH 3
(484) (465) (322)

j
OH~
~~
(485)R=THP (330)
(486)R=H

u~o RO,CCU,~O
0
II
+ (CH 3 0),PCH 2 C0 2 CH 3
0 0
(355) (196c) (488) R=H
(489) R=CH 3

I
/
CH 302~
CHO
-- CH302C~ CH302~
C0 2 H CHO
(490b) (487) (490a)

The C 7 -phosphonium bromide (499) was made by the reaction of the

hydroxy aldehyde (319b) with phosphorus tribromide followed by treatment
of the corresponding bromide with triphenylphosphine [247], and the C 7 -
phosphonate (500) was prepared by the Arbuzow reaction ofthe corresponding
primary bromide [211].
 VI. Total Syntheses 407

/CH(OC2Hsh

HOCH2~ '* + HC(OC2Hsh ---+ HOCH2~ '*

(I 58 a) (416) (491)

j
HOCH 2~
I tH(OC2Hsh
(493) (492)

1
HOCH2
~CHO
I ~C02CH3 OH
(319b) (494) (495)

j
e I
BrCH 2~
E&
Br (C6 H 5 ),PCH 2 ~
C0 2CH 3 ~ ~ C0 2CH 3

(497) (496)

l
(C6 H 5 ) 3 P===CHC0 2CH 3
AcOCH2~C02CH 3
AcOCH 2
~CHO
_.....;

(498) (391) (352)

Br9 (C6 H 5 ),~CH2~CHO ~ ~C0

(C2H 50).PCH 2 "<::::::
"<::::::
2C 2H 5
(499) (500)
408 H. MAYER and 0. ISLER

5. C 8 -Components
The C 8 -acetylenic acetal (381) (Section B.13) required for a new synthesis
of rhodoxanthin (209) was prepared by vinyl ether condensation of the C 6 -acetal
(470) with ethyl vinyl ether (250a) in the presence of zinc chloride [167].

6. C 9 -Components
The synthesis of the C 9 -triene ketone (31 0) and the C 9 -acetylenic alcohols
(273) and (506) was achieved starting from crotylideneacetone (501). Treatment
with lithium or calcium acetylide gave the ethynyl carbinol (502), which was
partially hydrogenated over a palladium-calcium carbonate catalyst. The
resulting vinyl carbinol (503) was isomerized with 0.05% sulphuric acid to the
fully conjugated isomer (504), which, on oxidation with acetone and aluminium
t-butoxide or phenoxide, or with manganese dioxide, yielded the ketone (310)
[48, 318]. When shaken with sulphuric acid, (502) underwent an allylic rear~
rangement to (273) [318].

~[SJ
~0
(501)
- ~~
(502)
OH ~ - HO~~
(273)
~

/
~
(503)
- HO~
(504)
- 0~
(310)

1
'*
~CH 2 0H
(506)
-- h~
?> OH
(505)

7. C 10 -Components
The acyclic C 10 -components described in this section have proved to be
very valuable building units for the synthesis of various types of symmetrical
and unsymmetrical C 40 -carotenoids, of apo-, diapo- and of retro-diapo-
carotenoids, and several well established routes have been developed towards
their synthesis.
 VI. Total Syntheses 409

Two key intermediates, namely the acetoxy aldehyde (511) and the hydroxy
ester (518), have gained particular importance for the simplification of polyene
synthesis and for the preparation of new potential food colourants.
A Grignard reaction of trans-3-methyl-2-penten-4-yn-1-ol (158 a) with the
ethyl enol ether of methylmalonaldehyde (289), and acidic hydrolysis of the
reaction product (508), furnished the acetylenic hydroxy aldehyde (510), which
was partially hydrogenated over Lindlar catalyst giving, after stereomutation
and acetylation, the acetoxy aldehyde (511) [166, 249]. Alternatively, (510) was

?;- OH~CHOC2 H,
ROCH,~ + I"
(I 58 a) R=H (289)
(507) R=C(CH 3 h0CH 3

IH ,._CHOC,H, /
~CHO

ROCH2~ "*
?:-~ I
ROCH2~ ----.

(508) R=H (510) R=H

(509) R=C(CH 3 h0CH 3 (512) R=Ac

AcOCH 2
~CHO
I I
/ HOCH 2
~,
~~
~~~ I

(511)
~(51~

! !
~CHO ~C02R
RCH, I I HOCH, I I
(513) R=OH (517) R=H
(514) R=Br (518) R=C 2 H 5

!
(515)
410 H. MAYER and 0. ISLER

acetylated first, giving the acetylenic acetoxy aldehyde (512), which was then
partially hydrogenated [247]. Instead of (158 a) the corresponding acetal (507)
has been condensed with (289) in the presence of lithium amide in liquid
ammonia yielding the intermediate (509), which was transformed into the
acetoxy aldehyde (511) as mentioned [319].
Alkaline hydrolysis of (511) gave the hydroxy triene aldehyde (513). This
was converted into the corresponding bromide (514), which on treatment with
triphenylphosphine furnished the C 10 -phosphonium bromide (515) [247].
Silver oxide oxidation of either (511) or (513) led to the hydroxy acid (517)
[320], which could also be obtained by silver oxide oxidation of the acetylenic
acetoxy aldehyde (512) followed by partial hydrogenation of the resulting
hydroxy acid (516) [320].
Acetalization of (511) gave the acetoxy acetal (519), which, on alkaline
hydrolysis and manganese dioxide oxidation of the product, yielded th~
triene dialdehyde monoacetal (520) [247]. Acid hydrolysis of the latter furnished
the dialdehyde (520a) [321]. The same reaction sequence was performed with
the acetylenic acetoxy aldehyde (512). The intermediate acetylenic acetoxy
acetal (521) was hydrolysed and oxidized to give the monoacetal (522) [249].
The synthesis of the hydroxy ester (518) and the corresponding bromides
(529) and (526), respectively, has been achieved by three different procedures:
1) The C10 -hydroxy acid (517) was esterified with diethyl sulphate in the
presence of tris(2-hydroxypropyl)amine [320].
2) Condensation of ethyl 4-oxo-2-pentenoate (523) with lithium acetylide
gave the acetylenic carbinol (524). This, after etherification with dihydropyran,
partial hydrogenation, lithium aluminium hydride reduction and manganese

~CH(OCH,)z ---+ OH~CH(OCH 3 ) 2

Ac0CH 2 I I
(519) (520)

1
OH~CHO

(520a)

~CH(OCzHsh ~CH(OCzHsh

?:-- I ~ I
Ac0CH 2~ ---+ OH~
(521) (522)
 VI. Total Syntheses 411

dioxide oxidation, yielded the aldehyde (525). Subsequent Wittig reaction with
(carbomethoxyethylidene)triphenylphosphorane (402, Table 2) and treatment
of the condensation product with hydrobromic acid gave the bromide (526),
which was immediately converted into the C 10 -phosphonium bromide (527)
[298].
3) Reaction of the C 7 -hydroxy aldehyde (319b) with the phosphonium
bromide (405) [247], with the phosphonate (406) [247] or with the phosphorane
(404) [249] (Table 2) yielded the hydroxy ester (518), which was either oxidized
with manganese dioxide to the corresponding aldehydic ester (528) or trans-
formed into the bromide (529) by treatment with phosphorus tribromide.
From this bromide the C 10 -phosphonium bromides (530), (531) and (532)
as well as the C 10 -phosphonate (533) were prepared by reaction with triphenyl-
phosphine, tris( dimethylamino )phosphine, his( dimethylamino )phenyl phos-
phine and triethyl phosphite, respectively [247].

T ~/CHO
~ OH
O~C0 2 C2Hs -----+ " ~C0 2C2Hs ~?~
'l"

(523) (524) (525)

l
~C02CH3
+-- BrCH 2 I I
(527) (526)

~C0 2 C2Hs OH~C0 2 C 2 H 5

HOCH2 I I
(518) (528)

~C0 2 C2Hs
l ~C02C2Hs
BrCM2 I I RCH2 I I
Ell
(529) (530) R=Br 9 (C 6 H 5 lJP
Ell
(531) R=Br 9 [(CH 3 ) 2 N] 3 P
Ell
(532) R = Br 9 [(C 2 H 5 ),N] 2 (C 6 H 5 )P
(533) R =(C 2 H 5 0),0P
412 H. MAYER and 0. ISLER

Three different reaction paths have been devised for the synthesis of the
useful C10-building units (304), (536) and (308):
1) The C 7 -aldehydic ester (317 a) was first converted into the corresponding
acetal (534) by treatment with triethyl orthoformate in the presence of ammo-
nium nitrate and ethanol. Subsequent propenyl ether condensation led to the
tetraethoxy ester (535), which could also be obtained by propenyl ether con-
densation of the triethoxy ester (475). Acid hydrolysis of the compound (535)
with benzenesulphonic acid then furnished the desired C 10-aldehydic ester
(304) [36, 316].
2) The C 5 -component (440) reacted with the phosphonate (437) in the
presence of sodium methoxide giving the acetal ester (537), which on acid
hydrolysis furnished the Cwaldehydic ester (536) [294].
The C10 -acetoxy aldehyde (308) was readily obtained from (304) by the
following reaction sequence: acetalization, lithium aluminium hydride reduc-
tion, acetylation and removal of the acetal grouping [36].
3) Utilizing the triene dialdehyde monoacetal (520), its transformation into
(536) was effected by silver oxide oxidation followed by esterification and
hydrolysis of the acetal grouping [294].
The C10-aldehydic esters (304) and (536) were required for the preparation
of the C10 -phosphonate (540) [211] and the Cwphosphonium bromide (541)
[294], which were obtained in the usual way by sodium borohydride reduction,
followed by reaction with phosphorus tribromide and treatment of the resulting
bromides (538) and (539) with triethyl phosphite and triphenylphosphine,
respectively.
The C10-tetraethoxy compound (542) used for syntheses of vitamin A and
related compounds was prepared by the condensation in the presence of zinc
chloride of 1-ethoxy-3-methyl-1,3-butadiene (177) with the ethoxy acetal (436)
[124].
The frequently used Cwbuilding unit geranyltriphenylphosphonium
bromide (544) was readily obtained by treatment of geraniol (543) or linalool
(545) with triphenylphosphonium bromide [58, 322, 323] or by reaction of
linalool (545) with phosphorus tribromide, and treatment of the intermediate
geranyl bromide (546) with triphenylphosphine [60, 324].
The C10 -Wittig reagents (550), (557), (558), (566), (569) and (575) are now
comparatively readily available by total synthesis as was demonstrated by the
teams of Weedon and Surmatis.
The synthesis of the C10-Wittig salt (550) used for the synthesis of e.g.
3,4,3',4'-tetradehydrolycopene (17) [323] was achieved in four steps starting
from 2-methyl-3-butyn-2-ol (78). Condensation with diketene and pyrolysis of
the resulting acetoacetate gave the methylheptadienone (547). Condensation
of (547) with lithium acetylide in liquid ammonia resulted in the formation of
3,7-dimethyl-4,6-octadien-1-yn-3-ol (548), which was selectively hydrogenated
over Lindlar catalyst yielding 3,7-dimethyl-1,4,6-octatrien-3-ol (549). Conden-
sation of(549) with triphenylphosphonium bromide then gave the phosphonium
bromide (550) [323].
 VI. Total Syntheses 413

(C 2H 5 0),HC
~C02C2H
"<:::::: "<::::::
5

(534) (535)

j
OHC~CH 2 0Ac OHC~C0 2 R
(308) (304) R=C 2 H 5
(536) R=CH 3
(303) R=H

(CH 3 0),HC~
I CHO I
(440)
I I C02CH,
+ (CH 3 0) 2 HC~
(537)

~ ~C0 2 CH 3
(C 2H 5 0),PCH 2 "<::::::
(437)

BrCH 2
~C0 2 R
"" "" ""
~ ~C0
(C 2H 5 0),PCH 2 "<::::::
"<:::::: "<::::::
2 C2Hs
(538) R=C 2 H 5 (540)
(539) R=CH 3

1
~ /-'.._ l_
(C2H,0)2HC' ~ I ~
,CH20C2Hs

OC2Hs
(541) (542)

A Reformatskii-type reaction of acetone with propargyl bromide in the

presence of aluminium gave the carbinol (551), which, on treatment with
methyl iodide in dimethyl sulphoxide, was transformed into the corresponding
414 H. MAYER and 0. ISLER

~CH2 0H

/ 1
~ OH
-- ~CH2 Br
(545) (546)

+.~
(78)
-- ~0
(547)
-- ~~ (548)
OH ~

l
~$
~ ~ ~ CH 2 P(C6 H 5 )J Br e
--- ~
(549)
OH
(550)

methyl ether (552). The lithium derivatives of (551) and (552) reacted with
methyl vinyl ketone (465) to give the diol (553) and the alcohol (554), respec-
tively. These were reduced with lithium aluminium hydride yielding the trans
ethylenes (555) and (556). Treatment with triphenylphosphonium bromide then
afforded the Wittig salts (557) and (558) [325].
Another synthesis of the triphenylphosphonium salt (558) was based on
6-methyl-5-hepten-2-one (12). The introduction of a methoxy substituent was
achieved by stirring (12) in a solution of sulphuric acid and methanol at room
temperature giving 6-methoxy-6-methyl-2-heptanone (559). Reformatskii reac-
tion with ethyl bromoacetate followed by dehydration with phosphorus
oxychloride in pyridine, or Wittig reaction with (carbethoxymethylene)-
triphenylphosphorane (394) [326], yielded the methoxy ester (560), which was
converted into the unsaturated ester (561) by reaction with N-bromosuccin-
imide followed by dehydrobromination with dimethylaniline. Subsequent
lithium aluminium hydride reduction led to the corresponding primary alcohol
(562), which on treatment with triphenylphosphonium bromide gave the desired
Wittig compound (558) [323].
 VI. Total Syntheses 415

0~ RO,I
~r
0~ OH

(551) R=H (465) (553) R=H

(552) R=CH 3 (554) R=CH 3

j
RO~
OH
(557) R=H (555) R=H
(558) R=CH 3 (556) R=CH 3

r
CH,O,I I CH,O,I I
~CH 2 0H ~C02C2Hs

(562) (561)

r
CH,O,I I
~C02C2H,

(394) (560)

~0
CH,O,I I
~0
(559) (12)

To synthesize OH-chlorobactene (53) and rhodopin (56) the phosphonium

bromide (566) was required [327]. Treatment of 6-methyl-5-hepten-2-one (12)
with 35% sulphuric acid gave 6-hydroxy-6-methyl-2-heptanone (563) [328],
which by a Horner reaction with methyl (diethylphosphono)acetate (196b) was
converted into the hydroxy ester (564). Subsequent reduction with lithium
aluminium hydride gave the glycol (565). Partial bromination of the latter and
reaction of the product with triphenylphosphine yielded (566) [327].
416 H. MAYER and 0. ISLER

0
HO,I I HO,I I
~0 + (C2H,0)2~CH2C0 2CH 3 ~C0 2 CH 3

(563) (196b) (564)

j
HO...._I I
~CH 2 0H

(566) (565)

1
~CH2 ~(C6H5 ). Br6
(544)

Alternatively, (566) was obtained in one step from geranyltriphenylphos-

phonium bromide (544) by simply refluxing its aqueous solution for 20 hours
[323].
6-Methoxy-6-methyl-2-heptanone (559) was also applied to the synthesis
of the C10-Wittig salt (569) [323]. Ethynylation with sodium acetylide in
liquid ammonia gave 7-methoxy-3,7-dimethyl-1-octyn-3-ol (567), which was
selectively hydrogenated to 7-methoxy-3,7-dimethyl-1-octen-3-ol (568). The
salt (569) was prepared by stirring (568) and triphenylphosphonium bromide
in methanol for forty-eight hours.
Alternatively, (568) was first transformed into the bromide (570), which
was then treated with triphenylphosphine [309].
The C10 -phosphonium bromide (569) was also prepared by lithium
aluminium hydride reduction of the methoxy ester (571), which was obtained
from (559) and the phosphorane (394) or the phosphonate (196a), followed
by treatment of the resulting alcohol (572) with triphenylphosphonium
bromide [326, 329].
Recently, an elegant one-step synthesis of (569) was reported starting from
linalool (545) or geraniol (543) and triphenylphosphonium bromide with
simultaneous acid-catalysed addition of methanol to the isopropylidene
double bond [329].
The C10 -Wittig compound (575) required for a synthesis of spheroidenone
(182) [309] was prepared by reaction of methyl P-formylcrotonate (426) with
the C 5 -phosphorane (456) giving the keto ester (573), which was reduced with
lithium aluminium hydride to the glycol (574). The latter was then converted
into (575) by stirring with triphenylphosphonium bromide in methanol [309].
 VI. Total Syntheses 417

CH,O...._I I j_
, . . . ., . . . .,
CH 3 0':>l_ /'-..
~0 6;-~

{559) {567)

j
CH,O..._I
~CH 2 P(C.Hsh Br
I <il e +- CH 30~
OH
(569) (568)

r j
CH,O...._I I CH,O...._I I
~CH 2 0H ~CH 2 Br

(572) (570)

CH,O...._I I
~co.c.H,

(571)

(C6 H 5) 3 P=CHC02 C 2 H 5 1 or (C 2 H 5 0).iCH 2 C0 2 C2 H 5

(394) (196a)

CH,O...._I I
~0
(559)

The CwWittig compound (575 e) used for the synthesis of lycoxanthin

tetrahydropyranyl ether (1001) [330] was prepared by the reaction of levulin-
aldehyde (575 a) with the phosphorane (404). The resulting keto ester (575 b),
after ket!ll formation, was reduced with lithium aluminium hydride to the
corresponding primary alcohol, which was converted into the tetrahydro-
pyranyl ether (575 c). Chain extension by two carbon atoms through a Horner
reaction with the phosphonate (196a) gave the protected hydroxy ester (575d),
which was transformed into the phosphonium salt (575 e) by successive
lithium aluminium hydride reduction, reaction with phosphorus tribromide
and quaternization [330].
Carotenmds 27
 418 H. MAYER and 0. ISLER

01
CH,O'>l
~ y~ CH P(C6 H 5 ). + ~'- C02CH3
OHC~

(456) (426)

, y~
CH 30'>l ~
~ ~C0 2CH 3
~ CH,O~~A/ 2
~H "<:::;,_ "<:::;,_ CH 0H
0
(573) (574)

l
cu,o~,;,c,u,), 8 ,,

(575)

OHC~O +
r"·
(C6 H 5 ).P= CC02C2Hs ---+-
C2Hs02C~O
(575 a) (404) (575b)

0
II
(C2 H 50),PCH 2C0 2C2 H 5 +
THPOCH2~0
(575 c)

THPOCH 2~
~'- ~ C02C2Hs ___..
THPOCH 2
~CHl~(CoHsh Br9
"""' """'

(575 d) (575 e)

For the synthesis of some conjugated polyene diketones and for com-
parison with capsorubin (205) the two Cwketones (577) and (582) were
required. A Darzens reaction between methylheptenone (12) and ethyl
 VI. Total Syntheses 419

~
~
(12)
-- ~ -- H~ (577)

-- ~ -- ~co.R 0~
0
II
CIC~
C0 2CH 3 co.cH 3
(578) (579) (580) R=CH 3
(581) R=H

l
1\ 1\

~ ~ 0
(585) (582)

r
~co.cH. +--
~
CIC~CO.CH•

(584) (583)

a-chloropropionate gave the ketone (576), which was hydrated with sulphuric
acid to yield the hydroxy ketone (577) [328]. Reaction of the half ester acid
chloride (578) of a,a-dimethylsuccinic acid with di-n-propylcadmium gave the
keto ester (579), which was converted into the corresponding ketal (580).
Alkaline hydrolysis then gave the acid (581), which on treatment with methyl-
lithium afforded the C 10 -ketone (582) [328].
The Cw-ketone (585) was used for the introduction of the 2,6,6-trimethyl-
3-oxo-1-cyclohexen-1-yl end group into the carotenoid skeleton. It was
synthesized start~ng from the half ester acid chloride (583) of a,a-dimethyl-
glutaric acid. Reaction with diethylcadmium furnished the keto ester (584),
which was transformed into (585) by the following reaction sequence: acetaliza-
tion, alkaline hydrolysis and treatment of the product with methyllithium [331].
 420 H. MAYER and 0. ISLER

8. C 11 -Components
The C 11 -acetylenic alcohol (506) (Section C.6) was prepared by ethynylation
of the C 9 -triene ketone (310) followed by acid rearrangement of the resulting
acetylenic carbinol (505) [332].

9. C 12 -Components
The intermediate acetylenic Cw-acetoxy acetal (521) was also used for
the synthesis of the C 12 -phosphonium bromide (589).
An enol ether condensation with ethyl vinyl ether (250a) gave the
acetylenic C12 -acetoxy aldehyde (586), which was partially hydrogenated to
the C 1 racetoxy aldehyde (587). This was hydrolysed to the corresponding
hydroxy aldehyde (588), which was converted into (589) as usual: reaction
of (588) with phosphorus tribromide followed by quaternization of the resultiQg
bromide with triphenylphosphine [247].
A base-catalysed condensation with malonic acid was applied to the
preparation of the C 12 -acid (591) [333] and the C 12 -aldehydic ester (594) [334]
starting from the aldehydes (590) and (592), respectively. Thus, condensation
of (592) with malonic acid and esterification of the initial product furnished
the acetylenic aldehydic ester (593), which was then partially hydrogenated
[334]. Interestingly, the condensation occurred at both ends of the molecule
when the triene dialdehyde (619, Section D.3) was used instead of (592).

~CH(OC2H 5 }, ~CHO

AcOCH 2~
~ I
Ac0CH2~ "'* I
(521) (250a) (586)

l
~CHO ~CHO
HOCH2 I I Ac0CH2 I I
(588) (587)

l
(589)
 VI. Total Syntheses 421

CH 30CH2~C02H
CH2(C02Hl2
~ ~ ~I /CHO
CH,OCH2/ ~ ~

(590) (591)

~CHO CH2(C02Hl2
~C0 2 CH 3

OHC~?;- OHC ~
"*' I
(592) (593)

j
OHC~C02CH,
(594)

10. C 15 -Components
Farnesyltriphenylphosphonium bromide (597) has been prepared by treat-
ment with triphenylphosphine of farnesyl bromide (596), which could be
obtained readily by reaction of trans-nerolidol (595) with phosphorus tri-
bromide [296, 303, 335]. The conversion of farnesyl bromide (596) into
farnesal (598) was effected by the nitropropane route [296].

~CH 2 Br
~
(595) OH
----+

(596)

~CHO
/ 1
(598) (597)

~CH2~(CoHs), X
~
(599) OH
----+ 8

(600) X=Br
(601) X=CI
422 H. MAYER and 0. ISLER

~CHO ~CHO

Ac0CH2~
~ I
Ac0CH 2 ~ '* I I

(586) (602)

J
~CHO
HOCHl I I I
(604) (603)
9 ® ~CHO ~CHO
Cl (CoHshPCHl '<:::::I + (CoHshP=CH '<::::::1
(449) l (450)

e ® ~CHO ® ~CHO
Cl (C6 H 5 hPCH 2 '<:::::I "<:::::: '<:::::I + Cl
9
(CoHshPCHl I
(605) (449)

l
(606)

(607)

The synthesis of the (pseudoionylideneethyl)triphenylphosphonium halides

(600) and (601) was based on pseudoionone (85) [153, 166]. Ethynylation
with lithium acetylide in liquid ammonia followed by partial hydrogenation
of the product gave the vinyl alcohol (599), which was reacted with triphenyl-
phosphine in the presence of methanolic hydrochloric acid or with triphenyl-
phosphonium bromide.
Two different procedures have been worked out for the synthesis of the
Cwtriphenylphosphonium halides (604) and (606):
1) Chain lengthening by three carbon atoms of the acetylenic acetoxy
aldehyde (586) by the propenyl ether condensation led to the C 15 -acetoxy
aldehyde (602). Partial hydrogenation and alkaline hydrolysis of the product
 VI. Total Syntheses 423

HOCH2
~CHO
I I + (C6 H 5 ),P=CH ~
~OCH 3

l
(513) (456)

HOCH 2 ~ ~ ~

(608)

gave the hydroxy aldehyde (603), which was transformed into (604) in the
usual way [247].
2) Refluxing of a solution of the C 5 -phosphonium chloride (449) and the
corresponding phosphorane (450) in acetonitrile furnished a solution of the
Cw-phosphonium chloride (605), which was again reacted with (449) in the
presence of sodium methoxide. The resulting C 15 -Wittig salt (606) was not
isolated but immediately converted into the corresponding phosphorane
(607) [336].
The C 15 -phosphonium bromide (609) was required for a synthesis of
spheroidenone (182) [309]. Reaction of the Cw-hydroxy aldehyde (513) with
the C 5 -phosphorane (456) gave the hydroxy ketone (608), which was trans-
formed into (609) by treatment with triphenylphosphonium bromide [309].
The C 15 -ester (611) and the corresponding aldehyde (614), intermediates
in a synthesis of y-vitamin A, were synthesized by three different procedures
[296]. Application of the methoxyacetylene method to pseudoionone (85)
gave the acetylenic ether (610), which on treatment with sulphuric acid led
to (611)! The same product was obtained by reaction of the Wittig reagent
(345) or the ph.osphonate (347 a) with citral (88).
Reduction of (611) with lithium aluminium hydride gave the alcohol (613),
which was oxidized with manganese dioxide to the aldehyde (614). The alcohol
(613) could also· be made by base-catalysed condensation of citral (88) with
methyl P-methylcrotonate (425) giving the acid (612), which was reduced with
lithium aluminium hydride.
424 H. MAYER and 0. ISLER

~0
(85)
--~~ OH ~
(610) ""-ocH 3

1
~CH2 0H +--- ~C02CH 3
(613) (611)

1
~CHO ~C02H
(614) (612)

/ ~C02CH3 (425)

~CHO
(88)

Br
6 (!)
(C6 H 5 ),PCH2
~C02CH
"'
3 ~ ~C02CH3
(C 2H 5 0j,PCH 2 '-":::::
(345) (347 a)

~C02CH3
(611)

11. C 16 -Components
The acetylenic carbinol (615) and the C 16 -hydrocarbon (617) were applied
to a synthesis of lycopene (19) according to the scheme C 16 + C 8 + C 16 = C4 0
[337, 338].
 VI. Total Syntheses 425

~0-- l . l l '*
~
(85) (615) R=H OR
(616) R=Ac

l
~'*
(617)

j
I I I ~
~
(618a)

A Reformatskii-type reaction ofpseudoionone (85) with propargyl bromide

in the presence of aluminium [338] or magnesium [339] furnished the acetylenic
carbinol (615), which was dehydrated with phosphorus oxychloride and
pyridine to give (617) in about 50% yield [340]. Better yields were obtained
by first converting (615) into the corresponding acetate (616), which was then
treated with potassium t-butoxide [338].
The acetylenic carbinol (618) and the corresponding hydrocarbon (618a),
used for syntheses of 5,6,5',6'-tetrahydrolycopene (906) were prepared in an
analogous way [341].

12. C 11 -Components
The synthesis of the C 17 -phosphonium salt (623), required for a stereo-
chemically controlled synthesis of natural methylbixin (266a) has recently
been reported [288]. Reaction under suitable conditions of the C 7 -phosphonium
bromide' (497) at one end of the triene dialdehyde (619) furnished the aldehydic
ester (621), which was selectively reduced with sodium borohydride to the
hydroxy ester (622). The latter was also prepared by partial reduction of(619)
with sodium borohydride, followed by a Wittig condensation between the
resulting hydroxy aldehyde (620) and the phosphonium bromide (497). The
Wittig salt (623) was obtained by reaction of the hydroxy ester (622) with
triphenylphosphonium bromide.
426 H. MAYER and 0. IsLER

OHC~CHO _..L ~ ~
HOCH2~--~ ~ I
/CHO

(619) (620)

(497)

,...._ ,...._ """- ,...._ ,, ,...._ C02CH 3

OHC ""-' '-' "' '-' " "'
(621)

l
C0 2CH 3
HOCH 2 ~ ~ ~ ~ ~ ~

(622)

l
® C0 2 CH 3
Br 8 (C 6 H 5 ),PCH 2 ~ ~ ~ ~ ~ ~

(623)

13. C 20 -Components
The C 20 -aldehydes and phosphonium salts discussed in this chapter have
proved to be valuable building units for the synthesis of carotenoid hydro-
carbons, such as phytoene (32), phytofluene (30) and neurosporene (22) [296].
A Horner reaction of farnesal (599) with the phosphonate (347 a) gave the
ester (624), which was reduced with lithium aluminium hydride to the alcohol
(625). Oxidation with manganese dioxide then gave the C 20 -aldehyde (626)
[296].
The C 20 -Wittig reagent (627) was obtained from the alcohol (625) by
conversion into the corresponding bromide followed by treatment of the
product with triphenylphosphine ['296].
 VI. Total Syntheses 427

~
~CHO + ~ ~C0 2 CH 3
(C 2H 5 0),PCH 2

1
(599) (347 a)

C0 2CH 3

"""
(624) """ """ """
t
CH 20H

"""
(625)
""" """ """ """

I
CHO

"""
(626) """ """ """
(j)
CH2P(CoHsh
"""
(627)
""" """ """ Br 6

"""
(628)
""" ::?'

~
CH 2Br

"""
(629) """
I
(j)
CH2P(CoHsh
"""
(630) """ Br 6

CHO
"""
(631)
"""
428 H. MAYER and 0. ISLER

Treatment of all-trans-geranyllinalool (628) with phosphorus tribromide

gave geranylgeranyl bromide (629), which was converted into the corresponding
C 20 -triphenylphosphonium bromide (630). Geranylgeranyl bromide was also
converted by the nitropropane route into geranylcitral (631) [296, 335].

D. Syntheses of Symmetrical Central Components

Symmetrical central components possessing a carbon skeleton identical

with that of the central portion of /3-carotene offer numerous routes for the
synthesis of many classes of carotenoids.
Acetylene and diacetylene have been used as convenient C 2 - and C 4 -central
components, and the important C 8 -, C 10 -, C 12 -, C 14 - and C 20 -intermediates
are now comparatively readily available by total synthesis. In this sectiop
only syntheses of central components with up to 18 carbon atoms are described.
Syntheses of symmetrical building units possessing 20 or more carbon atoms
will be dealt with in the section on diapo-carotenoids.

I. C 4 -Components
The C 4 -bisphosphonium chloride (631 a) and bromide (631 b) and the
C4 -diphosphonate (631 c) (Table 4) were prepared by treatment of 1,4-dichloro-
2-butene and 1,4-dibromo-2-butene with triphenylphosphine and triethyl
phosphite, respectively. The bisphosphonium bromide (631 b) could also be
obtained from 2-butene-1,4-diol by treatment with triphenylphosphonium
bromide or triphenylphosphine and HBr.

Table 4. C 4 -Components

Compound References
Ell Ell
X 8 (C 6 H 5 ) 3 PCH 2 CH=CHCH 2 P(C 6 H 5 )J X 6 [58,61, 139,342,343]
(63la) X=Cl; (631b) X=Br

(C 2 H 5 0h0PCH 2 CH=CHCH 2 PO(OC 2 H 5 h [21]

(631 c)

2. C 8 -Components

a) C6 +Cz=Cs
3,5-0ctadiene-2,7-dione (638) and 4-octene-2,7-dione (639), used success-
fully for the synthesis of IX-carotene (5), /3-carotene (3), y-carotene (8),
e-carotene (10), lycopene (19) and carotenoids with aromatic end groups
[e.g. isorenieratene (13)], were prepared by a Grignard reaction of the
 VI. Total Syntheses 429

I C 6 +C 2 =C 8 1 C 4 +C 4 =C 8 1

~~
(632)
"">
+ OHCCH 3
-- ~~ (633) ~JoH

l
HO~~ ~
+ OHCCH 3
-- RO~~ ~
y OH

(636) (634) R=H

(635)R=THP

l
0~0
(638)
~~~~OH
THPO~---~ ~
(637)
I

~~ HO~~ ~
HO ~

(641)
~
"-..,_ ~
+ (640)
_1_ ~ /'... ~0
o:.~~ ~
(639)
l
~
y OH
~JoH

acetylenic carbinol (632) with acetaldehyde [344]. The resulting glycol (633)
was treated with dilute sulphuric acid to give the glycol (634), which was also
obtained from the C 6 -alcohol (636) and acetaldehyde. Treatment of the
tetrahydropyranyl derivative (635) with LiAIH 4 gave the corresponding diene
(637) which, however, was converted into the dione (638) in poor yield only.
Reduction of(638) with zinc in acetic acid then furnished the desired C 8 -dione
(639) [345].
430 H. MAYER and 0. ISLER

b) C4 +C4=Cs
The synthesis of the C 8 -diketone (638) could also be achieved by oxidative
dimerization of 3-butyn-2-ol (640) in the presence of oxygen and cupric
chloride [346, 347], reduction of the resulting glycol (641) with LiAIH 4 followed
by oxidation of the product with manganese dioxide [344].

c) C3 +C2+C3=Cs
When 1,2-dichloro-1,2-diethoxyethane (642) was reacted in a Reformatskii-
type reaction with propargyl bromide in the presence of aluminium and
mercuric chloride, the diyne (643) was obtained. Subsequent hydration gave
the diane (644), which was hydrolysed to the C 8 -diketone (638). In the fmal
step (638) was reduced with zinc in acetic acid yielding the desired C 8 -diketone
(639) in a total yield of up to 50% [338].

d) C1 +C6+Ct=Cs
4-0ctene-2,7-dione (639) could also be synthesized directly in 65% yield
starting from trans-dihydromuconic acid dichloride (645) by the introduction
of two methyl groups with methylzinc iodide [338].
According to the same scheme the C 8 -diketone (638) has been made by
condensation of glyoxal with acetoacetic acid under strictly controlled con-
ditions, but the yield could not be raised above about 4% [345].

"
C2H,O OC2H,

"
HC=CCH 2Br + CH-CH / + BrCH 2c=cH
CI/ Cl

'"+,.
~ (642)

OC2Hs
~ 0)-_;t"~/'
~ ~C2 H, r
-

(643) (644) (638)

0
l
II
CIC~ ~ /"...
""-./ ~ 'CCI
(643) &
-- 0~0
(639)
 VI. Total Syntheses 431

3. C 10-Components
2,7-Dimethyl-2,4,6-octatrienedial (619) and the C10 -bisphosphonium bromide
(668) as well as their central-acetylenic analogues (592) and (666) are important
central building units for the synthesis of many C40-carotenoids, apo-
carotenoids and C 20 -components. They have been prepared according to
the following reaction schemes.

a) C6 + C4= Cto
RUegg et al. [280] synthesized the bis(diethyl acetal) (646) of the Cw-yne
dialdehyde (592) by a Nef-type reaction of the C 6 -acetal (470) with methyl-
malonaldehyde enol ether (289).

OHC~
1
::::::r OC2 H 5

(470) (289)

j
I ~CH(OC2Hsh
~'*'
(C2H 0),HC '<::::::
5
I
(646)

b) C 5 +C 5 =C10
In a process patented by the Badische Anilin- & Soda-Fabrik [348]
1,1-dimethoxy-2-methyl-3-butyn-2-ol (422) was dimerized in the presence of
oxygen and cupric chloride giving the diyne glycol (647), which was hydro-
genated over Raney nickel to the glycol (648). Subsequent dehydration to the
triene (649) with thionyl chloride followed by acid hydrolysis yielded the
C10-dialdehyde (619).
By oxidative dimerization of 1-methoxy-2-methyl-3-butyn-2-ol (464)
Inhoffen et al. [349] and Ahmad and Weedon [333] obtained the Cw-glycol
(650) in ~bout 70% yield. Reduction with lithium aluminium hydride produced
the diene glycoJ (651) [333], which on dehydration and subsequent treatment
with mineral acid was transformed into the methoxy aldehyde (652).
According to the same scheme Inhoffen et al. [349] reported the pre-
paration of the C10 -diester (653) by a Wurtz-type reaction of methyl y-bromo-
a-methylcrotonate (444) (Section C.2f) followed by bromination and dehydro-
bromination.
432 H. MAYER and 0. ISLER

(CH 30),H~s,
I + ~ fcH(OCH,h
OH""
(422)

(CH,O),H~"
OH ~ ' - '> fcu(OCHJ, ~
/

(647)

j
OH~CHO __.l ~CH(OCH h
(CH,O),HC~ .~ ~
../.>o._

I
......,.,_
3

(619) (649)

CH 3 0CH2 .1 . .
'6~ &
+
(464)

CH30CH2~s,
/
OH "" '--._ "'- ,J~
~ TCH20CH,

(650)

l
_1__ ../.>o.. ../.>o.. ~C0 2CH 3 CH30CH2~CHO
CH,02C~--~ ~I
(653) (652)
 VI. Total Syntheses 433

c) C4 +C 2 +C4=C 10
The first syntheses of the C 10 -key intermediates (619) and (592) were
performed by the schools of Inhoffen [350, 351] and Weedon [352].
Methacrolein (654) was condensed with acetylene in a Grignard reaction to

~
,;:;-- CHO
+ HC=CH + OHC'Y-7
I
(654) (654)

~CH 2 Br

BrCH
~'*
~
2
I

(655) (657)

l
~CH 2 0H

HOCH2~--~ ~
~ ~ ~
I
,CH 20H
HOCH 2 ~
I '* I
(658) (656)

l l
~CHO
OHC~CHO OHC~'*
(619) (592)

l
~ ~ ~
,C0 2 R
R02C' ~~I

(661) R=H (659) R=H

(662) R=CH 3 (660) R=CH 3

Carotenotds 28
434 H. MAYER and 0. ISLER

give 2,7-dimethyl-1,7-octadien-4-yne-3,6-diol (655). This was either rearranged

directly to 2,7-dimethyl-2,6-octadien-4-yne-1,8-diol (656) or first converted
into the dibromide (657), which on treatment with potassium acetate and
subsequent saponification yielded the same dial (656). Manganese dioxide
oxidation of the latter then led to the acetylenic C10 -dialdehyde (592).
Partial hydrogenation of the acetylenic dial (656) gave the triene dial (658),
which was oxidized with manganese dioxide in acetone and isomerized to
the Cw-dialdehyde (619). The latter compound was also obtained by partial
reduction of the acetylenic C10 -dialdehyde (592).
Chromic acid oxidation of the C10 -dialdehyde (592) gave the diacid (659),
which was also obtained by direct oxidation of the glycol (656). Esterification
with diazomethane gave the dimethyl ester (660). Partial hydrogenation of
the latter followed by isomerization then afforded the triene diester (662),
which was hydrolysed to the corresponding diacid (661) [352].
Surmatis and Ofner [353] used the acetylenic dial (655) for the preparation
of both Cw-phosphonium salts (666) and (668). Partial hydrogenation over

~ OH
(655) (663)

j l
'*
~CH 2 Br

I I .1. _.. ,.,_ _.. ,.,_ ~CH 2 Br

BrCH2~
(664)
BrCH 2~,~ ~
(665)
l

l j

Ell
(666) R= P(C 6 H 5h Br 9 (668)
(667) R=PO(OC 2 H 5h
 VI. Total Syntheses 435

Lindlar catalyst afforded the ethylenic diol (663), from which the dibromide
(665) was formed with hydrobromic acid. Treatment of the latter with tri-
phenylphosphine then gave the Wittig compound (668) in good yield. The
acetylenic analogue (666) was made by a very similar procedure but starting
from the acetylenic diol (665) via the dibromide (664). When this was heated
with triethyl phosphite, the acetylenic diphosphonate (667) was formed [21].

d) C 3 +C4+C3=C1o
The enol ether condensation has been used for the synthesis of the C 10 -
dialdehyde (619) by Wittig and Pommer [21, 61], by Isler et al. [354] and
by Makin et al. [355]. Malealdehyde bis(diethyl acetal) (669) was condensed
in the presence of zinc chloride or boron trifluoride with 2 moles of propenyl
ether (415) to give the C 10 -diacetal (670), which was hydrolysed by acid treat-
ment to the Cw-dialdehyde (619).

(
OC2H 5
+

(415) (669) (415)

/
OC 2 H 5
~ ./"'- ~ • ~CH(OC 2 H 5 ),
(C 2 H 5 0),HC' ~ ~ ~
OC 2H 5
(670) (619)

The application of the Wittig reaction to the synthesis of Cw-intermediates

has been studied by Pommer [21]. When the bisphosphonium bromide (631 b)
reacted with either hydroxyacetone (671) or the keto acetal (421 b), the
yields ofthe reaction products (658) and (672), respectively, could not be raised
above 10%. By the use of the diphosphonate (631 c), however, good yields of
the desired C 10 -compounds could be obtained.
The preparation of the C 10 -diacids (673) and (674) according to the scheme
c3 + c4 + c3 = clO was achieved by alkylation of diethyl methylmalonate
with 1,4-dibromo-2-butyne and 1,4-dibromo-2-butene, respectively, saponi-
fication of the resulting tetraesters and decarboxylation [356]. Esterification
of the diacid (674) followed by bromination with N-bromosuccinimide to the
dibromide (676) and subsequent dehydrobromination then afforded the
already described Cw-diester (662).
 436 H. MAYER and 0. ISLER

_1_ ~ ~
I
/CH 20H
HOCH2~--~ ~
(658)

1
HOCH 2~0 + + OJ__CH 20H
(671) (631b) (671)

(C 2H 5 0),HC ~0 +
o
II
(C 2H 5 0),PCH 2
~CH2P(OC2Hsh
II
+ 0
)__CH(OC2Hsh
(421 b) (631 c) (421 b)

l
_1_ ~ ~
I
/CH(OC 2H 5 ) 2
(C2H,O),H~--~ ~
(672)

~C0 2 H Br

~'* ~ ~ ~ ~ ~
H0 C
2
I
R02C/ '-./' ~ I
_,...__ /C0 2R
- CH,02C/ I ~ I /C0 2CH 3

Br
(673) (674)R=H (676)
(675) R=CH 3

4. C 12 -Components
a) C6 + C6 = C 12
The C 12 -hydrocarbon (678), used as central building unit in a synthesis
of {J-carotene by Isler et al. [305], was synthesized from 1-bromo-3-methyl-
2-penten-4-yne (679). This bromide was transformed either into the phos-
phonium bromide (453 a) or into the aldehyde (677) by the nitropropane
route. Condensation of the phosphonium bromide with the aldehyde under
the conditions of the Wittig reaction then afforded the hydrocarbon (678) [305].
The analogous C 12 -hydrocarbon (680) was made by a Wurtz-type dimeriza-
tion of the bromide (679) using zinc or the Zn-Cu couple [357, 358, 358a].
 VI. Total Syntheses 437

+ OHC~ '*'
(453 a) (677) (678)

~CH2Br l_ /'--. ~ / '*'

'*' (679)
~~~~~ ~I
(680)

HOCH2~ +
~

(158 a)

l
OHC~CHO OHC~
~
"'-..;,._
(683) (682) ~yrno

The retro-C 12 -dialdehyde (683) possessing a polyene system identical with

that of the central part of retro carotenoids was a key intermediate for a total
synthesis of rhodoxanthin (209) [44]. Oxidative coupling of trans-3-methyl-
2-penten-4-yn-1-ol (158 a) with oxygen in the presence of cuprous chloride-
ammonium chloride gave the trans-C 12 -diol (681), which on manganese
dioxide oxidation furnished the corresponding dialdehyde (682). Partial
hydrogenation and subsequent isomerization yielded the desired all-trans-
retro-C12-dialdehyde (683).

b) C 5 +.C2+C 5 =C 12
The C 12 -bis(enol ether) (687), an intermediate for the synthesis of P-carotene
(3), 3,4,3',4' -tetradehydro-{J-carotene (1) and zeaxanthin (1 004) [359], was
obtained by a Grignard reaction of two moles of tiglaldehyde (684) and
acetylenedimagriesium bromide to give the glycol (685), which, after allylic
rearrangement and manganese dioxide oxidation, yielded the diketone (686).
Subsequent ketalization and elimination of alcohol then led to (687).
438 H. MAYER and 0. ISLER

~CHO + HC:::::CH + OHC~

(684) (684) (685)

(686)

c) C 2 +Cs+C2=C12
The C 12 -building unit (688) similar to (678) and (680), used for the prepara-
tion of 15,15'-dihydro-P-carotene (899), was made from 2,7-octanedione and
two moles of lithium acetylide [360].

(688)

dJ cl + C10+ cl = C12
An alternative route to the diketone (686) starting from the Cw-dialdehyde
(592) was followed by the introduction of two methyl groups with two moles
of methylmagnesium bromide and subsequent manganese dioxide oxidation
[361].

5. C14-Components
C 2 + C 10 + C 2 = C 14 . The C14-dialdehyde (689) and its central-acetylenic
analogue (693), central components in some of Pommer's P-carotene syntheses
[21, 61, 97, 98, 129, 139] have been prepared by enol ether condensation
from the diacetals (672) and (646), respectively, and ethyl vinyl ether (250a)
in the presence of zinc chloride and/or BF3 etherate [21, 61, 245, 355].
The C 14-dialdehyde (689) and the Cwdiester (690) have also been made
by chain lengthening at both ends of the C 10-dialdehyde (619) with the phos-
phoranes (394a) [267] and (394) [343], respectively.
 VI. Total Syntheses 439

.1. . . ,._ . . ,._ ~CH(OCzHsh . . ,._ ..1. . . ,._ . . ,._ . . ,._ ~CHO
(C 2Hs0hH~--~ ~ ~
(672) (250a)
OHC~--~ ~ ~
(689)
I ~

OHC ~ ~ ~
~ ~ I
_CHO (CoHshP=CHCOzCzHs
- - - - - - - + C2Hs02 ~
(619) (394) (690)

1 CH 2 (C02Hh

~CHO
. . ,._ ..1. . . ,._ . . ,._ ~ ~C0 2R
OHC~
I "'*- I
R0 2C~-~~ ~ ~ ~ ~
(691) R=H (693)
(692) R=CH 3

Starting from the same dialdehyde (619), a condensation of the Doebner-

type furnished the Cwdiacid (691) and its dimethyl ester (692) [334].

6. C 18 -Components

a) C 4 + C10+ C4= C1s

A Doebner-type condensation has also been used for the preparation of
dinor-crocetin (695) and its dimethyl ester. Condensation of diethyl ethylidene-
malonate (694) with the Cw-triene dialdehyde (619) followed by hydrolysis
afforded a heptaene tetraacid which on decarboxylation led to the diacid (695)
[334].

.1. . . ,._ . . ,._ ~CHO

(CzHsOzC)2C=CHCH 3
(694)
+ ouc---~ ~
(619)
1 + CH 3 CH=C(C0 2 C2 Hsh
(694)

j
H02
(695)
440 H. MAYER and 0. ISLER

b) C 3 +C12+C3=C1s
The retro-C 18 -diketone (696), an intermediate for the synthesis of retro-
diapo-carotenoids, was readily obtained by the Wittig reaction at both ends
of the retro-C 12 -dialdehyde (683) with the phosphorane (410) [362].

I C3+C,z+C3=C,s I
0
CH,&:H-P(CoHsh + OHC~CHO +
(410) (683) (410)

j
0
o:::"' ::;:?" ::;:?" ::;:?" ::;:?" ::;:?" ::;:?" :::"""

(696)

E. Syntheses of Apo-carotenoids

The apo-carotenoids, such as {3-apo-8'- and -12'-carotenal and the cor-

responding apo-carotenoic acid esters, represent a group of widely distributed
natural pigments possessing considerable provitamin A activity. They have
been considered as possible intermediates in the in vivo conversion of {3-
carotene into vitamin A arising by the formal successive removal of fragments
from one end of the C 40 -carotenoid skeleton.

1. {3-Apo-carotenals
a) {3- Apo-14'-carotenal
rx) C20 + C 2 = C22 • {3-Apo-14'-carotenal (698) was obtained by a Horner
reaction of retinal (325a) with diethyl (cyanomethyl)phosphonate (193) giving
the C 22 -nitrile (697) as initial product, which was reduced with diisobutyl-
aluminium hydride [192, 363].

{3) C 22 = C 22 • /3-Apo-14'-carotenal (698) has also been prepared from

methyl {3-apo-14'-carotenoate (699) by diisobutylaluminium hydride reduction
to the corresponding primary alcohol followed by manganese dioxide oxidation
[192].

b) {3-Apo-12' -carotenal
{3-Apo-12' -carotenal (262) is a key intermediate for the synthesis ofthe higher
members of the {3-apo-carotenal series. It could be synthesized by four different
routes.
 VI. Total Syntheses 441

I Czo+Cz=Czz/

'<:::,
CHO ~
(C 2 H,O),PCH 2 CN '<:::, '<:::, '<:::, '<:::,
CN
"""' """' """'
(193)
(325 a) (697)

l
C0 2CH 3 CHO
'<:::, '<:::, '<:::, '<:::, '<:::, '<:::,
"""' """'
(699) (698)

rx) C15 + C10 = C 25 . In a process patented by the Badische Anilin- &

Soda-Fabrik AG [364, 365] /3-apo-12'-carotenal (262) and the corresponding
15,15'-didehydro compound (701) have been made by the Wittig reaction at
one end of the C 10 -dialdehydes (619) and (592), respectively, with one mole of
(/3-ionylideneethyl)triphenylphosphonium sulphate (206) or chloride (204)
(Table 5).

Table 5. I c15 + c!O = c25

CwComponents C 10 -Components P-Apo-12' -carotenals Refer-
ences

~CHO
+ ~'*
OHC """' ~ 15,15'-Didehydro- [364,
xe (592) P-apo-12' -carotenal 365]
(701)
~CH,~(C6H 5 ) 3

(206) X=HS0 4 + OHC~CHO ~ P-Apo-12'-carotenal [364,

(2Q4) X=Cl (619) (262) 365]

Br8

~
OH~CHO
lX'*
"""' CH 2 ®
P(C6 H,j,
+ ~ 3-Hydroxy- [123]
7,8-didehydro-
(619) P-apo-12' -carotenal
HO (165) (794)
442 H. MAYER and 0. ISLER

/3) C19 +C 6 =C 25 . 15,15'-Didehydro-{3-apo-12'-carotenal (701), an inter-

mediate in a technical synthesis of /3-apo-8'-carotenal (248), was prepared from
the f3-C 19 -aldehyde (285) by a Grignard reaction with the C 6 -acetal (470)
followed by acid-catalysed dehydration of the resulting hydroxy acetal (700)
and hydrolysis of the acetal grouping [280]. The further conversion of (701)
into /3-apo-12'-carotenal (262) was achieved by selective hydrogenation
followed by isomerization.

I Ct9+C6=C251

c:P' c:P' ~CH(OC2Hsh

c:P' CHO +
"*'
(285) (470)

J
~CH(OC2Hsh

c:P' c:P' c:P' "*'

H

(700)

l
~CHO
~ ~ ~ "*'

(701)

y) C 20 + C 5 = C 2 5 . /3-Apo-12'-carotenal (262) has also been synthesized

by the C 20 + C 5 = C 25 route starting from retinal (325 a) as C 20 -component
(Table 6).

~) C 21 + C 4 = C 25 . Condensation of the Grignard derivative of the acety-

lenic C 21 -carbinol (387) with methylmalonaldehyde enol benzoate (419)
gave the dehydro ester (703), which was transformed into 15,15'-didehydro-
/3-apo-12'-carotenal (701). Subsequent partial hydrogenation and isomerization
afforded all-trans-{3-apo-12'-carotenal (262) [280].
 VI. Total Syntheses 443

C 20 -Component C 5 -Components Refer-

ences

+ [366]

~CHO P-Apo-12' -carotenal

0
(262)
(325a)
+ ~CH(OCH 3 ), [367]

(702)

Lithium aluminium hydride reduction of (262) gave the corresponding

primary alcohol which was characterized as the corresponding acetate [188].
The P-apo-10'-, -8'-, -6'-, -4'- and -2'-carotenals discussed below have been
converted into the corresponding primary alcohols and characterized as the
corresponding acetates in an analogous way [188].

I c21 +C4=Czsl

?;- 0
?"' ?"' ?"' + OHC"r II
?"' OCC6H 5
OH

(387) (419)

1
OH

?;- ~~ OCC6H 5
?"' ?"' ?"'
OH

(703)

j
?;-
~CHO
~ ~ ~

(701)
444 H. MAYER and 0. ISLER

e) C 25 =C 25 • /3-Apo-12'-carotenal (262) has also been prepared by

diisobutylaluminium hydride [368] or lithium aluminium hydride [368a]
reduction of methyl {3-apo-12'-carotenoate (875) followed by manganese
dioxide oxidation of the resulting primary alcohol.

c) 3-Hydroxy-7,8-didehydro-{3-apo-12'-carotenal
C15 + C10 = C25 • 3-Hydroxy-7,8-didehydro-{3-apo-12'-carotenal (794, Table 5),
required for a synthesis of 7,8, 7',8'-tetradehydrozeaxanthin (1 003), was prepared
by condensation of the C 10-dialdehyde (619) at one end with the C 15 -Wittig
compound (165) [123].

d) {3-Apo-10'-carotenal
rx) C13 + C 14 = C 27 • 15,15'-Didehydro-{3-apo-10'-carotenal (704) was syn-
thesized by the condensation of the acetylenic C 14-dialdehyde (693) at on~
end with one mole of {3-ionyltriphenylphosphonium chloride (95) [364].

~CHO
I I
~~C.U.h"'
q,
+ OHC~

(95) (693)

(704)

{3) C 20 + C 7 = C 27 • A patent granted to F. Hoffmann-La Roche & Co. Ltd.

[249] describes the synthesis of the diethyl acetal (705) of 11',12'-didehydro-
{3-apo-10'-carotenal by a Wittig reaction of {3-retinyltriphenylphosphonium
chloride (367) with the C 7 -dialdehyde monoacetal (492).
Alternatively, {3-apo-10'-carotenal (256) was prepared according to the
same scheme by condensation of retinal (325 a) with the C 7 -phosphorane
(499a), which was obtained from the corresponding phosphonium salt (499,
Section C.4) [366].
 VI. Total Syntheses 445

I C20 +C,=C2 7 1

_.......CH(OC.H 5h
'*'
(!)
CH.P(C.Hsh
~ ~ ~
ae + OH~

(367) (492)

l
_.......CH(OC2 H 5 h

~ ~ ~ ~ ~
'*'

(705)

CHO
~ ~ ~ + (C6 H 5 hP=CH~CHO

(325a) (499a)

l
CHO
~ ~ ~ ~ ~ ~

(256)

y) C 25 + C 2 = C 27 . Starting from the diethyl acetal of 15,15'-didehydro-

P-apo-12'-carotenal (706), P-apo-10'-carotenal (256) was obtained by the vinyl
ether condensation yielding as an intermediate 15,15'-didehydro-{J-apo-10'-
carotenal (704), which was partially hydrogenated and isomerized [188].
P-Apo-10'-carotenal (256) could also be obtained directly by vinyl ether
condensation starting from the diethyl acetal of P-apo-12' -carotenal (707) [369].

e) P-Apo-8' -carotenal
rx) C 20 +C 10 =C 30 • Four different procedures have been devised for the
synthesis of P-apo-8'-carotenal (248) by the combination of a C 20 - and a
C10 -building unit as depicted in Table 7.
 ~
Table 7. I C2o + clO :c:30J
C 20 -Components Cw-Components References

~H,;(C6H,),CI 9 + OHC~CH(OCH,). [247]

(367) (520)
--
., ;I:
+ Br9 (C6H,),PCH,~CH(OCH 3) 2 [247]
(708)
--
~CHO i8.
(325a) P-Apo-8' -carotenal
CI9 (C6H,),;CH,yHo 9
(248)
(449)

+
I -- [336]
5-
(C6H,),P==CH~CHO
I (450)

OHC ""'=: ""'=: ""'=: ""'=: ""'=: ""'=: ""'=: CHO + Br9
.,'Yl
(C6H,),PCH,~ [364]
(267) (27)
--
 VI. Total Syntheses 447

I C2s+C2=C211

~CH(OC 2 H,),

'<::::::: '<::::::: '<::::::: '*

(706)

1~0C2H,
(250a)

~CHO
'<::::::: '<::::::: '<::::::: '*
(704)

CHO
'<::::::: '<::::::: '<::::::: '<::::::: '<::::::: '<:::::::

(256)

~~OC2Hs
(250a)

CH(OC 2H 5 ),
'<::::::: '<::::::: '<::::::: '<::::::: '<::::::: '<:::::::

(707)

The phosphonium salt (708) was obtained from the corresponding Wittig
compound (515, Section C.7) by acetalization [247].
The Wittig compound (449) was first condensed with the phosphorane (450),
followed by re~ction of the product with retinal (325a) in the presence of
sodium methoxide [336].

/3) C 25 + C 5 = C 30 • According to this scheme /3-apo-8'-carotenal (248) was

synthesized from {3-apo-12'-carotenal (262) and the C 5 -components listed in
Table 8.
 448 H. MAYER and 0. ISLER

Table 8. I c25 + Cs = C3o

C 25 -Component C 5 -Components Refer-

ences

[366]

""':: ""':: CHO + (C.,H,),P=CH

~CHO
I [366]
P-Apo-
(262) (450) 8' -carotenal
(248)

+ ~CH(OCH3)2 [367]

(702)

y) C 27 + C 3 = C 30 . An industrial process for P-apo-8'-carotenal (248)

according to the scheme C19 +C 6 =C 25 +C 2 +C 3 =C 30 (F.Hoffmann-La
Roche & Co. Ltd.) [188] starts from the P-Cwaldehyde (285), which, by
successive chain lengthening by six and two carbon atoms, was transformed
into 15,15'-didehydro-P-apo-10' -carotenal (704) as described (Sections E.l b p
and E.ldy). Chain lengthening of the latter by three carbon atoms was
achieved by propenyl ether condensation of the corresponding diethyl
acetal (709) yielding 15,15'-didehydro-p-apo-8'-carotenal (710), which was
partially hydrogenated and isomerized to P-apo-8'-carotenal (248).

f) P-Apo-6'-carotenal
a.) c25 + C7= C32. P-Apo-6'-carotenal (713) was prepared from P-apo-
12'-carotenal (262) by condensation with the C 7-phosphonium salt (715) [247],
which was obtained from the Wittig compound (499, Section C.4) by acetali-
zation.

P) C 30 + C2= C 32 . Vinyl ether condensation of 15,15'-didehydro-P-apo-8'-

carotenal (710) yielded 15,15'-didehydro-P-apo-6'-carotenal (711) as an inter-
mediate, which was partially hydrogenated and isomerized to P-apo-6'-
carotenal (713) [188].

g) P-Apo-4'-carotenal
a.)C 25 +C 10 =C 35 . Starting from P-apo-12'-carotenal (262), P-apo-4'-
carotenal (714) was prepared by the Wittig reaction with the C 10 -phosphonium
bromide (708) [247].

PJ C 32 +C 3=C 35 . Propenyl ether condensation of 15,15'-didehydro-P-

apo-6'-carotenal (711) gave 15,15'-didehydro-P-apo-4'-carotenal (712), which
 VI. Total Syntheses 449

(709)

~CHO

'* l I

~OC 2 H,/ (710)

(250a) /

(711)

OC 2 H, CHO
~ ~ ~ ~ ~ ~ ~ ~

~(415) (248)

~CHO
~ ~ ~ '*
(712)

CHO
~ ~ ~ ~ ~ ~ ~ ~ ~

(713)

CHO
~ ~ ~ ~ ~ ~ ~ ~ ~ ~

(714)

Carotenmds 29
 450 H. MAYER and 0. ISLER

(262)
+ +

(715) (708)

(713)

(714)

was partially hydrogenated and isomerized to all-trans-fJ-apo-4' -carotenal (714)

[188].

h) P-Apo-2'-carotenal
rx) C3o+ C7= c37' P-Apo-2'-carotenal (233) has been obtained from
P-apo-8' -carotenal (248) by reaction with the phosphonium bromide (715)
under the condition of the Wittig reaction [247].

p) c35 + c2 = c37. P-Apo-2'-carotenal (233) was also synthesized by the

vinyl ether condensation of 15,15'-didehydro-fJ-apo-4'-carotenal (712) followed
by partial reduction and isomerization of the intermediate 15,15'-didehydro-
fJ-apo-2'-carotenal (716) [188].

2. rx-Apo-12'-carotenal
C15 + C 10 = C 25 . rx-Apo-12' -carotenal (792), an intermediate for the syn-
thesis of c5-carotene (11), was synthesized by a controlled Wittig reaction at
one end of 2,7-dimethyl-2,4,6-octatrienedial (619) with (rx-ionylideneethyl)-
triphenylphosphonium bromide (224) [166] (Table 9).
 VI. Total Syntheses 451

(248)
+
e <ll ~CH(OCH 3 ),
Br (C6 H 5 ),PCH 2 ~~ ~

(715)

(233)

(716)

(712)

3. Apo-carotenals with aromatic end groups

IX) c20 + Cw = c30. Isorenieral (798) and renieral (799), intermediates for
the synthesis or' chlorobactene (15) and okenone (181), respectively, were
prepared by the C 20 + C 10 = C 30 route starting from crocetindialdehyde (267)
as shown in Table 9.
 Vl
"""'
N

Table9. I C15 +C 10 =C25 I I C2o+Cw=C3o I [ c2s + c!O = c3, I

Cw and C 20 -Components Cw-Components Apo-carotenals References

~@2 a-Apo-12'-carotenal (792) [166]

~ -- - CH P(C6 H,), Bra + OH~CHO
(224) (619)
-
;I:
~
>
+ Bra (C.H,),~CH,y Isorenieral (798) [327] ~
(49)
- "'p..::;
OHC """' """' """' """' """' """' """' CHO 9......
(267)
5
+ Bra (C.H,),~CH, (Y Renieral (799) [369a]
(52) ~ -
@
(RO),H ~~~~~~~~~ ,CHO + (C.H,),PCH, y Apo-4'-okenal (809) [297]
Bra

(52)
-
(716a)
 VI. Total Syntheses 453

/3) C 25 +C 10 =C 35 . Apo-4'-okenal (809), used for another synthesis of

okenone (181), was made by the condensation of the C 25 -dialdehyde mono-
acetal (716a) and the Wittig salt (52) [297] (Table9).

4. Apo-lycopenals
a) Apo-15-lycopenals
y-Retinal (719), an important C 20 -intermediate, has been synthesized by
three different procedures according to the schemes c10 + c10 = c20' c15 +
C 5 =C 20 and C 16 +C 4 =C 20 .

OHC~CH20Ac
(308)

(717) R=CH 2 0H
(718) R=C0 2 CH 3

(719)

~CHO+ ~ ~colcH.
(C2 H,O),PCH2""<::::::
(614) (347 a)

""<:::::: CHO

(722)
~CH 2 0Ac
CH 30~ + I I ~
OHC~
(720) 0 (602)
454 H. MAYER and 0. ISLER

rx) C10 + C10 = C 20 . Condensation of geranyltriphenylphosphonium bro-

mide (544) with the C 10 -acetoxy aldehyde (308), followed by hydrolysis of the
product, afforded y-vitamin A (717), which was oxidized with manganese
dioxide to y-retinal (719) [166].

/3) C15 +C 5 =C 20 . Condensation of the C 15 -aldehyde (614) with the

phosphonate (347 a) in the presence of sodium methoxide led to the C 20 -ester
(718), which was reduced with lithium aluminium hydride toy-vitamin A (717).
Subsequent oxidation with manganese dioxide gave y-retinal (719) [296, 335].
2-Keto-1-methoxy-3,4-didehydro-1,2-dihydroapo-15-lycopenal (722), em-
ployed for the synthesis of 2-ketorhodovibrin (1021), was made by the base-
catalysed condensation of 3-methoxy-3-methyl-2-butanone (720) with the C 15 -
acetoxy aldehyde (602), followed by partial hydrogenation of the resulting
alcohol (721) and manganese dioxide oxidation [370].

y) C16 +C 4 =C 20 . A patent granted to Eastman Kodak Co. [371] de-

scribes the synthesis of y-retinal (719) by reaction of the Grignard complex of
the acetylenic carbinol (615) with 4,4-dimethoxy-2-butanone (181) followed by
partial hydrogenation and hydrolysis of the initial acetylenic diol acetal (723).

b) Apo-12'-lycopenals
C15 + C10 = C 25 . Apo-12'-lycopenal (793) and its 7,8-dihydro analogue (737)
have been synthesized by the C15 + C10 = C 25 route as depicted in Table 10.

/~~ "-.../~
/'-.
~ ~ 6~
/>-. 1 --~ + 0~CH(OCH3h
(615) H (181)

j
I CH(OCH3h
'*~
OH

(723) OH

l
~ ~~~~~CHO
(719)
 VI. Total Syntheses 455

CwComponents Cw-Component Apo- Refer-

12'-lycopenals ences

+ ----+ Apo-12'-lycopenal [166]

(793)
(600)
OHC~CHO
(619)
+ ----+ 7,8-Dihydroapo- [309,
12'-lycopenal 372]
(597)
(737)

c) Apo-8'-lycopenals
a) C 20 + C 10 = C 30 . The apo-8' -lycopenals listed in Table 11 are important
C 30 -intermediates for the synthesis of various carotenoids according to the
scheme C 30 + C 10 = C 40 . They were synthesized starting from crocetindial-
dehyde (267), its central-acetylenic analogue (741) and 2-keto-1-methoxy-3,4-
didehydro-1,2-dihydroapo-15-lycopenal (722) by condensation at one end
with the C 10 -Wittig compounds listed in Table 11.

f3) C 30 = C 30 . 7,8-Dihydroapo-8'-lycopenal (800, Table 35) was synthesized

from methyl 7,8-dihydroapo-8'-lycopenoate (884) by lithium aluminium
hydride reduction to the corresponding primary alcohol followed by manganese
dioxide oxidation [309, 372].

d) Apo-4' -lycopenals
C 20 + C15 = C 35 . 2-Keto-1-methoxy-3,4-didehydro-1,2-dihydroapo-4'-lyco-
penal (873) was made by condensation of the C 20 -apo-lycopenal (722) with
the C 15 -triphenylphosphonium bromide (604) (Table 11) [370].

5. /3-Apo-carotenoic acid esters

The synthesis of the C 22 -, C 25 -, C 27 -, C 30 -, C 32 -, C 35 - and C 37 -f3-apo-
carotenoic acid esters, of which ethyl /3-apo-8'-carotenoate (879) is a commercial
product, was achieved by the application of the Wittig, Horner and aldol
condensations. Numerous building principles have been followed, and the
necessa,ry cyclic and acyclic components used as well as the various synthesized
products are compiled in Tables 12, 13 and 14.

6. /3-Apo-carotenones
This group of carotenoids comprises the naturally occurring compounds
citranaxanthin (237), 8'-hydroxy-7' ,8' -dihydrocitranaxanthin (239), sintaxanthin
(244) and several citranaxanthin vinylogues (Table 15).
 Table II J C2o+Cw=C3o \ .j:>.
V'l
C\
C20 -Components C 10-Components Apo-8' -lycopenals References

+ ~CH2 ~(C6H,), Bre 15,15'-Didehydroapo- [373]

~~CHO
- 8'-lycopenal (871)
(544)
OH '*'
(741)
+ HO~Iil
""'::: CH,P(C6 H 5), Br6 1-Hydroxy-15,15'-didehydro- [373]
- 1,2-dihydroapo-8'-lycopenal
(566)
(872)

+ ~CH2 ~(C.H 5 ) 3 Br6 Apo-8' -Iyeo penal [327,374] ;z:

- (254)
(544) ~
iii
HO~Iil I»
+ ""'::: ""'::: H,P(C6 H 5) 3 Br6 1-Hydroxy-3,4-didehydro- [325] "'::s
P-
- 1,2-dihydroapo-8'-lycopenal
(557) 9
(801)
HO~Iil
"!;:;'
CHO + ""'::: H 2 P(C.H,), Br6 1-Hydroxy-1,2-dihydroapo- [327,374]
OH ""'::: ""'::: ""'::: ""'::: ""'::: ""'::: ""':::
-"'
- 8'-lycopenal (802)
(566)
(267)

+ THPOCH,~CH,;(C.H,), Bre 16-(Tetrahydropyranyloxy)- [330]

apo-8'-lycopenal (806a)
(575e)
-
CH,O~"' e 1-Methoxy-3,4-didehydro- [309]
""'::: ""'::: CH 2 P(C.H,), Br
- 1,2-dihydroapo-8'-lycopenal
(558) (803)

CH,O
+ '"~CHO 2-Keto-1-methoxy- [370]
(C.H,),PCH 2 ""'::: ""'::: ""':::
- 3,4-didehydro-1,2-dihydroapo-
0 Br6 8'-lycopenal (804)
(722) (515)
 I C2o+C1s=C3s \

C20 -Component CwComponent Apo-4' -lycopenal Reference

CH 3 0
"":, "":, "":, "":, "":, CHO + --+ 2-Keto-1-methoxy- [370]
"
(C 6 H 5 ) 3 PCH,~CHO
Bre I I I 3,4-didehydro-1,2-dihydroapo-
(722) 4' -lycopenal (87 3)
(604)

Table 12. C2o +C2 =C22 C20+Cs=C2s

I I I C2s +C2 =C21 I
Cyclic components Acyclic components P-Apo-carotenoates References ;:;
>-l
0
~CHO [
+ Br" (C 6 H 5 ),PCH,CO,CH 3
" Methyl P-apo-14'-carotenoate (874) [375, 376] (/)
- '<
::;
(325a) (390) ;.
"'"'~
+ ~co,c,H, [377]
P-Apo-12'-carotenoic acid (876)

(443b)
-
~CHO
(325a) + ?
(C 2 H,O),PCH 2 ~CO,CH,
"":, Methyl P-apo-12' -carotenoate (875) [368]
(196b)
-
~CHO
+ (C6 H 5 ),P=CHCO,CH 3 Methyl P-apo-10'-carotenoate (877) [378]
(391)
~~ - ~ Ul
(701) P-Apo-10'-carotenoic acid (260) [378]
"""'
-.J
 .j:>.
Table 13. I C2o+C10=C3o C2s +Cs =C3o Vo
I I 00

Cyclic components Acyclic components {3-Apo-8' -carotenoates References

CH,P(C H,), Cl 6
OH~CO,C,H,
~® e +
I [247]
(367) (528)

e ~COCH
Br9 [(CH,),N],PCH, "'> "'> "'> 2 2 '
+
(531) I [247]
Ethyl {3-apo-
- 8'-carotenoate (879)
ti:
9 e ~co,c,H, ~
~CHO + Br (C6 H,)[(C2 H,j,NJ,PCH, "'> "'> "'> >
[247] ~
iOj
(532)
(325a) §
p..
e ~CO,C,H,
+ Br9 (C6 H,),PCH 2 "'> "'> "'> 9......
[375]
(530) "'
5
Br9 (C6 H,),PCH,
e ~C0 2 CH 3
+ Methyl 15,15'-didehydro- [298]
"*" - {3-apo-8'-carotenoate
~~ (447)
( ) (878)

~~~CHO + yo,c,H, ---+ {3-Apo-8'-carotenoic [368a]

acid (250)
(262) (443b)
 I C27+C3=C3o I
Cyclic component Acyclic component fJ-Apo-8' -carotenoates References

TH'
+ (C6 H 5 ) 3P=CC02 CH 3 ---+ Methyll5,15'-didehydro- [378]
(402) fJ-apo-8' -carotenoate
(878)

t
~~0 Methyl fJ-apo- [378]
"""' '* 8'-carotenoate (251)
(704)
TH'
+ (C6 H 5 ) 3 P=CC0 2 CzHs ---+ Ethyl fJ-apo- [13] ;::;
o-j
(404) 8' -carotenoate (879)
:aE.
~
g.
ri
"'

.j::o.
VI
\C)
 Table 14. CJo +Cz =C32 C 25 + C 10 = C 35 C 30 +C 5 =C3 5 C 32 +C3=C3s ~
0

Cyclic components Acyclic components P-Apo-carotenoates References

+ (C,H,) 3 P==CHC02 CH 3 -----+ Methyl P-apo- [378]

~,~' (391) 6' -carotenoate
(880)
(710)

~,~rno
;:c
s::
>
(701) -<
"' ~co,c,H,
+ Br8 (C6 H,),PCH 2 ""::: ""::: -----+ Ethyl P-apo- [247] I"'
"'I»
4' -carotenoate ::s
(530) p..
(882)
9
......
"'t""
(262) I"'
"'
~ _CHO + ~C0 2 CH 3 -----+ P-Apo-4' -caro- [368a]
tenoic acid
(443a) (234)

~H,
+ (C6 H,) 3 P==CC0 2 CH 3 -----+ Methyl P-apo- [378]
~,~"' (402) 4' -carotenoate
(881)
(711)
 [ C3o+C7=C3~~C-=-+~:-;~

Cyclic components Acyclic components P-Apo-carotenoate References

~ ~0 ..

Br9
(710)
[298]
+ (C,H,),PCH,~CO,CH,
" -----+

(497) Methyl P-apo-

~~~~~~~ ,CHO
2' -carotenoate
(883) ;5
(248)
>-l
0
+ (C,H,) 3 P={;HC0 2CH 3 [378] e.
tzl
~ ~0 .. (391)
~
~
(712) ~
"'

~
-
 Table15.Citranaxanthinandvinylogues I c~~'"'SJ I C3o+Ct=C31 I I C3o+Cs=C35 I ~
n=20, 25, 27, 30, 32, 35

Refer-
ences

C23 [75,
261,
- 336]
(724)
p::
a::>
c2s. [13, ~
382] II>
C3o '"""" '"""" '"""" p..
=
(726) (727) 9......
"'
H
5
c3, ,.
I iJ -- -- -- -T -- -T [381]
-
c33 ~~~~~~~ [379,
380]
- (237)
 VI. Total Syntheses 463

• r""1
....,N
-L.....J....,
00 -
5'
....,
L.....J

f
f
f
.....
~

...... f

--
!:::-
f ;;;-
......
!:::-
f
f

~
~

V)
f ~
...... ......
;::;- !:::- ~
f s...... ~ ......
!:::- !:::-
f f
f f
f f
f

.,
= .,
.;; "' ...
r.Jr.J r.J r.J r.J
464 H. MAYER and 0. ISLER

a) Cn+ C 3 = Cn+J (n=20, 25, 27, 30, 32, 35)

Citranaxanthin (237) [379] and the vinylogous ketones listed in Table 15
were readily prepared by the base-catalysed condensation of retinal (325 a) and
the appropriate P-apo-carotenals, respectively, with acetone.
The C 23 -ketone (724) has also been made from vitamin A in the presence
of acetone under the conditions of the Oppenauer oxidation [261].
Reduction of the ketone (724) with potassium borohydride gave the cor-
responding secondary alcohol (725) [75].
When P-apo-8'-carotenal (248) was condensed with acetone at 0 °C 8'-
hydroxy-7',8'-dihydrocitranaxanthin (239) could be isolated as an intermediate.
Treatment of the latter with hydrochloric acid in chloroform then gave citra-
naxanthin (237) [380], which was further transformed into citranaxanthol
(729) by sodium borohydride reduction [379].

b) C30 +C1 =C 31
Sinthaxanthin (244) was obtained by treatment of P-apo-8'-carotenal (248)
with methyllithium followed by manganese dioxide oxidation of the initially
formed sinthaxanthol (728) [381].

c) C3o+ Cs= C3s

The P-apo-carotenone (733) and the 15,15'-didehydro analogue (732),
required for a synthesis of echinenone (148), were prepared by the base-catalysed
condensation of isopropyl methyl ketone with P-apo-8'-carotenal (248) and
15,15'-didehydro-P-apo-8'-carotenal (710) [310], respectively.
The ketones (734) and (735) were obtained similarly from 15,15'-didehydro-
apo-8'-lycopenal (871) and 1-hydroxy-15,15'-didehydro-1,2-dihydroapo-8'-
lycopenal (872), respectively [373].

7. Apo-lycopenoic acid esters

The synthesis of methyl y-retinoate (718) has been described (Section E.4 a p).
Methyl 7,8-dihydroapo-8'-lycopenoate (884) and methyl apo-6'-lycopenoate
(243) have been prepared by the C 25 + C 5 = C 30 and by the C 30 + C 2 = C 32 route,
respectively, as depicted in Table 16.

F. Syntheses of Diapo-carotenoids

The vinylogous series of diapo-carotenedials and of diapo-carotenedioic

acid esters are of special interest as potential yellow, red and violet food
pigments. They represent derivatives in which the carbon skeleton has been
shortened by the formal removal of fragments from both ends of a C40 -carot-
enoid. By the aid of key intermediates all members of the series have been
synthesized by relatively simple procedures.
~
g
0
~
::5

Table 16. l Czs +Cs =C30 C3o +Cz =C32

I I I
C25 • and C 30 -Components C2 • and C 5 -Components Apo-lycopenoates References

0
II ~CO,CH,
+ (C2 H,O),PCH 2 """"
;5
>-:1
~""-../""""
CHO
(196b)
- Methyl 7,8-dihydroapo- [309]
0
§:
"""" """" """" """" """" 8'-lycopenoate (884) en
(737) '<
"' ~C0 2 CH 3
+ (C6 H,),PCH 2 """" a
:r
Br8 0

(447)
- l "'0
"'

~""-../~~"""" CHO + (C 6 H,) 3 ~HC0 2 CH 3 Methyl [383]

apo-6' -lycopenoate (243)
"""" """" """" """" """" """" (391)
(254)
-

~
Vl
466 H. MAYER and 0. ISLER

1. Diapo-carotenedials
a) C 20 -Diapo-carotenedials
Crocetindialdehyde (267) and its central-acetylenic (741) and diacetylenic
(746) analogues (Table 17) representing the central 20 carbon atoms of the
carotenoid skeleton proved to be very valuable symmetrical C 20 -building
units for the synthesis of a great variety of carotenoids. Several well established
routes have been developed so that today these compounds are readily available
by total synthesis.
Crocetindialdehyde (267), 15,15'-didehydrocrocetindialdehyde (741) and
11,12,11',12'-tetradehydrocrocetindialdehyde (746) have been synthesized ac-
cording to the building schemes illustrated in Table 17. 11,12,11',12'-Tetra-
dehydrocrocetindialdehyde (746) was used in the first total synthesis of an
oxygenated carotenoid [315, 384].

b) C 24-, C 30 - and C 34 -Diapo-carotenedials

Diapo-6,6' -carotenedial (886), diapo-4,4' -carotenedial (748) and diapo-2,2'-
carotenedial (887) have been prepared utilizing mainly the C 10 -dialdehyde (619)
and its central-acetylenic analogue (592). The various components, central and
acyclic, and their interactions are gathered together in Table 18.

2. Diapo-carotenediones
The C 26 -diapo-carotenedione (743) was obtained by Warren and Weedon
[328] in the course of an investigation on capsorubin (205), and the C 30 -diapo-

I C3+C2o+C3=C261
0

0
"""" """" """" """" """" """" """" """" """"
(743)

I Cs+C2o+Cs=C3o I

"""" """" """" """" """" """" """" """"

(744)

(745)
 Table 17. I Cw+Cw=Czo I C6+Cs+C6=Czo C 5 +C 10 +C 5 =C 20 C3 +C,4 +C3 =Czo
I
Acyclic compone!lts Central components Diapo-carotenedials References

~CHO ® ~CH(OCH3),
(CH,O),H + Br 9 (C 6 H,),PCH 2 "<::::, "<::::, "<::::, Crocetindialdehyde (267) [247]
(520)
(708)
-
THPOCH 2
~¢
"<::::, + 0~0 11,12,11',12'-Tetradehydro- [315]
- crocetindialdehyde (746)
;5
(738) (639) '"'!
0
E
Vl
~"' 2
CH P(C H,), Cl 6 9
'<
2
(C H,O),H + OHC~CHO Crocetindialdehyde [302] a
::r
bis(diethyl acetal) (885)
(283b) (619)
- "'"'
"'"'

+ (C 2 H,0) 2 HC~CH(OC2 H,) 2 ---+ Crocetindialdehyde (267) [21, 60]

,,H, (739)

(415) I
-:::- ~CH(OC,H,),
+ (C,H,O),HC~ I 15,15' -Didehydrocrocetindialdehyde (741) [21, 60]
(740)

0\
"""'
-.J
 ~
00

Table 18. C7+Cw+C7=C24 I Cz +Czo+Cz=C24 I Cw+Cw+Cw=C30 I c12 +Cw+C12=C34

Acyclic components Central components Diapo-carotenedials References

(CH30),H~CH,;(C6H,) 3 [247]
Bra + OH~HO
(619)
Diapo-6,6' -carotenedial
(886)
OH~-$~CHO [309] ;:r:
OHCCH=P(C6 H,), + ~~
(394a) (741)
~
[
+ OH~CHO ----+-
Diapo-4,4' -carotenedial [247,385] 9
(748)
(619)
(CH 30),HC~CH,;(CoH 5) 3
Bra
5
(715) I '*~CHO
+ OHC~ I ----+-
15,15'-Didehydrodiapo- [247,385]
4,4' -carotenedial (788)
(592)

(CH30),H~CH,;(C6H,) 3 + OH~CHO Diapo-2,2' -carotenedial [247]

Bra - (887)
(742) (619)
 VI. Total Syntheses 469

carotenediones (744) and (745) were used as intermediates for a synthesis of

canthaxanthin (193) [310]. They were synthesized by an aldol-type condensa-
tion of crocetindialdehyde (267) and 15,15'-didehydrocrocetindialdehyde (741)
with acetone and isopropyl methyl ketone, respectively.

3. Diapo-carotenedioic acid esters

Diapo-8,8' -carotenedioic acid (crocetin, 269) and the corresponding di-
methyl ester (dimethylcrocetin, 270), 6'-methyl hydrogen 9'-cis-diapo-6,6'-
carotenedioate (bixin, 265) and dimethyl 9-cis-diapo-6,6' -carotenedioate
(methylbixin, 266a) are C 20- and C 24-dicarboxylic acids and esters, respectively,
which have been used as natural food-pigments for a long time. Bixin (265) the
principal pigment in seeds of Bixa orellana, was the first carotenoid of which
geometrical isomers have been encountered. It is the monomethyl ester of a
C 24-dicarboxylic acid, and stereochemical investigations have shown it to
contain a 9-cis double bond [386]. Isomerization of the natural product gave
the all-trans isomer, and the first synthetic efforts were directed towards the
preparation of the related all-trans dimethyl ester (all-trans-methylbixin, 266)
[315]. Recently, Weedon et al. [27, 288] succeeded in synthesizing dimethyl
9-cis-diapo-6,6'-carotenedioate (natural methylbixin, 266a) by a stereo-con-
trolled condensation of the C 17 -Wittig salt (623) with the C 7 -aldehydic ester
(490a) under carefully controlled conditions (Table 20).
The synthesized C 20-, C 24-, C 30- and C 34 -diapo-carotenedioic acid esters
and the necessary acylic and central components are compiled in Tables 19, 20
and 21.

4. Retro diapo-carotenoids
Retro diapo-carotenoids represent a new class of symmetrical carotenoids
which possess the retro structure found in retro C 40 -carotenoids, such as
rhodoxanthin (209) and eschscholtzxanthin (84). They were synthesized by the
usual chain-lengthening methods according to the symmetrical schemes out-
lined in the accompanying chart [362].
Horner reaction of the retro-C 12 -dialdehyde (683) at both ends with the
C 5 -phosphonate (347 a) gave dimethyl retro-diapo-7,7'-carotenedioate (751).
Horner reaction of the retro-C 18 -diketone (696) with the phosphonate (193)
yielded the C 22 -dinitrile (749), which was reduced with diisobutylaluminium
hydride to retro-diapo-7,7'-carotenedial (750). Wittig reaction of the latter with
the phosphorane (410) produced the retro-C 28 -diketone (752), which was
further transformed into the C 32 -dinitrile (753). Reduction of this dinitrile with
diisobutylaluminium hydride afforded retro-diapo-3,3'-carotenedial (754). Hor-
ner reaction of the retro-C 22 -dialdehyde (750) at bod). ends with the C5 -phos-
phonate (347 a) produced dimethyl retro-diapo-3,3'-carotenedioate (755), which
could also be prepared by chain lengthening of the retro-C 12 -dialdehyde (683)
at both ends with the C 10 -Wittig compound (541).
 Table 19. I CIO+CIO=C20 C~+C 10 +C 5 =C 20 I C3+C14+C3=C2o I
Acyclic components Central components Diapo-carotenedioic acid esters References -.1
"""'
0

0
~HO II ~co,c,H,
c,H,o,c """"' """"' ""=: + (C,H,0) 2 PCH2 Diethylcrocetin (888) [247]
(528) (533)
-
~CO,CH, 15-cis-Dimethylcrocetin (890) [350, 361, 387]
BrCH,
OH
+
N "* ~CHO - ~
(444) (592)
Dimethylcrocetin (270) [350, 361, 387]

~CH2 P(C
® 6 H5 ) 3 Bre p::
CH,O, [299] =::
>
(447)
+ OH~CHO
- ~
101
Dimethylcrocetin (270) P>
0 t:l
(619) p..
Nil H,P(OC2H 5 ),
CH 30 2 [21] 9
!;;'
(448)
- ~
101

~~CHO 15,15'-Didehydrodimethylcrocetin (891) [388]

+ OH "* - 15,15' -Didehydrodiethylcrocetin (892) [388]
R02C~(C6H5)3 (693)
(402) R=CH 3
(404) R=C 2 H 5
1 + OHC~CHO Dimethylcrocetin (270) [262]
- Diethylcrocetin (888) [262]
(689)

CH,O,C
)SP(OC,H,), + OH~CHO all-trans- + 9-cis-Dimethylcrocetin [12, 21]
(407) (689)
- .'
~
9-cis-Dimethylcrocetin (889) [12]
 Table20. C11+C7=C24 C 5 +C14 +Cs =Cz4 C 2 +Czo+Cz=Cz4

Acyclic components Central components Diapo-carotenedioic Refer-

acid esters ences

CH 3 0 2 C "
CH,P(C.H 5 ), Methylbixin (266a) [27, 288]
~ + OHC~
Br 9 - (dimethyl 9-cis-diapo-
(623) 6,6' -carotenedioate)
)O,CH,
(490a)

CH 30 2C~(C 6 H 5 ) 3 + OHC~CHO all-trans-Methylbixin (266) [306,

307]
(689)
(455)
- ;:;...,
~CHO 0

~~~~ all-trans- Methylbixin (266) [315]

e.
'<
CH 2 (C0 2 H), + OHC """ (746)
- "'::>
Er
C'O
"'C'O
~CHO
"'
~~ 15-cis-Methylbixin (893) [387]
BrCH,C0 2 CH 3 + OHC """ """ """
(741)
- ~
all-trans- Methyl bixin (266) [387]
0
II
(C 2 H 5 0),PCH 2C0 2 C,H, (196 a) [247]

'11-"""-Di"hylnod,;,in
(C6 H 5 ),P==CHC02 C 2 H, (394) (894) [262]

+ OHC """
CHO
l-
""" """ """ """ """ """ [262]
(C6 H 5 ),P==CHC0 2 CH 3 (391) (267) .j).
-.]
all-trans-Methylbixin (266)
0
II
(C 2 H,O),PCH,C0 2CH 3 (l96b) l- [21]
-
 ~
N

Table 21. Cw +Cw +C 10 =C3o C7 +Czo +C7 =C34 Cz+C3o+Cz=C34

Acyclic components Central components Diapo-carotenedioic Refer-

acid esters ences

~
(C,H,O),PCH,~CO,C,H, OHC~CHO Diethyl diapo- [247]
+
- 4,4' -carotenedioate
(533) (619) (895)
;r:
:::::
>

[389]
~
+ OHC """ """ """ """ """ """ """ CHO §

C,H,O,~CH,~(C.H,), Bre
(267)
- ""9
......
(747) OHC~-?-~CHO 5"'
+ Diethyl diapo- [389]
(741) - 2,2' -carotenedioate
(896)

~ """ ,CHO
C,H,O,CCH,P(OC,H,), + OHC'""" [247]
(196a)
""" """ """ """(748)""" """ """ """ """ -
 VI. Total Syntheses 473

c12 OHC~CHO
(683)
~
(C2 H 5 0),PCH 2
~C0
~
2 CH 3
(347 a)

0
o""' c::? c::? c::? c::? c::? c::?
C,s
(696)
"""

0II
(C 2 H 5 0),PCH 2CN
(193)
j
c22 R
c::? c::? c::? c::? c::? c::? c::? c::? R

(749) R=CN (751) R=C0 2CH 3

(750) R=CHO

(C6 H 5 ),P==CHCCH 3
(410)
0II j
0
o""' c::? c::? c::? c::? c::? c::? c::? c::? c::? c::?
c2s
(752)
"""

0II
(C 2 H 5 0},PCH 2 CN
j w JJO,CH,
(C,H,O),P3
(193) (347 a)

c32 R
c::? c::? c::? c::? c::? c::? c::? c::? c::? c::? c::? c::? R

(753) R=CN (755) R=C0 2CH 3

(754) R=CHO

l
2CH 3 0 2C~CH P(C H 2
<±>
6 5 ),
+ OHC~CHO
(541) Br8 (683)
474 H. MAYER and 0. ISLER

G. Syntheses of C 40 -Carotenoids

1. Symmetrical building schemes

a) C2o+ C2o= C4o
a) /3-Carotene. The synthesis of /3-carotene (3) by the combination of two
C 20 -building units has been achieved by six different procedures.
1) Wittig Reaction. The synthesis developed in the laboratories of the
Badische Anilin- & Soda-Fabrik AG [21, 246, 390, 391] was based on the
condensation of /3-retinyltriphenylphosphonium chloride (367) or sulphate
(369) with retinal (325 a) in the presence of methanolic potassium hydroxide,
sodium methoxide or ethylene oxide to yield all-trans-/3-carotene (3) in good
yield. Vitamin A (307 a) [246], vitamin A acetate (309a) [390], anhydro-
vitamin A (370) [246, 392] and vitamin A methyl ether (366) [248] have been
employed for the preparation of the Wittig salts, which were not usually
isolated but directly treated with retinal and base. The boron trifluoride adduct
of /3-retinyltriphenylphosphonium hydroxide has also been used [393].
Three variants of this procedure have been reported recently:
a) Instead of all-trans-retinal (325 a) the 11-cis (325 e) and 13-cis (325 b)
isomers were condensed with /3-retinyltriphenylphosphonium sulphate (369)
to give 11-cis- (3 a) and 13-cis-f3-carotene (3 b), respectively, which were con-
verted into all-trans-/3-carotene by thermal isomerization [219].
b) Condensation of the Wittig compound (369) with the keto aldehyde
(756)-readily accessible from the C 20 -diol (332) (see Section B.12d) by manga-
nese dioxide oxidation-resulted in the new type of keto carotenoid (757).
Subsequent sodium borohydride reduction and dehydration with hydrogen
bromide at 25-30 oc furnished all-trans-/3-carotene [252]. When the dehydra-
tion was performed at 0°C, 4,4'-didehydro-4,7-dihydro-/3-carotene (758) was
formed, which could be easily rearranged to all-trans-f3-carotene.
c) The triphenylphosphonium salts (367) and (369) could be replaced by
the C 20 -phosphonate (374) (Section B.12j) in a Horner reaction yielding, after
isomerization, all-trans-/3-carotene in good yield [254, 255].
Another variant was described in a patent assigned to Eastman Kodak Co.
[253] claiming the formation of /3-carotene by a Wittig condensation of the
retro-C 20 -triphenylphosphonium bromide (371) (Section B.12i) and retinal
(325 a).
2) Grignard Reaction. Retinal (325 a) was reacted with the Grignard com-
plex of the C 20 -acetylenic carbinol (364) giving the unsymmetrical C 40 -diol
(759), which on treatment with lithium aluminium hydride was transformed
into /3-carotene [245].
3) Dimerization Reactions:
a) Oxidative Dimerization. /3-Carotene could be obtained when /3-retinyl-
triphenylphosphonium iodide (368) was converted into the ylide and treated
with oxygen [394].
 VI. Total Syntheses 475

I C2o+C2o=C4o I

$ e
CH2 P(C 6 H 5 ), X
~ ~ ~ + OH ~ ~ ~ ~

(325a)

l
(367) X=Cl
(369) X=HS04

~ ~ ~ ~ ~ ~ ~ ~

(3)

1
0

~ ~ ~ ~ ~ -::7

(757)

r
®
~ ~ ~ CH 2 P(C6 Hsh
+ OH ~ ~
HSO~
(756)
(369)
 476 H. MAYER and 0. ISLER

I Czo+Czo=C4o I

~
CHO +
""""" """"" """"" """"" """""
(364)
(325a)

l
H

~
""""" """"" """""
""""" """""
(759)

l
P-Carotene
(3)

b) Reductive Dimerization. P-Carotene was prepared by treating retinal

(325 a) with phosphorus pentasulphide in pyridine [395]. In another procedure,
retinal (325 a) was treated with hydrogen sulphide to form the 2-({J-ionylidene-
methyl)-4-methyl-2H-thiopyran (760), which on treatment with zinc amalgam
in pyridine was converted into P-carotene [396]. Similar results were obtained
with 13-cis-retinal (325 b) and diisobutyl phenylphosphonite as desulphurizing
agent [397].

""'-::: ""'-::: CHO

--- P-Carotene

(325a) (760) (3)

(761)
 VI. Total Syntheses 477

c) Pinacol Formation. When retinal (325a) was reacted with zinc amalgam
in pyridine the pinacol (761) was formed, which on treatment with phosphorus
triiodide gave P-carotene [398]. Treatment of (761) with HCl resulted in
4,4'-didehydro-P-carotene (retro-dehydrocarotene, 36).
d) Other Dimerizations. The formation of P-carotene by dimerization of a
vitamin A compound was first observed when vitamin A p-toluenesulphonate
was treated with sodium iodide to give mainly anhydrovitamin A (370) and a
very low yield of P-carotene [399]. Treatment of P-retinyltriphenylphos-
phonium salts with aqueous alcoholic potassium hydroxide resulted in about
5% P-carotene [21]. When P-retinyltriphenylphosphonium sulphate (369),
however, was reacted with N,N-dimethyl-p-nitrosoaniline in the presence of
diethylamine the yield of P-carotene was improved to about 50% [141].

p) Other carotenoid hydrocarbons. Besides P-carotene a large number of

other carotenoid hydrocarbons and oxygenated carotenoids listed in Tables 22
and 23 have been synthesized by the C 20 +C 20 =C 40 route.
Racemic 15-carotene (11) and e-carotene (903) were synthesized by the conden-
sation of (J(-retinyltriphenylphosphonium chloride (343) withy-retinal (719) and
()(-retinal (344), respectively, in the presence of sodium ethoxide [166].
y-Retinal (719) on treatment with phosphorus pentasulphide or hydrogen
sulphide in pyridine furnished lycopene (19) [339, 395].
The colourless carotenoids neurosporene (22), unsymmetrical (-carotene
(25), phytofluene (30) and phytoene (32)-now generally accepted as inter-
mediates in carotenoid biosynthesis-have been synthesized by Weedon's
group in the course of an investigation of their structure and stereochemistry
[296, 335]. Thus reaction of y-retinal (719) with the Wittig reagent (627) gave
all-trans-neurosporene (22). Unsymmetrical all-trans-(-carotene (25), phyto-
fluene (30 a) and phytoene (32 a) were obtained by condensation of all-trans-
(geranylgeranyl)triphenylphosphonium bromide (630) withy-retinal (719), with
the C 20 -aldehyde (626) and with geranylcitral (631), respectively.
Lycopersene (34) was synthesized from geranylgeranyl bromide (629) in a
Wurtz-type reaction by treatment with sodium [ 400] or lithium in ether [23].

y) Oxygenated carotenoids. The synthesis of a number of keto carotenoids

by the C 20 + C 20 = C 40 route has recently been reported by Surmatis et al. [167].
Thus (4-acetoxy-P-retinyl)triphenylphosphonium sulphate (379) was condensed
with retinal (325 a) yielding isocryptoxanthin (40), which was oxidized to
echinenone (148) by the Oppenauer method [401]. Alternatively, 4-acetoxy-
retinal (375) was reacted with P-retinyltriphenylphosphonium sulphate (369)
furnishing echinenone (148) by a similar sequence of reactions [167]. Condensa-
tion of (4-acetoxy-p-retinyl)triphenylphosphonium chloride (380) with 4-acet-
oxyretinal (375), followed by saponification of the product, resulted in iso-
zeaxanthin (71);which was oxidized to canthaxanthin (193) by the Oppenauer
method [256]. Treatment of the acetate of isocryptoxanthin with 68% hydro-
bromic acid at -45 °C gave 4,4'-didehydro-P-carotene (retro-dehydrocarotene,
 Table22. I C2o+C2o=C4o I .j:o.
-.J
00

C 20 -Components Carotenoids References

+ ---+ .:>-Carotene (11) [166]

CH 2 P(C6 H 5 ), (719)
~@ Cl 8
# (343)

+ OHC~ ---+ e-Carotene (903) [166]

(344)
;:c
~
>
+ OHC ---+ Lycopene (19) [339, 395] -<
'"""' :0
"'
(719) "'::sp..
CHO

'"""' '"""' ""' '"""'

'"(719) '"""' '"""' 9
+ (C6 H,j,PCH 2
" ---+ Neurosporene (22) [296, 335]
Br8
(627)
-
"~'
:0

+ ---+ '-Carotene (25) [296, 335]

(unsymm.)
(719)

CH 2 P(C6 H,),
"
+ ---+ Phytofluene (30a) [296, 335]
'"""' '"""' '"""' '"""' Br8
(630)
(626)

+ ---+ Phytoene (32a) [296, 335]

'
(631)
""" """ """ """ CH,Br ---+ Lycopersene (34) [23,400]
+ BrCH, """ """ """ """
(629)
(629)

~~~CH,P(C,H,), "
+ Isocryptoxanthin (40) [401]
HSOf'
OH~
(325a)
OAc
(379)
Echinenone (148) [401]

t
~~~ ,CHO Yl --+ Isocryptoxanthin (40) [167]
+ (C6H 5 ) 3 "'PCH,~
HSOf' ;:5
(369)
...,
0
OAc
(375) E.
C/)
'<
a
::r
"
""'"'
~~~~CH,P(C,H,) 3 "' + OHC'~~~~ ---+ Isozeaxanthin (71) [256]
CI 9

(375)
OAc
~
(380)
Canthaxanthin (193) [256]

~~~CH,P(C,H,), " ~YI

+ OHC~ 4,10'-Diketo-7',10'-di- [167]
HSOf'
hydro-P-carotene -.J
(756) \0
"""
(1023)
OAc
(379)
480 H. MAYER and 0. ISLER

(36), which, on reaction with N-bromosuccinimide in chloroform and acetic

acid, yielded isozeaxanthin diacetate (1008). Saponification, followed by oxida-
tion with aluminium isopropoxide, then yielded canthaxanthin (193) [167].
The cross-conjugated keto carotenoid (1023), in which the keto function is
substituted on the unsaturated chain, was prepared by the condensation of the
Wittig compound (379) with the keto aldehyde (756) followed by Oppenauer
oxidation of the product [167].
Several patents assigned to Eastman Kodak Co. [339, 397, 402, 403]
describe the formation of zeaxanthin diacetate (1 005), isozeaxanthin diacetate
(1008), isozeaxanthin dimethyl ether (1009) and of canthaxanthin (193) by
dimerization of the suitably substituted retinals (762), (375), (763) and (764),
respectively (Table 23).

C 20 -Components Carotenoids References

~CHO Zeaxanthin diacetate

(1005)
[339,402,403]

AcO (762)

Isozeaxanthin diacetate [339, 402, 403]

(1008)

OAc
(375)

Isozeaxanthin dimethyl [339, 402, 403]

ether (1 009)

(763)

'"""' ""':: "'>: CHO --+- Canthaxanthin (193) [339,402,403]

(764)

b) C19+C2+C19=C4o

a.) f3-Carotene. The manufacturing procedure for /3-carotene elaborated in

the laboratories of F. Hoffmann-La Roche & Co. Ltd. follows the scheme
C 19 + C 2 + C 19 = C 40 . It was based on the first synthesis of Inhoffen et al. [206]
and was adapted to technical requirements by Isler et al. [186]. Condensation
of two moles of f3-C 19 -aldehyde •(285) with acetylenedimagnesium bromide
 VI. Total Syntheses 481

I c,9+C2+C,9=C4 o I

,::? ,::? ,::? + Hc=cH + OH

CHO ,::? ,::? ,::?

(285)
(285)

l
H
,::? ,::? ,::?
,::? ,::? ,::? '*'
OH
(765)

(766)

(767)

l
all-trans-{J-Carotene
(3)

Carotenoids 31
482 H. MAYER and 0. ISLER

gave the C 40 -diol (765), which was readily converted into 15,15'-didehydro-
P-carotene (766) by allylic rearrangement and simultaneous dehydration.
Subsequent partial hydrogenation over Lindlar catalyst gave mainly 15-cis-
P-carotene (767), which was isomerized to all-trans-P-carotene (3) by treatment
in high-boiling petroleum ether.

P) Other carotenes. The syntheses of 3,4,3',4'-tetradehydro-p-carotene (1)

[260, 404], 3,4,3',4'-tetradehydro-7,7'-dihydro-P-carotene (897) [245], 4,4'-
didehydro-P-carotene (36) [245] and 7,7'-dihydro-P-carotene (37) [245] were
all based on the same reaction scheme starting from the appropriate C 19 -
aldehydes (293), (292) and (285), respectively (Table 24). When the C 40 -diol
(765) was treated with lithium aluminium hydride, the central acetylenic
grouping was reduced giving 7,7'-dihydro-P-carotene (37) [245].

C 19-Aldehydes Carotenoids References

~CHO 3,4,3',4'-Tetradehydro-P-carotene (I) [260,404]

I
(293) 3,4,3',4'-Tetradehydro-7, 7' -dihydro- [245]
P-carotene (897)

~CHO --+- 4,4'-Didehydro-P-carotene (36) [245]

(292)

~CHO --+- 7,7'-Dihydro-P-carotene (37) [245]

(285)

y) Oxygenated carotenoids. Zeaxanthin (1004) [41, 187], physalien (1006)

[ 41 ], zeaxanthin dimethyl ether (1 007) [ 102], astaxanthin dimethyl ether
(1026) [102], isozeaxanthin (71) [208, 405] and canthaxanthin (193) [208]
have also been prepared by the c19 + c2 + c19 = c40 route starting from the
appropriate C 19-aldehydes (295), (295 a), (296) and (292), respectively (Table 25).

c) Cts+C4+Cts=C4o
P-Carotene. P-Carotene has been prepared from P-C 18 -ketone (275) by
means of a Grignard reaction with diacetylene. The resulting diacetylenic
glycol (768) was partially hydrogenated and the product treated with phos-
phorus diiodide to give P-carotene in low yield [ 406].
 VI. Total Syntheses 483

C 19-Aldehydes Carotenoids References

~CHO--
AcO (295)
Zeaxanthin (I 004) [41, 187]
Physalien (I 006) [41]

~Ho--
Aco (295a)

~CHO-- Zeaxanthin dimethyl ether

(1007)
[102]

CH,O (296) Astaxanthin dimethyl ether [102]

(1026)

~CHO --
Isozeaxanthin (71)
Canthaxanthin (193)
[208, 405]
[208]
(292)

Following the same scheme Pommer [21] synthesized P-carotene by the

condensation of a C 18 -triphenylphosphonium halide, e.g. (282a), with male-
aldehyde or fumaraldehyde.

d) C16+Cs+C16=C4o
rx.) P-Carotene. The early syntheses of P-carotene following the scheme
C 16 + C 8 + C 16 = C 40 were accomplished by Karrer and Eugster [2] and by
Inhoffen et al. [5] in 1950. The condensation of the Grignard complex of the
acetylenic c16-carbinol (237) with 4-octene-2,7-dione (639) yielded the c40-
tetrol (769), which was partially hydrogenated to the tetrol (770). Subsequent
dehydration and isomerization afforded P-carotene in rather low yield.
When the C40 -tetrol (769) was dehydrated prior to partial hydrogenation,
the sterically hindered 11,11'-di-cis-P-carotene (3 c) was obtained [407].
A variant of the procedure using the acetylenic C 16 -hydrocarbon (238) has
been reported [3]. As was shown by Eiter et al. [338, 408], however, the retro
isomer (239) (Section B. 8 b) had actually been employed in the synthesis, which
explains the very low yields obtained by this approach. When the pure C 16 -
hydrocarbon (238) was condensed with the C 8-diketone (639), the yield of pure
P-carotene could be raised to about 32%. The initial diacetylenic glycol (771)
was reduced with lithium aluminium hydride and the product dehydrated
 484 H. MAYER and 0. ISLER

(275)
(275)

(768)

l
P-Carotene
(3)

l
(282a)
(282a)

with N -bromosuccinimide in benzene. The glycol (771) could also be dehydrated

with phosphorus oxychloride in pyridine giving 11,12,11',12'-tetradehydro-P-
carotene (784), which was then selectively hydrogenated and isomerized.
A closely related procedure, which used the isomeric acetylenic C 16 -car-
binol (247) (or 247 a?) (Table 26~ was briefly reported [6].

p) Other carotenes. The following symmetrical carotenes (listed in Table 26)

have been prepared similarly from suitable acetylenic C 16 -carbinols and the
C 8 -diketone (639): e-carotene (903) [179, 337], (- )-(6S,6'S)-e-carotene (904)
[107, 179a], (+)-(6R,6'R)-e-carotene (10) [107, 179a], isorenieratene (13)
[180-182], renierapurpurin (16) U82], lycopene (19) [337, 338] and 5,6,5',6'-
tetrahydrolycopene (906) [341].
 VI. Total Syntheses 485

~~+0~+~~
(639) (237)
(237)

(769)

l
(770)

P-Carotene (3)

0~0
(639) (238)
(238)
486 H. MAYER and 0. ISLER

Table 26. I c16 + Cs + c16 = c40

C8 =0~0
(639)

C 16 -Cornponents Carotenoids References

~¢ ?'
[3]
#
(239)

~¢
(247)
H - P-Carotene (3) [6]

~¢ (?) [6]

(247 a)

~¢
(243)
- e-Carotene (903) [179, 337]

~¢
#
(244a)
H
- (- )-(6S, 6' S)-e-Carotene (904) [107, 179a]

~¢
(244b)
- (+ )-(6R, 6' R)-e-Carotene (10) [107, 179a]

~¢
""""
">:,..

(246)
H
- Isorenieratene (13) [180-182]

:()+¢
">:,..
"""" H
- Renierapurpurin (16) [182]

(246a)

(Continued)
 VI. Total Syntheses 487

Table 26. (Continued)

Carotenoids References

~~

~)
OH
(615) Lycopene (19) [337, 338]

(617)

~~

~)
OH
(618) 5,6,5',6'-Tetrahydrolycopene (906) [341]

I I I ¢
~
(618a)

Table27. I c.6+Cs+Ct6=C40

C,=O~O
(639)

Carotenoids References

Ld,r~ ~~ (243)
- rae.
IX-Carotene
(900)
[409, 410]

(d,r* ~~ (244a)
- (- )-(S)-
IX-Carotene
(901)
[107]

Ld,r¢ ~~ (244b)
- (+)-(R)-
IX-Carotene
(5)
[107]

~- - y-Carotene
(8)
[411]

(615)

~~ (246)
--+ Renieratene
(14)
[182]
 ~
I C1s+C10+Cts=C4 0
00
00

+ OH~CHO ,8-Carotene
(3)
'"'
(619)
-
2 CH2P(C.H,),
~ CI 8

(204)
15,15'-Didehydro-
I '*~CHO ------+- ,8-carotene
+ OH~ I (766)
(592)
p::
s:::
~
[
'"' / ,8-Carotene
'"' H 6 5 ), Bre
+ Br 8 (C 6 H 5 ),PCH 2/
~~~~
~~~
~ CH2P(C 9
- (3)
(668) [
'"'
Bre (C H ) PC'"' I .q;. ~CH2P(C.H,h Bre
+ 6 53 H2~ I -
2~CHO (666)
15,15'-Didehydro-
(180) ,B-carotene
(766)
0

o II
(C H 0) II I .q;. ~CH2P(OC2Hsh -
+ 2 5 2PCH2~ I
(667)
 VI. Total Syntheses 489

Unsymmetrical carotenoid hydrocarbons such as rae. oc-carotene (900) [ 409,

410], (- )-(S)-oc-carotene (901) [107], ( + )-(R)-oc-carotene (5) [107], y-carotene
(8) [411] and renieratene (14) [182] have also been prepared by the
reaction of 1:1 mixtures of two different C 16 -components with the C 8 -diketone
(639) (Table 27). Mixtures were always obtained from which the unsymmetrical
carotenoid could be separated in low yield by chromatography.

e) cl5 + clO + c15 = c40

oc) {3-Carotene. The synthesis of all-trans-{3-carotene according to the
scheme c15 + clO + c15 = c40 was realized by two different procedures:
1) Two moles of ({3-ionylideneethyl)triphenylphosphonium chloride (204)
were condensed with 2,7-dimethyl-2,4,6-octatrienedial (619) [159, 161,207, 412]
in the presence of sodium methoxide or sodium acetylide in dimethylformamide
or acetonitrile. When a C10 -dialdehyde with a central-cis double bond was
used, 15-cis-{3-carotene (767) was formed [412].
2) Two moles of {3-ionylideneacetaldehyde (180) were condensed with
the C 10 -Wittig compound (668) in the presence of phenyllithium [353]. When
the symmetrical central components (619) or (668) were replaced by the
central-acetylenic analogues (592) [161, 412] and (666) [353] or (667) [21],
respectively, 15,15'-didehydro-{3-carotene (766) was obtained, which could
readily be transformed into all-trans-{3-carotene as mentioned (Section G. 1 boc).

f3) Other carotenes. The following symmetrical carotenes listed in Table 28

have been synthesized in an analogous way from the appropriate c15-tri-
phenylphosphonium salts and the Cw-dialdehyde (619): a-carotene (903) [166],
lycopene (19) [166], (-carotene (symm.) (26) [296, 335] and 15-cis-(-carotene
(26a) [296, 335].

y) Oxygenated carotenoids. 9,9'-Di-cis-alloxanthin (1002) [12, 27], zea-

xanthin (1004) [109, 123], 3,4,3',4'-tetradehydro-16,16'-lycopenedial (1017)
[247] and 2,2' -diketospirilloxanthin (208) [308] have been prepared accordingly
from the C 15 -phosphorane (166) and the C 15 -Wittig compounds (165), (231),
(715) and (609), respectively (Table 28).
Surmatis et al. [167] synthesized a number of symmetrical and unsym-
metrical oxygenated carotenoids utilizing the Wittig compound (668) as
symmetrical central building unit. Thqs condensation of (668) with two moles
of the substituted C 15 -aldehydes (226) and (227) afforded the 3-oxygenated
carotenoids (772) and (773), respectively. When (772) was hydrolysed with
10% aqu'eous sulphuric acid, the product consisted of 4,4' -dihydrorhodoxanthin
(772a) and two minor products of structure (774) and (775). Hydrolysis with
36%HC1 gave (775) as the main product, contaminated with (772a) and (774).
Hydrolysis of (773) by either of the two methods yielded (775) as the main
reaction product.
A mixture of the f3-C 15 -aldehydes (226) and (180) reacted with the C10-
compound (668) according to the same scheme giving a condensation product
490 H. MAYER and 0. ISLER

.1. . . .,._ . . .,j._

c,.=OHC' ~ ~
,CHO

(619)

C15 -Components Carotenoids References

----+ e-Carotene (903) [166]

~H2 ~(C.,H,) 3 ----+ Lycopene (19) [166]

Cle
(601)

(597)
- all-trans-
'-Carotene (symm.) (26)
15-cis-,-Carotene (26a)
[296, 335]

[296, 335]

- 7,8,7',8'-Tetradehydro-
zeaxanthin (1003)
[123]

~
Zeaxanthin (1004) [123]

~~ CH=P(C.,H,), ---+ 9,9'-Di-cis-alloxanthin [12,27]

(1002)
HO (166)

----+ Zeaxanthin (1004) [109]

----+ 3,4,3',4'-Tetradehydro- [247]

16, 16'-lycopenedial
(1017)

CH,o~e
'<::::: '<::::: '<::::: '<::::: CH 2 P(C6 H 5), ----+ 2,2'-Diketospirillo- [308]
xanthin (208)
(609) Br"

~CHO
I ~ I
*C1o=OH~
(592)
 VI. Total Syntheses 491

2 ~CHO
oM +

(.o (226)

l 0~
0

(772)

"""' """' """' """' """' """' """'

o:r
(772 a)

"""' """' """' """' """' """' """'

(774)

"""' """' """' """' """' """' """'

o:r
(775)

l
C2H 5 0
(773)

2 ~CHO +

C 2H 5 0
A)l
(227)
492 H. MAYER and 0. ISLER

+ +

(776)

(777)

which was hydrolysed to the unsymmetrical carotenoid (776). Purification

of this compound led to the isolation of another keto carotenoid, shown to
be 4,4'-didehydro-{3-caroten-3-one (777) [167].

f) C14+C12+C14=C4o
rx) {3-Carotene, 3,4,3',4'-tetradehydro-{3-carotene, zeaxanthin, 7,7'- and 15,15'-
dihydro-{3-carotene. Two different procedures have been devised for the syn-
thesis of the carotenoid skeleton by the c14 + c12 + c14 route:
1) Enol Ether Condensation. Condensation of the diethyl acetal of the
f3-C 14 -aldehyde (250) with 2,9-diethoxy-3,8-dimethyl-1,3,7,9-decatetraen-5-yne
(687) yielded the acetylenic C 40 -diketone (778). Reduction with aluminium
 VI. Total Syntheses 493

~(OC,U,), (C,U,Oj,H~
(250) (250)

+ +

(778)

(779)

l
15,15'-Didehydro-P-carotene ----+- P-Carotene
(766) (3)
494 H. MAYER and 0. ISLER

C14-Components Carotenoids References

~CH(OC,H,), --+ 3,4,3',4'-Tetradehydro-P-carotene (1) [359]

(780)

~CH(OC,H,), - Zeaxanthin (1004) [359]

AcO)__)l
(781)

~CH(OC,H,), - Zeaxanthin (1004) [359]

o"" (782)

isopropoxide led to the corresponding glycol (779), which on dehydration

gave 15,15'-didehydro-P-carotene (766) [359]. The latter was converted into
all-trans-P-carotene as mentioned earlier (Section G. 1 hoc).
Starting from the diethyl acetals (780), (781) and (782), 3,4,3',4'-tetradehydro-
P-carotene (1) [359] and zeaxanthin (1004) [359], respectively, have been pre-
pared by a very similar sequence of reactions (Table 29).
2. Nef-Type Reaction. Condensation of the dilithium derivative of 3,8-
dimethyl-3,5,7-decatriene-1,9-diyne (678) with two moles of the P-C 14-aldehyde
(123) [305, 413] gave the initial diacetylenic C 40 -glycol (783), which on acid-
catalysed dehydration yielded 11,12,11',12'-tetradehydro-P-carotene (784).
Subsequent selective hydrogenation over Lindlar catalyst led to the sterically
hindered 11,11'-di-cis-P-carotene (3 c), which was thermally isomerized to
all-trans-P-carotene.
3,4,3',4'-Tetradehydro-P-carotene (1) and zeaxanthin (1 004) were synthesized
in the same way from 3,4-didehydro-P-C 14-aldehyde (139) [413] and 3-acetoxy-
P-C14-aldehyde (145) [413], respectively (Table 30).
7,7'-Dihydro-P-carotene (37) [414] and 15,15'-dihydro-P-carotene (899)
[360] were prepared by the condensation of the C12 -hydrocarbon (680) and
the C12 -glycol (688), respectively, with two moles of P-C14 -aldehyde (123)
(Table 30).
 VI. Total Syntheses 495

(123) (678) (123)

J
~H ~~~
~~~~
~
~ 6u (783)

1
Ill
~~~~~
~~
(784)

1
11,11 '-Di-cis-P-carotene
(3c)

1
All-trans-P-carotene
(3)
496 H. MAYER and 0. ISLER

Table30. I c14+C12+C14=C40
c 12 = ..&
?'-
~~
(678)

C 14-Aldehydes Carotenoids References

~CHO - 3,4,3',4'-Tetradehydro-P-carotene (1) [413]

(139)

AcO
~CHO - Zeaxanthin (1004) [413]

(145)

- 7,7'-Dihydro-p-carotene (37)* [414]

~CHO
(123)
- 15,15'-Dihydro-P-carotene (899)** [360]

* Cu =¢ ~~ ** Cu =¢ ~~
(680) (688)

p) Rhodoxanthin. A total synthesis of rhodoxanthin (209) has been reported

[44] by the condensation of the retro-C14-phosphorane (156) with the retro-
C12-dialdehyde (683).

g) c13 + c14 + c13 = c4o

The condensation of P-ionyltriphenylphosphonium chloride (95) or
bromide (9.4) with the c14-dialdehyde (689) in the presence of methyllithium
or sodium methoxide yielded a mixture of P-carotene isomers from which all-
trans-P-carotene could be obtained by iodine-catalysed isomerization and
chromatography [61, 97]. When the central-acetylenic C 14-dialdehyde (693)
was employed, a mixture of 15,15'-didehydro-P-carotene isomers (mainly the
9,9'-di-cis isomer) was formed from which the all-trans isomer (766) was obtained
by crystallization [97, 98].

h) C10 +C 20 +C 10 =C 40
Crocetindialdehyde (267) and its central-acetylenic analogue (741) have
been widely used as symmetrical central components for the synthesis of a
 VI. Total Syntheses 497

~CH=P(C6H5) 3
0
~ (156) (156)

+ +

OH~CHO
(683)

(209)

great variety of carotenoids by the C 10 +C 20 + C10 =C 40 route. Reaction of the

C 20 -dials, at both ends, with the appropriate C 10 -components was effected by
the Wittig, Horner and aldol condensations.
Thus condensation of two moles of P-cyclogeranyltriphenylphosphonium
bromide (27) with crocetindialdehyde (267) yielded P-carotene [129, 415].
When the C 20 -acetylenic dialdehyde (741) was employed, 15,15'-didehydro-P-
carotene (766) was formed [60], which was converted into P-carotene as
mentioned.
The various carotenoids synthesized from crocetindialdehyde (267) and
the appropriate C10 -components are listed in Table 31.
The aldol-type condensation was successfully applied by Warren and
Weedon [331] to the first unambiguous synthesis of canthaxanthin (193).
The 2,6,6-trimethyl-3-oxo-1-cyclohexen-1-yl end groups were elegantly intro-
duced into the carbon skeleton by condensation of crocetindialdehyde (267)
at both ends wjth the C 10 -monoketal (585) in the presence of alkali yielding
the nonaene dione (785). The latter compound, after acid hydrolysis of the ketal
grouping, was then cyclized to canthaxanthin (193) in the presence of alkali.
The reaction ·was also used to establish the structure and configuration
of natural capsorubin (205) [69]. Condensation of crocetindialdehyde (267)
with the trans keto alcohol (61) gave racemic capsorubin (1027) thus proving

Carotenoids 32
 1 cl3 + c:,4 + [c,;+ C~o +C10 = C4o
.j:>.
c,3 :-c::-1 \0
00

/3-Carotene /3-Carotene
(3) (3)

r r
OHC~CHO CHO
OHC'~~~ ~ ~ ~ ~
(689)
(267)
+ + ;:r:
+ +
~
>
$
$111 aCH2P(C6 H 5 h Br 8 ~
~$ Cle (C.H,),P~
8
6 5
~ P(C H hCl
Br8
$'(1
(C H ),PCH2~
~
6 5 9
(95) (95) (27) (27)

+ +
i
+ +

"':::CHO
?;-
~~ OHC~?;-~CHO
OHC "'::: "'::: (741)
(693)

1 1
15, 15'-Didehydro-p-ca rotene 15, 15'-Didehydro-p-ca rotene
(766) (766)
 VI. Total Syntheses 499

Table 31.

C20 = OH

(267)

C 10-Components Carotenoids References

-- Isorenieratene (13) [66]

A) H P(C H,), Br6

~e 2

(52)
6
> Renieratene (14)

Renierapurpurin (16)
[66]

[66]

3-Isorenieratenol (52) [67, 68]

3,3'-Isorenieratenediol [67, 68]

(79)

(52 d)

~e
"""" """" """" CH 2 P(C6 Hsh Br

(550)
6
-- 3,4,3',4'-Tetradehydro-
lycopene (17)
[323]

~CH,;(C6H,), Br6
(544)
•
- Lycopene (19) [59, 60,
416]

HO~"
"""" """" CH2 P(C6 H,), Br

(557)
e
- 1,1'-Dihydroxy-
3,4,3',4' -tetradehydro-
1,2,1 ',2' -tetrahydrolycopene
[325]

--
(1013)

HO~ e 6 1,1'-Dihydroxy- [323]

"""" CH2 P(C6 H,), Br
1,2,1',2' -tetrahydrolycopene
(566) (81)

I I ~CHO
*C 20 also 0HC~'* I I
(741)
(Continued)
500 H. MAYER and 0. ISLER

Table 31. (Continued)

C 10 -Components Carotenoids References

Spirilloxanthin (108) [323, 325]

(558)

CH,O~"'
"""

(569)
CH,P(C.H,), Br
9
- 3,4,3',4'-Tetrahydro-
spirilloxanthin (110)
[323, 326]

(CH 3 0),HC
~.,
"""
2 """

(708)
""" CH P(C6 H,), Br
e
- 3,4,3' ,4'-Tetradeh ydro-
16,16'-lycopenedial (1017)
[247]

~CH,~(C6H,), Br9
C2 H 5 0 2 C
(530)
- 3,4,3',4'-Tetradehydro-
16,16'-lycopenedioic acid
[247]

C 2 H 50 2C
~II
""" """ """
0

CH 2 P(OC2 H,), 1 diethyl ester (1 033)

(533)

I C10+Czo+C10=C4o I

g
II
+ OHC ~ ~ ~ ~

(267)
~ ~ ~
CHO +
:Q (585)
(585)

1
0
I I0

~ ~ ~ ~ ~ ~ ~ ~ ~

u
(785)

1
Canthaxanthin
(193)
 VI. Total Syntheses 501

Table 32.

(267)

C 10 -Components Carotenoids References

~ 0
----+ 6,6' -Diketo-5,6,5',6' -tetrahydrolycopene (1025) [309]

(576)

H0~- 1,1 '- Dihydroxy-6,6' -diketo- [328]

0 1,2,5,6,1',2',5',6' -octahydrolycopene (I 030)
(577)

u (63) ----+ rae. 3,3'-Dideoxycapsorubin (1024) [70]

-t! (61) ----+ rae. Capsorubin (1027) [69]

t7- bH
(60) ----+ rae. Capsorubin epimer (1028) [69]

HO
u 0

(66) ----+ rae. Capsorubin isomer (1029) [71]

the trans relationship of the oxygen substituents at the cyclopentane ring

(Table 3,2). When the cis keto alcohol (60) was employed, a capsorubin epimer
(1028) was obtained that was clearly different from the natural product.
Some other· carotenoids prepared by this method are listed in Table 32.

i) Cs+C3o+Cs=C4o
Aldol-type condensations have also been used for carotenoid syntheses
according to the C 5 + C 30 + C 5 = C 40 building scheme.
 502 H. MAYER and 0. ISLER

(788)
+ +

RO~
~R
(786) R=H
(787) R=CH 3
j
RO '<::::::: '<::::::: '<::::::: '<::::::: '<::::::: '<:::::::
'<::::::: '<::::::: '<::::::: '<::::::: '<::::::: '<::::::: '* OR

0
(789) R=H
(790) R=CH 3

j
RO
'<::::::: '<::::::: '<::::::: '<::::::: '<::::::: '<::::::: '<::::::: '<::::::: '<::::::: '<::::::: '<::::::: '<:::::::
OR
0
(207) R= H
(208) R=CH 3

1
CHO
OH '<::::::: '<::::::: '<::::::: '<::::::: '<::::::: '<::::::: '<::::::: '<::::::: '<::::::: '<::::::: '<:::::::

+ (748) +
0
RO~
0
~OR
(786) R=H
(787)R=CH 3
 VI. Total Syntheses 503

Thus reaction of 15,15'-didehydrodiapo-4,4'-carotenedial (788) at both

ends with 3-hydroxy-3-methyl-2-butanone (786) or 3-methoxy-3-methyl-2-
butanone (787) in the presence of methanolic potassium hydroxide gave
15,15'-didehydrophillipsiaxanthin (789) and 2,2' -diketo-15,15'-didehydrospi-
rilloxanthin (790), respectively. Subsequent partial hydrogenation over Lindlar
catalyst and isomerization furnished phillipsiaxanthin (207) and 2,2'-diketo-
spirilloxanthin (208) in good yields [385]. Much lower yields were obtained
in these condensations when the fully conjugated C 30 -dialdehyde (748) was
used as symmetrical central component [385].
Condensation of the central-acetylenic C 30 -diketone (745) at both ends
with a large excess of 1-diethylamino-3-pentanone methiodide (462) in the
presence of alcoholic potassium ethoxide yielded 15,15'-didehydrocantha-

l
"'(~~~~
~ .1 ~ _,l /~~
0 (745)

+ +

~~ CH2~(C2Hsh
CH,
(462) (462)

(791)

1
Canthaxanthin
(193)
504 H. MAYER and 0. ISLER

xanthin (791) [310], which was transformed into all-trans-canthaxanthin (193)

by partial hydrogenation followed by isomerization [208]. Again, much lower
yields were obtained when the fully conjugated diketone (744) (Section F.2)
was used as starting material.

2. Unsymmetrical building schemes

a) ell+ c19 = c40
The unsymmetrical building scheme C 21 +C 19 =C 40 was devised as a
variation of the already discussed c19 + c2 + c19 = c40 route that allowed the
synthesis of both symmetrical and unsymmetrical carotenoids. In principle,
a C 2 cacetylenic carbinol [e.g. (387)], prepared from an appropriate C 19 -alde-
hyde, was condensed with the same or a different C 19 -aldehyde to give an
acetylenic C 40 -diol such as (765) (Section G. 1 biX). This was converted into the
desired carotenoid either by direct reduction with lithium aluminium hydrid~
according to the reaction of Nayler and Whiting [ 417] or by the following
sequence of reactions: dehydration with simultaneous allylic rearrangement,
partial hydrogenation and isomerization of the resulting cis to the all-trans
isomer.
The various carotenoids that have been prepared by the combination of
C 2 c and C 19 -components are displayed in Table 33.
A total synthesis of rhodoxanthin (209) following the C 21 + C 19 = C 40
building principle has recently been reported by Surmatis et al. [167] (Table 33).
The Wittig reaction of the acetylenic C 21 -keto aldehyde (383) with the C 19 -
triphenylphosphonium bromide (300) in the presence of sodium methoxide
yielded 10,11,10',11'-tetradehydrorhodoxanthin (1031). Subsequent partial
hydrogenation with Lindlar catalyst followed by thermal isomerization
afforded all-trans-rhodoxanthin (209). Rhodoxanthin could also be obtained
directly by the condensation of the fully conjugated C 2 cketo aldehyde (384)
with the Wittig compound (302) [111].

bJ c2s + c1s = c4o

Many unsymmetrical carotenoids are synthetically accessible today by
the Wittig synthesis according to the scheme c25 + c15 = c40. P-Carotene (3),
for example, has been prepared by the condensation of 15,15'-didehydro-P-
apo-12'-carotenal (701) with (fJ-ionylideneethyl)triphenylphosphonium bromide
(205) yielding 15,15'-didehydro-P-carotene (766), which was transformed into
all-trans-P-carotene (3) as mentioned (Section G. 1 biX) [153]. The carotenoids
that have been prepared by this method are shown in Table 34 together with
the C 25 - and C 15 -components used.

c) C30 + C10 = C40

This building principle has been extensively used for the synthesis of a
large number of unsymmetrical carotenoids, especially by the teams led by
Weedon, Isler and Surmatis. The construction of the C40 -carbon skeleton was
 VI. Total Syntheses 505

achieved by two different methods, namely by the Wittig reaction and by the
aldol-type condensation.
1) Wittig Reaction. Suitable apo-8'-carotenals and C 10 -triphenylphos-
phonium halides were condensed in the presence of sodium methoxide or
ethoxide, propyl- or butyl-lithium, or anhydrous potassium carbonate to
yield the desired carotenoids directly. When 15,15'-didehydroapo-8'-carotenals
were employed, the resulting 15,15'-didehydrocarotenoids were converted into
the desired end products by successive selective hydrogenation and isomeri-
zation. The various carotenoids synthesized by this method are listed in Table 35.
2) Aldol-Type Condensations. The first unambiguous synthesis of echi-
nenone (148) was achieved by Warren and Weedon [331]. Similar to a syn-
thesis of canthaxanthin (193) (Section G. 1 i), the 2,6,6-trimethyl-3-oxo-1-
cyclohexen-1-yl end group was introduced by the base-catalysed condensation
of P-apo-8'-carotenal (248) with the C 10 -monoketal (585) yielding the decaene
ketone (807). Acid-catalysed hydrolysis of the ketal grouping followed by
intramolecular aldol-condensation then led to echinenone (148).

(248) (585)

(807)

(148)
 VI
Table33. I c2. +C.g=C40 I ~

C 21 -Components C19 -Components Carotenoids References

+ P-Carotene (3) [186, 418]

OH~ -
(285)

+ OH 3,4-Didehydro-P-carotene (2) [260] p::

~ (293)
- s::>
~
g:
Po
9.....
+ rae. at-Carotene (900) [118]
,?' OH~ "'
~~ H
- i
(387) (287c)

+ 4,4'- Didehydro-4, 7-dihydro-p-carotene [245]

OH~ - (758)
(292)

+ OH rae. Cryptoxanthin (908) [187]

~ (295)
-
 + OH 3,4,3',4'-Tetradehydro-P-carotene (1) [260]
~ (293)
-
+ 3,4-Didehydro-p-carotene (2) [260]
,:7 OH~ -
~~ H
(285)
(388)

+ 3,4-Didehydro-7, 7' -dihydro-p-carotene [245] <

~
OH~ - (898)
>-l
(285) 0
§:
en
'<
" 0 10,11,10',11'-Tetradehydro- [167] a
+ (C6 H,),PCH 2 Y" ~
Br9 ~ rhodoxanthin (1031)
if
rl
0 ~~~ .., ,:7 ,:7 HO ,:7 ,:7
- "'
(383)
(300)
j
+ (C6 H,h;CH 2 Rhodoxanthin (209) [111, 167]
~CHO Br9
#
0 (384)
~- (302)

9
 ...,
0
Table 34. C2s +Cis =C4o
I I 00

C 25 -Components C 15 -Components Carotenoids Refer-

ences

+ Br6 (C,.H,),PCH,
®~ ""'=:: ""'=:: -----+ P-Carotene (3) [153]

(205)

+ Br6 -----+ rae. a-Carotene (900) [153]

(C6H,),~CH,~ ;:z:
~~rno ~ ~
(224) >
"""" ~
(701) I»
+ Br6 (C6H,),~CH,~ -----+ y-Carotene (8) [153] I:S
(l.

(600) 9
......
"'r;;
+ Br6 (C6H 5 ),;CH 2~ -----+ P-Zeacarotene (9) [153] "'
(597)

+ Br6 (C6H,),~CH,~ -----+ ( + )-(R)-a-Carotene (5) [106]

(224a)
~~~-CHO

(262) -----+ P-Zeacarotene (9) [28]

+ Br6 (C.H,),;CH,~
(597)
 + Bra (CoHsh~CH,~ ot-Zeacarotene (12) [28]

~~~~yCHO
(597)
-
(792) + Bra (C6H 5 h~CH,~ .5-Carotene (11) [166]
(600)
-
<!
+ Cia (C.H 5 hPCH,
$~ """" """" y-Carotene (8) [166]
- >-l
-
(204) g
til
'<
;.
=
~~~~~~~~~CHO + Cia .5-Carotene (11) ""'
(C6H,h;CH,~ [166] ""'
(793) (225)
-
+ Bra (C6H 5h;CH 2~ ~eurosporene(22) [296,
335]
(597)
-
~~~~~~~~yCHO + Bra (C.Hsh~CH,~ Spheroidenone (182) [309]
OCH, -
(737) (609)
U'o
0
\0

(Continued)
 Table 34. (Continued)

C 25 -Components C 15 -Components Carotenoids Refer-

ences
Ul
~
OH 0
X! 9-cis-Crocoxanthin [12]
+ (C,H,),P=r .
--- (909)
y~
(166)

7,8-Didehydro- (123]
- cryptoxanthin (907)

~CHO p::
D:~ 7,8,7',8'-Tetradehydro- (123] s::>
HO I (794)
~X!OH --- zeaxanthin (1003) ;:j
~
~ l>l
+ Br9 (C,H,),~CH,~ ::s
Zeaxanthin (1004) [123] ~

(165) 7,8-Didehydro- [123]

9
--- zeaxanthin (1 003 a) ;
7,8-Didehydrolutein [123]
- (1010)

rae. Cryptoxanthin [109]

(262) I - (908)
e ., ~"
+ Br (C,H,),PCH,

(231) rae. Zeinoxanthin [109]

--- (1000)
(792)
 Table 35. I c30 + c10 = c40 I
C 30-Components C10-Components Carotenoids Refer-
ences

®
Br9 (C.H,),PCH P-lsorenieratene [66]
+ , I
(49)
n - (6)

e Ell P- Renierapurpurin [66] ;:S

+ Br (C6 H,),PCH 2 ...,
(902)
0
(52)
¥ - g,
{I')
Ell ~
+ Br9 (C6H,),PCH,~ y-Carotene (8) [153] ;.
(544)
- "'~
~~~~~~~ -CHO

(248) 1'-Hydroxy- [327]

·+ Br9 (C6H,),~CH,~OH - 1',2'-dihydro-
(566) y-carotene (48)

Bre "
(C6H,),PCH,~CH(OC,H,)z Torularhodin- [247]
+
- aldehyde (142)
(797)

+ Br9
..
(C6H,),PCH,~CO,CH 3 Torularhodin [298]
- methyl ester (212) VI
(527) Torularhodin [23, -
ethyl ester (1032) 247]

(Continued)
 Vl
.....
N

Table 35. (Continued)

C 30 -Components C 10 -Components Carotenoids Refer- p::

ences ~
>
><:
~
"'
Ill
::;
~,.~" """ '-":::: ----+ 0.
+ Br 9 (C 6 H,),PCH
@~ 2 y-Carotene (8) [153]
9
(544) ......
(710)
"'t"'
~
"'

+ Br6 (C6 H 5 ),PCH

@~ 2 """ '-":::: ----+ Chlorobactene [327]
(15)
(544)
(798)

~~~~ ,CHO
+ Br6 (C,H,) 3 ~CH,~OCH ----+ 4' -Deoxookenone [369a]
3
(1014)
(569)
(799)
~
Okenone (181) [369a]
 + Br9 (C6 H5 J,PCH,
" ---+ Chlorobactene [327]
I
~
(49)
¥ (15)

+ Bre (CoH,),;CH,y ---+ y-Renierapurpurin [327]

(905)
(52)

+ "~
Bre (C6 H 5),PCH 1 ""'I ""'I _____.. Lycopene (19) [327] <
t-'
>-1
(544) 0
[
~ ~~~~~~~~":::, ,CHO

(254) ~:r
+ Br9 (C.H,J,~CH,~ ---+ 3,4-Didehydro- [325] "'"'
I /"oH rhodopin (55) "'"'
(567)

+ Br9 (C6H5 J,;CH,~H ---+ Rhodopin (56) [327,

374]
(566)

+ "~
1
Br9 (C6H,J,PCH,
l"ocH, ---+ Anhydrorhodovibrin [374]
(97)
(558) Ul
.....
\,;.>

(Continued)
 Table 35. (Continued)

C 30 -Components C 10 -Components Carotenoids Refer-

VI
ences .......
""'"
+ Br8 (C6H,),~CH 2~0H Chloroxanthin [309,
-- (58) 372]
(566)

~~~~~~~~y~yCHO + Br8 (C6 H,),PCH,

®~ """" """" Spheroidene [308,
OCH,
(800) (99) 309]
(558)
--
OH

+ ®~
Br8 (C6 H,),PCH,
I I Spheroidenol [308,
OCH, - (1016) 309] :r:
(575)
t a::
)>
-<
Spheroidenone [308, ~
(182) 309]
"'
I'>
::s
0..

HO
9
CHO ......
+ ®~
Br8 (C,H,),PCH, ""'1 """" - OH-Spirillo- [325] "'t"'
OCH, ~
"""" """" """" """" """" """" """" """" I """" """" I xanthin (105) "'
(801) (558)

+ ®
Br e (C6 H,),PCH 2 - OH-Chloro- [327]
bactene (53)
(49)
v
HO, I
CHO
+ Br8 (C 6 H 5 ),PCH,
®~ """" """" Rhodopin (56) [327]
"""" """" """" """" """" """"
"""" """" """" (802) -
(544)

+ @~
Br e (C,H,),PCH, """" ~- Rhodovibrin [374]
OCH,
(106)
(558)
--
 ~y~yCHO + Br8 (C 6 H 5 ),PCH,
®~ """ """ - Anhydro- [309]
(803) rhodovibrin (97)
(544)

+ Br8 (C6 H 5 ),PCH,

®~ """ H 2-Ketorhodo- [370]
CH,O, I
- vibrin (1021)
I (566)

0 2-Ketorhodo-
(804) + Br8 (C6 H 5 ),~CH,~OCH, [370]
- vibrin methyl
(569) ether (1022)

~,.~" + ®~
Br8 (C6 H 5 ),PCH,
"""1 OH 4,1 '-Dihydroxy- [373]
I """ - 15,15' -didehydro-
(566) 1',2'-dihydro-
~
I ...,
OH 0
(805) y-carotene (I 0 11)
§:.
[/)
'<
1:1
+ Br8 (C6H 5 ),~CH,~ 4-Keto- [373] :;.
- y-carotene (151)
~CHO (544) "'"'~
~~~~ '*'
+ Br 8 (C,H,),PCH,
®~ """ 1'-Hydroxy-4-keto- [373]
OH
0
- 1',2'-dihydro-
(806) (566) y-carotene (160)

~ Lycoxanthin [330]
THPOCH,/ ~'--../~~~~~~y~yCHO + tetrahydropyranyl
(806a) ®~
Br8 (C6 H 5 ),PCH, """ """ ether (1001)
(544)
CHO + ------> Methyl [330a]
CH,O,C
""" 16-lycopenoate
Vl
""" """ """ """ I """ """ I
""" """ """ (806b)
(1001 a) ......
Vl
1
Lycoxanthin (62) [330a]
 516 H. MAYER and 0. ISLER

C30 -Components C10-Components Carotenoids Refer-

ences

+ ---+-3'-Deoxy- [70]
cryptocapsin
(1019)
(63)

(248)

--+-rae. Cryptocapsin [419]

(1020)

--6-
(61)

-., -., -., -., -., CHO +

---+- Capsanthin [28]
(170)
(249) ~0
(61)

Similarly, rae. cryptocapsin (1020) [419] and capsanthin (170) [28] were
prepared by the condensation of the trans keto alcohol (61) with P-apo-8'-
carotenal (248) and natural P-citraurin (249}, respectively. This proved the
position and the trans configuration of the two oxygen substituents on the
five-membered rings in the natural products.
The carotenoids prepared by the aldol condensation are listed in Table 36.

d) C3s+Cs=C4o
Wittig and aldol condensations have also been applied to the synthesis
of carotenoids by the C 35 +C 5=C 40 route. Thus condensation of P-apo-4'-
carotenal (714) with (3,3-dimethylallyl)triphenylphosphonium bromide (452)
or·ethyl cx-methylcrotonate (443 b) gave torulene (7) [303] and torularhodin
(211) [368a], respectively. Base-catalysed condensation of 15,15'-didehydro-
P-apo-4'-carotenal (712) with 3-hydroxy-3-methyl-2-butanone (786) led to
2'-dehydroplectaniaxanthin (162), which could be reduced to plectaniaxanthin
(76) by treatment with lithium aluminium hydride [321]. When the methoxy
ketone (787) was employed, the corresponding methyl ether of plectania-
xanthin (1012) was obtained [321].
Besides P-apo-4'-carotenals the c35-apo-carotenones listed in Table 37
were used as starting materials for unsymmetrical 3-oxygenated carotenoids.
The introduction of the oxygenated end group was again achieved by base-
catalvsed condensation with 1-d1ethylamino:3-pentanone methiodide (462).
 VI. Total Syntheses 517

Interestingly, much better yields could be obtained when C 35 -15,15'-dide-

hydroapo-carotenones were employed as was observed in the case of echinenone
(148) and 15,15'-didehydroechinenone (1018) [310]. The various carotenoids
prepared by the C 35 + C 5 = C 40 route together with the necessary C 35 - and
C 5 -components are recorded in Table 37.

e) C 37 +C 3 =C 40
15,15'-Didehydro-/J-apo-2'-carotenal (716) was the starting material for
a number of carotenoids synthesized by the C 37 +C 3 =C 40 building scheme
(Table 38). Chain lengthening by three carbon atoms was effected by the
propenyl ether condensation to give 15,15'-didehydrotorularhodinaldehyde,
which was partially hydrogenated and isomerized to all-trans-torularhodinal-
dehyde (142) [188]. Subsequent lithium aluminium hydride reduction and
acetylation gave the corresponding primary alcohol and acetate, respectively
[188]. Treatment of (716) with the phosphorane (402) furnished 15,15'-dide-
hydrotorularhodin methyl ester, which was similarly converted into all-trans-
torularhodin methyl ester (212). Saponification of the latter then yielded torula-
rhodin (211) [378].

H. Syntheses of Carotenoids with Less than or More than 40 Carbon Atoms

The carotenoids discussed in this section comprise the isoprenologous
series of symmetrical P-carotene analogues with 30, 50 and 60 carbon atoms
and carotenoids with 20, 30, 34 and 37 carbon atoms possessing acylic and
aromatic end groups.

1. C 20 -Carotenoids
The C 20 -hydrocarbon (811) as a model of phytoene (32) has been synthesized
by Davis et al. [296] by the condensation of geranyltriphenylphosphonium
bromide (544) with citral (88).

+ OHC~
(544) (88)

l
(811)
 Vl
Table 37. I C 35 +C 5 =C4o 00
I -
C 35 -Components C 5 -Components Carotenoids References

+ Br8 (C6 H,),~CH,~ - Torulene (7) [303]

(452)
~~~~~y~y~yCHO
~co,c,H,
(714) + - Torularhodin (211) [368a]

(443 b) p::
3:::
>
...:
0
~
"'
Ill
::;
+ - 2'-Dehydroplectaniaxanthin [321] 0..
~OH (162) p
(786) ......
~"~rno ~ "'t"'
~
"'
""" Plectaniaxanthin (76) [321]
(712) 0

+ - Plectaniaxanthin [321]
methyl ether (I 012)
~OCH 3
(787)

~~~~~~CHO + LiCH 2~OCH, - Okenol (1015) [12]

(809) (810)
~
Okenone (181) [12]
 ~~~~~~~~ + ----+- Echinenone (148) [310]
(733)

~4~ + • ----+-
1
15,15'-Didehydroechinenone [310]
~ ~ (1018)
(732)
~ ~H, CH,rJ{C,H,),
Ie

(462) ----+- y-Caroten-4-one (151) [373]

h ~~~""'~ +'
~
~~~
<
!"""'
...,
HO h ""' ""' + ----+- 1'-Hydroxy-1',2'-dihydro- [373]
~~Y~I
~ ~ ~ s.a:.
y-caroten-4-one (160) en
~
&
0
~
"'
Table 38. I C37 + C3 = C4o I
C 37 -Component C 3-Components Carotenoids References
,H,
+ ----+- Torularhodinaldehyde (142) [188]
'
(415)
~·~0
(716) TH'
+ (C6 H 5 ) 3P=CC0 2 CH 3 ----+- Torularhodin [378]
(402) methyl ester (212) VI
,.....
\0
~
Torularhodin (211) [378]
 Table 39. I ct6+Ct4=C3o=C15 +Cts I c14 +C2 +Ct4=C3o=C13 +C4 +C13 I Cw+Cw+Cw=C3o=Cs +C2o+Cs ' VI
IV
0
References

~~ + [420]

(237)
-- ?'
OHC~ '<:::::

(123)
~-~ (812)
'<:::::
~~ + -- J [420]
~
~
(238) >
-<
:<'
"'
"'::sp..
~CHO + [395, 403]
OH~ -- 9
!i;'
(180) !"'
(180) !:l

+ H<=CH --I ~ [420,421]

2 ~CHO
(123) a~~~"""
(813)

+ Hc=c-c=cH [406]
2 ~0 --
(89)

CH2 P(C6 H,), Br

e e + OH~CHO [61]
2 -- J
ex (27)
(619)
 "'
~CH 2 P(C 6 H 5 ), Br8 I '*~CHO
2~ - + OHC~ I
~*~ [61]

(27) (592) (812)

2
I
~CH 2 P(C,H,),
I "'
+ OH~CHO ~ ~
Br 9
[296]
(544) (619)
- (814)

2 ~CH 2 ~(C,H,),
+ ~ ~ ~ ~ ~ ~ ~ ~ rno ~~
Br8
(453) (267) (815) [296]
;s
-1
:aE.
r/l
8
;.
"'~

Vl
N
......
522 H. MAYER and 0. ISLER

2. C 30 -Carotenoids
Several syntheses of the C 30 -/]-carotene isoprenologue (813) and its central-
acetylenic analogue (812) as models for /]-carotene have been performed by
Inhoffen's group. The various routes and building units used are assembled
in Table 39. The hydrocarbon (813) could also be obtained from the C 15 -
aldehyde (180) by reductive dimerization with phosphorus pentasulphide [395]
or hydrogen sulphide in pyridine [ 403] (Table 39). A patent granted to Badische
Anilin- & Soda-Fabrik AG [61] describes the preparation of (813) and (812)
by the Wittig reaction of the C 10 -dialdehydes (619) and (592), respectively,
with two moles of cyclogeranyltriphenylphosphonium bromide (27).
The C 30 -hydrocarbon (814, Table 39) as a model of (-carotene (26) was
obtained by the Wittig reaction of the C 10 -dialdehyde (619) at both ends with
geranyltriphenylphosphonium bromide (544) [296].
The related C 30 -hydrocarbon (815, Table 39) has been synthesized by
reaction of crocetindialdehyde (267) with isopentyltriphenylphosphoniuin
bromide (453) [296].

3. C 34 - and C 37 -Carotenoids
In the course of the synthesis of renieratene (14) and related arylpolyenes,
Weedon's group [66] synthesized the diphenyl analogue (817) of /]-carotene
by the condensation of benzyltriphenylphosphonium bromide (816) with
crocetindialdehyde (267) as shown in Table 40. Compound (817) had earlier
been obtained in low yield by the condensation of the acetylenic carbinol
(817 a) with the C 8 -diketone (639) [421 a].
The phenyl analogue (818) of /]-isorenieratene (6) and /]-renierapurpurin
(902), respectively, was readily obtained by reaction of /]-apo-8'-carotenal (248)
with benzyltriphenylphosphonium bromide (816) in the presence of propyl-
lithium [66] as shown in Table 40.

4. C 50 -Carotenoids
Three different routes have been devised for the synthesis of the interesting
C 50 -/]-carotene isoprenologue decapreno-/]-carotene (1035), which differs from
/]-carotene by two isoprene units in the chain structure.

a) C21+Cs+C21=Cso
The first synthesis of decapreno-/]-carotene (1 035) was carried out by
Karrer and Eugster [258]. By analogy with their synthesis of /]-carotene
(Section G.1da) they used as building units the acetylenic C 2 ccarbinol (385)
and the C 8 -diketone (639) (Table 41).
According to the same scheme decapreno-B-carotene (1 036) was synthesized
starting from the acetylenic carbinol (386) [259].
 VI. Total Syntheses 523

Refer-
ences

" Bre [66]

20
~CH 2 P(C,.H,),
'"""' '"""' '"""' CHO

(816) (267)

l
(817)

r
~4 + + ~~ ,? ,?
:::::,.1
[421 a]

(817a) (639) (817a)

[66]
+

(248) (816)

(818)

b) Czo+C10+C2o=Cs0
In a synthesis developed by Surmatis and Ofner [353] two moles of retinal
(325a) were condensed with the Wittig compounds (668) and (666) in the
presence of phenyllithium to give decapreno-P-carotene (1035) and 19,19'-
didehydrodecapreno-P-carotene (1034), respectively (Table 41). The latter
compound could be converted into (1035) by selective reduction and iso-
merization [353]."
The C 50 -diketone (1037) was prepared by the reaction of the C10 -Wittig
salt (668) with two moles of the C 20 -keto aldehyde (756, Table 41) [167, 422].
 VI
Table 41. N
C21 +Cs +C21 =Cso C2o+Cto+C2o=Cso ~
! I I I c.9+Ct2+C.g=Cso I
Cyclic Components Central Components Carotenoids References

"'::, + Decapreno-P-carotene (1 035) [258]

~~ H -
(385)
0~0
(639)
"'::, + ---+ Decapreno-e-carotene (1036) [259]
~~ t:t:
# (386) H =:::
>
;:i
!»
"'::s
(C.,H,),;CH, ~eCH 2 P(~ 6 H,), Po
+ ---+ Decapreno-P-carotene (1035) [353]
9 Br
Br
9......
(668)
"'t""
~CHO "'"'
(325a)
..
~.& ~CH 2 P(C 6 H,), 1
e • B~
+ (C.H,),PCH2 "'::, ---+ 19,19'-Didehydrodecapreno-p-carotene (1034) [353]
Br9
(666)

EO ~CH,;(C.H,),
~CHO + (C.,H,),PCH, e ---+ 10,10'-Diketo-7,10,7',10'-tetrahydro- [167, 422]
9 Br
Br decapreno-p-carotene (1 037)
(668)
(756)
 ~ r,H,
~CH(OC1H,) 1 + ~~~ - - 19,19' -Didehydrodecapreno-{1-carotene (1 034) [359]
OC1 H,
(819) (687) i
Decapreno-{1-carotene (1 035) [359]

Table 42. I C26 + C 8 + C26 = C6o -\ I C2o + C2o + C20 = C60 ;:;

Ol
Cyclic Components Central Components Carotenoid References [
Vl
~
~~~~
~
0?
~~~- &
+ ...._..... ~ yo [261] ~
OH
(389) (639)
--1
Dodecapreno-p-carotene
(1038)
~CH1 ;(C6H,), + OH"'::,"'::,"':::,"':::,"'::,"':::,"'::,CH0--
~ HSO~ toLm [321]
(369) (267)

~
VI
526 H. MAYER and 0. ISLER

c) C19+C12+C19=Cso
Isler et al. [359] employed the enol ether condensation of the P-C 19 -acetal
(819) with the C 12 -bis(enol ether) (687, Table 41) in analogy to one of their
P-carotene syntheses (Section G.lfoc) to obtain 19,19'-didehydrodecapreno-P-
carotene (1 034), which by partial hydrogenation and isomerization gave
decapreno-fl-carotene (1 035).

5. C 60 -Carotenoids
Dodecapreno-fl-carotene (1 038) contammg 60 carbon atoms and 19
conjugated double bonds represents the highest member of the isoprenologous
P-carotene series synthesized thus far. It has been obtained by two different
routes as shown in Table 42.

a) C26+Cs+C26=C6o
The compound was first synthesized by Karrer and Eugster [261] by the
reaction of the Grignard complex of the acetylenic carbinol (389) with 4-octene-
2,7-dione (639). The reaction sequence followed was similar to that used for
a synthesis of P-carotene by the same authors (Section G.1doc).

b) C2o+C2o+C2o=C6o
Dodecapreno-fl-carotene (1 038) has also recently been obtained by the
Wittig reaction of crocetindialdehyde (267) with two moles of P-retinyltri-
phenylphosphonium sulphate (369) in the presence of sodium methoxide [321].

I. Syntheses of Related Compounds

1. Analogues with altered carbon skeleton
This section contains brief accounts of some polyenes that are in some way
related to the carotenoids mentioned previously in Sections E, G and H. The
following types of compounds have been synthesized as shown in Tables 43
and 44:
a) Symmetrical P-carotene and canthaxanthin analogues in which the
central portion has been shortened or lengthened [formulae (823), (826),
(831), (837) and (846)]. The diketone (826) was prepared as a model of cantha-
xanthin (193) [331]. A novel type of reaction was used for the synthesis of the
C 44 -analogue (846) of P-carotene by treatment of retinal (325 a) with butadiene
(845) in the presence of(cyclopentadienyl) 2 Ni [423].
b) Unsymmetrical analogues of phytoene and P-carotene with 30, 33, 35
and 40 carbon atoms [formulae (822), (827), (828) and (834)].
c) Unsymmetrical compounds possessing aromatic end groups [formulae
(821), (824) and (830)].
d) Carotenoids with additional alkyl substituents [formulae (836), (839),
(841), (843) and (844)].
 VI. Total Syntheses 527

e) Demethylcarotenoids [formula (833)].

t) Isoprenologues of fJ-apo-12'-carotenoic acid [formulae (847) and (848)]
and retinoic acid [formulae (850), (851) and (852)] (Table 44).

2. Abscisic acid
Abscisic acid [ 430, 431 ], previously known as abscisin II [ 432] or dormin
[433, 434], is a naturally occurring plant growth inhibitory factor controlling
such processes as dormancy, abscission, germination and senescence. It was
first isolated in crystalline form from young cotton fruit [ 435] and sycamore
leaves [433, 434]. Its structure [436-438] and absolute stereochemistry [439]
were established as shown in formula (857)*.
Four different routes have been devised for the total synthesis of this
compound.
The first synthesis was performed by Cornforth et al. [ 438] in 1965. cis-
3,4-Didehydro-{J-ionylideneacetic acid (212b) [164] was irradiated with
visible light in an atmosphere of oxygen in the presence of eosine as a photo-
sensitizer yielding the epidioxide (853). This was rearranged into racemic
abscisic acid (855 b) by heating at 100 oc for 7.5 minutes in 0.07 N aqueous
sodium hydroxide followed by acidification.

~ ~
u ~02R
(212b) R=H
~
(853) R=H
~02R
(211 b) R = CH 3 (854) R=CH 3

'L:l
~~10
C0 2 H
0
(857)

The same reaction sequence was carried out with the all-trans-C 15 -acid
(212 a) affording the all-trans isomer (855 a) of abscisic acid.
Resolution of the synthetic racemate (855 b) into the enantiomers was
achieved by fractional crystallization of the brucine salt followed by additional
fractional crystallizations of the partly resolved enantiomeric free acids [ 439].
The synthesis of racemic abscisic-10_1 4 C acid and all-trans-abscisic-10- 14 C
acid has been reported recently [ 440].
Mousseron-Canet et al. [65] reported the photosensitized oxidation in the
presence of Rose Bengal of methyl cis-3,4-didehydro-{J-ionylideneacetate
* The numbering follows the current carotenoid convention and therefore differs from that
in which C0 2 H=C-l.
528 H. MAYER and 0. IsLER

Table43.

~CHO +
(325 a) (820)

"""" "'
CH,P(C,H,), Br8
+ OH~
(630) (88)

2 ~CHO
{123)

~CHO
(325 a) (75)

+ OH~CHO +
(825) (585)
(585)

+ CI 8 (C,H,),~~
(95)
~CHO
(325 a)

(204)

NOCH,

+ OHC~
(77)
 VI. Total Syntheses 529
Related compounds

References

[392]

[296]
(822)

[406]

[73]

(824)

[331]

(826)

[392]

(827)

[75]

OCH,

[392]

(Continued)
Carotenoids 34
530 H. MAYER and 0. ISLER

Table43.

~CHO +

(325 a) (282 b)

2 ~CHO +

(832)

2 u
~@
'~'" CH P(C,H
2 5
), CI
9
+ OH~CHO

(204) (520a)

(387)

~CHO
u --- + Hc==c-c=cH

2
(285)

0~0
2D:+~ +
(639)

(840)

2 ~CHO + HC=cH

(835) R = C 2 H 5
(842) R = CH 2 CH(CH 3 ),

2 ~CHO + ~
(845)
(325 a)
 VI. Total Syntheses 531

(Continued)
References

[75]

[424]

[425]

[426]

[406]

[427]

[428]

(841)

[426]

(843) R=C 2 Hs
(844) R=CH 2 CH(CH 3 h

[423]
 Table 44. Related compounds

Refer-
VI
ences w
IV

~co,c,H,
~CHO + ~CO,H [377]
- "
(180) (443 b) (847) n= 1
xe (848) n=3,4, 5

~H,~(C.J1,) 3 + OH~C02 R -l [246]

(369) X=HS0 4 (354) R=C 2 H 5

(367) X=Cl (353a) R=H
p::
=:::
OH~C0 2H >
-<
~~(C.J15),CI6 + [159]
- !il
(316) !»
(282b)
::s
' /
"-
~CO,R 9
~
-
:<1
6
(850) R = C 2 H 5
~CH,~(C.H,), Br + OHC~CO,H (851) R=H [159,
(303) 207,
(205)
- 429]

~~(C6H,), CI 6 + OH~CO,H --+-I [98,

(849) 129]
(95)

OHC~C02C2H, C0 2 C 2H 5
~CH 2 ~(C6H 5) 3 +
""".' """ """ """ """ """ """ [246]
HSO~ (304)
(369)
- (852)
 VI. Total Syntheses 533

(211 b) leading to the epidioxide (854). This, on base-catalysed rearrangement

with basic aluminium oxide, gave the ester (856), which was saponified with
N/10 sodium hydroxide. Similar treatment of methyl all-trans-3,4-didehydro-
fJ-ionylideneacetate (211 a) furnished all-trans-abscisic acid (855 a).
The synthesis developed by Roberts et al. (J. R. Reynolds Tobacco Co.)
[441] started from !X-ionone (101), which on oxidation with t-butyl chromate
gave a 4:1 mixture of 6-hydroxy-3-keto-IX-ionone (118) and 3-keto-!X-ionone
(858). The pure diketone (118), obtained by chromatography and crystallization,
was treated with (carbethoxymethylene)triphenylphosphorane (394) in a
Wittig reaction yielding a 1: 1 mixture of the cis-trans-isomeric esters (859 b)
and (859a). Subsequent hydrolysis of this mixture with methanolic potassium
hydroxide produced abscisic acid (855 b) and its all-trans isomer (855 a) which
could be readily separated by crystallization from ether.

(101)

l
~0
0~
+

(118) (858)

\ (C 6 H 5 )JP==CHC02C2H,

\ (394)

C02C2Hs
+ H
0
(859b) (859a)

l
~ ~co~
U to2R +

(220b) R=C 2 H 5 (220a) R=C 2 H 5

(860b) R=H (860a) R=H
534 H. MAYER and 0. ISLER

~0 + ~~ ~ ~~H,OH
\:-a
CH2 0H

(5) (I 58 b) (861)

--
Alternatively, IX-ionone (101) was first reacted with the phosphorane (394),
with the phosphonate (196 c) [ 442] or with ethyl bromoacetate [ 443] giving
a mixture of cis-trans-isomeric ethyl IX-ionylideneacetates (220b) and (220a).
Interestingly, the Wittig reaction gave equal amounts of (220b) and (220a),
whereas the Horner reaction gave predominantly (220a). Subsequent oxidation
of the ester mixture with t-butyl chromate, however, yielded only 5 % of the
esters (859a) and (859b). Hydrolysis of the esters (220a) and (220b) gave the
corresponding acids (860a) and (860b), respectively, the cis isomer (860b)
being almost as active as abscisic acid in accelerating abscission and inhibiting
growth [ 443].
Two alternative syntheses of the diketone (118) have already been mentioned
(Section B.5 f).
In the Roche synthesis [ 444] the diketone mono ketal (5) was condensed in
the presence of lithium amide in liquid ammonia with cis-3-methyl-2-penten-
4-yn-1-ol (158b), an intermediate in the technical Roche vitamin A synthesis.
The resulting acetylenic diol (861), on treatment with lithium aluminium
hydride and subsequent acid hydrolysis of the product, gave the diol (862).

(864)

0
~
~0/1'-./
_co,H

(865)
ua
~
(866)
bo,H
 VI. Total Syntheses 535

Oxidation of the latter with manganese dioxide in methylene chloride furnished

the aldehyde (863), which was transformed into abscisic acid (855 b) by alkaline
silver oxide oxidation.
Ohkuma [ 445, 446] described the synthesis of some analogues of abscisic
acid, namely of the 1-cis-9-trans isomer (855 c), of the lactone (864) and of the
spiro compound (865). The synthesis of the epoxy acid (866) and the cor-
responding ethyl and methyl esters, exhibiting similar plant growth inhibiting
properties as abscisic acid, has been reported recently [ 447].

3. Grasshopper ketone
For a detailed review of the chemistry of the allenic ketone from grass-
hoppers see Chapter V.
A synthesis of the allenic ketone (870), tentatively proposed for the grass-
hopper ketone, has recently been reported [ 448] as outlined in the accom-
panying scheme. The lithium salt of 3-butyn-2-ol (103) was added to the
silylated ketone (867) yielding the protected acetylenic diol (868), which on
lithium aluminium hydride reduction and subsequent hydrolysis gave the triol
(869). Manganese dioxide oxidation then furnished the ketone (870).

No ,. . "*AoH
TMSO~OTMS
OH
TMSO OTMS
(867) (103) (868)

!
N·~o N·~YOH
HO~OH HO~OH
(870) (869)

J. Syntheses of Labelled Compounds

Carotenoids, vitamin A and related compounds labelled with 14 C or 3 H

are indispensable tools for the investigation of their absorption, distribution,
storage and metabolism. P-Carotene and lycopene labelled with deuterium at
the central double bond were used for the elucidation of the fragmentation
pattern of these carotenoids in mass spectrometry [ 425]. Table 45 shows the
labelled positions and the specific activities of some of these compounds which
have mainly been prepared by Dr. J. Wiirsch in the laboratories of F. Hoff-
mann-La Roche & Co. Ltd., Basle.
536 H. MAYER and 0. ISLER

Table 45. Labelled compounds

Compound Labelling Specific activity References
j!Ci/mg
(!lCifmmole)
Vitamin A
all-trans 6,7-t4c2 43.7 [449]
14-14C (296) [244, 450]
15-14C 29.6 [449]
15- 3 H 26,400 [449]
all-trans (acetate) 11,12- 3 H 2 300 [449]
7-cis 14-14C [244]
13-cis 14-t4c [244]
all-trans-Vitamin A acid 6,7-14c2 20 [449]
14-14C (314) [244,450]
15-14C 37.2 [449]
10,11- 3 H 2 352 [449]
P-Carotene 6,6'-14C2 0.014 [451]
15.15'-14C 2 { 1.0
15
[418]
[449]
15,15'- 3 H 2 1,093 [449]
15,15'-02 [425]
Lycopene 15,15'- 3 H 2 715 [449]
15,15'-02 [425]
Canthaxanthin 15,15'- 3 H 2 1,615 [449]
P-Apo-8' -carotenal 15,15'- 3 H 2 590 [449]
Ethyl P-apo-8' -carotenoate 15,15'- 3 H 2 1,666 [449]
Torularhodin ethyl ester 15,15'- 3 H 2 750 [449]
Abscisic acid 10-14C 36 [449]
all-trans-Abscisic acid 10-t4c 36 [449]

K. Annex. Synthetic Carotenoids (Tables)

Table 46. Synthetic apo-carotenoids

a) Apo-carotenals

(698) P-Apo-14'-carotenal

~CHO
Horner [192, 363]

(262) P-Apo-12' -carotenal

CHO
'"""' '"""' '"""' '"""' '"""'
c1s + clO = c2s Wittig [364, 365]
C19+C6=C2s Grignard [280]
C2o+Cs=C2s Wittig [366]
Aldol [367]
C21 +C4=C2s Grignard [188,280]

(Continued)
 VI. Total Syntheses 537
Table 46. (Continued)

(794) 3-Hydroxy-7 ,8-didehydro-fl-apo-12'-carotenal

~CHO
De~
HO
c.s + clO = c2s Wittig [123]

(704) 15,15'-Didehydro-fl-apo-1 0' -carotenal

~CHO

~~"""
C13+C14=C27 Wittig [364]
C2o+C7=C27 Wittig [249]

(256) P-Apo-1 0' -carotenal

CHO
""" """ """ """ """ """
C2o+C1=C21 Wittig [366]
C2s +C2 =C21 Enol ether [188, 369]

(248) /l-Apo-8' -carotenal

HO
""" """ """ """ """ """ """
C2o+C10=C3o Wittig [247, 336, 364]
C2s+Cs=C3o Wittig [366]
Aldol [367]
C21+C3=C3o Enol ether [188]

(713) P-Apo-6' -carotenal

CHO
""" """ """ """ """ """ """ """
C2s+C1=C32 Wittig [247]
C3o+C2=C32 Enol ether [188]

(714) p!Apo-4' -carotenal

CHO
""" """ """ """ """ """ """ """ """
c2s + clO = C3s Wittig [247]
C32 +C3 =C3s Enol ether [188]

(Continued)
538 H. MAYER and 0. ISLER

Table 46. {Continued)

(233) P-Apo-2' -carotenal

CHO
"""' """' """' """' """' """' """' """' """' """' """'
C3o+C1 =C37 Wittig [247]
C3s +C2 =C37 Enol ether [188]

(792) IX-Apo-12' -carotenal

CHO
"<:::: "<:::: "<:::: "<:::: "<::::
"""'
c1s + clO = c2s Wittig [166]

(798) Isorenieral

CHO
"<:::: "<:::: "<:::: "<:::: "<:::: "<:::: "<::::

C2o+CIO=C3o Wittig [327]

(799) Renieral

"<::::
HO
"<:::: "<:::: "<:::: "<:::: "<:::: "<::::

C2o+C10=C3o Wittig [369a]

(809) Apo-4' -okenal

CHO
"<:::: "<:::: "<:::: "<:::: "<:::: "<:::: "<:::: "<:::: "<::::

Wittig [397]

(719) y-Retinal

CHO
"<:::: "<:::: "<:::: "<:::: "<::::
Wittig
"""' [166]
clO + clO = c2o
Cts+Cs=C2o Wittig [296, 335]
Ct6+C4=C2o Grignard [371]

(722) 2-Keto-1-methoxy-3,4-didehydro-1,2-dihydroapo-15-lycopenal

(Continued)
 VI. Total Syntheses 539

Table 46. (Continued)

(793) Apo-12'-lycopenal

Wittig [166]

(737) 7,8-Dihydroapo-12' -lycopenal

Wittig [309, 372]

(871) 15,15'-Didehydroapo-8' -lycopenal

~CHO

'* I I
[373]

(872) 1-Hydroxy-15,15' -didehydro-1 ,2-dihydroapo-8' -I yeo penal

HO ~CHO

""' ""' ""'[373]'*

Wittig
C2o + Cw = C3o

(254) Apo-8' -lycopenal

CHO
""' ""' ""' ""' ""' ""' ""' ""' ""' ""'
Wittig [327, 374]

(80 1) 1-Hydroxy-3,4-didehydro-1,2-dihydroapo-8' -lycopenal

HO

Wittig [325]

(802) 1-Hydroxy-1,2-dihydroapo-8' -lycopenal

HO

Wittig [327, 374]

(806a) 16-(Tetrahydropyranyloxy)apo-8' -lycopenal

THPOCH 2

Wittig [330]

(Continued)
540 H. MAYER and 0. ISLER

Table 46. (Continued)

(803) 1-Methoxy-3,4-didehydro-1,2-dihydroapo-8' -lycopenal

CH3 0

Wittig [309]

(804) 2-Keto-1-methoxy-3,4-didehydro-1,2-dihydroapo-8' -lycopenal

(873) 2-Keto-1-methoxy-3,4-didehydro-1,2-dihydroapo-4' -lycopenal

HO

b) Apo-carotenoic acids and esters

(874) Methyl P-apo-14'-carotenoate

~co.cH,

Wittig [375, 376]

(875) Methyl P-apo-12' -carotenoate

co.cH,
"""' """' """' """' """'
C2o+Cs=C2s Homer [368]

(876) P-Apo-12'-carotenoic acid

0 2H
"""' """' """' """' """'
Aldol [377]

(877) Methyl P-apo-10'-carotenoate

Wittig [378]

(Continued)
 VI. Total Syntheses 541
Table 46. (Continued)

(260) P-Apo-10'-carotenoic acid

Wittig [378]

(878) Methyl15,15'-didehydro-{J-apo-8' -carotenoate

~CO,CH,

"""
'* I I

Czs+Cs=CJo Wittig [298]

Cz7+CJ=CJo Wittig [378]

(251) R=CH 3 Methyl P-apo-8'-carotenoate

C 27 + C 3 =C 30 Wittig [378]
(879) R=C 2H 5 Ethyl P-apo-8'-carotenoate
C 20 + C 10 = C 30 Wittig [247, 375]
C 27 +C 3 =C 30 Wittig [13]

(250) R= H P-Apo-8' -carotenoic acid

C 25 + C 5 = C 30 Aldol [368a]
C 27 + C 3 = C 30 Wittig [378]

(880) Methyl P-apo-6' -carotenoate

[378]

(881) R=CH 3 Methyl P-apo-4'-carotenoate

c32 + c3 = c35 Wittig [378J
(882) R = C 2H 5. Ethyl P-apo-4' -carotenoate
C 25 + C10 = C 35 Wittig [247]

(234) R=H ./J-Apo-4'-carotenoic acid

C 30 +C 5 =C 35 Aldol [368a]
C3z+C3=C3s Wittig [378]

(Continued)
542 H. MAYER and 0. ISLER

Table 46. (Continued)

(883) Methyl {J-apo-2' -carotenoate

CO,CH,

'"""' '"""' '"""' '"""' '"""' '"""' '"""' '"""' '"""' '"""'
C3o+C7 =C37 Wittig [298]
C3s+C2=C31 Wittig [378]

(884) Methyl 7,8-dihydroapo-8'-lycopenoate

CO,CH,

'"""' '"""' '"""' '"""' '"""' '"""' '"""' '"""' '"""'

C2s +Cs =C3o Wittig, Horner [309]

(243) Methyl apo-6'-lycopenoate

CO,CH,
'"""' '"""' '"""' '"""' '"""' '"""' '"""' '"""' '"""' '"""' '"""'
C3o+C2=C32 Wittig [383]

Table 47. Synthetic diapo-carotenoids

(267) Crocetindialdehyde

C 10 + clO = c2o Wittig [247]

C 3 +C,4 +C3=C2 0 Enol ether [21, 60]

(885) Crocetindialdehyde bis(diethyl acetal)

Wittig [302]

(741) 15,15'-Didehydrocrocetindialdehyde
I I "*~CHO
OH~
C 3 + C 14 + C 3 = C 20 Enol ether [21, 60]

(746) 11,12,11',12'-Tetradehydrocrocetindialdehyde

[315]

(Continued)
 VI. Total Syntheses 543

Table 47. (Continued)

(886) Diapo-6,6' -carotenedial

CHO
OHC
""" """ """ """ """ """ """ """ """
C7+Cw+C7=C24 Wittig [247]
C2 +C2o +C2 =C24 Wittig [309]

(748) Diapo-4,4' -carotenedial

CHO
OHC
""" """ """ """ """ """ """ """ """ """ """
Wittig [247, 385]

(788) 15,15'-Didehydrodiapo-4,4' -carotenedial

Wittig [247, 385]

(887) Diapo-2,2' -carotenedial

OHC """

Wittig [247]

R0 2C """ """

(270) R = CH 3 Dimethylcrocetin
Reformatskii [350, 361, 387]
Wittig [299]
Horner [21]
Wittig [262]
Horner [12, 21]
(888) R = C 2H 5 Diethylcrocetin
Cw + Cw = C2o Horner [247]
C3 +C14 +C3=C2o Wittig [262]

(889) 9-cis- Dimethylcrocetin

C0 2 CH 3

""" """ ~ """ """ """ """ C0 2CH 3

Horner [12]

(Continued)
544 H. MAYER and 0. ISLER

Table 47. (Continued)

(890) 15-cis-Dimethylcrocetin

CH,O,C
Reformatskii [350, 361, 387]

~co,R
I I -¢ 1 I
RO,~
(891) R=CH 3 15,15'-Didehydrodimethylcrocetin
C3 +Ct4 +C3=C2o Wittig [388]
(892) R=C 2H 5 15,15'-Didehydrodiethylcrocetin
C3 +Ct4 +C3=C2o Wittig [388]

R02 ...,., ...,., ...,., ...,., ...,., ...,., CO,R

R=CH 3 Methyl bixin

(266) all-trans
Cs+Ct4 +Cs=C24 Wittig [306, 307]
C2+C2o+C2=C24 Doebner [315]
Reformatskii [387]
Wittig [262]
Horner [21]
(893) 15-cis
C2 + c2o + c2 = C24 Reformatskii [387]
(266a) 9-cis
C17 +C 7 =C24 Wittig [27, 288]
(894) R=C 2H 5 all-trans-Diethylnorbixin
c2 + c2o + c2 = c24 Wittig [262]
Horner [247]

(895) Diethyl diapo-4,4'-carotenedioate

Horner [247]

(896) Diethyl diapo-2,2'-carotenedioate

C1 + C2o + C1 = C34 Wittig [389]

c2 + C3o + c2 = c34 Ho~r [247]
 VI. Total Syntheses 545

Table 48. Synthetic C40 -carotenoids

(1) 3,4,3',4'-Tetradehydro-p-carotene

c19 + c2 + c19 = c40 Grignard [260,404]

c14 + c12 + C14 = c4o Enol ether [359]
Nef [413]
Grignard [260]

(2) 3,4-Didehydro-P-carotene

Grignard [260]

(3) All-trans-P-carotene

C2o+C2o=C4o Wittig [21,219,246,248, 252,253,390-393]

Horner [254, 255]
Grignard [245]
Dimerization [21, 141,394-399]
C19+C2 +C19=C40 Grignard [186, 206]
Cta+C4 +Cts=C4o Grignard [406]
Wittig [21]
Ct6+Cs+Ct6=C4o Grignard [2, 3, 5, 6, 338, 408]
Cts+Cto+Cts=C40 Wittig [21, 159, 161,207,353,412]
C14 +C12 +C14=C4o Enol ether [359]
Nef [305, 413]
C13+C14 +C13=C40 Wittig [61, 97, 98]
Cto+C2o+C10=C4o Wittig [60, 129, 415]
C21 +C19=C4o Grignard/Nef [186,418]
C2s+C1s=C4o Wittig [153]

(3a) 11-cis-P-Carotene
C2o+C2o=C40 Wittig [219]

(3b) 13-cis-P-Carotene
c2o +C2o ,;,c40 Wittig [219]

(767) 15-cis-P-Carotene
C19+C2+Ct9=C40 Grignard [186, 206]
C1s + Cm + C1s = C4o Wittig [412]

(Continued)
Carotenmds 35
 546 H. MAYER and 0. ISLER

Table 48. (Continued)

(3c) 11,11'-Di-cis-P-carotene
C16+Ca+C16=C40 Grignard [407]
c14 +ell+ c14 = c4o Grignard [305, 413]

(899) 15,15'-Dihydro-P-carotene

"""' """' """' """' """' """' """'

c14 +ell+ c14 = c40 Nef [360]

(900) rae. ex-Carotene

"""' """' """' """' """' """' """' """'

C16 +Ca +C16 =C40 Grignard [409,410]

C21 +C19=C40 Grignard [118]
C2s+C15=C40 Wittig [153]

(S) ( + )-(R)-cx-Carotene

"""' """' """' """' """' """' """' """'

C16+Ca+C16=C40 Grignard [107]

C2s+C1s=C40 Wittig (106]

(901) (- )-(S)-cx-Carotene

"""' """' """' """' """' """' """' """' """'

c16 + Ca + c16 = C4o Grignard (107]

(6) P-Isorenieratene

"""' """' """' """' """' """' """' """'

C3o+C10=C40 Wittig [66]

'
(Continued)
 VI. Total Syntheses 547

Table 48. (Continued)

(902) P- Renierapurpurin

"""' """' """' """' """' """' """' """'

C3o+Cto=C40 Wittig [66]

(7) Torulene

"""' """' """' """' """' """' """' """' """' """' """'

C3s +Cs =C40 Wittig [303]

(8) y-Carotene

"""' """' """' """' """' """' """' """' """'

Ct6+Cs+Ct6=C40 Grignard [411]

C2s+C1s=C40 Wittig [153, 166]
C3o+C10=C40 Wittig [153]

(9) P-Zeacarotene

"""' """' """' """' """' """' """'

c2s + Cu = c40 Wittig [28, 153]

(903) B-Carotene

"""' """' """' """' """' """' """' """'

c2o + c2o = C4o Wittig [166]

Ct6+Cs+Ct6=C4o Grignard [179, 337]
Cts+Cto+Cts=C40 Wittig [166]

(10) ( + )-(6R, 6' R)-B-Carotene

"""' """' """' """' """' """' """'

Ct6+Cs+Ct6=C40 Grignard [107, 179a]

(Continued)
548 H. MAYER and 0. ISLER

Table 48. (Continued)

(904) (- )-(6S, 6'S)-e-Carotene

""""" """"" """"" """"" """"" """"" """"" """""

Ct6+Ca +C16=C4o Grignard [107, 179a]

(11) Ci-Carotene

""""" """"" """"" """"" """"" """"" """"" """"" """""

Czo + Czo = C40 Wittig [166]

Czs+Cts=C4o Wittig [166]

(12) cx-Zeacarotene

""""" """"" """"" """"" """"" """"" """"" """""

Czs + c1s = c40 Wittig [28]

(13) Isorenieratene

""""" """"" """"" """"" """"" """"" """"" """""

Ct6+Ca +C16=C4o Grignard [180-182]

CIO + Czo + CIO = c40 Wittig [66]

(14) Renieratene

""""" """"" """"" """"" """"" """"" """"" """"" """""

Ct6+Ca +C16=C40 Grignard [182]

c!O + c2o + c!O = c4o Wittig [66]

(15) Chlorobactene

""""" """"" """"" """"" """"" """"" """"" """"" """""

C3o+Cto=C4o Witti~ [327]

(Continued)
 VI. Total Syntheses 549

Table 48. (Continued)

(905) y-Renierapurpurin

(16) Renierapurpurin

Ct6+Cs +Ct6=C4o Grignard [182]

Cw + C2o + Cw = C4o Wittig [66]

(17) 3,4,3',4'-Tetradehydrolycopene

Wittig [323]

(19) Lycopene

C2o+C2o=C4o Dimerization [339, 395]

Ct6+Cs+Ct6=C4o Grignard [337, 338]
Cts+C10+Cts=C4o Wittig [166]
C10+C2o+Cto=C4o Wittig [59, 60, 416]
C3o+C10=C4o Wittig [327]

(906) 5,6,5',6'-Tetrahydrolycopene

Grignard [341]

(22) Neurosporene

G2o+C2o=C4o Wittig [296, 335]

C2s+C1s=C40 Wittig [296, 335]

(25) (-Carotene (unsymm.)

"'""""
Wittig [296, 335]

(Continued)
550 H. MAYER and 0. ISLER

Table 48. (Continued)

(26) (-Carotene (symm.)

"""' """' """' """' """' """' """' """'

c1s + ciO + c1s = c40 Wittig [296,335]

(26a) 15-cis-(-Carotene
C1s+C1o+C15=C4o Wittig [296, 335]

(30a) All-trans-phytofluene

C2o+C2o=C40 Wittig [296, 335]

(32a) All-trans-phytoene

C2o+C2o=C40 Wittig [296, 335]

(34) Lycopersenc

c2o + c2o = c40 Wurtz [23, 400]

(36) 4,4'-Didehydro-P-carotene

,:7 ,:7 ,:7 ,:7

C2o+C2o=C40 Wittig [167]

Dimerization [398]
C19+C2+C19=C40 Grignard [245]

(897) 3,4,3',4'-Tetradehydro-7, 7' -dihydro-P-carotene

,:7 ,:7 ,:7 ,:7 ,:7 ,:7 ,:7

C19+C2 +C19=C40 Grignard [245]

(898) 3,4-Didehydro-7, 7'-dihydro-P-carotene

,:7 ,:7 ,:7 ,:7 ,:7 ,:7 ,:7 ,:7

C21 +C19=C40 GrigiV~rd [245]

(Continued)
 VI. Total Syntheses 551

Table 48. (Continued)

(758) 4,4'-Didehydro-4, 7-dihydro-P-earotene

,?' ,?' ,?' ,?' ,?' ,?' ,?' ,?'

C1o+C1o=C40 Wittig [252]

ell+ c19 = c40 Grignard [245]

(37) 7, 7'- Dihydro-P-earotene

,?' ,?' ,?' ,?' ,?' ,?' ,?'

c19 + cl + c19 = C40 Grignard [245]

c14 +ell+ c14 = c40 Grignard [414]

(907) rae. 7,8-Didehydroeryptoxanthin

"""" """" """" """" """" """" """" """"

it~
HO

Cls +Cts =C4o Wittig [123]

(908) rae. Cryptoxanthin

"""" """" """" """" """" """" """" """"

HO

C11 +C19=C40 Grignard [187]

C1s+C1s=C40 Wittig [109]

(40) lsocryptoxanthin

"""" """" """" """" """" """" """" """"

OH

C1o + C1o = C40 Wittig [167, 401]

(Continued)
552 H. MAYER and 0. ISLER

Table 48. (Continued)

(909) 9-cis-Crocoxanthin

* Ill

"""' """' """' """' """' """' """' """'

c2s + c1s = c4o Wittig [12]

(1000) Zeinoxanthin

"""' """' """' """' """' """' """' """'

HO

C2s +Cts =C4o Wittig [109]

(48) 1'-Hydroxy-l',2' -dihydro-y-carotene

"""' """' """' """' """' """' """' """' """' """' OH

C3o+Cw=C4o Wittig [327]

(52) 3-Isorenieratenol

"""' """' """' """' """' """' """' """' """'

HO

Cw+C2o+Cw=C4o Wittig [67, 68]

(53) OH -Chlorobactene

"""' """' """' """' """' """' """' """' """' OH

Wittig [327]

(55) 3,4-Didehydrorhodopin
HO

Wittig [325]

(Contmued)
 VI. Total Syntheses 553

Table 48. (Continued)

(56) Rhodopin
HO

""" """ """ """ """ """ """ """ """ """ """
C3o+Cw=C4o Wittig [327, 374]

(58) Chloroxanthin
HO

""" """ """ """ """ """ """ """ """

Wittig [309, 372]

(62) Lycoxanthin

HOCH, """

Wittig [330a]

(1001) Lycoxanthin tetrahydropyranyl ether

THPOCH, """ """

Wittig [330]

(1002) 9,9' -Di-cis-alloxanthin

YYOH

4~
Wittig [12, 27]

(1 003) 7,8, 7',8'-Tetradehydrozeaxanthin YYOH

q,~
J:c~
HO

c,s + clO + c,s = C4o Wittig [123]

C2s+C1s=C4o Wittig [123]

(Continued)
554 H. MAYER and 0. ISLER

Table 48. (Continued)

(1003 a) 7,8-Didehydrozeaxanthin
OH

;):~ """' """' """' """' """' """' """' """'

HO

Cls+Cts=C40 Wittig [123]

(1004) Zeaxanthin
OH

"""' """' """' """' """' """' """' """'

HO

Ct9+C2+C19=C4o Grignard [41, 187]

Cts+CIO+Cts=C40 Wittig [109, 123]
Ct4 +Cu +C14=C40 Enol ether [359]
Nef [413]
C2s+Ct,=C40 Wittig [123]

(1005) Zeaxanthin diacetate

C2o+C2o=C40 Dimerization [339,402,403]

(1006) Zeaxanthin dipalmitate (Physalien)

C19+C2 +C19=C40 Grignard [41]
C14 + Cu + C14 = C40 Enol ether [359]

(1007) Zeaxanthin dimethyl ether

c19 + cl + c19 = c4o Grignard [102]

(71) Isozeaxanthin

OH

C2o+C2o=C40 Wittig [256]

Ct9+C2+Ct9=C40 Grignard [208, 405]

(I 008) Isozeaxanthin diacetate

C2o+C2o=C40 Dimerization [339,402,403]

(1009) Isozeaxanthin dimethyl ether

Dimerization [339, 402, 403]

(Contmued)
 VI. Total Syntheses 555

Table 48. (Continued)

(1010) 7,8-Didehydrolutein OH

j)(~ """ """ """ """ """ """ """ """

HO

C2s +CIS= c40 Wittig [123]

(1011) 4,1 '-Dihydroxy-15, 15'-didehydro-1 ',2' -dihydro-y-carotene

~ """ """ """ """ """ OH

""" """ """
OH

C3o+Cw=C4o Wittig [373]

(76) Plectaniaxanthin
H

""" """ """ """ """ """ """ """ """ """ OH

C3s +Cs =C40 Aldol [321]

(1012) Plectaniaxanthin methyl ether

C3s +Cs =C4o Aldol [321]

(79) 3,3' -Isorenieratenediol

OH

""" """ """ """ """ """ """ """ """

HO

Cw + C2o + Cw = C4o Wittig [67, 68]

(1013) 1,1 '-Dihydroxy-3,4,3',4' -tetradehydro-1,2, 1',2' -tetrahydrolycopene

HO

""" """ """ """ """ """ """ """ """ """ """ """ """ OH

C1o+C2o+Cw=C4o Wittig [325]

(81) 1,1 '-Dihydroxy-1 ,2, 1',2' -tetrahydrolycopene

HO

""" """ """ """ """ """ """ """ """ """ """ OH

Cw + C2o + Cw = C4o Wittig [323]

(Continued)
556 H. MAYER and 0. ISLER

Table 48. (Continued)

(83) Lycophyll

"""' /CH,OH
HOCH, """'
"""' """' """' """' """' """' """' """' """' 1 """' """' 1
Cto+C2o+C10=C4o Wittig [330a]

(1014) 4'-Deoxookenone
j
~~~~~~~~ 'OCH,

C 30 +C 10 =C 40 Wittig [369a]

(97) Anhydrorhodovibrin

CH,O
-
"""' """' """' """' """' """' """' """' """' """' """' """' """'
C3o + clO = c40 Wittig [309, 374]

(99) Spheroidene

CH,O
- y~
"""' """' """' """' """' """' """'
C3o+Cto=C40 Wittig [308,309]

(1015) Okenol

~~~~~y~y~~ I'OCH,
~

C3s+Cs =C40 Alkyllithium [12]

(1016) Spheroidenol

"""' """' """' """' """' """' """' """' """' """' """' """' OCH,

c3o + clO = c40 Wittig [308, 309]

(105) OH-Spirilloxanthin

HO._j

"""' """' """' """' """' """' """' """' """' """' """' """' """' OCH,

C 30 +C 10 =C 40 Wittig [325]

(Continued)
 VI. Total Syntheses 557

Table 48. (Continued)

(106) Rhodovibrin
HO

OCH 3
Wittig [374]

(108) Spirilloxanthin
CH3 0

OCH 3
Wittig [323, 325]

(110) 3,4,3',4'-Tetrahydrospirilloxanthin
CH 3 0

OCH 3
Wittig [323, 326]

(142) Torularhodinaldehyde

C3o+Cto=C4o Wittig [247]

C37+C3 =C40 Enol ether [188]

(I 017) 3,4,3',4'-Tetradehydro-16, 16'-lycopenedial

Cts+CIO+Cts=C4o Wittig [247]

Cto+C2o+Cto=C4o Wittig [247]

(777) 4,4'-Didehydro-P-caroten-3-one

Wittig [167]

(776) P-Caroten-3-one

Wittig [167]

(Continued)
558 H. MAYER and 0. ISLER

Table 48. (Continued)

(1018) 15, 15'-Didehydroechinenone

Mannich [310]

(148) Echinenone

C2o+C2o=C40 Wittig [167, 401]

C3o+C10=C40 Aldol [331]
C3s+Cs=C40 Mannich [310]

(757) 10-Keto-7,10-dihydro-P-carotene

,:9'" ""::: ""::: ""::: ""::: ""::: ""::: "":::

0

C2o+C2o=C40 Wittig [252]

(lSI) y-Caroten-4-one

""::: ""::: ""::: ""::: ""::: ""::: ""::: ""::: "":::

C3o+Cm=C40 Wittig [373]

C3s+Cs=C40 Aldol [373]

(1019) rae. 3'-Deoxycryptocapsin

""::: ""::: ""::: ""::: ""::: ""::: ""::: "":::

Aldol [70]

(Continued)
 VI. Total Syntheses 559

Table 48. (Continued)

(1020) rae. Cryptocapsin

C3o+C10=C40 Aldol [419]

(160) 1'-Hydroxy-l ',2' -dihydro-y-caroten-4-one

""" """ """ """ """ """ """ """ """ OH

C3o+C10=C40 Wittig [373]

C3s+Cs=C40 Mannich [373]

(162) 2'-Dehydroplectaniaxanthin

""" """ """ """ """ """ """ """ """ """ """ OH

C3s+Cs=C40 Aldol [321]

(170) Capsanthin

-A-
OH

""" """ """ """ """ """ """ """ ""o

Aldol [28]

(181) Okenone

OCH 3

C3o+C10=C40 Wittig [369a]

C 35 +C 5 =C40 Alkyllithium [12]

(Continued)
 Mannich [310]

(193) Canthaxanthin

Wittig [256]
Dimerization [339, 402, 403]
c19 + C2 + c19 = c4o Grignard [208]
C10 + c2o + clO = c40 Aldol [331]
Cs+C3o+Cs=C4o Mannich [310]

(Continued)
 VI. Total Syntheses 561
Table 48. (Continued)

(I 023) 4, 10'-Diketo-7',1 0' -dihydro-P-carotene

C2o + C2o = C40 Wittig [167]

(774) 4,6'-Dihydrorhodoxanthin
0
<?

""" """ """ """ """ """ """ """

c1s + clO + c1s = c4o Wittig [167]

(775) 6,6'-Dihydrorhodoxanthin
0
<?

""" """ """ """ """ """ """ """

c1s + cto + c1s = C4o Wittig [167]

(1024) rae. 3,3'-Dideoxycapsorubin

""" """ """ """ """ """ """ """ """

Aldol [70]

(I 025) 6,6'-Diketo-5,6,5',6' -tetrahydrolycopene

clO + c2o + clO = C4o Aldol [309]

(1026) Astaxanthin dimethyl ether

OCH,

CH,O
0

C19+C2 +Ct9=C4o Grignard [102]

(Continued)
Carotenouls 36
562 H. MAYER and 0. ISLER

Table 48. (Continued)

(1027) rae, Capsorubin

t?-' OH

clO + c2o + clO = c4o Aldol [69]

(1028) rae. Capsorubin epimer

-8-' JH
clO + c2o + clO = c4o Aldol [69]

(1029) rae. Capsorubin isomer

Aldol [71]

(207) Phillipsiaxanthin

HO

OH
0

Aldol [385]

(1 030) 1,1 '-Dihydroxy-6,6' -diketo-1,2,5,6, 1',2' ,5',6' -oetahydrolyeopene

0

OH

(Continued)
 VI. Total Syntheses 563
Table 48. (Continued)

{208) 2,2' -Diketospirilloxantliin

'"""" '"""" '"""" '"""" '"""" '"""" '"""" '"""" '"""" '"""" '"""" '"""" OCH 3

Cts + Cto + Cts = C40 Wittig [308]

Cs + C3o + Cs = C40 Aldol [385]

{1031) 10,11,10',11'-Tetradehydrorhodoxanthin

0
~·~~ ~ ~ ~ ~ ~
~ ~

C21 +C19=C40 Wittig [167]

{209) Rhodoxanthin
<70

~ ~ ~ ~ ~ ~ ~ ~

C14 + Cu + c14 = C4o Wittig [44]

Czt +Ct9 =C40 Wittig [111, 167]

{211) Torularhodin

C02 H
'"""" '"""" '"""" '"""" '"""" '"""" '"""" '"""" '"""" '"""" '""""

C3s+Cs=C40 Aldol [368a]

C37+C3=C40 Wittig [378]

{212) Torularhodin methyl ester

C02CH 3

'"""" '"""" '"""" '"""" '"""" '"""" '"""" '"""" '"""" '"""" '""""

C3o+C10=C40 Wittig [298]

C31+C3=C40 Wittig [378]

{1032) TorularhQdin ethyl ester

002C2Hs
'"""" '"""" '"""" '"""" '"""" '"""" '"""" '"""" '"""" '"""" '""""

Wittig [23, 247]

{Continued)
 564 H. MAYER and 0. ISLER

Table 48. (Continued)

(1033) 3,4,3',4'-Tetradehydro-16,16'-lycopenedioic acid diethyl ester

C,H,O,• ~~~~~~"""" """" """" """" """" """" """" ""'- _..co,c,H,
C 10 +C 20 +C10 =C 40 Wittig [247]
Homer [247]

Table 49. Synthetic carotenoids with more than 40 carbon atoms

(1034) 19,19'-Didehydrodecapreno-/1-carotene

~ -~

Czo+ClO+Czo=Cso Wittig [353]

C 19 +C 12 +C 19 =Cso Eno! ether [359]

(1035) Decapreno-fl-carotene
........_".. 1
"""" """" """" """" """" """" """" """" """" """"
Cz, +Cs+Cz, =Cso Grignard [258]
Czo+ClO+Czo=Cso Wittig [353]
C 19 +C 12 +C 19 =Cso Eno! ether [359]

(1036) Decapreno-s-carotene

"""" """" """" """" """" """" """" """" """" """" Y'
Cz, +Ca+Cz, =Cso Grignard [259]

(1037) 10,10'-Diketo-7,10,7',10'-tetrahydrodecapreno-fl-carotene

~~~'
Czo + CIO + Czo = Cso Wittig [167, 422]

(1 038) Dodecapreno-p-carotene

~~~~~~~~~~~~~'

Cz6+Ca+Cz6=C60 Grignard [261]

Czo+Czo+Czo=C60 Wittig [321]
 VI. Total Syntheses 565

References

[1] 0. Isler, W. Huber, A. Ronco and M. Kofler, Helv. Chim. Acta 30, 1911 (1947).
[2] P. Karrer and C. H. Eugster, Helv. Chim. Acta 33, 1172 (1950).
[3] H. H. Inhoffen, H. Pommer and F. Bohlmann, Ann. Chern. 569, 237 (1950).
[4] H. H. Inhoffen, F. Bohlmann, K. Bartram, G. Rummert and H. Pommer, Ann. Chern. 570,
54 (1950).
[5] H. H. Inhoffen, H. Pommer and F. Westphal, Ann. Chern. 570, 69 (1950).
[6] N.A. Milas, P. Davis, I. Belil~ and D. Fie~. J. Amer. Chern. Soc. 72, 4844 (1950).
[7] 0. Isler, H. Kliiui and U. Solms, in The Vitamins, ed. by W. H. Sebrell, Jr., and R. S. Harris,
Vol.l (Academic Press, New York 1967), p.101.
[8] U. Schwieter and 0. Isler, in U/lmanns Encyclopiidie der technischen Chemie, ed. by W. Foerst,
Vol.18 (Urban & Schwarzenberg, Munich 1967), p.225.
[9] 0. Isler, U. Solms and J. Wiirsch, in Fat-Soluble Vitamins, ed. by R.A. Morton (Pergamon
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[10] U. Schwieter and 0. Isler, in The Vitamins, ed. by W.H. Sebrell, Jr., and R.S. Harris, Vol. I
(Academic Press, New York 1967), p. 5.
[11] L. Zechmeister, Fortschr. Chern. Org. Naturst. 18, 223 (1960).
[12] B. C. L. Weedon, Pure App/. Chern. 20, 531 (1969).
[13] 0. Isler, R. Riiegg and U. Schwieter, Pure App/. Chern. 14,245 (1967).
[14] H. H. Inhoffen and F. Bohlmann, Fortschr. Chern. Forsch. 1, 175 (1949).
[15] H. H. lnhoffen and H. Siemer, Fortschr. Chern. Org. Naturst. 9, 1 (1952).
[16] 0. Isler, H. Lindlai", M. Montavon, R. Riiegg, G. Saucy and P. Zeller, Chern. Soc. Spec. Pub/.
4, 47 (1956).
[17] 0. Isler and P. Zeller, Vitamins Hormones 15, 31 (1957).
[18] O.Isler and M.Montavon, C.R.31e Congr. Internat. Chim. Ind.,Liege 1958, pub!. in Ind.
Chim. Beige 24, Suppl. Vol. 2, 683 (1959).
[19] 0. Isler and M. Montavon, Chimia (Switz.) 12, 1 (1958).
[20] E. Jucker, Angew. Chern. 71, 253 (1959).
[21] H. Pommer, Angew. Chern. 72, 911 (1960).
[22] 0. Isler, R. Riiegg and P. Schudel, Chimia (Switz.) 15, 208 (1961).
[23] 0. Isler, R. Riiegg and P. Schudel, in Recent Progress in the Chemistry of Natural and
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572 H. MAYER and 0. ISLER

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 VI. Total Syntheses 573

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 VI. Total Syntheses 575

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 577

VII. Biosynthesis
T. W. GOODWIN
Department of Biochemistry, University of Liverpool, Liverpool L69 3BX, England

A. General Basic Pathway . . . . 578

B. Formation of C 20 -Intermediate . 583
C. Formation of First C 40 -Polyene 584
Detailed stereochemistry of phytoene biosynthesis . 585
D. Desaturation of Phytoene-Higher Plants, Algae, Fungi 589
Mechanism of desaturation of phytoene 592
Cis carotenes . . . . . . . . . . . . . 593
E. Cyclization of Acyclic Precursors . . . . . 594
1. Evidence for cyclization of neurosporene . 594
2. Evidence for cyclization of lycopene . . . 596
3. Mechanism of cyclization . . . . . . . 596
F. Xanthophyll Formation in Higher Plants, Algae and Non-Photosynthetic Bacteria. 600
Interconversions ofxanthophylls . . . . . 602
G. Site of Carotenoid Synthesis in Green Tissue . 603
H. Carotenoid Metabolism in Senescing Leaves 605
I. Carotenoid Formation in Fruit . 606
1. Ripening . . . . . . . . . . . . . . 606
2. Genetic studies . . . . . . . . . . . . 607
3. Epoxides of acyclic carotenes in tomato fruit 610
1. Regulation of Synthesis in Green Tissues. . . 610
K. Special Aspects of Synthesis in Algae and Fungi. 612
1. Production of extraplastidic carotenoids . 612
2. Biosynthesis in red yeasts . . 612
L. Bioinduction of Carotenogenesis 614
1. Trisporic acid . . . . . . . 614
2. Other factors . . . . . . . 615
M. Biosynthesis in Non-Photosynthetic Bacteria . 616
1. Biosynthesis of C 45 - and C 50 -carotenoids 618
a) Introduction . . . . . . . . . . . . 618
b) Biosynthetic pathways . . . . . . . . 618
N. Photoinduction of Carotenogenesis in Non-Photosynthetic Bacteria . 620
0. Biosynthesis of Carotenoids in Photosynthetic Bacteria 620
1. Introduction . . . . . . . . . . . . . . . . . . . 620
2. General pattern of biosynthesis in the Athiorhodaceae . 622
3. Formation in the Athiorhodaceae. 626
4. Rhodomicrobium vannie/ii . . . . . . . . . . . . 627
5. Chlorobacteriaceae . . . . . . . . . . . . . . . 627
6. Formation ofxanthophylls in photosynthetic bacteria 627
P. Formation of K,eto Carotenoids in Crustacea . 628
References . . . . . . . . . . . . . . . . . . . . . 629

Carotenoids 37
578 T.W. GOODWIN

A. General Basic Pathway

Carotenoids are tetraterpenes, and their overall biosynthesis must be

expected to follow the general pattern for all terpenoids. The biological iso-
prene precursor is now known to be isopentenyl pyrophosphate (IPP, 1), and
there is clear evidence that it is involved in carotenoid biosynthesis. However,
the first specific terpenoid precursor is mevalonic acid (2), and experiments

(1)

H 3 '\_ PH
H0,(:,6 ,....~:C,....<;H20H
H2 H2
(2)

with this compound have been responsible for revealing many details of ter-
penoid (including carotenoid) biosynthesis. Even before mevalonic acid was
discovered and shown to arise from three acetyl-CoA units, experiments with
[1- 14 C]- and [2- 14C]-acetate had shown that a mechanism such as envisaged
in Scheme 1 was probably functioning. In the carotenoid field the utilization
of acetate for P-carotene (3) synthesis was first demonstrated by Schopfer and
Grob [1-4] in Phycomyces blakesleeanus; Grob and Butler [5] then degraded
the P-carotene synthesized by Mucor hiemalis in the presence of [l 4 C]acetate,

C-C+C-C

l
e-c-c-e + c-c•

l
c-~---c-c

c•

Scheme 1. Pattern of production of a C 5 -unit from three C 2 -units

 VII. Biosynthesis 579

• • •~ •~
~ ~ ~ ~ ~ ~
• • X X

• •
• Carbon atoms from the methyl group of acetate
x Carbon atoms from the carboxyl group of acetate
Scheme 2. The pattern of labelling in P-carotene formed from [ 14 C] acetate [5]

and the location of the label, indicated in Scheme 2, was consistent with the
repeating pattern of a C 5 -unit envisaged in Scheme 1. Similar results were
obtained with Phycornyces blakesleeanus [6], Euglena gracilis [7] and carrot
root slices [6, 8]. The formation of 3-hydroxy-3-methylglutaryl-CoA (HMG-
CoA, 3) from three units of acetyl-CoA (Scheme 3) was clearly a possibility in

0 0 0 0
II II II II
2 CH,-c-s--coA CH,-C--cH 2 -C-S--coA + CH 3-C-S--coA

CoA-SH

Co A-SH

OH 0
I II
CH,-?--cH 2-C-S--coA

?H2
C0 2 H

Scheme 3. Formation of HMG-CoA from acetyi-CoA

the biosynthetic pathway. HMG-CoA has not been examined in carotenoid-

synthesizing systems but HMG is reported to be active in preparations from
Phycornyces blakesleeanus [9, 10]. This implies the existence of a HMG-acti-
vating enzyme which is not of common occurrence. In E. gracilis [ 14 C]HMG
is taken into the cells but not metabolized [7].
Then came the discovery of mevalonic acid (MVA, 2), which turned out
to be of great i.mportance in the elucidation of the basic biosynthetic pathway
to all terpenoids. MV A was isolated as the acetate-replacing factor for a
mutant of Lactobacillus acidophilus [11]. Its structural resemblance to HMG
prompted experiments which demonstrated that racemic [2- 14 C]MVA was
converted into sterols with an efficiency of 43% [12]. This indicated an almost
quantitative conversion of the naturally occurring enantiomer, which has the
 580 T.W.GooDWIN

3R configuration (2) [13]. (See Chapter V for a discussion of the R and S

convention.) There is no evidence that the 3S enantiomer has any biological
activity. The conversion of HMG-CoA into MV A (Scheme 4) was quickly

? H 0
II ?H 0
II
CH,-y--cH2-C-S--coA + HS-Enz CH 3-y--cH2-C-S-Enz + CoA-SH
CH 2--c0 2H CH2--c02H

1l NADPH, H®

OH OH
~ / \i. NADPH, H®
~H /0~

"
CH 3-,--cH2-CH2 Enz-SH CH 3- --cH2-CH Enz
CH2--c02H CH 2--c02H S/

j
CH 3-~~H2--cH20H + HS-Enz
CH2--c0 2H

Scheme 4. Mechanism of formation of MVA from HMG-CoA

demonstrated in yeast and liver [14-16] although it has yet to be observed

in higher plants. The reaction is essentially irreversible and involves an enzyme-
bound intermediate charge transfer complex at the same oxidation level as
mevaldic acid. Although mevaldic acid is reduced by a specific reductase it
is not considered a normal intermediate [16]. It is the carboxyl group of
HMG esterified with coenzyme A which is reduced, so the absolute con-
figuration of naturally occurring HMG-CoA is 3S (3). The incorporation of
[2- 14C]MVA into carotenoids has been demonstrated many times and is now
a laboratory routine. The first demonstrations were made with Phycomyces
blakesleeanus [6, 9, 17, 18], Mucor hiemalis [19], Blakeslea trispora [20],
Neurospora crassa [21], carrot root preparations [6, 22], tomato fruit [6,
23-25], Euglena gracilis [7] and with a mutant of Staphylococcus aureus
[26, 27]. The general pattern of labelling observed in P-carotene synthesized
by Phycomyces in the presence of [2- 14C]MVA is indicated in Scheme 5. This
•
•
____._,

'-v-'
•
•
• Carbon atoms from C-2 of MV A
Scheme 5. The pattern of labelling in P-carotene formed from [2- 14C] mevalonate [6]
 VII. Biosynthesis 581

pattern, as will be seen later, is to be expected from the further metabolism

of MV A to IPP. Incorporation of MVA into carotenoids is, however, not
always easily demonstrated, mainly for reasons of membrane permeability;
for example only slight incorporation is observed in E. gracilis v. bacillaris [7],
E. gracilis strain Z [28], Chlorella pyrenoidosa [20] and various etiolated
seedlings [29-31]. This problem is discussed later (p. 610).
IPP was recognized as the fundamental C 5 -terpenoid precursor by Bloch
[32, 33] and by Lynen [34], and its stepwise formation is indicated in Scheme 6.

?"
CH,-?--cH2--cH20H ATP ADP

CH2--c02H
'\.../

OH ADP ATP OH
CH,-?--cH2--cH2-o-®-®
'=-../ CH,-t--cH2--cH2-o--®
CH2--c02H tH2--c02H

\ ATP ADP, P,
~---~~~L~--------
t
CH 3-C--cH2--cH2---o--®-®
~H2
C0 2

Scheme 6. Formation of IPP from MVA

MVA kinase has been demonstrated in a number of plant tissues including

Hevea latex [35, 36], pea seeds [37], seedlings of Cucurbita pepo [38], Pinus
radiatus [39], leaf tissue of Phaseolus vulgaris [40, 41], plant tissue culture
[42] and in Staphylococcus aureus [43]. Mn 2 Eil is more effective than Mg 2 Eil
for the plant enzyme [36, 38]. Only in S. aureus has MV A 5-phosphate been
converted into carotenoids [ 43].
The formation of MV A 5-pyrophosphate has been demonstrated in yeast
[32, 33], and it has been identified as a metabolite in a system capable of
converting MV A into carotenes [ 44].
As the enzymic conversion ofMVA 5-pyrophosphate into IPP is being dealt
with in detail later, it is only necessary here to state that the enzyme, MV A
5-pyrophosphate anhydrodecarboxylase, has been purified from yeast [32-34,
45] and' that IPP is found in enzymic systems capable of synthesizing carotenes
[46]. The conyersion of [1- 14C]IPP into carotenoids has been achieved in
homogenates of tomato fruit [ 47] and Staphylococcus aureus [ 46]. A recent
study with a partly purified soluble system from acetone-dried powders of
tomato fruit plastids has demonstrated the conversion of IPP into lycopene
[48]. A similar soluble enzyme system has been isolated from spinach lea-
ves [49].
 582 T. w. GooDWIN

Under certain circumstances HMG-CoA can arise from the branched

chain amino acids leucine and valine. The pathway from leucine is indicated
in Scheme 7 [50].1'his explains the original observation that leucine and valine

CH 3 NH 2 CH 3 0
'-CH---cH I Transamination '-CH---cH2-C---c02H
II
2-CH---c0 2H
/ /
CH 3 CH 3

CoA-SH{
C0 2
2H

CH 3 0 CH 3 0
"c==CH-~-s---coA '-CH---cH -C-S---coA
II
2
/ /
CH 3 CH 3
Dimethylacrylyl-CoA Isovaleryl-CoA

'C02'l

CH 3 0 H 20 CH 3 0
"c==CH-~-s---coA '-
Ho-C---cH II
2-C-S---coA
/ /
CH 2-C02 H CH2-C0 2 H

3-Methylglutaconyl-CoA HMG-CoA

Scheme 7. The formation of HMG-CoA from leucine [50]

are extremely carotenogenic in Phycomyces blakesleeanus [51]. Experiments

with [l- 14 C]-, [2- 14 C]-, [3- 14 C]-, [4- 14C]- and [5,5'- 14C 2 ]-leucines [52-58]
c
indicated that the 'c unit was effectively incorporated but that the two car-
e/
bans arising from C-2 and C-3 were not. The probable explanation is that
HMG-CoA is converted into acetoacetate and acetyl-CoA (from C-2 and
C-3) by the HMG cleavage enzyme and that in resynthesis of HMG-CoA,
which involves acetoacetyl-CoA and acetyl-CoA, the acetyl-CoA mixes with
a much larger endogenous pool than does the acetoacetyl-CoA (Scheme 8).
The mechanism indicated in Scheme 8 can also explain why 14 C0 2 is fixed
into P-carotene in P. blakesleeanus only when it is metabolizing leucine [6, 59].
As indicated from the pathway outlined in Scheme 7 the C0 2 fixed in the
HMG-CoA molecule is located in the position (C-1) which is lost on conver-
sion of HMG-CoA into IPP via MVA. Scheme 8 indicated how some of this
label can be conserved in C-5 of MVA.
 VII. Biosynthesis 583

cH. o CH 3 0
'\. II Cleavage
HQ-C-cH2-C-S-coA "c=O + CH 3-~-s-coA
/ enzyme /
CH2-C02H CH2-C02H

1
CoA-SH ........_I Activating
enzyme

CH 3
Condensing '\.
HQ-C-cH2---c02H
enzyme /
CH 3 cH-c-s-coA
'\. 2 II
C=O 0
/
CH-C-S-coA
2 g
Scheme 8. The breakdown and resynthesis of HMG-CoA postulated to account for the incorpora-
tion of [ 14 C]leucine and 14 C0 2 in the presence of unlabelled leucine into P-carotene [51-59]

[U- 14C]Leucine is also incorporated into carotenoids in green tissues [60],

but the amino acid is not carotenogenic in Rhodospirillum rubrum [61], Coryne-
bacterium poinsettiae [62] or Sarcina lutea [63]. In Rhodotorula shibatana
although leucine was active valine, which is active in P. blakesleeanus and which
undergoes changes similar to those observed for leucine, was not [64]. Of the
intermediates indicated in Scheme 7 only dimethylacrylic acid has been
examined, and then only in its free form, not as the CoA ester. In the original
investigation in Phycomyces blakesleeanus dimethylacrylic acid was not stim-
ulatory [51], but in later experiments from another laboratory it was claimed
to be carotenogenic [65]. [3- 14 C]Dimethylacrylic acid was incorporated into
carotenoids in Euglena gracilis [7], Chlorella pyrenoidosa [20] and Blakeslea
trispora [20], but not in tomato fruit under conditions where MV A was being
actively incorporated [20]. This may mean that the C0 2 -fixing step (Scheme 7)
is absent from tomato fruit.

B. Formation of C20 -lntermediate

The way by which the higher terpenoid precursors are built up from IPP
is now well understood. The first step is the isomerization of IPP to dimethyl-
allyl pyrophosphate (DMAPP) [66, 67], which acts as the starter for chain
elongation (Scheme 9). Iodoacetamide inhibited the conversion of MV A into
caroten6ids in a cell-free system from Phycomyces [68]; because the DMAPP-
IPP isomerase is a-SH enzyme this is circumstantial evidence for DMAPP
being an obligatory intermediate in carotenoid biosynthesis as it is for other
terpenoids.
The enzyme prenyl-transferase transfers one IPP molecule to a DMAPP
molecule to form geranyl pyrophosphate (GPP) (C 10) [66, 69-71]. Further
transfer of one IPP molecule to GPP leads to farnesyl pyrophosphate (FPP)
584 T. W. GOODWIN

j
"(!)c-cH---cH2----o--®-®
CH 3 H CH 3

/
CH 3
,r:l
"
/
CH 3
C=CH---cH2----o--®-®

Scheme 9. Formation of DMAPP from IPP

(C 15 ) [72-76], and transfer of IPP to FPP yields geranylgeranyl pyrophosphate

(GGPP) [72, 77-79].
Direct utilization of G PP for carotenogenesis remains to be demonstrated
but it is formed by preparations from pea seedlings [37] and from Pinus
radiatus [39]. Free geraniol is, however, incorporated into P-carotene in carrot
root slices [80]. It is well known that unlike animal and fungal tissue, higher
plants appear to be able to pyrophosphorylate higher terpene alcohols
and thus use them for biosynthetic purposes [81]. FPP is incorporated into
P-carotene and other c40-polyenes in a cell-free extract of Phycomyces [68],
and in plastid systems from carrots and tomatoes [82, 83].
Early work implicated GGPP as a carotenoid precursor; for example, the
presence of MVA was mandatory for conversion of FPP into phytoene in
cell-free systems from Phycomyces [68], and in a plastid system from tomatoes
IPP was necessary for the conversion of FPP into phytoene [79]. More
recently, however, direct demonstration of the conversion of GGPP into
phytoene has been obtained with an enzyme system from tomato plastids
[84], Phycomyces blakesleeanus [85] and chloroplasts [85a].

C. Formation of First C40 -Polyene

If the reaction of two molecules of GGPP to form the first C 40 -polyene

were entirely analogous to the formation of squalene from two molecules of
FPP [86] then the compound formed would be lycopersene (34) and NADPH
would be required for the reaction. It is now generally agreed that the first
C 40 -compound is phytoene (15,15'-didehydrolycopersene, 32). Early work sug-
gested that lycopersene was present in mycelia of Neurospora crassa [87] and
in certain, but not all, carrot strains examined [88] and that it was formed
from GGPP in extracts of N. crassa [77]. However, it is probable that the
early techniques were inadequate, and many later investigations have failed
to reveal lycopersene as a natural product [89-92]; furthermore in the
presence of diphenylamine, which inhibits carotenogenesis (see p. 589) and
 VII. Biosynthesis 585

causes the massive accumulation of phytoene, no labelled lycopersene was

detected with [2- 14 C] MV A as substrate, although large amounts of labelled
phytoene accumulated [89]. Experiments with enzyme preparations also
demonstrate phytoene but no lycopersene formation. In chloroplasts isolated
by non-aqueous techniques MV A is effectively incorporated into phytoene
but no lycopersene is detected even in the presence of NADPH, which stimu-
lates squalene synthesis in the preparations [93]. A cell-free system from pea
fruit synthesizes phytoene from MV A without any reported synthesis of lyco-
persene [94]. Furthermore no lycopersene was detected in the products of
incubation of [ 14 C]GGPP with a soluble system from tomato fruit plastids
which synthesized phytoene [84]. In a Mycobacterium species, cultured under
anaerobic conditions in the absence of a hydrogen acceptor [95], phytoene
accumulates; clearly the formation of lycopersene prior to phytoene would
require an oxidizing step to convert lycopersene into phytoene.
Additional evidence in support of phytoene as the first C 40 -compound is
provided by mutant studies. Many mutants have been obtained which accu-
mulate phytoene, for example in Chiarella vulgaris [96], C. pyrenoidosa [97],
Neurospora crassa [98, 99] and Rhodopseudomonas spheroides [100], but none
has been described which accumulates lycopersene.

Detailed stereochemistry of phytoene biosynthesis

The synthesis of various stereospecifically labelled species of MV A by
Cornforth and Popjak and their colleagues [101, 102] enabled them to settle
most of the subtle stereochemical problems involved in the conversion of
MV A into squalene [86]. The conversion of MV APP into IPP involves a
trans elimination of the carboxyl and 3-hydroxyl groups as indicated by the
formation of [4-cis- 2 H]IPP from (2R)-[2- 2 H]MVA (4). The mechanism indi-
cated in Scheme 10 is consistent with this observation and with the observa-

eO) 0
'(~
C"~ •••••••CH 3
HnonoC-C--cH ---cH IJi"l IJi"l
r/
A

(AOH 2 2~
(4)

(H0) 2 P__['Io-®--®--Actenosine
g

l
"
H CH 3

"
C=C /
D/ CH 2 ---cH 2 -o--®--® + ADP + C0 2 + H 3 P0 4

Scheme 10. Mechanism of conversion of MVAPP into IPP [86]

586 T. w. GooDWIN

tions that deuterium does not appear in the terminal methylene group of IPP
when the reaction is carried out in the presence of D 2 0, that the oxygen at
C-3 of MV A appears in the inorganic orthophosphate formed and that C0 2
and ADP are liberated simultaneously [103].
The next step involves the addition of a proton to a vinylic carbon and
the elimination of a proton from C-2 of IPP. It is not known from which side
the proton approaches the methylene group but from experiments with (4R)-
[4-3H]MVA (5) and (4S)-[4- 3H]MVA (6) it was shown that the pro-R hydro-

H•\. pH H.e DH
HOle....._ __..'!....._ __..eHlOH HOle....._ Y~ __..eHlOH
e e f\
l
~
/\
D H
Hl Hl 'r
(4) (5)

gen* (HR) is lost from C-2 of IPP; this is the hydrogen which, by the R
and S rules, is the Hs hydrogen of C-4 of MVA (Scheme 11) [101]. The

"e=e
eH 3 CHl--o--®-®

"
/
/3
eH Hs

Scheme 11. Stereochemistry of hydrogen removal from C-2 ofiPP when it is isomerized to DMAPP

prenyl transferase reaction is similar to the isomerase reaction except that,

instead of a proton, an allylic residue (DMAPP, GPP or FPP) is added to
the vinylic carbon of IPP. The hydrogen eliminated is the same in all cases
(i.e. HR from C-2 of IPP). It was also shown from experiments with (2R)-
[2-2H]MVA (4) and (2S)-[2- 2H]MVA that the allylic residues are added to
the vinylic carbon of IPP from the side of the double bond from which the
four substituents -CH 2CH 20--®-®, -CH 3, -H and -D appear in a
clockwise order [102].
Experiments with (4R)-[2- 14C-4- 3H]MVA and (4S)-[2- 14C-4- 3H]MVA
indicated that in phytoene biosynthesis in plants and microorganisms the
situation was the same as in squalene biosynthesis in animals, that is the H 8
hydrogen is lost from C-4 of each molecule of MVA incorporated [104]. The
allylic double bonds which are formed in squalene and phytoene have the
* The pro-R or pro-S hydrogen is that hydrogen which when replaced by tritium or deuterium
would confer the absolute configuration R or S at the carbon atom involved.
 VII. Biosynthesis 587

trans configuration; the corresponding double bonds in rubber are cis, and in
this case it is the HR proton which is lost from C-4 of MVA [105]. Long chain
polyprenols also exist which have a mixed biogenetic origin, some of the
allylic double bonds being cis whilst others in the same molecule are trans
[106].
The mechanism of the carbon-carbon bond formation in this chain elonga-
tion was further defined when experiments with (5R)-[5- 2 H]MVA showed
that at each condensation there was inversion of configuration around C-1
of the allylic pyrophosphate [102], as can be seen by comparing the configura-
tion of the starting MV A with that of the squalene synthesized from it
(Scheme 12). The overall reaction resulting in the formation of the C-C bond

R R s
••••••D

Scheme 12. The absolute configuration of squalene synthesized from (5 R)-[5- 2 H] MV A [86]

cannot be considered as a concerted reaction because it would lead to the

elimination of the wrong hydrogen atom or the addition of the allylic group
to the wrong side of the double bond. Consequently the two-step mechanism
indicated in Scheme 13 was proposed [86]: firstly the trans addition of the

8 X-A

TH3 17
H8 C ~H2-o--®-®
'\.~'-../
. . F"-
H H H*
CH 3

/
t CH 2 .-C.6--®-®
R-CH2
~
T/
H

Scheme 13. The overall stereochemistry of the formation of C-C bonds in chain elongation in
terpenoid biosynthesis [86]

allylic residue and X (probably an enzymic residue) to the double bond of

IPP followed by a trans elimination of X and H 8 to form the new double bond.
588 T. w. GooDWIN

The exact mechanism involved in squalene biosynthesis from two mole-

cules of FPP is still not settled but it is clear that one hydrogen atom (H 8 )
is lost from C-1 of one of the participating farnesyl pyrophosphates and is
replaced by a hydrogen (4-pro-S) from NADPH; this incoming hydrogen
takes up the pro-R position. There is also inversion of configuration at the
carbon atom arising from C-1 of the other FPP molecule involved [101]. This
situation is indicated in Scheme 12.
In phytoene biosynthesis no NADPH is required and one hydrogen is lost
from C-1 of each of the participating GGPP molecules. With the use of
(5 R)-[2- 14 C-5- 3 H]MV A and [2- 14 C-5,5- 3 H 2 ]MVA it has been shown in

" ..~·"
CH 3 Enz
/
~s
H3y
c
/~/I~
c'
R T ?..-' T
H@-®

"
--
CH 3 Enz
HJc;:: (~::r/ H H3c;:: T H
cI
/~/1'/~/
l ..T I
o··~ c R t R
t
/~/~/~/
C

R ?~)\.?
H T H CH 3
R ? HT T
H CH 3
trans- Phytoene

Ill
"
CH 3 Enz
H3y (.Se/ J H3C T T

c '· · ·
/ ~ /c~···"
--+
/~/
CI " C=C/
" /
CH3
R ~C e-T CH R T /C=C"
I " C==C /
"
3
H H H R
H/ R cis- Phytoene

Scheme 14. Possible mechanism tor phytoene biosynthesis from GGPP f1071
 VII. Biosynthesis 589

tomato fruit slices and in isolated chloroplasts that the two Hs hydrogens
are lost [107, 108]. A possible mechanism ofphytoene synthesis can be derived
from these observations (Scheme 14). In this scheme, previously considered in
a slightly different form for squalene biosynthesis [101], the GG residues are
linked via a sulphonium ylide, which requires a mechanism involving a thio-
ether grouping, such as a methionine residue, at the active site of phytoene
synthetase. The thio-ether group displaces the pyrophosphate from one mole-
cule ofGGPP by an SN 2 substitution, which involves inversion of configuration
at C-1 of the GG group, to give a sulphonium ion. This is stabilized by the
loss of the Hs proton, and the resulting compound tends to ionize to form an
ylide, which alkylates a second molecule of GGPP, again with inversion of
configuration at C-1, to yield a lycopersenyl sulphonium ion. The central
double bond is then introduced by a trans elimination of the S-enzyme and
a proton. In this way the loss of two pro-S hydrogens leads to the formation
of a cis double bond at the centre of the molecule; there exists good evidence
that the central double bond of naturally occurring phytoene in higher plants
is cis [109, 110] as elimination of one pro-S and one pro-R hydrogen would
yield a trans double bond at the centre of the molecule. Recently it has been
stated that the central double bond of phytoene from Flavobacterium dehydro-
genans is trans [111].

D. Desaturation of Phytoene-Higher Plants, Algae, Fungi

Following studies with mutant strains of tomatoes Porter and Lincoln

[112] proposed a step-wise dehydrogenation of a saturated precursor (tetra-
hydrophytoene) to form lycopene. Considering that at the time this was put
forward no one knew the structure of the proposed intermediates or the basic
mechanism of terpenoid biosynthesis, it was in general terms very near the
truth. The pathway now envisaged is indicated in Scheme 15. A similar accu-
mulation of intermediates has been observed in Chlorella [96, 113-115]
mutants and in many organisms in the presence of the inhibitor diphenyl-
amine (DPA) [116-119] {Table 1). When DPA is washed out of a culture and

Table 1. Accumulation of partly saturated polyenes in presence of diphenylamine in

Blakeslea trispora

Polyene Total Concentration %of total

/18 Jlg/g dry weight polyene present

Phytoene 14,200 2910 99.52

Phytofluene 27.5 5.6 0.19
P-Carotene 20.0 4.1 0.14
(-Carotene 10.8 2.2 0.076
y-Carotene 5.4 1.1 0.038
Lycopene 5.1 1.05 0.036

Normal cultural conditions; but with 0.3 ml of diphenylamine solution (0.572 g/100 ml in
ethanol) added to each 100 ml of medium.
 590 T.W. GOODWIN

""'::: ""'::: ""'::: ""'::: ""':::

Phytoene

1-2H

""'::: ""'::: ""'::: ""'::: ""'::: ""'::: ""':::

Phytofluene

1-2H

""':::
(-Carotene

1-2H

""'::: ""'::: ""'::: ""'::: ""'::: ""'::: ""'::: ""'::: ""'::: ""'::: ""':::
Neurosporene (22)

1-2H

""'::: ""'::: ""'::: ""'::: ""'::: ""'::: ""'::: ""'::: ""'::: ""'::: ""'::: ""':::
Lycopene (19)

Scheme 15. Pathway of conversion ofphytoene into lycopene in higher plants. algae and fungi [112]

the culture resuspended in an appropriate medium then more saturated carote-

noids are synthesized at the expense of the accumulated intermediates [6, 116,
117]. One of the difficulties in interpreting these experiments was that the very
large quantities of phytoene which accumulated during the inhibitory phase
did not disappear completely after the removal of DPA. This can now be
explained by assuming that when the end product of synthesis (e. g. /)-carotene)
 VII. Biosynthesis 591

is removed it ceases to exert feed-back inhibition at the first specific C 40 step,

and phytoene thus accumulates. However, the situation may be more com-
plicated than this because in the presence of DPA and P-ionone, which stim-
ulates P-carotene synthesis without itself being incorporated into the molecule,
Phycomyces blakesleeanus accumulates both P-carotene and phytoene [117],
and the same situation exists in the presence of DPA and either riboflavin
or AMP [120].
The mechanism of DPA action is still a mystery. In an ingenious proposal
it was shown that DPA models can lie snugly on the middle of the trans-
phytoene model; thus it could compete with phytoene for the active site of
the dehydrogenase enzyme [121]. Unfortunately cis-phytoene rather than
trans-phytoene may be the intermediate in some organisms.
The specific activities of the proposed intermediates in ripening tomatoes
injected with [2- 14 C]MV A are not consistent with the phytoene pathway [25],
but supporting data were obtained when the system was simplified using
isolated tomato plastids [122]. However, in complex systems with varying
pool sizes of endogenous polyenes, specific activity measurements are frequently
oflittle significance and, as just indicated with DPA inhibition, experimental
conditions can undoubtedly be arranged so that a large accumulation of
phytoene does not disappear when synthesis of coloured carotenoids is stim-
ulated. Such an experiment has recently been reported: if [2- 14C]MVA is
added to 2.5 day old continuous cultures of Neurospora crassa, labelled phyto-
ene accumulates. If the nutrients are removed and unlabelled MVA added,
the cells become coloured, and the pigments have specific activities 20-100
times greater than that of phytoene [123]. The author's conclusion is that
phytoene is not a precursor of carotenoids, whilst other investigators consider
that what has been demonstrated is that under the particular experimental
conditions employed a pool of phytoene accumulates which is metabolically
inert [124]. This possibility is discussed further on p. 623.
A few early reports exist which claim the conversion of phytoene into
coloured carotenoids; extracts of Staphylococcus aureus [26] are said to con-
vert phytoene into 15-carotene (11) and P-carotene is said to be formed from
phytoene in extracts of Sporobolomyces shibatanus [125, 126]; maize chloro-
plasts are reported to convert phytofluene into P-carotene [127].- The problem
in all this type of work is the difficulty in preparing both starting material and
final product in a radiochemically pure form.
The conversion of phytoene into phytofluene has been demonstrated in
tomato plastids [128]; NADPED was not required, and the reaction went
equally well in air or under nitrogen; the hydrogen acceptor, whatever its
nature, must have been present in sufficient amounts in the plastid preparation.
Conversion of phytofluene into (-carotene or any further step has not yet
been directly demonstrated. The phytofluene-(-carotene step is particularly
important because it probably involves both dehydrogenation and isomeriza-
tion. However, a purified enzyme system from spinach leaves will convert
IPP into phytoene, phytofluene and lycopene [129]. Recently an enzyme
 592 T. w. GooDWIN

system which converts phytoene into lycopene and cyclic carotenes has been
obtained from tomato plastids [129a]. A complex set of co-factor require-
ments are reported; light is mandatory and maximal activity is achieved
anaerobically in the presence of NADP, FAD and a boiled extract of 'leaf
supernatant solution'. In the absence of FAD radioactivity accumulates in
phytoene and decreases in lycopene, which indicates that FAD is involved in
the desaturation process. However, with a cell-free system from Phycomyces
blakesleeanus, no NADP or FAD co-factors were required to convert GGPP
into phytoene, phytofluene, P-carotene, neurosporene and lycopene [85].

Mechanism of desaturation of ph ytoene

As indicated by the experiments discussed above desaturation of phytoene
can go on under anaerobic conditions, and this is also obvious from the
presence of coloured carotenoids in obligately anaerobic photosynthetic bac-
teria. Furthermore a Chiarella mutant which accumulates phytoene when the
organism is cultured anaerobically in the dark converts phytoene into carote-
noids when the cells are illuminated under anaerobic conditions [113-116].
Experiments with (2R)-[2- 3 H]MVA, (2S)-[2- 3 H]MVA, (5R)-[5- 3 H]MVA
and [5,5- 3 H 2 ]MVA were undertaken to attempt to reveal the stereochemistry
of hydrogen removal in converting phytoene into lycopene, because four
hydrogens are removed from four carbons arising from C-5 of MVA and
four from four carbons arising from C-2 of MV A (Scheme 16). It has now
HsR H2s
I;I~RHss H2R
"'<:::::: "'<:::::: H
H2R H;~ H2s 5R
H2s "sR
Phytoene

l
HsR
H
"'<:::::: "'<:::::: "'<::::::
H2R
H2s H2R H2R HsR Hss Hss
Lycopene

Scheme 16. The probable stereochemistry of the desaturation of phytoene to lycopene (hydrogen
atoms indicated 2 arise from hydrogens at C-2 of MY A and those indicated 5 arise from hydrogens
at C-5 ofMVA) [130, 131]

been shown in a number of systems that it is the pro-R hydrogens which are
removed from the carbons originating from C-5 of MVA [107, 108, 130], but
it has not yet been possible to show which hydrogen is removed from the
adjacent carbon atoms in forming the four double bonds. Experiments with
 VII. Biosynthesis 593

(2R)-[2- 3 H]MV A and (2S)-[2- 3 H]MV A gave indecisive results, in which

much 'scrambling' of the label had taken place, but the indication was that
the 2-pro-S hydrogen was removed [ 131 a]; on this assumption the stereo-
chemistry of the reaction is as indicated in Scheme 16. This scrambling of label
at C-2 of MVA has been observed in higher plants, algae and fungi but is not
detectable in animal systems [132]. The probable reason for this is the slug-
gishness of the prenol transferase in plants and protista, which permits the
IPP-DMAPP isomerase, which is the only easily reversible step in the basic
biosynthetic pathway, to scramble the label at C-4 of IPP as indicated in
Scheme 17. The configuration can either be retained or reversed or the label
can be lost completely.

H~~
H
IPP
4'
H~~ =~~~
T
IPP T DMAPP

T~~
H
IPP

Scheme 17. The probable mechanism involved in scrambling the stereospecific label at C-4 of
IPP [132]

Cis carotenes
Poly-cis isomers of the phytoene series are known in certain fruit and
petals. For example proneurosporene and prolycopene are present in anum-
ber of fruit, in particular tomato mutants [133], and also in the Chiarella
mutant 5/520 grown in the dark [96]. Until the exact structures of these
compounds are known it is difficult to assess their biosynthetic significance.
The available information is also somewhat inexplicable. For example, pro-
neurosporene and prolycopene in Chiarella mutant 5/520 are converted into
their all-trans forms when the cells are illuminated, light absorbed by chloro-
phyll being effective for this conversion [115, 134]. This may be a photoreaction
rather than an enzymic reaction, because in solution the isomerization easily
occurs ort illumination. On the other hand tangerine tomatoes produce large
amounts of prolycopene which appear to be unaffected in situ by light [133].
The shift from trans to cis in carotenoid synthesis is controlled by one gene
in tomatoes: tangerine tomatoes are homozygous for the recessive allele t, the
normal red fruit p·ossess the dominant allele t+ [134, 135]. Preliminary studies
with (4R)-[4- 3 H]MVA indicate that prolycopene arises from all-trans-GGPP
[107].

Carotenoids 38
594 T. w. GooDWIN

E. Cyclization of Acyclic Precursors

So far only acyclic carotenes have been considered, but in green tissues,
algae and many fungi the major carotenoids are the cyclic compounds /3-
carotene (3) with two {3-ionone residues and oc-carotene (5) with one
/3-ionone residue and one oc-ionone residue. Others exist, e.g. y-carotene (8),
with a ring at one end and an open chain (pseudoionone residue) at the other.
The basic problems of cyclization are: (i) at what stage does cyclization take
place and (ii) what is the mechanism of forming the oc- and {3-ionone ring
systems?
There is no evidence that cyclization occurs before the neurosporene (22)
stage; a cyclic (-carotene, for example, has not been detected, but the discovery
of oc-zeacarotene (12) and /3-zeacarotene (9) [136] makes it clear that cycliza-
tion of neurosporene can occur. The evidence accumulating suggests that
cyclization at both the neurosporene and lycopene (19) level can take placB.
The possible pathways of cyclization from neurosporene and lycopene are
indicated in Scheme 18.

1. Evidence for cyclization of neurosporene

Apart for the existence of oc- and /3-zeacarotenes, just mentioned, and the
accumulation of the latter by Rhodotorula [137] and Phycomyces blakesleea-
nus [138], in the presence of inhibitors, and the failure of lycopene to accu-
mulate under similar conditions, the evidence for neurosporene as a precursor
is based mainly on experiments which appear to rule out lycopene. For example,
/3-carotene synthesis but not lycopene synthesis is inhibited by ripening at
temperatures above 30 °C [139, 140]. However, in a tomato phenotype carry-
ing the gene B obtained by back crossing a Lycopersicon esculentum x L. hir-
sutum hybrid to L. esculentum, the usual high levels of lycopene are replaced
by equivalent amounts of {3-carotene [112, 141], and the synthesis of this /3-
carotene is temperature-sensitive [141]. Furthermore, the fruit of the I\ genera-
tion of the high-/3 strain contain more y-carotene than normals, andy-carotene
is clearly the only intermediate possible between lycopene and {3-carotene
[142]. None of these observations can be taken as conclusive proof for either
pathway, but do strongly suggest that there are two physically separate path-
ways of carotenoid biosynthesis in tomato fruit. This is supported by the
observation that dimethylsulphoxide inhibits lycopene synthesis but not /3-
carotene synthesis in tomatoes [143] (see also p.606).
The aquatic fungus Rhizophlyctis rosea synthesizes lycopene during the
early stages of growth whilst older cultures contain both lycopene and y-
carotene. If [2- 14C]MVA is added to the medium at the time of inoculation
and removed when the lycopene level is maximal, then they-carotene eventually
produced is essentially unlabelled and the lycopene strongly labelled. If, on
the other hand, addition of [2- 14C]MVA is delayed until the lycopene level
has reached its peak, the y-caroten'e.eventually isolated is strongly labelled and
 VII. Biosynthesis 595

'<:::::: '<:::::: '<:::::: '<:::::: '<:::::: '<:::::: '<::::::

P-Carotene (3)

r
'<:::::: '<:::::: '<:::::: '<:::::: '<:::::: '<:::::: '<:::::: '<:::::: '<::::::

y-Carotene (8)

1
'<:::::: '<:::::: '<:::::: '<:::::: '<:::::: '<:::::: '<::::::

I
P-Zeacarotene (9)

'<:::::: '<:::::: '<:::::: '<:::::: '<:::::: '<:::::: '<::::::

(19) +-- (22)
#

\ •-C•"""' (5)

'<:::::: '<:::::: '<:::::: '<:::::: '<:::::: '<:::::: '<::::::

IX-Zeacarotene (12)

1
'<:::::: '<:::::: '<:::::: '<:::::: '<:::::: '<:::::: '<:::::: '<::::::

<>-Carotene (11)

1
'<:::::: '<:::::: '<:::::: '<:::::: '<:::::: '<:::::: '<::::::

t:-Carotene (10)
Scheme 18. Possible pathways of cyclization of neurosporene (22) and lycopene (19); cf. Scheme 15
596 T. W. GOODWIN

the lycopene only slightly labelled [144]. This looks like strong evidence
against lycopene as a precursor in this case, but invocation of compartmenta-
lization could again explain the situation in favour of lycopene.

2. Evidence for cyclization of lycopene

Evidence for the participation of lycopene in cyclization is much more

positive. Labelled lycopene has now been reported to be converted into cyclic
carotenes in chloroplasts [145, 146] and by soluble preparations from spinach
chloroplasts and tomato plastids [147]. With spinach chloroplasts maximum
activity was observed with illuminated chloroplasts at 25 °C under nitrogen
and in the presence of NADP$; in soluble preparations from both chloro-
plasts and plastids an absolute requirement for FAD was noted as well as a
partial requirement for NADP$. In addition mutants of Phycomyces blakes-
leeanus have been obtained which accumulate large amounts of lycopene and
no /3-carotene, the major pigment of the wild strain [148].
Three compounds have recently been found to inhibit {3-carotene synthesis
and cause the accumulation of lycopene. Pumpkin cotyledons treated with
cycocel [(2-chloroethyl)trimethylammonium chloride] accumulate lycopene
instead of cyclic carotenoids [149]. Treatment of apricot, prune, peach and
non-lycopene producing tomato fruit, roots of carrot and sweet potato, and
mycelia of Blakeslea trispora and Phycomyces blakesleeanus with CPTA [2-
(p-chlorophenylthio )triethylamine hydrochloride] causes the inhibition of /3-
carotene synthesis and the accumulation of lycopene [150]. In Mycobacterium
species, nicotine inhibits {3-carotene synthesis and lycopene accumulates [151];
a similar effect is observed with Flavobacterium species and P. blakesleeanus
[152]. When the nicotine is washed out of the Mycobacterium culture and the
cells resuspended {3-carotene is formed at the expense of lycopene [151]. The
same thing happens with the Flavobacterium species, but in this case the con-
version can go anaerobically [152].

3. Mechanism of cyclization

Experiments with (4R)-[2- 14 C-4- 3 H]MVA in a number of systems indi-

cated that the /3-ionone residue is formed by a mechanism formally involving a
carbonium ion, but almost certainly an enzyme-substrate complex (Scheme 19).
The 14 CPH atomic ratio of phytoene (taken as 8: 8) fell to 8:6 in /3-carotene
although that of lycopene remained at 8:8. This indicated the loss of a tritium
atom from both C-6 and C-6', which arise for C-4 ofMVA, during the cyclization
process [153]. The corresponding ratio in a-carotene is 8:7, which indicated
that the a-ionone ring did not arise from the /3-ionone ring [154]. Experiments
with [2- 14 C-2,2- 3 H 2 ]MVA showed that the a-ionone ring was formed pre-
sumably by a similar mechanism but that the carbonium ion was stabilized
by expulsion of a proton (Hf) from C-4 (Scheme 19). The expected distribu-
tion of 14C and 3 H in a- and {3-carotene if the {3- and a-rings arose separately
 VII. Biosynthesis 597

~..-P-Ring

~
H!'
v
~.. ··
a-Ring

Scheme 19. Basic mechanism for biosynthesis of a- and P-rings in carotenoids [155]

is given in Scheme 20. The expected ratios, with phytoene taken as 8: 6 are
{j-carotene 8:12 and a-carotene 8:11. These were experimentally observed
[155]; if the {j-ionone ring had been formed from the a-ionone ring then both
a-carotene and {j-carotene would have had the same 14 C/ 3 H atomic ratio of
8:10. Thus there is no significant isomerization of the formed ring systems. It
was suggested that the same enzyme-substrate intermediate was involved in
forming both rings but recent work on the absolute configuration of a-carotene
[156] indicates that the situation may be more complex. If the 1{1-methyl,

T T
T
T
T

P-Carotene

T
T

T T

a-Carotene.

Scheme 20. The expected labelling of a-carotene and P-carotene from [2- 14C-2,2- 3 H 2 ] MY A if the
a- and P-ionone rings arise separately
598 T. W. GOODWIN

Chair folding

a) IfB·l'P\
(below)
~
I
"'

\
H2S H2 R

Ill

b)

H2s (6R)-e<-Rmg

(c··
H2 s H2 R B-Rmg

Scheme 21. The stereochemistry of cyclization to produce oc- and fJ-rings

which is equatorial, arises from [2- 14 C]MV A [an assumption based on the
view that trisporic acid C, a carotenoid-stimulating compound in the Muco-
rales (seep. 614), which has the absolute configuration (7), arises biosynthetically
from /)-carotene] [157], then cyclization via the usually accepted chair form
•••••.H
H

(7)
 VII. Biosynthesis 599

Boat folding

c)

~(below)

--if~
(6S)-e<-Rtng

.. ··~H2R

H4R H2s

d)

Ill

Scheme 21. [Continued]

with the loss of the axial hydrogen at C-6 will yield /)-carotene of the right
configuration around C-1 (Scheme 21 a). If the same carbonium ion yields
a-carotene by loss of the axial hydrogen at C-4, which would be the 2-pro-S
hydrogen of MV A, then the compound formed is the enantiomer of natural
a-carotene. Only a folding in the boat form (Scheme 21 d) can give a carbonium
ion which will yield a-carotene with the right configuration and this would
involve loss of the 2-pro-R hydrogen of MVA. Experiments with (2 R)- and
(2S)-MVA in appropriate systems should solve this problem; preliminary
experiments with J-carotene synthesis by Del tomatoes (see p. 609) suggests
that it is the 2-pro-S hydrogen which is lost [158].
600 T.W.GooDWIN

F. Xanthophyll Formation in Higher Plants,

Algae and Non-Photosynthetic Bacteria

The most commonly encountered xanthophylls in the green tissues of

higher plants are those with hydroxyl groups at 3 and/or 3' (e.g. lutein, 73)
and with an epoxide group at 5,6 (e.g. violaxanthin, 135). It is generally agreed
that the insertion of the hydroxyl and epoxide groups is a late stage in the
biosynthetic process. This conclusion is based on circumstantial evidence that
oxygen-containing precursors (e.g. of phytoene--+ lycopene) have never been
detected in green tissues, and on experiments with a Chlorella mutant [159].
Mutant 5/520 synthesizes mainly phytoene when cultured in the dark hetero-
trophically; on illumination in the absence of oxygen coloured carotenes are
formed which appear to be converted into xanthophylls if the cultures are
put in the dark and allowed access to oxygen.
The claim that labelled rx- and j3-carotenes have been converted into
lutein (73) and zeaxanthin (67), respectively [160], can be criticized on tech-
nical grounds. However, under certain anaerobic conditions a Flavobacterium
species which normally synthesizes zeaxanthin [152], will accumulate P-
carotene which on aerating the culture causes the j3-carotene to disappear
with a simultaneous accumulation of zeaxanthin [152]. Further investigations
on this problem are in progress.
Stereochemical studies on the hydroxylation leading to lutein, j3-crypto-
xanthin and zeaxanthin has been made in maize leaves and also in the fruit
of Physalis alkekengi and a Flavobacterium species [152, 161]. As C-3 of P-
carotene arises from C-5 of MVA, experiments with (5R)-[2- 14C-5- 3 H]MVA
and [2- 14C-5,5- 3 H 2 ]MVA showed that the 5-pro-R hydrogen is lost and the
5-pra-S hydrogen retained at C-3 of the xanthophylls [152, 161]. This demon-
strates that no ketonic intermediate is involved; furthermore, if the reasonable
assumption is made that hydroxylation involves a typical mixed-function
oxidase [162] with the hydroxyl group being inserted without inversion of
configuration, as in sterol hydroxylations, then these results bear out the
absolute configuration R at C-3 established chemically for j3-cryptoxanthin
and zeaxanthin [163]. The fact that j3-carotene contains two more tritium
atoms than zeaxanthin when (5R)-[2- 14C-5- 3 H]MVA is the precursor [161]
eliminates the suggestion that carotenes are formed in chloroplasts by reduc-
tion of xanthophylls [164].
Experiments with 18 0 2 and H 218 0 indicated that the hydroxyl oxygen of
lutein, violaxanthin and neoxanthin arises from oxygen and not water in
Chiarella [ 165 -167].
The epoxide oxygen of antheraxanthin and violaxanthin arises from 18 0 2
in Phaseolus leunatus as does that of violaxanthin in Chiarella [165-167].
There is, however, a report that the epoxide oxygen of Chlorella comes from
water, but the techniques used are open to criticism [168].
A major xanthophyll in chloroplasts of higher plants is neoxanthin (122).
No biosynthetic investigations have. been reported on this compound, but a
 VII. Biosynthesis 601

HO:trOH

-? •"""
HO
(122)

Scheme 22. A possible mechanism for the biosynthesis of neoxanthin (122) from violaxanthin (135)

OH

"--"'"""' "--"'"""' "--"'"""' "--"'"""' "--"'"""' "--"'"""'

HO
(119)

1-H®

OH

~ ~ ~ ~ ~ ~ ~ ~

HO OH

1-H 2 0

OH

~ ~ ~ ~ ~ ~ ~ ~

HO
(84)

Scheme 23. A possible mechanism for the biosynthesis of eschscholtzxanthin (84) [170]
602 T. w. GooDWIN

plausible synthesis would involve violaxanthin (135) as a precursor (Scheme 22).

Many new types of xanthophylls exist in flower petals, fruit and certain algae,
and all pose interesting biosynthetic problems very few of which have been
studied.
The retro carotenoid eschscholtzxanthin (84) [169], produced in petals of
Eschscholtzia californica, appears to be derived from a P-carotene derivative
because with (4R)-[2-14C-4- 3 H]MV A as substrate, eschscholtzxanthin retains
all the chain tritium atoms found in P-carotene [170]. A possible mechanism
for the formation of eschscholtzxanthin from antheraxanthin (119) is given in
Scheme 23. Compounds which have not yet been studied biosynthetically are
those with primary hydroxyl groups [e.g.lycoxanthin (62) and siphonaxanthin
(174)], tertiary hydroxyl groups [fucoxanthin (190)], keto groups [fucoxanthin,
siphonaxanthin, rhodoxanthin (209)], five-membered rings [capsanthin (170)],
5,8-epoxy groups [auroxanthin (141)], and those containing acetylenic linkages
[alloxanthin (65)] (see Chapter XII for further examples). A new group of 1,2.
epoxides of acyclic carotenes found in tomatoes [171, 172] are analogous to
squalene epoxide, which is an intermediate in the biosynthesis of sterols. There
is no indication yet that these compounds are biosynthetically important
(see p. 610).

Interconversions of xanthophylls
It is clear from many reports that epoxidation and de-epoxidation occur
in chloroplasts. The mechanism involved is not at all clear and subtle differen-
ces between different experimental systems appear to exist.
Sapozhnikov [173-176] and his colleagues were first to observe reversible
epoxidation but it was claimed that the reaction resulted in the conversion
of zeaxanthin into lutein (Scheme 24) [173-178]. Improved techniques have
revealed that this is not so but that a cycle is involved (Scheme 25) [179-182].
Furthermore experiments with (4R)-[2- 14 C-4- 3 H]MVA showed that in maize

~-··
HOA)l_
(a)/
/ - H20

'-...;:-H 2 0
(b)--...,._

Scheme 24. First ideas on the intercdnversion of xanthophylls in green tissues [173]
 ---
VII. Biosynthesis 603

Photoproducts in the dark

D J:i J::Xo-0 ~ooX

-----...
HO

--- --- OH HO ~OH HO~ ~OH

Photoproducts in the light

Scheme 25. Recent views on the interconversion of xanthophylls in green tissues [181]

seedlings there was no interconversion of the rx- and P-rings of lutein and zea-
xanthin [161]; this agrees with the results at the hydrocarbon level (p. 597)
[153-155].
In Euglena gracilis de-epoxidation of antheraxanthin is a dark enzymic
reaction which requires a source of reducing power such as NADPH or
FMNH 2 [182]; similar results have been observed with Typhanium and Arum
[183], Spinacea aleracea [184, 185] and Chiarella vulgaris [184, 185]. In iso-
lated spinach chloroplasts de-epoxidation requires the presence of ascorbic
acid and light; it can be inhibited by DCMU, indicating that photosystem II
is concerned in the reaction, which presumably requires ATP produced dur-
ing photophosphorylation; indeed ATP will replace light in the reaction. The
reaction also takes place in the absence of both light and ATP if the pH of
the suspension of chloroplasts and the soluble de-epoxidase is reduced to 5.0.
This reaction is not inhibited by uncouplers of photosynthetic phosphoryla-
tion. Rather similar results have been obtained with Chiarella vulgaris [184,
185].
The epoxidation of zeaxanthin is not simply reversal of the de-epoxidase
reaction under appropriate conditions. In Euglena the reaction requires oxygen
and light and was thought to be non-enzymic [186]. These experiments were
carried out on lyophilized tissues, and it is claimed that under these circum-
stances the changes observed represent merely photochemical destruction of
zeaxanthin [187]. However epoxidation has been demonstrated in intact Chia-
rella and needles of Taxus baccata immediately after very strong illumination
and is stimulated somewhat by exposure to pure oxygen or dim light. Either
pure 0 2 or dim light is mandatory for the reaction to occur in intact Spinacea
aleracea. No enzymic epoxidation has been observed in isolated cell frag-
ments [185-187].

G. Site of Carotenoid Synthesis in Green Tissue

In photosynthetic tissues carotenoids exist in the chloroplast, and the ques-

tion arises whether or not they are synthesized in these organelles. Although
chloroplasts exhibit a considerable degree of autonomy there is no evidence
that they can carry out the steps of glycolysis leading to pyruvate and even-
tually acetyl-CoA, the primary precursor of the carotenoids. A possible source
604 T. W. GOODWIN

*?H20H
*~HNH 2
• 0 2H
- *~H 2 0H
* 0
•to2H - *~H 2 0H
* HOH
•to2H

j j
*CH2 *CH 20H
:~:· ·~-o--®
:i:~
+-- +--
•co2H •co2H

~,1
*?"·
•co-s---coA

Scheme 26. Possible route for formation of acetyl-CoA in chloroplasts ( + glyoxisomes)

of pyruvate is glycolate [188-191], an early product of C0 2 fixation, involv-

ing the pathway indicated in Scheme 26; all the intermediates are actively
incorporated into P-carotene by maize seedlings [188]. The possibility that
the step involving glycolic oxidase involves glyoxisomes, discrete organelles
closely associated with chloroplasts, cannot be ignored. The enzyme is present
in these organelles but apparently absent from pure chloroplasts [192-194].
If pyruvate is formed in this way, there is still no evidence that the chloroplasts
can convert it into acetyl-CoA [195]. It may be that the mitochondrion carries
out this step but then the acetyl-CoA has to penetrate the mitochondrial and
chloroplast membranes, which are considered impermeable to this compound
[196]. Other possibilities have been considered [195, 197] but more work is
urgently required.
 VII. Biosynthesis 605

There is no evidence for the conversion of acetyl-CoA into MV A in chloro-

plasts but certainly all the steps from MV A to phytoene can take place therein,
because non-aqueously prepared chloroplasts will convert MV A into phytoene
[198]; on the other hand, conventionally prepared chloroplasts are not active,
the reason for this is not yet clear. Enzymes phosphorylating MVA have
also been demonstrated in chloroplasts [ 40-42]. However, recent work sug-
gests that in spinach preparations phytoene synthesis from IPP occurs in a
supernatant fraction from which chloroplasts have been removed; the enzyme
might of course have been liberated from the chloroplasts during preparation
[129]. Further experiments showed that the 0-80% (NH 4 hS0 4 precipitate
was active, that light was mandatory for activity, and that the addition of
boiled supernatant was stimulatory. With non-aqueous chloroplasts no con-
version of phytoene into more saturated carotenes has been observed [93,
198]; however, addition of an extract of conventionally prepared chloroplasts
from spinach to the supernatant and boiled supernatant just described results
in conversion ofiPP into phytofluene and lycopene in the presence ofNADPEil,
FAD and light [129]. As indicated on p. 596, isolated chloroplasts can cyclize
lycopene to cyclic carotenes.
Although the recent evidence for the conversion of phytoene into coloured
carotenoids in either chloroplasts or chloroplasts plus supernatant is impres-
sive, genetic evidence suggests that the desaturation of phytoene is under
nuclear control. The albino maize mutant (W -3) accumulates phytoene (32)
[199, 200]; the block is due to a gene on maize chromosome 2 [201]. Two
other maize mutants accumulate (-carotene (26) and lycopene (19) [202];
however, the block is not complete because phytofluene (30), (-carotene and
trace amounts of neurosporene (22) are present; both mutations are inherited
in normal Mendelian fashion.
A yellow mutant of Helianthus annuus tends to accumulate xanthophylls
rather than carotenes [203-205]. This might mean that the nucleus controls
a step in carotene synthesis which is not involved in xanthophyll synthesis, a
possibility which is against present biochemical views ofbiosynthesis ofxantho-
phylls. Further information on the nature of these xanthophylls and their
intracellular location is required. Etiolated seedlings [206] produce xantho-
phylls rather than carotenes, as do etiolated cotyledons of beans [207, 208];
a possible explanation is that the pigments in the yellow mutant are synthesized
extraplastidically by a pathway similar to that in the plastids but that regula-
tion of the oxidation of carotenes is 'set' to produce almost entirely xantho-
phylls, and this characteristic shows Mendelian inheritance and is controlled
by a sirtgle recessive factor.

H. Carotenoid Metabolism in Senescing Leaves

The yellow colour of autumn leaves is due to the more rapid oxidative
degradation of chlorophylls than carotenoids as chloroplasts degenerate and
finally disintegrate during leaf senescence. The mechanism involved in the
606 T. w. GooDWIN

break-down of carotenoids is not known but the formation of 5,6-epoxides

is the first step [209-213]. Rather unexpectedly the carotenes disappear more
quickly than the xanthophylls [211, 214, 215]; this may be due to the fact that
the xanthophylls are protected to some extent by becoming esterified as they
are liberated into the cytoplasm on disintegration of the chloroplasts [211,
216, 217]. The esterified xanthophylls were first thought to be autumn caro-
tenes by Berzelius in 1837 [218], because of their lack of polarity owing to
the esterification of the free hydroxyl groups at C-3.

I. Carotenoid Formation in Fruit

1. Ripening
In many fruits ripening is accompanied by massive synthesis of carotenoids
as the chloroplasts change into chromoplasts. These morphological change;
have been demonstrated in Sambucus racemosa, Physalis alkekengi and Sorbus
aucuparia [219], Solanum capsicastrum [220], Capsicum annuum [221, 222],
navel oranges [223] and tomatoes [224]. It is also probable that such fruits have
a functional set of carotene-synthesizing enzymes because they can be ripened
effectively off the vine; for example a stored tomato can synthesize as much
as 1.2 mg of lycopene per day [225]. It has also been shown that isolated
pericarp discs of Capsicum annuum will ripen and form the characteristic red
carotenoids (capsanthin etc.) when cultured aseptically in an aerated liquid
medium [226].
The ripening of fruit is usually accompanied by the loss of chlorophyll as
the chloroplasts degenerate but this is not mandatory. There are tomato and
pepper mutants in which considerable amounts of chlorophylls are present
in mature fruit (see next section), and in grape fruit active synthesis of
carotenoids begins before the chlorophylls start to disappear [227, 228].
A characteristic of the ripening process is that it frequently involves the
synthesis of large amounts of 'non-chloroplast' pigments such as lycopene
(19) (tomato), capsanthin (170) (red peppers) or greatly enhanced synthesis of
one chloroplast carotenoid without concomitant synthesis of the others [e.g.
zeaxanthin (67) in Physalis alkekengi]. The synthesis of non-chloroplast carote-
noids as red peppers ripen is indicated in Table 2 [229].
Clearly the massive changes both morphological and biochemical which
are triggered off by a ripening hormone, possibly ethylene, involve (i) repres-
sion of some enzymic activity e. g. cyclization of lycopene (19) in tomatoes,
(ii) either derepression or synthesis or activation of others as in zeaxanthin syn-
thesis, and (iii) derepression or stimulation of synthesis of a new enzyme as
in the presumed conversion of antheraxanthin into capsanthin (Scheme 27).
It is more than possible that a physically separate pathway of synthesis arises
in developing fruit and that this is superimposed on the chloroplast pathway,
which may continue at its normal rate or gradually disappear. For example,
in normal tomatoes lycopene synthesis is inhibited if the fruit are held at
 VII. Biosynthesis 607

Table 2. The major carotenoids of leaves, unripe and ripe fruit of Capsicum annuurn v. lycopersici-
forrne rubrurn [229]

Pigment Amount (mg/100 g fresh weight)

Leaves Unripe fruit Ripe fruit

P-Carotene 7.92 0.095 2.35

Cryptoxanthin 0.45 0.027 1.10
Lutein 13.99 0.276
Zeaxanthin 1.75
Antheraxanthin 1.14 0.031 0.99
Violaxanthin 8.27 0.042 0.70
Neoxanthin 5.66 0.058
Capsanthin 9.60
Capsorubin 1.46

temperatures above 30 °C, whilst {3-carotene (3) synthesis is not [230-232];

the effect is not permanent and on lowering the temperature lycopene synthesis
continues [233]. On the other hand, in high-/3 tomatoes, in which lycopene
is replaced by {3-carotene (seep. 608), the synthesis ofthis additional {3-carotene
is also temperature-sensitive [141, 234]. Furthermore, in normal tomatoes
dimethylsulphoxide (DMSO) inhibits the synthesis of lycopene but not
{3-carotene [235].
Electron microscope studies also indicate separate sites of accumulation,
if not synthesis, in tomatoes. {3-Carotene is found in lipid droplets during the
early stages of chromoplasts development, whereas lycopene bodies develop
in association with rigid membranes, which do not appear in the chromoplasts
of tomatoes kept at 32 °C, when no lycopene is synthesized [224]. Finally the
pattern of incorporation of substrates such as glyoxylate into the carotenoids
of ripening fruit is very much less than into carotenoids of chloroplasts [236].

~·~
HO~~t ---- V v G

OH
. .·
~
H'"
OH

Scheme 27. Possible mechanism for conversion of antheraxanthin into capsanthin

2. Genetic studies
A considerable number of genetic studies on tomatoes has produced
interesting information which in general fits in with the accepted pathway of
biosynthesis. The pigment distribution in the various tomato phenotypes is
given in Table 3. Normal red tomatoes possess the dominant allele r+ whilst
yellow tomatoes are homozygous for the recessive allele r. The r+ /r gene
controls the total amount of pigment formed, the rr genotype synthesizing
608 T. W. GooowiN

Table 3. Carotenoid distribution in various tomato phenotypes

Genotypes Pigments and Total concentration References

colourless polyenes* Jlg/g fresh weight
Total Pigments
polyenes only

Red (normal) 1, 2, 3, 4, 5 87
50}
69 [234]
High pigment (r+ r+) 1, 2, 3, 4, 5 88
Tangerine (r+ r+ tt) 2, 3, 4, 5, 6 158 90
Yellow (rrtt) 1, 2, 4, 5 4 2 [253]
Yellow Tangerine (rrtt) 1, 2, 4, 5, 7 24 14 [238]
Apricot (atat) 1, 2, 4, 5 13 11 [238]
Yellow-} 2, 4, 5 (trace) 2 [238]
A . t (rrtt at at)
pnco
Tangerine} ) 1, 2, 4, 5, 6, 7 29 16 [238]
Apricot tt atat

Yellow- }
Tangerine- (rrtt atat) 4, 5 10 0 [238]
Apricot
295 1 [244]

%)
Ghost (ghgh) 1, 4, 5
Intermediate-P (B+ B+) 1, 2, 3, 4, 5 50
High-/1 (B+ B+ Moa Moa) 1, 2, 3, 4, 5 80 70
[243]
Delta (DeJ+ Del+) 1, 2, 3, 4, 5, 7, 8, 9, 10 84 68
High-delta (Del+ Dei+ hphp) 1, 2, 3, 4, 5, 8, 9, 10 55 46

*Abbreviations used: 1. Lycopene, 2. P-Carotene, 3. y-Carotene, 4. Phytoene, 5. Phytofluene,

6. Prolycopene, 7. (-Carotene, 8. a-Carotene, 9 . .5-Carotene, 10. Neurosporene.

only 5% of that formed by the r+ r+ genotype [237]. In apricot fruit, which

are homozygous for the recessive allele at, the synthesis of the acyclic series
is considerably inhibited whilst that of /3-carotene is unaffected [238]. A third
recessive gene hp tends to increase the content of all compounds [142].
Qualitative changes are controlled by another series of genes. Tangerine
tomatoes carry the recessive t, whilst the dominant allele t+ is carried by
normal fruit [239]. Tangerine tomatoes produce prolycopene, a poly-cis-
lycopene of unknown structure, at the expense of lycopene; the corresponding
proneurosporene is also present [112, 240]. The effects of rand tare reinforced
by at [238].
An orange phenotype obtained by back crossing a L. esculenturn and L.
hirsuturn hybrid has /3-carotene and not lycopene as its major carotenoid [241].
The fruit of these high-/3 tomatoes are either homozygous or heterozygous
for the dominant allele B+, whilst normal fruit are homozygous for the reces-
sive B [140]. The expression of the B+ /B gene is partly controlled by an
independently inherited modifier gene Mot/MoB [140]. In the presence of
Mot in either the homozygous or heterozygous form, /3-carotene which rep-
resents over 90% of the total carotenes in the presence of B+, is reduced whilst
that of lycopene is increased so that about equal amounts of each pigment
are present. In the presence of BB (normal fruit) MoB has no effect on lycopene
 VII. Biosynthesis 609

production. A very similar gene to B+ /B and one which is tightly linked to

the self-pruning locus exists in an orange fruited L. pimpine/lifolium from the
Galapagos Islands [242]. The allele Def+, which produces J-carotene (11) and
related cyclic carotenoids at the expense of lycopene [243], is either incom-
pletely dominant or dominant but affected by a modifier gene [234].
It is interesting that in plants containing both Band Del genes, less amounts
of carotenoids with a-rings are formed than in plants containing Del gene
only; this confirms the conclusion from experiments with stereospecific MV A's
(p. 597) that a- and fJ-rings of carotenoids are not interconvertible but probably
arise by a parallel pathway from a common precursor.
The ghost phenotype appears spontaneously in normal tomato lines.
Seedlings homozygous for the recessive allele gh germinate with green cotyle-
dons which rapidly lose chlorophyll as they grow. Grafting of normal scions
on to ghost stock results in the production of milky white unripe fruit which
become yellowish on ripening [244]. Phytoene accumulates in considerable
amounts, the yellow colour of the mature fruit being due to an alkali-soluble
pigment. A striking difference of the gh gene compared with others is that
it also affects the carotenoid pattern in the leaves, which also produce
phytoene but no coloured carotenoids. Other genetic information relating
to carotenoids in fruit has been discussed by Goodwin and Goad [245].
Although, as indicated above, tomato plastids can synthesize carotenoids
from primary precursors, their production appears to be under nuclear con-
trol because most of the mutations just discussed are inherited in a Mendelian
fashion [246]. One can tentatively propose the site of action of these genes:
(i) r+ /r, at+ fat, hp+ jhp act before phytoene in controlling quantitatively the
flow of precursors into phytoene; (ii) gh+ /gh controls the desaturation of
phytoene; (iii) B+ /B controls the cyclization oflycopene at both ends; (iv) De[+ I
Del may control mainly either the cyclization of lycopene at one end or the
cyclization to produce a-rings rather than fJ-rings; (v) the effect of Mof;/Mo 8
is difficult to assess at the moment, but it may regulate the activity of the
cyclizing enzyme; (vi) t+ /t appears to inhibit the later stages of desaturation,
because (-carotene, neurosporene etc. occur in comparatively large amounts,
and also the stereochemistry of the final molecule, i.e. prolycopene and not
lycopene is formed. Little more can be said until the structures of the cis
derivatives are known.
As already indicated red peppers as they ripen produce the characteristic
carotenoids capsanthin (170) and capsorubin (205). An orange-yellow mutant,
in which the character is due to a single recessive Mendelian factor [247],
clearly makes less pigment but the details are not known, whilst in yellow
strains the chlqroplast carotenoids are the major components with no syn-
thesis of capsanthin or capsorubin being observed [248]. The synthesis of
these pigments in peppers appears to be controlled in a somewhat similar way
to that functioning in tomatoes.
It has long been known that a characteristic of citrus fruit is the presence
of the apo-carotenoids such as fJ-apo-10' -carotenal (256), fJ-apo-8' -carotenal

Carotenmds 39
610 T. W. GOODWIN

(248) and P-citraurin (249); indeed Triphasia trifolia contains 30 mg of carote-

noid per 100 g fresh weight, of which the major component is semi-p-carote-
none (213) [249]. Studies of new citrus hybrids have recently revealed the
presence of the unique methyl ketones sintaxanthin (244), citranaxanthin (237)
and reticulataxanthin (238) in the trigeneric hybrid of the oval kumquat (For-
tunella margarita) with the Rusk citrange (Poncirus trifoliata x Citrus sinensis)
[250]. The interesting fact emerges that these pigments only appear in signi-
ficant amounts on hybridization; neither parent produces more than traces,
if any, of these pigments. The synthesis of these compounds in such large
amounts alongside the more conventional apo-carotenoids poses interesting
problems of the pathway of degradation of carotenoids.
The red-fleshed (lycopene) Carica papaya, which differs from the normal
yellow (/J-carotene) type by a single gene [251], is by analogy with the tomato
just discussed homozygous for the recessive genet. A similar situation probably
exists between the orange-fleshed watermelon (/J-carotene) and the norm&
pink-flesh type [252].

3. Epoxides of acyclic carotenes in tomato fruit

The discovery of phytoene 1,2-epoxide (116) in ripe tomatoes [253] led
eventually to the demonstration of the presence of the corresponding derivative
of phytofluene, '-carotene and lycopene [254]. These are analogues of squalene
2,3-epoxide, the immediate precursor of triterpenes. The carotene epoxides
probably do not have any biosynthetic significance; they are possibly formed
by the rather unspecific squalene epoxidase when the carotenes are liberated
into the cytoplasm as the chromoplasts degenerate.

J. Regulation of Synthesis in Green Tissues

Etiolated seedlings produce only traces of carotenoids, the majority of
which are xanthophylls [255-257]; in response to light 'chloroplast carote-
noids ', including considerable amounts of P-carotene, are synthesized as func-
tional chloroplasts are formed [256, 258, 259]. The mechanism involved in
the triggering off of chloroplast development by light is not known but a
possible means of regulation of carotenoid synthesis during this complex pro-
cess has been suggested [260, 261]. If etiolated seedlings are excised from their
leaves, placed in [2- 14C]MVA and allowed to green up then relatively little
activity is found in the chloroplast terpenoids: the carotenoids and the side
chains of the chlorophylls, plastoquinones, tocoquinones and menaquinone,
although the extraplastidic terpenoids: squalene sterols, P-amyrin and the side
chain of ubiquinone, are very strongly labelled. On the other hand, if the same
experiment is carried out with 14C0 2 the situation is reversed: the chloroplast
terpenoids are highly labelled and the extraplastidic terpenoids only slightly
labelled. Thus regulation of terpenoid synthesis in developing seedlings may
be achieved by compartmentalization with two separate sites of terpenoid
synthesis from MV A onwards, one inside and the other outside the chloro-
 VII. Biosynthesis 611

plast. The basic reactions of synthesis of the C 5 -unit from MVA and chain
elongation by condensation of C 5 -units can occur at both sites whilst certain
specific reactions can occur only inside or outside the chloroplast. The situation
is summarized in Scheme 28.

Extrachloroplastidic reactions Common reaction Chloroplast reactions

Storage material-----+ Acetate - - - - - - - - C0 2

t
Mevalonic acid

t
Isopentenyl--®---@

t
Geranyl--®---@

t
~ Farnesyl--®---@
~ t Tocopherols K 1 Chlorophyll

Squalene Geranylgeranyl--®---@ ~ j/
/ \ t ~Phytol
Other triterpenoids Sterols Solanesyl--®---@ Phytoene

Ub.....,.. ·=.,l=~,,_ ""~ c~L,. .

geranyl--®---@
Plastoquinone

Scheme 28. Compartmentalization of terpenoid synthesis in green tissues of higher plants

In addition the chloroplast membrane appears to be relatively impermeable

to MV A and the intermediates following it on the terpenoid biosynthetic path-
way. This prevents this substrate penetrating to the plastids when the seedling
is germinating and prevents its moving out from the plastids as they are being
converted into <;hloroplasts on illumination. This ensures that MV A is where
it is required at the appropriate time.
The problem of regulation in such organisms as Chiarella and Scenedes-
mus which are green and produce functional chloroplasts in the dark still
remains to be investigated. Cotyledons from pine seedlings germinated in the
dark are green, but the MVA/C0 2 pattern of incorporation of label into
612 T.W.GooowiN

chloroplast terpenoids appears to be the same as in tissues greening in the

light [262].
As chloroplasts mature the incorporation of 14C0 2 into carotenoids drops
almost to zero [31] as the turn-over of photosynthetic pigments comes to a
halt; the mechanism by which this is achieved is unknown.

K. Special Aspects of Synthesis in Algae and Fungi

1. Production of extraplastidic carotenoids

Under certain environmental conditions, particularly poor nutritional sta-
tus involving nitrogen deficiency, many algae turn red owing to the massive
synthesis of extraplastidic carotenoids (see [263] for full list). In this they tend
to resemble ripening carotenogenic fruit. In some cases the pigment is /3-
carotene (3) but more frequently keto carotenoids [e.g. echinenone (148),
canthaxanthin (193) and astaxanthin (203)] are formed [263]. The mechanism
involved in this synthesis is not known but presumably the keto carote-
noids are formed from {3-carotene as in Crustacea (see p. 628). It has been
suggested that these pigments are synthesized in the chloroplast from products
of the breakdown of chlorophyll and then diffuse into the fat vesicles [264];
there is no evidence to support this view.

2. Biosynthesis in red yeasts

A characteristic of red yeasts is that they synthesize carotenoids with one
additional conjugated double bond compared with the usual eleven in higher
plant carotenoids; these are torulene (3',4'-didehydro-y-carotene, 7) and torula-
rhodin (211). Rhodotorula glutinis synthesizes mainly /3- andy-carotenes when
cultured at 5 oc; at 25 oc the levels of these carotenes are reduced, and toru-
lene aud torularhodin accumulate [265]. Recent work suggests the biosynthetic
sequence outlined in Scheme 29, in which y-carotene is considered the branch
point and the desaturation to torulene the temperature-sensitive step [266].
The cyclization is considered to involve neurosporene rather than lycopene
(p. 594) because under no conditions could lycopene be detected. This path-
way is supported by mutant studies on R. mucilaginosa, from which it was
concluded that pigment 'X' (probably /3-zeacarotene) andy-carotene are pre-
cursors of torulene and torularhodin [267]. Other mutants of R. mucilaginosa
have been described [267] as have mutants of R. rubra [268].
Work with 18 0 2 indicated that one oxygen atom from 0 2 is incorporated
into the carboxyl group of torularhodin [269]. This result is consistent with
the CH 3 -+ C0 2 H reaction involving first the oxidation to CH 2 0H via a
mixed function oxidase, which would require 0 2 , followed by dehydrogenation
reactions: -. CHO-. C0 2 H. Detection of the possible intermediates has
recently been reported [270].
The biological oxidation of the terminal methyl group to C0 2 H allowed
investi~ation of the question whether the terminal methyl groups in carotenoids
 VII. Biosynthesis 613

'-'::::::: '-'::::::: '-'::::::: '-'::::::: '-'::::::: '-'::::::: '-'::::::: '-':::::::

Neurosporene

j Cyclization

P-Zeacarotene

1-2H

y-Carotene

j -2H
~··· P-Carotene

Torulene (7)

1 Oxidation

1 Oxidation

C0 2 H

Torularhodin (211)

Scheme 29. Probable biosynthetic route for formation of torularhodin (211) in red yeasts [266]
614 T. W. GOODWIN

retained their individuality as they do in diterpenes (e.g. gibberellins) and

triterpenes (soyasapogenol A). Experiments with [2- 14 C]MV A and
[2- 14 C-2,2- 3 H 2 ]MVA showed that the individuality is retained and that it is
the methyl group arising from C-2 of MVA, which is oxidised to the carboxyl
group of torularhodin [271]. Scheme 30 shows the expected labelling from
T T
T
(a)
'-'::::: 1' T
T

T T T
'-':::::
•
C02 H(b)
1'

T
T
(c)
T
T C0 2 H

•=t4c
T= 3 H
Scheme 30. Expected labelling pattern from [2- 14C-2,2- 3 H 2 ]MVA: (a) torulene, (b) torularhodin
if the carboxyl group at C-1' originated from C-2 of MVA, (c) torularhodin if the methyl group at
C-1' originated from C-2 of MVA [271]

[2- 14C-2,2- 3 H 2 ]MVA in torulene and in torularhodin in which the carboxyl

group or the methyl group arose from C-2 of MVA. Experiments showed that
torularhodin contained two tritiums fewer than torulene, and thus the carboxyl
group originated from C-2 of MVA (Scheme 30b). This was confirmed by
showing that the carboxyl group was labelled from C-2 of MVA [270].

L. Bioinduction of Carotenogenesis
1. Trisporic acid
When ( +) and (- ) strains of the heterothallic fungus Choanephora cucur-
bita are cultured together some twenty times more carotenoid is synthesized
than in (-) or ( +) strains cultured separately [272]. The same phenomenon
is observed with ( +) and (-) strains of Blakeslea trispora [273-277]. The
stimulation is so great under appropriate cultural conditions that industrial
production of P-carotene (3) is feasible [273]. The biostimulator (so-called
P-factor) has been isolated from mated cultures of B. trispora and consists of
 VII. Biosynthesis 615

a series of organic acids, of which trisporic acid C (7) (p. 598) is the major
component [278-280]. Trisporic acid B (8) [279] and 'compound 3' [280]
are probably identical. Trisporic acid only stimulates carotenogenesis in the
(-) strain [277, 282] (Table 4). [ 14C]Trisporic acid is not incorporated into
Table 4. Stimulation of carotenogenesis in ( +) and (-) strains of Blakes lea trispora by trisporic
acid [277]
Culture Carotenoid concentration (Jlg/100 ml medium)

Phytoene P-Carotene (-Carotene y-Carotene Lycopene

Untreated (-)strain 28 20 12
(-) strain plus trisporic acid* 1760 2300 240 50 30
Untreated ( +) strain 20 16 10
( +) strain plus trisporic acid* 40 24 16

* 24-hr cultures treated with trisporic acid and incubated for a further 48 hr before harvesting.

P-carotene by B. trispora [276] and is thus not a precursor of P-carotene. On

the other hand, it has recently been claimed that trisporic acid itself is formed
from P-carotene via retinal [282]. The trisporic acid effect is inhibited by
actidione [283], a specific inhibitor of protein synthesis, which means that
trisporic acid probably derepresses synthesis of an enzyme or enzyme complex
concerned with carotenogenesis. In the presence of D PA trisporic acid-treated
(-) B. trispora produces relatively enormous amounts of phytoene (32) [277].
As ( +) B. trispora but not (-) B. trispora produces trisporic acid when
cultured with the homothallic fungus Zygorhynchus moelleri it is concluded
that trisporic acid is synthesized only by the (+)form; this view is supported
by kinetic studies on trisporic acid formation. Although there is a suggestion
otherwise [281], it appears that physical contact is not required for trisporic
acid production but that it is stimulated by a 'pro gamone' released from the
(-) strain [280].

""o

(8) (9)

2. Other factors
Effects somewhat similar to that observed with trisporic acid have been
known for some time. In the early 1950's it was shown that P-ionone (9) stim-
ulated carotenogenesis in Phycomyces blakesleeanus [284-286] without itself
being incorporated into the carotene molecule [287]; furthermore P-ionone
did not dilute out the incorporation of [ 14C]glucose into P-carotene in P.
blakesleeanus [288]. A similar stimulatory effect of P-ionone was observed on
carotene production in heterothallic cultures of Blakeslea trispora [289]. The
mechanism involved is not yet clear although the failure of chloramphenicol
 616 T. w. GooDWIN

-2H -2H
Phytoene (32) Phytofluene (30) ---------------------~

j-2H

"'<::::, "'<::::, "'<::::, "'<::::, "'<::::, "'<::::, "'<::::,

(-Carotene (26)

j-2H
~"
Neurosporene (22) "
"'<::::, "'<::::, "'<::::, "'<::::, "'<::::, "'<::::, "'<::::, "'<::::,
"] -->-

j -2H

"'<::::,
Lycopene (19)
"'<::::, "'<::::, "'<::::, "'<::::, "'<::::, "'<::::, "'<::::, "'<::::, "'<::::, "'<::::, "'<::::, "'<::::,
--
Scheme 31. Probable pathways for biosynthesis of C 45 - and C 50 -carotenoids [297]

to inhibit the effect was construed as indicating that induction of enzyme

synthesis was not involved [290]. However, chloramphenicol was not signi-
ficantly active in inhibiting the action of trisporic acid in B. trispora or the
incorporation of [ 14C]leucine into protein either, although as indicated above
actidione is active in both ways [283]; it is important to know the effect of
actidione on P-ionone-stimulated carotenogenesis in P. blakesleeanus. A fur-
ther difference is that P-ionone, unlike trisporic acid, stimulates synthesis in
B. trispora only in mated strains and not in the (-) or ( +) strain [290].

M. Biosynthesis in Non-Photosynthetic Bacteria

The most interesting developments in this aspect of carotenoid biosynthesis

involve the formation of the carotenoids with 45 and 50 carbon atoms and the
photoinduction of carotenoid synthesis.
 VII. Biosynthesis 617

~ ~ ~ ~ ~ ~ ~ ~
Unsymmetrical (-carotene (25)
I
I
I
I
I
...

HOH2 C ~

Nonaprenoxanthin (218)
I
I
: -2H
I
...

HOH2 C ~ ~ ~ ~ ~ ~ ~ ~

11',12'-Didehydrononaprenoxanthin (217)
-2H
...

[uou,c ~

P452 (216a)
~ ~ ~ ~ ~ ~ ~ ~ ~ ~

J
1
HOH 2 C ~ ~ ~ ~ ~ ~ ~ ~ ~

Deshydroxydecaprenoxanthin (223)

I
CH 2 0H
HOH 2 C ~ ~ ~ ~ ~ ~ ~ ~ ~ ~

Decaprenoxanthin (226)
Scheme 31. [Continued]
618 T. W. GOODWIN

1. Biosynthesis of C 45 - and C 50 -carotenoids

a) Introduction
The first C 50 -carotenoid, decaprenoxanthin (226), was discovered in 1966
in Flavobacterium dehydrogenans [291-293]; since then a number of additional
C 50 -pigments have been characterized (see Chapter XII). They all carry addi-
tional isopentenyl residues at C-2 and C-2' and can be either (i) acyclic (e.g.
bacterioruberin, 231), (ii) monocyclic as in C. p. 473 (221) from Corynebacte-
rium poinsettiae, or (iii) bicyclic with either two ex-rings as in decaprenoxanthin
or two /3-rings, as in C. p. 450 (220) from Corynebacterium poinsettiae [294].
A C 50 -carotenoid with one ex- and one /3-ring has not yet been reported.
Corynexanthin (227) first described in 1954 [295] is a monoglucoside of deca-
prenoxanthin [296]. c45-carotenoids, e.g. nonaprenoxanthin (218), have also
been observed in F. dehydrogenans [293, 296].
So far the C 45 - and C 50 -groups of carotenoids have been found only in
gram-positive aerobic bacteria, if one excludes Halobacterium species, which,
although gram-negative, has an unusual cell wall more allied to cell walls of
the gram-positive than to those of gram-negative bacteria [297].

b) Biosynthetic pathways
Under cultural conditions, such as limited aeration or in the presence of
diphenylamine, which inhibit the synthesis of decaprenoxanthin and allow the
accumulation of acyclic carotenes and saturated precursors, a biosynthetic
picture similar, but not identical, to that in other organisms is obtained.
Phytoene (32) and phytofluene (30) accumulate, but in this case they are the
all-trans isomers [298]; (-carotene (7,8, 7',8'-tetrahydrolycopene, 26) appears
[298] together with its unsymmetrical isomer 7,8,11,12-tetrahydrolycopene
(25) [298]. When diphenylamine is removed and the cultures resuspended
these compounds disappear and decaprenoxanthin is formed. From these
observations reasonable possibilities for the formation of nonaprenoxanthins
and decaprenoxanthins can be proposed (Scheme 31) [297]. Lycopene, formed
either via (-carotene or 7,8,11,12-tetrahydrolycopene [299], could add two
isoprene units sequentially to C-2 and C-2' to yield intermediately a hydro-
carbon corresponding to P 452 and eventually, after hydroxylation, P 452
(216a) itself.
Nonaprenoxanthin (218) is probably formed by addition of a C 5-unit to
7,8,11,12-tetrahydrolycopene (25) followed by hydroxylation. There are two
alternatives for the formation of 11',12'-didehydrononaprenoxanthin (217) and
P 452 (216a): either addition ofC 5 to neurosporene and lycopene, respectively,
followed by hydroxylation, or desaturation of the c45-hydrocarbon formed
from 7,8,11,12-tetrahydrolycopene (25), followed by hydroxylation. In the case
of decaprenoxanthin (226) lycopene is converted into the C 50 -hydrocarbon
corresponding to decaprenoxanthin by sequential addition of two C 5-units at
C-2 and C-2', respectively. Presumably hydroxylation at the C 45 -stage occurs
 VII. Biosynthesis 619

because from kinetic experiments P 452 would appear to be a precursor of

decaprenoxanthin [292].
The mechanism proposed for producing the cyclic C 45 - and C 50 -carote-
noids with oc- and /1-rings such as decaprenoxanthin (226) and C. p. 450
(220) is an extension of that originally put forward for forming the rings of
oc- and P-carotenes (p. 597), in that the isopentenyl residue replaces a proton
as nucleophilic reagent (Scheme 32).

\ H(t)
a-End group

P-End group

Scheme 32. Probable mechanism for forming C 45 - and C 50 -carotenoids

with a- and P-rings [297]

The discovery of acyclic C 50 -carotenoids such as bacterioruberin means

that the addition of a supernumerary isopentenyl residue to the C 40 -unit does
not necessarily result in ring formation. It may be that addition takes place
only after the C 40 -precursor becomes fully unsaturated. In Halobacterium
species this could be 3,4,3',4' -tetradehydrolycopene (17) and the additional
double bonds at 3 and 3' could inhibit cyclization, thus facilitating formation
of a branched compound (Scheme 33) [298].

HO'>l r ~~~/
~ ~

Scheme 33. Probable mechanism for forming acyclic C 45 - and C 50 -carotenoids [297]
620 T. w. GooDWIN

N. Photoinduction of Carotenogenesis in Non-Photosynthetic Bacteria

A number of non-photosynthetic bacteria and fungi will synthesize signi-

ficant amounts of carotenoids only after a short exposure to light and oxygen
[292, 300-307]. In Neurospora crassa the action spectrum suggested that
either a flavin or a carotenoid could be the receptor pigment [303]. In a Myco-
bacterium species it is probably a flavin, because the action spectrum has
maxima at 365 nm and 460 nm and a minimum at 404 nm [300, 304]; further-
more, photoinduction is inhibited by hydrosulphite, which would keep the
flavins in a reduced form [300]. In Mycobacterium marinum, on the other hand,
the action spectrum has its main maximum at 404 nm with additional minor
maxima at 493 nm and 577 nm [301], suggesting that a porphyrin is the photo-
receptor in this case. Photoinduction also occurs in Flavobacterium de-
hydrogenans [292, 297] and Fusarium aquaeductuum [306].
The obligatory requirement for oxygen has been well-documented in all
organisms examined. In the case of a Mycobacterium species it is thought that
oxygen participates directly in producing a compound (photooxidative pro-
duct) which then acts as inducer of the enzyme multiplex concerned with
carotenoid synthesis. The failure of K 3 Fe(CN) 6 to substitute for oxygen in
this reaction, although it is a satisfactory electron acceptor for this micro-
organism held under anaerobic conditions [300], supports the view that 0 2
acts directly. Photoinduction of carotenogenesis is also temperature-sensitive
[292, 297' 307].
After photoinduction, carotenogenesis is prevented by the inhibitors of
enzyme synthesis, chloramphenicol and actidione, which supports the view that
enzyme synthesis is involved [297, 304, 306]. However, in the case of the fungi
F. aquaeductuum and N. crassa it is thought that oxygen is required to maintain
the photoreceptor in the appropriate oxidation state [307]. In F. aquaeductuum
carotenogenesis is stimulated in the dark by addition of p-chloromercuri-
benzoate to the culture medium [308, 309]. However, the locus of this effect
is different from that of light, because in a mutant in which photoregulation
has been deleted and which makes large amounts of pigments in the dark,
addition of p-chloromercuribenzoate stimulates even greater synthesis in the
dark [310]. Incubation with buffered H 2 0 2 solution (10- 1-10- 2 M) in the
dark also stimulates photoinduction [310].

0. Biosynthesis of Carotenoids in Photosynthetic Bacteria

1. Introduction
The characteristics of the carotenoids of photosynthetic bacteria are (i) with
one or two notable exceptions, they are acyclic; (ii) they contain tertiary
hydroxyl and methoxyl groups at C-1; (iii) they contain double bonds at C-3,4
(e.g. spirilloxanthin, 108). Other less widespread characteristics are the appear-
ance of keto groups in conjugation with the polyene system (e.g. spheroide-
 VII. Biosynthesis 621

'-":::, '-":::, '-":::, '-":::, '-":::, '-":::, '-":::, '-":::, '-":::, '-":::, '-":::,
Lycopene (19)

1 +H 0 2

'-":::, '-":::, '-":::, '-":::, '-":::, '-":::, '-":::, '-":::, '-":::, '-":::, '-":::,
Rhodopin (56)

l-2H

'-":::, '-":::, '-":::, '-":::, '-":::, '-":::, '-":::, '-":::, '-":::, '-":::, '-":::,
3,4-Didehydrorhodopin (SS)

1 +CH 3

'-":::, '-":::, '-":::, '-":::, '-":::, '-":::, '-":::, '-":::, '-":::, '-":::, '-":::,
Anhydrorhodovibrin (97)

l +H 0 2

'-":::, '-":::, '-":::, '-":::, '-":::, '-":::, '-":::, '-":::, '-":::, '-":::, '-":::,
Rhodovibrin (106)

1-2H

'-":::, '-":::, '-":::, '-":::, '-":::, '-":::, '-":::, '-":::, '-":::, '-":::, '-":::, '-":::, '-":::,
Monodemethylated spirilloxanthin (lOS) OH
1 +CH 3

OCH 3

'-":::, '-":::, '-":::, '-":::, '-":::, '-":::, '-":::, '-":::, '-":::, '-":::, '-":::, '-":::, '-":::,
Spirilloxanthin (108)
CH 0
3

Scheme 34. The probable main pathway from lycopene (19) to spirilloxanthin (108) in Rhodospirillum
rubrum
 622 T. W. GOODWIN

none, 182), the saturation of the terminal 1,2-double bond (e.g. 1,2-dihydro-
lycopene, 21) and the appearance of an aromatic ring (e.g. chlorobactene, 15).

2. General pattern of biosynthesis in the Athiorhodaceae

The inhibition of spirilloxanthin synthesis in Rsp. rubrum by diphenyl-
amine [311, 312] first suggested that this pigment was formed by sequential
desaturation of phytoene (32). Kinetic studies suggested that the pathway
observed in higher plants viz, phytoene--+ phytofluene (30)--+ '-carotene (26)--+

"-':::: "-':::: "-':::: "-':::: "-':::: "-'::::

Phytoene (32)

j -2H
"-':::: "-':::: "-':::: "-':::: "-':::: "-':::: "-'::::
Phytofluene (30)

l-2H

"-':::: "-':::: "-':::: "-':::: "-':::: "-':::: "-':::: "-':::: "-':::: "-'::::
7,8,11,12-Tetrahydrolycopene (25)

-2H
l

Neurosporene (22)

j -2H
"-':::: "-':::: "-':::: "-':::: "-':::: "-':::: "-':::: "-':::: "-':::: "-':::: "-':::: "-'::::

Lycopene (19)

Scheme 35. The pathway from phytoene (3'2) to lycopene (19) in photosynthetic bacteria [317]
 VII. Biosynthesis 623

neurosporene (22)-+ lycopene (19) was the first stage, and this was followed
by the steps unique to photosynthetic bacteria viz, dehydrogenation at C-3,4,
hydration at C-1,2 and methoxylation (Scheme 34) [318]. The accumulation
of massive amounts ofphytoene in systems inhibited by DPA raised problems
of interpretation in early experiments, but it is reasonable to assume that the
accumulation is due to removal of feed-back control when synthesis of the
end product (spirilloxanthin) is inhibited. The intermediation of lycopene was
suggested earlier by its appearance in very young cultures of Rsp. rubrum [313,
314]. The methyl groups arise conventionally as Ccunits [315], presumably
via S-adenosylmethionine [316]. It is now clear however that the pre-lycopene
pathway is not that observed in higher plants because (-carotene is not present
in Rsp. rubrum; it is replaced by 7,8,11,12-tetrahydrolycopene (25) [317]. The
probable pathway to lycopene in most photosynthetic bacteria is thus
indicated in Scheme 35.
Spheroidene (99) is the major pigment of Rhodopseudomonas spheroides
when it is grown under anaerobic conditions [318-320], and kinetic studies
indicated a pathway from neurosporene outlined in Scheme 36 [321-323].

Scheme 36. Originally postulated pathway for biosynthesis of spheroidene (99) in

Rhodopseudomonas spheroides [323]
 624 T. W. GOODWIN

"""' """' """' """' """' """' """' """'

7,8,11,12-Tetrahydrolycopene (25)

j +H 2 0

OH

"""' """' """' """' """' """' """'

1·Hydroxy-1,2,7',8',11',12'-hexahydrolycopene (59)

l +CH 3

CH3

"""' """' """' """' """' """' """'

3,4,11',12' · Tetrahydrospheroidene (102)

l-2H

OCH 3

"""' """' """' """' """' """' """' """'

11',12'-Dihydrospheroidene (101)

l-2H

CH 3

"""' """' """' """' """' """' """' """' """' """'
Spheroidene (99)

Scheme 37. Probable pathway for biosynthesis of spheroidene (99) in

Rhodopseudomonas spheroides [325]

However, as later investigations revealed the presence of 11',12'-dihydro-

spheroidene (101) in Rps. spheroides [324], the more probable pathway is that
indicated in Scheme 37 [325]. This pathway does not involve 'demethylated
spheroidene ', a compound not yet detected in Rps. spheroides.
The co-occurrence of spheroidene and spirilloxanthin in Rhodopseudo-
monas gelatinosa, together with kinetic studies, indicated an alternative path-
way to spirilloxanthin in this organism (Scheme 38) [323]. The discovery of
 VII. Biosynthesis 625

~ ~ ~ ~ ~ ~ ~ ~ ~ ~
Neurosporene (22)

j (ef. Seheme 36)

OCH 3

~ ~ ~ ~ ~ ~ ~ ~ ~ ~
Spheroidene (99)

1
OCH 3

~ ~ ~ ~ ~ ~ ~ ~ ~ ~
I' -Hydroxy-l ',2' -dihydrospheroidene (107)
OH

OCH 3
1
~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~
Rhodovibrin (106)
OH

OCH 3
l
~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~
Monodemethylated spirilloxanthin (105)
OH

OCH 3
1
~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~
Spirilloxanthin (108)
CH 3 0
Scheme 38. Alternative pathway for spirilloxanthin biosynthesis in
Rhodopseudomonas gelatinosa [323]

spheroidene (99) and 1'-hydroxy-1',2'-dihydrospheroidene (107) in diphenyl-

amine-inhibited cultures of Rsp. rubrum [324, 325] suggested that this pathway
was also functioning alongside the normal pathway in this organism [325].
However, detailed investigation of the carotenoids in diphenylamine-inhibited

Carotenmds 40
626 T. w. GooDWIN

cultures of Rsp. rubrum has revealed the presence of hydroxy and methoxy
derivatives of phytoene (61, 104), phytofluene (60, 103) and 7,8,11,12-tetra-
hydrolycopene (59, 102) [325-327]. This means that under the metabolic
restraint imposed by diphenylamine, the enzymes responsible for 1,2-hydration
and 0-methylation will utilize the partly saturated polyenes as substrates. Thus
in Rsp. rubrum studies with diphenylamine can be used only with considerable
caution to define biosynthetic pathways.
A number of Athiorhodaceae cannot complete the biosynthetic pathway
to spirilloxanthin. Rsp. molischianum produces mainly lycopene and rhodopin
(56) [328], which means that although it can hydrate the terminal double
bond, it cannot desaturate at C-3 or methoxylate at C-1. A similar pattern
of pigment distribution was inferred [328] from the gross absorption spectrum
of Rsp.fulvum [329], and this was later demonstrated experimentally [330];
traces of 1,1'-dihydroxy-1,2,1',2'-tetrahydrolycopene (81) were also found. Bio-
synthesis in Rhodopseudomonas viridis appeared to stop at neurosporene (22)
and lycopene (19) [331], but these are normally minor components, the main
pigments being 1,2-dihydrolycopene (21) and 1,2-dihydroneurosporene (23)
[332]; the corresponding more saturated polyenes 1,2-dihydrophytoene (33),
1,2-dihydrophytofluene (31) and 7,8,11,12-tetrahydrolycopene (25) are also
present. However, (-carotene is present but not its dihydro derivative; (-caro-
tene is not widely distributed in photosynthetic bacteria. The 1,2,1',2'-tetra-
hydro derivatives of neurosporene (28) and lycopene (24) have also been
detected [333]. Thus Rps. viridis has the unique ability to saturate the terminal
double bond of carotenoids, and this can occur as far back as phytoene. The
mechanism of hydrogenation is unknown, but may involve an enzyme inter-
mediate similar to that invoked in cyclization, with the final addition of H 6 ,
possibly from NADPH (Scheme 39).

H~_ ...

Scheme 39. Possible mechanism for the synthesis of 1,2-dihydro derivatives in

Rhodopseudomonas viridis

3. Formation in the Athiorhodaceae

A thorough survey of the carotenoids in a number of Athiorhodoceae has
shown that they can be divided into 3 main groups, those synthesizing (i) the
spirilloxanthin (108) type, (ii) the okenone (181) type and (iii) cross-conjugated
carotenals and carotenols, e.g. rhodopinal (144) [334-337] (see Chapter III
for full details). The biosynthetic problems concerned with spirilloxanthin
formation have just been discussed; in the case of the okenone series the major
unsolved problem is the mechanism of formation of the aromatic ring; nothing
is known experimentally but proposals have been made (Scheme 40). The
mechanism of formation of a keto group in strict anaerobes is also a challeng-
 VII. Biosynthesis 627

Scheme 40. Possible mechanism for synthesis of carotenoids with aromatic rings [294]

ing problem. The cross conjugated carotenals and carotenols represent a fur-
ther biosynthetic elaboration on the basic pattern of synthesis in the photo-
synthetic bacteria, and the elucidation of the mechanism by which oxidation
of an 'in-chain' methyl also involves isomerization of the adjacent double
bond from the trans to the cis configuration is awaited with interest.

4. Rhodomicrobium vannielii
Rhodomicrobium vannielii is unique in synthesizing not only spirilloxanthin
(108) but {J-carotene (3) [338-342], and recently a hybrid compound, 1'-
methoxy-3',4'-didehydro-1',2'-dihydro-y-carotene (96a), with one cyclic end
and one acyclic, 'spirilloxanthin-like', end has been isolated [342]. Nothing
is known of the pathway of formation of these pigments; that useful tool in
studies on carotenogenesis, diphenylamine, has no significant effect on Rps.
viridis.

5. Chlorobacteriaceae
Some members of the Chlorobacteriaceae synthesize carotenoids with an
aromatic ring, e.g. chlorobactene (15) from Chlorobium species [343, 344].
The mechanism involved is not known but a possibility has already been
indicated in Scheme 40.

6. Formation of xanthophylls in photosynthetic bacteria

The characteristic oxygen functions are methoxyl (at C-1) (e.g. spirilla-
xanthin, 108), ketone at the end of a polyene chain (e.g. spheroidenone, 182),
and ald~hyde by oxidation of an 'in-chain' methyl (e.g. rhodopinal, 144).
The insertiop of the hydroxyl group at C-1 appears to involve a hydration
across the 1,2-double bond, but no evidence exists for this conclusion; this
mechanism could clearly occur in strict anaerobes such as Chromatium spe-
cies. However, die insertion of oxygen as a keto group is an aerobic process
in Rps. spheroides. Cultured under anaerobic conditions this bacterium accu-
mulates mainly spheroidene (99); on allowing oxygen into the culture this
628 T. W. GOODWIN

pigment is rapidly converted into spheroidenone (182), in which the oxygen

arises from 0 2 ; the same change occurs in Rps. gelatinosa. In Rps. spheroides
'hydroxyspheroidene' (107) and spirilloxanthin (108) are similarly converted
into 'hydroxyspheroidenone' (185) and 2,2'-diketospirilloxanthin (208), respec-
tively. Rps. capsulata probably behaves in the same way. In spite of the rapidity
of these conversions in vivo, no convincing evidence for an in vitro reaction
has yet been presented. The keto group in okenone (181) is intriguing in that
it cannot arise from 0 2 because Chromatium okenii is a strict anaerobe;
a possible mechanism (Scheme 41) has been suggested. Nothing is known of
2H

·~ . ·--r
H 0
.··~··· L
2
OH
Scheme 41. Possible mechanism for insertion of a keto group into carotenoids in strict anaerobes

the oxidation of the 'in-chain' methyl to an aldehyde group in warmingone

(144), but again it must occur under anaerobic conditions.
A further interesting observation is the occurrence of P-cryptoxanthin in
Rm. vannielii cultured under anaerobic conditions [342]. Such an hydroxyla-
tion in higher plants and microorganisms requires 0 2 and is probably mediated
by a mixed-function oxidase.

P. Formation of Keto Carotenoids in Crustacea

Characteristic carotenoids of marine invertebrates are the keto carotenoids

echinenone (148), canthaxanthin (193) and astaxanthin (203) (see Chapter II).
Recently interest has focussed on the formation of these pigments in Crustacea
because they are clearly not provided in the diet.
In the Californian strain of the anostracan branchiopod Artemia salina,
P-carotene is converted into echinenone and canthaxanthin, and kinetic stu-
dies indicate that, as would be expected, echinenone is intermediate between
P-carotene and canthaxanthin [345-347] (Scheme 42). The recovery of
[ 14 C]echinenone and [ 14 C]canthaxanthin with essentially the same specific
e
activity as the 4 c]p-carotene administered, eliminates the possibility that
the P-carotene is first broken down to small molecules which are then hie-
synthesized into the keto carotenoids [347-348].

c;=·········~
0 0
P-Carotene Echinenone Canthaxanthin

Scheme 42. The probable transformation of P-carotene into canthaxanthin by Artemia

 VII. Biosynthesis 629

Obvious intermediates in the pathway are isocryptoxanthin (40), 4' -hydroxy-

{J-carotene-4-one (155) and isozeaxanthin (71). It would appear that they are
not involved in the pathway in A. salina. Neither isocryptoxanthin nor iso-
e
zeaxanthin can be detected after administration of 4 C]{J-carotene [346], and
dietary isozeaxanthin although absorbed is metabolically inert [346]. Ad-
ministered isocryptoxanthin is, on the other hand, converted into 4' -hydroxy-
{J-carotene-4-one but not into echinenone [346].
Although 4-hydroxy components are ruled out as intermediates in keto-
carotene biosynthesis in A. salina, they may well be involved in other Crusta-
cea. Isocryptoxanthin and zeaxanthin are converted into echinenone and
canthaxanthin, respectively, by the cladoceran branchiopod Daphnia magna
[349]; isocryptoxanthin is present in the anostracan branchiopods Branchipus
stt;tgnalis and Chirocephalus diaphanus, and isozeaxanthin is found in C. dia-
phanus and Tanymastix lacunae [350].

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 637

VIII. Metabolism
H. THOMMEN
Department of Vitamin and Nutritional Research,
F. Hoffmann-La Roche & Co. Ltd., Basle, Switzerland

A. Introduction . . . . . . . . . . . . . . . . . . . . 638
B. Degradation of P-Carotene: Glover-Redfeam Hypothesis 639
C. Conversion of P-Carotene to Keto Carotenoids . 641
D. Carotenoproteins . . . . . . . . . . . . . . . . . 656
638 H. THOMMEN

A. Introduction

Carotenoids undergo many metabolic reactions in plant and animal

organisms. There are, on the one hand, processes leading to total breakdown
of the molecule and, on the other, reactions that do not cleave the original
molecule but merely modify it chemically, for example by hydroxylation or
introduction of a ketonic function. Two such reactions are selected for a
summarizing discussion on the basis of numerous publications.
Glover and Redfearn postulated a degradation scheme for P-carotene (3)
in which it is broken down to vitamin A via aldehydes and carboxylic acids
[1, 2] (Scheme 1, a). The proposed reaction mechanisms correspond with the
oxidative processes known to occur with unsaturated and branched chain
fatty acids.

R=CH 2 0H: Retinol

R=C0 2 H: Retinoic acid

1
(a) Glover-Redfearn scheme
or (b) Central cleavage

P-Carotene (3)

j (c) Hydroxylation of the rings

and (d) Further oxidation of the rings

OH

Astaxanthin (203)

Scheme 1. Oxidations of P-carotene (3)

Oxidation at both rings of P-carotene (3) is a further reaction that has

been investigated in many invertebrates and also in birds especially [3-8]
(Scheme 1, c/d).
 VIII. Metabolism 639

B. Degradation of /1-Carotene: Glover-Redfearn Hypothesis

The fundamental scheme of Glover and Redfearn postulated a biological

degradation of P-carotene (3) to retinal or retinol and retinoic acid (Scheme 2).

14' 12' 10' 8'

"""" """" """" ~ """" """" """" ~

j End-position
oxidation
(Unknown intermediates)

P-Apo-8' -carotenal (248) + P-cyclocitral

j ,. P-Apo-8'-carotenoic acid (250)

I

P-Apo-10'-carotenal (256) ""

Central
cleavage
j K
t
..- P-Apo-10'-carotenoic acid (260)
I

P-Apo-12'-carotenal (262)

1 ~
/
..-- P-Apo-12' -carotenoic acid (263a)
I

P-Apo-14' -carotenal

j -
t
P-Apo-14'-carotenoic acid

Retinal

Retinol Retinoic acid

(Vitamin A)
Scheme 2. Glover-Redfearn degradation scheme

An oxidative mechanism beginning at one end of the polyene chain degrades

it through the homologous carotenals or carotenoic acids to the C 20 -compound.
The widespread distribution of the apo-carotenals in plants [9-15] supports
this view. It must be pointed out, however, that in plants it has only been
possible to isolate the apo-carotenals, but not the corresponding acids; the
former occur richly in the Gramineae and Papilionaceae particularly. In
 640 H. THOMMEN

animal cells, however, a quantitative conversion of an apo-carotenal to the

corresponding apo-carotenoic acid is indeed observed [2], but not any further
degradation. It has not yet been ascertained in what proportions P-carotene (3)
is degraded to vitamin A (retinol) through the apo-carotenals (p-oxidation)
and through central cleavage. Some clarification of the latter process is afforded
by the investigations of Goodman and Huang [17] and of Olson and Hayaishi
[18]. A P-carotene dioxygenase obtained from cell-free preparations of rat
duodenum, rat jejunum and rat liver was found to cleave P-carotene (3) to
vitamin A aldehyde (retinal). The ability of P-carotene dioxygenase to function
in the degradation according to Glover and Redfearn was shown experimentally
by submitting vinylogous apo-carotenals to the action of this enzyme. The
shorter the side chain, the faster the enzymic fission, e.g. it is eleven times
faster for P-apo-10'-carotenal (256) than for P-carotene (3) (Table 1). It is
suggested that, before oxidation, P-carotene and the apo-carotenals are
transformed to a carotene-lipoprotein complex [19]. In these experiments
vitamin A aldehyde was always the only end-product observed.
The biosynthetic formation of apo-carotenals and their corresponding
acids in the living cell cannot be absolutely excluded because definitive
experiment on this issue is lacking. Such a formation, however, is contra-
indicated in many findings from studies on the enzymic reactions in the

Table 1. Substrate specificity of the carotene cleavage enzyme (from Olson [123])

Substrate Specific Relative

activity activity

0.96 10.7

0.91 10.2

0.22 2.5

2.0
c4o

(142)

0.09 1.0
c4o
P-Carotene (3)
 VIII. Metabolism 641

biosynthesis of carotenes and carotenoids [20, 21]. Starting from mevalonic

acid, lycopene (19) is built up, with isopentenyl, geranyl and farnesyl pyro-
phosphates shown as intermediates. In further enzymic steps lycopene can be
cyclized. Normally the C 40 -skeleton is built up, and is then degraded enzymic-
ally to C 20 -fragments or smaller molecules.
To support the hypothesis of Glover and Redfearn, models have been
sought which offer the possibility of obtaining and identifying the various
degradation products. Of considerable interest in this connection, apart from
P-carotene (3), were the different apo-carotenals which have become available
in large quantities in recent years through the development of suitable syntheses
[22, 23]. Certain apo-carotenals as well as provitamins A are capable, after
oral administration to animals, of being stored, especially in the liver and
body-fat [24-31]. It was an obvious choice to use the growing chick or laying
hen as experimental animal for a study of the fate of P-carotene (3) and its
postulated degradation products. The growing chick was chosen above all for
those experiments in which the vitamin A activity was determined. The laying
hen appeared suitable since, after absorption of apo-carotenals, these or their
metabolites could be found in the egg yolk. Confining attention in such experi-
ments to the apo-carotenals, it is established that, after oral administration
of members of the homologous series, only P-apo-8'-carotenal (248) is retained
in the egg yolk in large amount, actually in the form of P-apo-8'-carotenoic
acid (250). This fact accounts for the application of the C 30 -aldehyde as a feed
additive for increasing the pigmentation of egg yolk.
Xanthophylls, certain keto carotenoids and other isolated examples of
the carotenoids are also capable of intensifying egg-yolk colouring. Since these
compounds are not included in the Glover-Redfearn degradation scheme
they are not discussed here.
The degradation scheme of Glover and Redfearn is regarded as entirely
acceptable, particularly in view of their demonstration of the formation
in vivo of P-apo-12'-carotenal (262) and the corresponding acid (263a) after
administration of 14C-labelled P-carotene to rats [2].

C. Conversion of JI-Carotene to Keto Carotenoids

In the animal kingdom carotenoids are the most widely occurring pigments
after the melanins. It is therefore not by chance that at the outset of carotenoid
research materials were sought from which the yellow to violet lipochromes
could b~ isolated. Many of these pigments occur, above all, in the plumage
of birds, and it was obviously possible, after the development of suitable methods
of isolation, to obtain and characterize such pigments. The occurrence of
carotenoids in widely different species of birds is described in numerous
publications especially those of VOlker and of Fox.
An early significant finding of general validity was the fact that the carot-
enoids of plumage have structures containing one or more hydroxy or keto

Carotenoids 41
642 H. THOMMEN

groups. The question arises as to whether the animal is capable of synthesizing

such compounds itself or whether its food source is solely responsible for the
feather pigmentation. Biosynthesis of carotenoids in the organism cannot be
excluded theoretically, but there are no examples of relevant experiments
with radioactively labelled precursors. However, on the basis of many observa-
tions made largely in zoological gardens, it may be regarded as almost excluded
that the bird itself can synthesize its pigments. Thus, if carotenoids are missing
from the diet, a definite fading of the plumage occurs already after the first
moult, while an increased dietary supplement of carotenoids gives rise to a
deeper colouring of the plumage and a significantly higher content of pigment
isolated from the feathers.
In the working up of feathered skins of the African oriole (Oriolus auratus)
a pigment could be isolated, identical with lutein (73), but only traces of P-
carotene (3) were present. This gives rise to the specific question whether the
African oriole receives no P-carotene but only lutein in its food, or whetnsr
P-carotene is transformed into lutein in the organism. It is known that tropical
orioles are defmitely insect feeders and that where carotenoids occur in insect
pigments, P-carotene is also found occasionally. It was then found in experi-
mental feeding that the African oriole requires dietary lutein quite specifically
and that although P-carotene is ingested, it cannot be absorbed. A conversion
of P-carotene to lutein is improbable since the transformation of the P- to the
tX-carotene (5) structure can be ruled out. Nevertheless, the plumage of the
canary contains a canary xanthophyll which is not a constituent of the admin-
istered food and is ascribed the basic structure of an tX-carotene. Canary
xanthophyll can be replaced by zeaxanthin (67).
In a number of tropical ornamented birds (Cotingidae) and fruit pigeons
(Megaloprepia and Ptilinopus) rhodoxanthin (209) occurs as feather pigment.
The fruit pigeons are mainly inhabitants of the Indonesian and Pacific archi-
pelagos. It would be an exercise full of interest to study the feeding habits of
these creatures, for such investigations would doubtless lead to significant
extensions of the natural occurrence of specific carotenoids. Rhodoxanthin
(209), originally isolated from the aril of the yew fruit (Taxus baccata), is
incorporated in the horny substance of the feathers resulting in colour tones
of deep violet and even blue-black.
Astaxanthin (203) was isolated in esterified form from the feathers of the
South African gonolek (red-bellied shrike, Laniarius atrococcineus). Astaxan-
thin is also present in the papillae surrounding the eye of the game pheasant
(Phasianus colchicus). It is known that the pheasant's food consists largely
of corn grain so that, in addition to P-carotene (3) and some trace carotenoids,
zeaxanthin (67) is mainly consumed. Zeaxanthin is transformed by the bird
into astaxanthin (203) (Scheme 3).
In our own experiments [32] on male and female pheasants (Phasianus
colchicus) we were able to establish that the papillae contain esterified asta-
xanthin ·(203). As ester moieties Cw- to C18 -carboxylic acids such as oleic,
linoleic and linolenic acid could be demonstrated. Surprisingly, the livers of
 VIII. Metabolism 643

:(("
(><Yx (67)
HOA)l

l
:(("
/
~OH :(("
~ (167)
HOY

HOY
~
~X (173)

l
~OH
~X
HOY
(203)

Scheme 3. Possible metabolic pathway of zeaxanthin (67) in the pheasant (Phasianus colchicus)
644 H. THOMMEN

the investigated animals which, without exception, were of game reserve stock
contained only traces of carotenoids in which no keto carotenoids whatever
could be found. All were well-fed specimens. It can be concluded from this
that, in contrast to other 'carotenoid-dependent' birds, the pheasant is unable
to store carotenoids in the liver as depot pigment. The conversion of zea-
xanthin (67) to astaxanthin (203) could therefore only take place in the papillae.
The intermediate compounds postulated in Scheme 3 could not be demon-
strated. For the present it remains speculative whether astaxanthin (203) is
formed through the corresponding tetrahydroxy compound crustaxanthin (93),
or partly through keto compounds as, for example, adonixanthin (167). Since
P-carotene (3) and zeaxanthin (67) are abundantly available in the bird's food,
the formation of astaxanthin from these two compounds may be regarded as
probable.
Normally plumage is not pigmented in a regular manner, and there is still
no explanation of the fact that, besides physical colour effects, colour differenCes
are brought about by differences not only in the concentration but also in the
composition of the plumage carotenoids.
A further carotenoid, canthaxanthin (193), isolated in 1950 by Haxo from
the chanterelle (Cantharellus cinnabarinus), soon gained from ;;til the studies
of feathers a key position. Canthaxanthin could be isolated from the cardinal-
grosbeak (Cardinalis cardinalis), the quetzal (or resplendent trogon) (Pharo-
machrus mocinno), the roseate spoonbill (Ajaia ajaja), the cock-of-the-rock
(Rupicola rupicola and Rupicola peruviana), the scarlet ibis (Eudocimus ruber,
Guara rubra) and above all from the flamingo (Phoenicopterus ruber, Phoeni-
coparrus jamesi and Phoeniconaias minor). [See colour-plates.]
In these bird species there were abundant supplies of material for study so
that tarsal skin, fat tissue and organs could be closely examined in addition to
the feathers. Surprisingly high levels of canthaxanthin (193) were found in the
livers of these birds. It may therefore be concluded that this liver depot is
responsible for the occasional intensive colouring of the plumage. Feather
colouring can be influenced as desired by supplementing the feed with synthetic
canthaxanthin; this finds practical application in some zoological gardens and
with bird breeders. A further step towards the clarification of the origin of
such carotenoids, in particular of canthaxanthin, was the investigation of the
carotenoid composition of the usual food. Not surprisingly, the muddy food
of flamingos, for example, in which there are large numbers of living and dead
animalcules, contained echinenone (148), lutein (73) and P-carotene (3) in
addition to canthaxanthin (193). In this connection mention may be made
especially of Copepoda, which are consumed in large quantities in the food
of the flamingo in its natural habitat. Numerous feeding experiments have
shown that these small creatures take up P-carotene from algae. The conclusion
may therefore be drawn that the possibility of oxidative processes in the bird
is not excluded a priori and that, through the Copepoda, algae are responsible
for the presence of canthaxanthin. For other birds in which canthaxanthin has
been found, similar relationships'may be assumed.
 VIII. Metabolism 645

The assertion that algae, as food source of fishes and Crustacea, only
provide P-carotene (3) or simple hydroxylated carotenoids must be revised
on the basis of more recent studies. In particular, secondary carotenoids, for
the most part containing keto groups, have been found in green algae (Chloro-
phyceae). Even though extreme conditions as, for example, nitrogen deficiency
are assumed for the formation of keto carotenoids in green algae, the possible
plant origin of this type of compound must not be excluded. Czygan [33-36]
especially has been concerned with such biosynthetic processes. Chiarella,
under nitrogen deficiency can, for example, synthesize so-called secondary
carotenoids, namely keto derivatives of P-carotene. The crustacean, Artemia
salina, was fed with Chiarella and subsequently the carotenoid composition
determined. A. salina serves as food for many fish and birds and can therefore
be regarded as a model organism. An interesting detail worthy of mention in
this connection is the fact that the formation of astaxanthin (203) is strain-
dependent. Scheme 4 presents the possible stages of the biosynthesis in Chiarella
and A. salina based on proposals of Czygan. In routine examination of a strain
of A. salina from the South of France, Thommen et al. [37] demonstrated the
presence of astaxanthin (203) in addition to echinenone (148) and cantha-
xanthin (193). The reaction scheme proposed by Czygan [38] is extended in
the sense that a hydroxylation essential for an appropriate stage need not pro-
ceed to completion before keto groups are formed. Thus, conversion of 4,4'-
dihydroxy-P-carotene (71) to 4-oxo-4'-hydroxy-p-carotene (155) or the hydrox-
ylation of echinenone (148) to 4-oxo-4'-hydroxy-p-carotene (155) is quite
conceivable.
From H aematacaccus pluvialis Flotow em. Wille Czygan [39] isolated
crustaxanthin (93) and also phoenicopterone (4-oxo-a-carotene) (149), the
latter a pigment found in the liver of the red flamingo, Phaenicapterus ruber
[40]. This was the first finding of a keto-oc-carotene in plant material, so that
consumption in food of carotenoids with a-carotene structure also appeared
possible. Speculation about the transformation from the P- to the a-carotene
structure was thereby rendered superfluous.
Dentice di Accadia et al. [41, 42] succeeded similarly in the isolation of
echinenone (148) and canthaxanthin (193) from another green alga, Dictyacac-
cus cinnabarinus. A further green alga, Pratasiphan batryaides·(Ktitz.) Klebs,
widely distributed on wet ground and at the edges of stagnant waters was
closely examined by Kleinig and Czygan [ 43]. This alga is also capable of
forming secondary carotenoids, but here too, nitrogen deficiency is required.
In addition to the already known secondary carotenoids, 4-oxo-4'-hydroxy-
(155) an'd 3,4-dioxo-4'-hydroxy-p-carotene (168) were isolated for the first
time. The hydr.oxy compounds are esterified with different combinations of
fatty acids.
The fact that the formation of secondary carotenoids runs parallel with the
degradation of chlorophyll in algae leads to the question whether chlorophyll
degradation products are starting materials for the biosynthesis of secondary
carotenoids and whether in some circumstances the P-carotene (3) present
646 H. THOMMEN

0::0 (3] ~ Q::0 (4()) ~ Q::0 (148)

OH l 0 l
c;(~ (71) ~ Q::Q (193)

H l:Q 0

:Q
l 0

Nx ~X
HOY
(85) ---+ (196)

l
HOY

OH 0 l

HOY
Nx
:(1" (93)

HOY
---+
:Ct"
Nx (203)

OH
i:GrOH
Nx (198)

HOY
0

Scheme 4. Possible pathway for the b\osynthesis of keto carotenoids in Chiarella and the
crustacean Artemia salina
 VIII. Metabolism 647

need not be oxidized at all. Keto carotenoids have been found, not only in
green algae but also in Cyanophyceae [ 44- 48]. Francis et al. [ 49] give the
carotenoid composition of Oscillatoria limosa shown in Table 2. Surprising

Table 2. Carotenoid composition of Osci/latoria limosa

Compound Percentage of
total carotenoid

P-Carotene (3) 17
Cryptoxanthin (39) -1
Echinenone (148) 23
Canthaxanthin (193) 7
Zeaxanthin-like 20
Myxol 2'-(0-methyl-5-C-methylpentoside) (91) 25
4-Ketomyxol 2' -(5-C-methylpentoside) (176) -1
Oscillol 2,2' -bis(O-methyl-5-C-methylpentoside) (96) 8

features are the high proportion of echinenone (148), the relatively small amount
of canthaxanthin (193) and the high proportions of a zeaxanthin-like carotenoid
and of the carotenoid glycoside (91); however, the latter compound contains
only one intact ring system.
These few examples from the class of Thallophyta should simply illustrate
that the presence of keto carotenoids, which occur principally as plumage
pigments, is not confined to animal organisms and that, above all, plants are
capable, albeit under rather extreme conditions, of introducing keto groups
into the ring system of P-carotene (3).
Before discussion of the Crustacea, which provide in their abundant variety,
especially of the small species, an inexhaustible source of food for higher
animals, the Echinodermata are worthy of mention. Their occurrence is more
limited, but because of their high pigment content and their possible considera-
tion as food, they ought not be ignored.
Fox and Hopkins [50] studied in detail the metabolism ofketo carotenoids
in echinoderms. They first attempted to explain certain differences in the
degradation and the bio-oxidation of P-carotene (3) through the fundamentally
different feeding habits of the animals. There followed first a classification into
an 'ambivalent' group which can take up carotenes and xanthophylls, and a
'xanthophyll' group which only absorb hydroxylated carotenoids [51, 52].
The first group which, as was originally assumed, 'is related to a herbivorous
diet', comprises Holothurioidea and Echinoidea; the second group, 'which is
related to a carnivorous diet', comprises Asteroidea and Ophiuroidea. Later
studies show, on the contrary, that this kind of simple classification is no
longer tenable [53-55]. One is now inclined to the view that, starting with
P-carotene (3), two principal reaction paths are possible in Echinodermata
(Scheme 5).
648 H. THOMMEN

/
uc ))
(3)

li )) ('<(
;Q
HO (39)
~
0
(148)

~ /
))
~X
HOY
(153)

1
~H
~X
HOY
(167)

1
~OH
~X
HOY
(203)

Scheme 5. Possible pathway of P-carotene (3) oxidation in Echinodermata

 VIII. Metabolism 649

In many cases the food of 'ketocarotenoid-dependent' birds consists

mainly of micro-crustaceans. Artemia salina is found in the natural habitat of
flamingos. Gilchrist and Green [56] found an abundance of esterified asta-
xanthin (203) in adult algae-fed Artemia salina. They showed that {3-carotene (3)
from algae is a precursor of astaxanthin (203). Hata and Hata [57], Hsu et al.
[58], Davies et al. [59] and Czygan [60], on the other hand, demonstrated
that {3-carotene (3) is converted mainly into echinenone (148) and cantha-
xanthin (193). The difference in the strains of Artemia salina, the stage of growth
and the sources of their food may account for the disagreement between these
results. Other micro-crustaceans of importance as a food source for flamingos
are Daphnia species [61, 62] and Cyclops species [63].
In extensive investigations Fox et al. have studied the composition of
carotenoids in flamingos [64]. After the occurrence of astaxanthin (203) (also
found in the flamingo by Volker [65-68]) had previously become a focal point
of interest, other keto carotenoids assumed growing significance in later
studies, above all as intermediate compounds. Because of its easy maintenance
in zoological gardens, the flamingo attracted the early interest of scientists. Since
the bird's colouring is typical for each species in the wild state and the plumage
pigment is often readily lost ·in captivity, it was natural to study the feeding
habits of this animal. Very soon the significance of the presence of carotenoids
in the food was recognized [69-79] and the possibility indicated how the bird
could transform, for example, {3-carotene (3) or zeaxanthin (67) into related
keto compounds. From the outset there were two opposing views on the
origin of the keto carotenoids. The main point of the first hypothesis is that
the flamingo is capable of transforming {3-carotene in its organism to keto
compounds. Opposed to this is the view that all carotenoids found in the bird
are ingested with the food. The most recent investigations of the feeding mud
from the shore area of Lake Nakuru in Kenya [80] appear rather to confirm
the hypothesis that the carotenoids found in the flamingo come primarily
from the food source. For the present it is not possible to decide to what
extent certain observed traces of carotenoids are artifacts or are formed through
bacterial action. However, as long as a conversion of {3-carotene (3) to asta-
xanthin (203) in the organism cannot be definitely excluded, the first hypothesis
cannot be rejected. The varied composition of the flamingo's natural food and,
above all, the considerable ecological differences perhaps explain why different
authors find different carotenoid compositions in flamingos and other carot-
enoid-dependent birds [81-83].
There are still extant six Phoenicopteridae species (Table 3), which have
all been examined qualitatively and quantitatively for their carotenoid compo-
sition [81, 82].
Scheme 6 depicts the intermediate compounds which come into considera-
tion for the bio-oxidation of {3-carotene (3) to astaxanthin (203). The carotenoids
found in larger amounts in the bird are underlined (broken line: trace-carot-
enoids). From this review it can be taken that in the bio-oxidations at the
ring systems astaxanthin (203) can be obtained in three different ways. Asta-
650 H. THOMMEN

Table 3. Flamingos (Phoenicopteridae)

Species Occurrence

Phoenicopterus ruber
Phoenicopterus ruber ruber
Phoenicopterus ruber roseus
Phoenicopterus ruber chilensis
Phoeniconaias minor
Phoenicoparrus andinus
Phoenicoparrus jamesi
Central America, Northern South America, Westindies
Southern Europe, Africa, Asia
Southern part of South America
Africa, Northwestern India
Andean area
Andean area

xanthin, which in the free state passes readily into astacene (198), is present
in the organs as a diester. Saturated C 6 - to C 18 -carboxylic acids and also oleic,
linoleic and linolenic acids appear as ester components.
Without definite proof to the contrary, the possible bio-transformation,)n
the bird, of [3-carotene to keto carotenoids should not be excluded, but this
may be regarded as improbable on the basis of many available experimental
findings.
Table 4 summarizes those keto carotenoids which have been identified in
the organs, tarsal skin, feathers and body fat of flamingos. The methods
consisted mainly of column- and thin layer chromatography and also to an
increasing extent, gas chromatography and mass spectroscopy. In many cases
it has been shown that the determination of partition coefficients only gives
correct values with quite pure compounds, but chemical reductions of very
small amounts of pigment are often unsuitable since mixtures and even artifacts
can arise.
In Scheme 7 are cited the carotenoids demonstrated in the scarlet ibis and
also a scheme for the probable formation of these carotenoids. a-Carotene (5)
was not shown to be present in the animal, so that the formation of 4-keto-a-
carotene (149) in the organism is not immediately explicable. A similar case is
that of 7,8-dihydro-[3-carotene, the natural occurrence of which is still not
verified. It is questionable whether a carotenoid hydrogenated in the 7,8-posi-
tion is ingested by the bird with food, in which bacteria and algae could be
involved. Similarly, the ingestion of exogenous guaraxanthin (204) is so far
not assured, since its detection in the liver has not been possible.
Thommen et al. [84] have repeated the work of Fox and Hopkins [83] on
the scarlet ibis under the same conditions, birds being flown from Venezuela to
Switzerland and examined in a fresh state. In these investigations the pre-
dominance of canthaxanthin (193) was confirmed as was the presence of P-
carotene (3), lutein (73) and astaxanthin (203). Phoenicopterone (149) and the
7,8(7',8')-hydrogenated keto compounds, on the contrary, could in no case be
definitely demonstrated although the purified crystalline fractions were exam-
ined by i.r., p.m.r. and m.s. In this case too it is indeed quite possible that such
different findings can result from the composition of the food, and in particular
the seasonal variations of carotenoid content of the plant and animal micro-
organisms in the birds' nutritio~. This is the more readily understood since
 VIII. Metabolism 651

X X
0 0
f
f
~~I-·
- ~· =R~I =R~
>Q= >Q= Qo
~ ······> ~ --+ ~
f
f
0 0 0
X X
f X

f i / i ~
f X X ..,
·-Qc =
0 0

~g
~

f
~~I 0 p
e -
:a=
=
>Q-!
"'><

Q=
f

>Q=
~

c
II X -
~ 0
~
0 0 0 E
X X X 2
"''?
i i i
""'-
._
0

q-.
=
.8
X X ~
0 0
...e

R~
<2

~~I r--,
~·
"'
...=
"'
E-<

Q= >Q
'-l:i

J:!r
~ --+
"e
...(------
~

"u
..c::
{/)

0 0
X X

i i i
Q<~' ~· Q<~l Q<~ q-· ~~

b J:!r >Q=
~ --+ ~ --+ ~ -<----··

>Q
~

0 0
X X
 Table 4. Carotenoids of lesser, Chilean and greater flamingos (from Fox et a!. [81])

Species Total Percentage of total carotenoids (with colour on column, and RF value)
carotenoids
mgj100g P-Carotene Echinenone Phoenico- Cantha- Phoenico- Astaxanthin Other
(3) (148) pterone xanthin xanthin (203) 0'\
Vl
(149) (193) (196) IV

Lesser (Phoeniconaias minor)

Feathers (wild, 1963) 53.4 68.3 15.0 14.6 2.0
(pink) (pink) (pink) (yellow;
unident.
zone 1)
Feathers (wild, 1966) 80.7 54.2 35.3 10.5
(orange: (orange: (pink:
0.453) 0.422) 0.367)
Blood (wild, 1966) 2.0 2.8 4.5 84.6 6.9 Trace 1.1
(yellow: (orange: (red-orange: (pink: (zeaxanthin)
0.727) 0.429) 0.312) 0.208; 0.136) (yellow:
0.104)
::r::
~lood (in Zoo 6 + yr) 1.75 1 18.0 80.2 0.8 Trace
(yellow: (orange: (orange: (pink: ;i
0
0.669) 0.405) 0.297; 0.264) 0.182) a::
Foot-web-skin (wild, 1966) 18.0 -5 (esters) 94.0 z~
-1 (free) (esters)

Chilean (Phoenicopterus chilensis) in Zoo 6+ yr

Feathers 27.5 12.8 60.0 17.0 10.3
(yellow: (deep orange: (orange: (pink -orange:
0.731) 0.671) 0.589) 0.555)
Blood 1.34 0.7 22.8 75.5 Trace? Trace?
(yellow: (yellow- (orange: (pale pink: (pale pink:
0.800) orange: 0.410) 0.269) 0.186)
0.531)

Greater (Phoenicopterus antiquorum) in Zoo 13/ 4 yr

Feathers 15.5 11.7 61.5 19.1 7.5
(yellow: (orange: (orange: (pink -orange:
0.695) 0.593) 0.458) 0.347)
Blood 3.16 0.4 17.2 78.0 2.4 2.0 Trace;
(pale yellow: (orange- (yellow- (pale pink: (yellow: unident.
0.862, yellow: orange: 0.181) 0.116) (yellow:
impure) 0.478) 0.290) 0.029)
 VIII. Metabolism 653

~~ ' (3)
~ ~X~ (5)

~ l
~~ 0
(148)
-- ----->-
~~
0
(149)

~ 0

~ ?
·····)>-
c;c:x~ 8

0 0
A

1 0
:?

HO
vx 0
~OH
(203)

,'
((':~ 8

f.
i

~OH
HO
~ 0
(204)

""" """ """ """ """ """/

X=_..."""
Scheme 7. Possible metabolic pathway in the scarlet ibis (Eudocimus ruber, Guara rubra)
654 H. THOMMEN

it is known that various keto carotenoids of the bird are indiscriminately

ingested and absorbed.
In addition to the oxidation products of P-carotene (3) the corresponding
derivatives of or:-carotene (5) can also be found in the flamingo. In Scheme 8

(5)

l
HO
(42)

l OH

HO
(73)

Scheme 8. Metabolism of oc-carotene (5) in flamingos

the metabolites of or:-carotene (5) are summarized, though they do not appear as
frequently as the P-carotene oxidation products. While zeinoxanthin (42) and
lutein (73) occur especially in plant materials, phoenicopterone (149) has so
far only been detected in a few bitds. Fox et al. [81] found, in the liver of the
 VIII. Metabolism 655

three-toed, yellow-legged Andean flamingo (Phoenicoparrus andinus), e-

carotene (10), doubtless ingested with food. Nothing is yet known of metabolites
of this carotene. A double-bond shift in the ring, that is, a possible conversion
from /3- to a- and e-carotene or the reverse, has also not been demonstrated
so far.
The investigations of Fox and Hopkins [83] of the scarlet ibis (Eudocimus
ruber, Guara rubra) have demonstrated canthaxanthin (193) as the predominant
carotenoid in the liver, plasma, foot-web-skin and feathers. In addition,
astaxanthin (203) and guaraxanthin (204) could also be detected in small
amounts in the feathers. Guaraxanthin is described for the first time by these
authors, suggesting that it is 7,8-dihydroastaxanthin. Guaraxanthin was not
found in the liver; instead there is an increase of 4-keto-a-carotene (149) and,
probably, 'bisdihydrocanthaxanthin'. The latter two compounds are also
reported to the present in plasma, skin and feathers.
Among the ornamented birds (Cotingidae) there is another representative
whose orange or deep red plumage indicates the presence of carotenoids. This
is the cock-of-the-rock, a tree-dweller native to South America, which feeds
mainly on fruit. The deep red species, Rupicola peruviana, inhabits the forest
areas of the Andes while the smaller orange-red Rupicola rupicola lives in the
northerly part of South America (see colour-plates). Thommen et al. [85] have
investigated the plumage, skin and beak of both species and established that
R. peruviana is pigmented by canthaxanthin (193) while lutein (73) is the plumage
pigment of the orange-red R. rupicola. For these first studies the internal organs
of both these wild species were, unfortunately, not available, so that nothing can
be said about possible intermediate compounds in the liver. It may be concluded
from the feeding habits of the cock-of-the-rock that it ingests lutein (73)
abundantly with fruit, and this is evidently stored in the feathers of R. rupicola.
In R. peruviana canthaxanthin (193) is the major carotenoid while lutein (73)
is not detectable. This selectivity in two closely related species is certainly
not exceptional and is also applicable, no doubt, to the varying carotenoid
composition in flamingos. It is highly probable that the skin and plumage of
R. rupicola contain phoeniconone (195) although it cannot be excluded that
this may be an artifact.
Table 5 shows the carotenoid contents for both cock-of-the-rock species.

Table 5. Carotenoid composition (J.1g/g) in Rupicola rupicola (A) and Rupico/a peruviana (B)

Compound Feathers Skin Skin (legs) Beak

A B A B A B A B

P-Carotene (3) 5
Echinenone (148) Traces 20
Canthaxanthin (193) 20 360 Traces 30 5 100 Traces 70
Lutein (73) 40 10
Phoeniconone (195) Traces Traces
656 H. THOMMEN

Closely related to the scarlet ibis (Eudocimus ruber) is the roseate spoonbill
(Ajaia ajaja) which lives in brackish waters of the coastal southerly regions of
North America, Mexico, Argentina and Chile. Fox [86] found the two keto
carotenoids canthaxanthin (193) and astaxanthin (203) in the plumage of this
bird. Ajaia ajaja differs from other carotenoid-dependent birds in that asta-
xanthin (203) is the predominant carotenoid in its blood plasma.

D. Carotenoproteins

The question arises as to the forms in which carotenes and carotenoids

generally function in metabolic processes. Above all a clarification is needed
of the manner in which carotenes and carotenoids are incorporated into cells.
Fundamentally, it may be maintained that .animal cells cannot synthesize
carotenes or carotenoids de novo but can only transform them. The different
plant cells, on the other hand, can synthesize this type of compound in almost
every variety with the aid of available enzyme systems.
The carotenes and carotenoids are found in the chromatophores (chloro-
and chromoplasts) in different physical forms, either in aqueous or oily solu-
tions, or as crystalloids and crystals. The pigments may be present in the free
or bound state. The manner of binding the different carotenoids to the organel-
les depends directly on their chemical structure. One cannot yet answer the
question whether specific enzymes in plants are responsible for the presence
of individual carotenoids. For example, it is of peculiar interest that eschscholtz-
xanthin (84) occurs in considerable amount in the orange-yellow petals of a
variety of Eschscholtzia californica or that rhodoxanthin (209) is found as the
main pigment in the red seed-coat of the berries of Taxus baccata. The specific
carotenoids found more recently in bacteria, algae and fungi also indicate that
specific biosynthetic reactions must be of decisive significance.
The further question arises whether the presence of such pigments is of
importance for particular metabolic processes or whether it is a freak of
nature. The suggestion that carotenes and carotenoids have a significant role
in photosynthesis can be demonstrated more or less convincingly by studies
on light-dependence.
It has long been argued that the carotenoids do not participate in biological
processes in the free state but as conjugates. On that basis the caroteno-
proteins acquire considerable importance. These are compounds in which carot-
enoid molecules are bound as prosthetic groups to protein in stoichiometric
proportions. Although more than 30 years have already passed since the
isolation of ovoverdin, the carotenoprotein of lobster eggs [87, 88], only
modest advances in the chemistry and biochemistry of carotenoproteins have
as yet been made. It is now known that carotenoproteins are especially widely
distributed in invertebrates. It may be assumed, however, that their distribution
is not limited to the invertebrates and that their occurrence is ubiquitous.
Verne [89, 90] and Lederer [91] were the first to review this class of compounds.
 VIII. Metabolism 657

In a more recent review by Cheesman et al. [92], the authors defme the different
types of carotenoproteins. 'True carotenoproteins' are compounds in which
carotenes and carotenoids are bound to proteins in stoichiometric proportions.
This involves the presence of specific sites for the attachment of carotenoid,
resulting in a considerable change in the absorption spectrum of the latter.
The authors postulate that a true carotenoprotein is present in a crude aqueous
extract if treatment with heat or organic solvents results in liberation of a
carotenoid. Under these circumstances the spectral characteristics of the free
pigments, which have not been evident previously, become visible.
So far, only three carotenoproteins have been isolated (Table 6). Crusta-
cyanin, the blue protein of the lobster carapace comprises, after removal of
the carotenoid, only amino acid residues [93-95]. The first carotenoprotein
isqlated was ovoverdin, the green storage protein of lobster eggs [96-99].
This has proved to be a lipoprotein with a large prosthetic group, rich in
phospholipids (approximately 22% of its weight). Furthermore the complex
contains a carbohydrate component (hexosamine and a non-amino-sugar
residue, amounting to approximately 5% of the whole). From the albumen
gland and the eggs of the gastropod Pomacea canaliculata a red protein,
ovorubin, was isolated by Cheesman [100] and Norden [101].

Table 6. Carotenoproteins isolated

Name Occurrence Colour Absorption max. Components of the

(inflections in carotenoprotein
Species Organ parentheses)
nm Carotenoid Protein

Crustacyanin Homarus Carapace Blue- (320), (370), 633 Astaxanthin Simple

vulgaris grey protein
Homarus Carapace Blue- (320), (370), 633 Astaxanthin Simple
americanus grey protein

Ovoverdin Homarus Eggs, Green Astaxanthin Glycolipo-

vulgaris ovaries { 464-468,
(330\(440\ ) protein
Homarus Eggs, Green (500), 650-670, Astaxanthin Glycolipo-
americanus ovaries higher maxima vary protein

Ovorubin Pomacea Albumen Red (330), (480), Astaxanthin Glyco-

canaliculata gland, 510, 545 protein
eggs

Cheesman et al. [92] propose a second class, the so-called lipoproteins with
a lipid-associated carotenoid. In such compounds the carotenoid is part of a
large lipid prosthetic group and does not show a specific interaction with the
protein. In such complexes the carotenoid includes a number of molecular
species in different proportions. The molecules seem to be 'dissolved' in the

Carotenmds 42
658 H. THOMMEN

lipid component. The lipoprotein of avian egg yolk has such characteristics
because its carotenoid composition differs with the diet of the birds. These
findings cannot be generalized, because it was shown that different fractions
of mammalian plasma lipoproteins are responsible for the binding of carot-
enoids, vitamin A and vitamin A ester [102]. P-Carotene (3) and lutein (73)
are differently distributed among the proteins of human plasma, as shown by
Krinsky et al. [103]. Oncley et al. [104] postulate that the carotenoid content
of plasma lipoproteins is not stoichiometric and that the observed selectivity
is only dependent on the choice of vehicle. The paper 'Carotenoproteins in
Invertebrates' by Cheesman et al. [92] gives a list of a large number of caroteno-
proteins, some of them regarded as relatively unspecific complexes, in the
absence of more precise analysis. Besides astaxanthin (203), xanthophylls, e.g.
lutein (73), and P-carotene (3) could be identified as partners of proteins. There
is no doub't that, particularly among the many species of crustaceans and even
insects, other keto carotenoids will be found. A recent observation made duri'ag
the isolation of canthaxanthin (193) in the Colorado potato beetle gave evidence
of a complex consisting of a still uncharacterized protein and canthaxanthin
[105]. Furthermore, keto carotenoids seem to be present as protein complexes
in the haemolymph of the cotton stainer (Dysdercus species) and in the yellow
mealworm (Tenebrio molitor) [106]. Cheesman et al. [92] observe that even
though a number of different xanthophylls may be present, the total carotenoid
content shows a remarkable constancy.
When a true carotenoprotein has a lipid prosthetic group, the carotenoids
are presumably present in non-stoichiometric amounts which may at least
equal in quantity the specific protein-bound carotenoid and may modify the
colour of the whole complex to a marked degree; such an observation has
been made by Lee [107] in the isopod ldothea granulosa.
Although we now have many indications that the carotenoproteins are
very widely distributed not only in invertebrates but generally in animals and
plants, it is still surprising how little is known of the composition and signifi-
cance of these complexes. For example, it would be interesting to know to
what extent the red carotenoproteins found in molluscs and Echinodermata
are the result of definite combinations between a carotenoid and a protein.
A further interesting fact is that in the carotenoproteins of invertebrates,
especially arthropods, astaxanthin (203) is the most frequent carotenoid.
More recent studies have shown, nevertheless, that in addition to astaxanthin
canthaxanthin (193) can appear as the carotenoid compo'nent [100, 101, 108].
Interesting relationships can be shown in insects, where, in addition to carot-
enoid, bile pigments or fragments thereof are incorporated into the complexes.
The isolation of carotenoproteins offers no difficulties since the colour of
the complex and an absorption maximum in the region of 280 nm (attributable
to the protein) provide useful aids. Carotenoproteins from eggs and soft tissue
are extracted by water or dilute saline, then precipitated by ammonium
sulphate and chromatographed on suitable small columns, chiefly of ion-
exchange cellulose [87, 88, 92, 98, 101, 107].
 VIII. Metabolism 659

To isolate stored carotenoproteins from crustaceans the carapace must

first be decalcified. The finely ground material is treated with a citrate buffer
(pH 5.0) [109], ethylenediaminetetraacetate (pH 7.5) [110] or ammonium
sulphate [111] to dissolve the protein. It is advantageous to carry out these
isolations in an air-conditioned room with exclusion of daylight. All caroteno-
proteins so far investigated can be split by an aqueous solution of acetone or
alcohol into the carotenoid and apoprotein, with the possibility, under mild
conditions, of obtaining the free carotenoid in pure form. In the case of crusta-
cyanin and ovorubin the carotenoproteins can be reconstituted by mixing an
acetone solution of the carotenoid with an aqueous solution of the protein
[112]. Carotenoprotein stabilities vary considerably. Since crustacyanin and
ovorubin are quite stable and can be stored indefinitely at low temperature,
most experiments on carotenoproteins have been performed with these two
complexes.
It is interesting that freeze-dried preparations of crustacyanin and ovoverdin
are red, from which it can be inferred that the presence of water is highly
important for the binding of carotenoid to protein. It is also known that
carotenoids bound to protein are much more stable towards photooxidative
processes than the free compounds. Crustacyanin is, so far, the only repre-
sentative obtained in crystalline form [95].
With regard to the determination of the molecular size of carotenoproteins,
only crustacyanin has undergone detailed physicochemical study. The native
form of the protein shows, as described by Gammack and Raper [110], a
molecular weight of about 360,000. The minimum molecular weight, based
on the carotenoid content, is estimated at approximately 36,000. Dialysis in
dilute solution yields a product called a' -crustacyanin. This is transformed,
after addition of salt, into a-crustacyanin. Prolonged dialysis of the latter
compound converts it to P-crustacyanin. The interconversions of 'X'-, 'Y.- and
P-crustacyanin can be followed by marked changes in the absorption spectra.
It is suggested that different apoprotein sub-units are responsible for the spectral
changes. Little is known concerning the molecular size of the other caroteno-
proteins.
Wyckoff [111] proposes, on the basis of sedimentation measurements,
a value of 300,000 for ovoverdin. This value was also obtained by gel-filtration
on Sephadex by Ceccaldi et al. [99]. The minimum molecular weight based
on the carotenoid content is about 170,000. In those complexes where the
carotenoid component contains a keto group (4- or 4'-positions) the carbonyl
function may possibly be responsible for the complex formation. If it is shown
to be true that the presence of a free carbonyl group is essential for the formation
of carotenopro!eins, there would be a real possibility of finding, with modern
analytical techniques, the recently isolated keto carotenoids functioning as
prosthetic groups.
Zagalsky et al. [112] examined a number of decapod crustacean caroteno-
proteins. The carapaces, mandibles and stomach walls of the decapods listed
in Table 7 have been studied. In all cases astaxanthin (203) was again detected
660 H. THOMMEN

Table 7. Absorption spectra of carotenoproteins in decapods (from Zagalsky et a!. [112])

Occurrence Form of Absorption maxima (nm)

carotenoprotein
Species Organ Main Others (inflections
in parentheses)
Aristeus antennatus Carapace IX (+salt) 593 370, 314, 278
IX (salt-free) 573 358, (314), 278
p 570 358, (318), 278
Mandibles IX (+salt) 595 369, 315, 277
IX (salt-free) 588 350, 310, 277
p 578 354, (312), 277
Stomach IX (+salt) 588 362, 310, 278
IX (salt-free) 578 358, (310), 278
p 575 358, (31 0), 278
Palinurus vulgaris Carapace IX (+salt) 560 (600), 326, 276
IX (salt-free) 560
Scyllarus arctus Carapace IX (+salt) 616 378, 320, 276
IX (salt-free) 595
p 589 378, 315, 276

Clibanarius erythropus Exoskeleton IX (+salt) 620 369, 320, 276

IX (salt-free) 602 365, (315), 276
p 593 364, (320), 276
Galathea strigosa Carapace IX (+salt) 589 367, (325), 275
IX (salt-free) 582
p 580
Eriphia spinifrons Carapace IX 536 (310), (340), (400), 278

Pachygrapsus marmoratus Carapace IX (+salt) 520 350, (312), 275

IX (salt-free) 510

,,
I
1.0
I
'2
·;:: 4 I

i
I
I
I
'2
I
:0
::::J
3 I
I 0
()
I fi
I 2
I
I
I
I
I
..!...

~ I
I
I
.f
c. I
I E.
~
<(

300
"' 400 500 600 700 nm
Fig. 1. Absorption spectra of crustacyanin and ovorubin (from Cheesman et a!. [92]). --Crusta-
~
<(

cyanin (1 mgjml) in 0.2 M Na K phosphate buffer, pH 7. ------ Ovorubin (1 mgjml) in water. The
prosthetic group of both proteins is astaxanthin, although that of ovorubin was erroneously
stated to be an astaxanthin derivative. The absorption spectrum of astaxanthin in petroleum
ether shows a single maximum in the visible range at 470-475 nm
 VIII. Metabolism 661

1.0

0.8

>- 0.6
=5
:;::::
e-
~ OA
<(

Q2

Fig. 2. Absorption spectra of ovoverdin and the egg protein of Eupagurus bernhardus (from Chees-
man et al. [92]). - - Ovoverdin (1 mgjml) in 0.05 M Na K phosphate buffer, pH 7. ------ Eupagurus
protein (uncertain concentration) in 0.1 M Na 2 HP0 4 . The prosthetic group of ovoverdin is
astaxanthin; that of the Eupagurus protein is an ester of astaxanthin. Both proteins are labile,
and the heights and positions of the absorption maxima differ significantly in different preparations

25

20

1.5

1.0

0.5

Fig. 3. Absorption spectra of two canthaxanthin-protein complexes (from Cheesman et al. [92]).
- - Carotenoprotein from Idothea granulosa [107]. ------ Carotenoprotein from Tanymastix
lacunae [124]. Both proteins are of unknown concentration in 0.05 M Na K phosphate buffer,
pH 7. The prosthetic group, canthaxanthin, shows an absorption maximum in petroleum ether
at 460-463 nm. The absorption maxima of the proteins in the region of 500 nm appear to be due
to lipid-associated xanthophylls

as prosthetic group. The proteins form a related group with similar amino acid
compositions and properties. The molecular size of the proteins varies from
40,000 to over a million. If the carotenoid is removed, the proteins dissociate
to units of moiecular weight 40,000-50,000 (oc-forms) and about 20,000.
Carotenoproteins show a characteristic spectral behaviour. As mentioned
at the outset, the absorption curve of the complex masks the typical carotenoid
maxima, which only appear characteristically after cleavage of the carotenoid-
protein link (Fig. 1-3). These physicochemical facts certainly provide no proof
662 H. THOMMEN

yet whether complexes of this kind are also, in fact, present as such in the cell
or whether they are only formed during extraction. The latter view is taken
by Ke et al. [113]. It is nevertheless possible to produce evidence for at least
the transient existence of genuine carotenoid complexes in the intact cell by
demonstration of a stable enzyme-substrate complex. Chromatophores of
Rhodospirillum rubrum [114, 115] or chloroplasts of Spinacia oleracea L. [116],
among others, served as experimental objects, and it was shown that viola-
xanthin (135) is bound to an enzyme protein. Violaxanthin can be liberated
from the buffer solution by addition of water; with a change of pH from 7.5
to 5.2 and simultaneous addition of ascorbic acid transformation to zeaxanthin
(67) follows. The enzyme can consequently be described as a de-epoxidase.
The reaction (Scheme 9) can be followed by absorption spectra, and it may
be assumed that especially in the plant cell, analogous phenomena occur.
Wolken [117] has found similar relationships with Phycomyces by spect~o­
photometry in intact sporangiophores; the conclusion can therefore be drawn
that in the phototropic area of the sporangiophore the light absorption of
carotenoid, which must be almost completely bound to proteins, becomes
effective.
These experimental findings provide evidence that carotenoids participate
to some degree in photosynthesis as carotenoproteins, with formation of

Enzyme-substrate complex (violaxanthin de-epoxidase

and violaxanthin)

(135)

l Violaxanthin de-epoxidase
and ascorbic acid

OH

HO
(67)

Scheme 9. Enzymic de-epoxidation; viofaxanthin (135)-+zeaxanthin (67) (from Hager [116])

 VIII. Metabolism 663

reversible or irreversible corresponding oxidation products. It is not clear at

present whether the appearance of canthaxanthin (193) or astaxanthin (203)
in invertebrates means that a corresponding hydroxylated precursor is bound
to the protein. Nor is it clear how far carotenoproteins have any general
physiological function in animals. An earlier study by Bamji and Krinsky [118]
dealt with the enzymic transformation of antheraxanthin into zeaxanthin
in the green alga Euglena gracilis. Although the authors have not discussed
protein binding of the carotenoid in detail, this work should be noted here since
the transformation certainly takes place in the cell through the carotenoid-
protein complex (Scheme 10).
Krinsky and Lenhoff [120], who studied the carotenoid composition in
Hydra littoralis, and Hydra pirardi, mention the possibility that a ketocarot-
enoid-protein complex may be responsible for the conversion (Scheme 11)
of canthaxanthin (193) into diketopirardixanthin (194), a compound with
saturated rings. The question is still open if such a reduction is related to photo-
synthetic reactions. In these experiments hydra were fed on a sole source of
food, Artemia salina. The carotenoid composition of A. salina is well known.
A recurring problem of growing urgency is the significance of caroteno-
proteins. Although abundant experimental data are available, attempts to
explain the almost ubiquitous occurrence of carotenoproteins are still only
suppositions and hypotheses. In addition to the comprehensive work of Chees-
man et al. [92] on carotenoproteins, the publications of Zagalsky et al. [112],
Thirkell and Hunter [121] as well as Herring [122] should be mentioned.
Further experimental evidence supports the view that carbonyl groups
participate in the formation of carotenoproteins involving 4-keto carotenoids.
Lee and Zagalsky [119] achieved the formation of complexes with such
carotenoids and the apoproteins of crustacyanin. It may be assumed that
non-enolized keto groups, as present in astaxanthin (203) and canthaxanthin
(193), are required for the formation of a ~omplex. Such keto groups, the
nucleophilic character of which is markedly reinforced by conjugation with
the polyene chain, are suitable for interaction with the basic groups in a
protein. Cheesman et al. [92] discuss in detail the nature of the carotenoid-
protein linkages and summarize the aspects which are still completely valid.
In conclusion, the properties and functions of carotenoid-protein com-
plexes may be summarized as follows:
1) There is mutual stabilization of the carotenoid and the protein. Cleavage
leads to a considerable increase in sensitivity of the carotenoid to oxidation
and, in the protein, to a change of configuration.
2) Carotenoproteins are used by the animal organism as camouflage. Some
animals, especially the lower ones, produce carotenoproteins of a colour to
match the environment.
3) Caroteno.proteins seem to have a definite role in the survival of the
embryo, particularly in egg-laying invertebrates.
4) Carotenoproteins may function in electron transport mechanisms.
 "'"'.!:>-

OH

~ l J
~~~~~~~
0

(119)
Malate NADP FMNH 2
G"oAA
o"m=,.JCAnPn)C FMN) ~ /'.. ,OH

/'-.. """' /~~~~~~~ ,_ /<-.. I

...,~
HO~
:r:
'-.../ I 'OH 0
J ~
~
(87?)
"'z
l-H,O
OH

~ TT
~~~~~~

HO~ (67)

Scheme 10. Enzymic de-epoxidation; antheraxanthin (119)--->zeaxanthin (67) (from Bamji and Krinsky [118]). Abbreviations: NAD P =nicotinamide adenine
dinucleotide phosphate (coenzyme II), NADPH=its reduced form, FMN=flavin mononucleotide (riboflavin 5'-phosphate), FMNH 2 =its reduced form
 VIII. Metabolism 665

Source of food
(Artemia salina)

l
""" """ """ """ """ """ """ """ """
0
(148)

(155)

l
""" """ """ """ """ """ """ """ I """
0

r
(193)

(Protein complex?)

""" """ """ """ """ """ """ """ """

0
(194)
Scheme 11. Formation of diketopirardixanthin (194) in Hydra pirardi (from Krinsky and Lenhoff
[120])
666 H. THOMMEN

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[87] R. Kuhn and N.A. S¢rensen, Ber. Deut. Chern. Ges. 71, 1879 (1938).
[88] R. Kuhn and N.A. S¢rensen, Z. Angew. Chern. 51, 465 (1938).
[89] J. V.erne, Les Pigments dans l'Organisme Animal (Doine, Paris 1926).
[90] J. Verne, Couleurs et Pigments des i!tres vivants (Colin, Paris 1930).
[91] E. Lederer, Les Carotiinoides des Animaux (Hermann, Paris 1935).
[92] D. F. Cheesman, W.L. Lee and P.F. Zagalsky, Bioi. Rev. Cambridge Phil. Soc. 42, 131
(1967).
[93] G. Wald, N. Nathanson, W. P. Jencks and E. Tarr, Bioi. Bull. 95, 249 (1948).
[94] W. P. Jencks and B. Butten, Arch. Biochem. Biophys. 107, 511 (1964).
[95] D. F. Cheesman, P.F. Zagalsky and H.J. Ceccaldi, Proc. Roy. Soc., Ser. B 164, 130 (1966).
[96] K.G. Stern and K. Salomon, Science 86, 310 (1937).
668 H. THOMMEN

[97] K.G. Stern and K. Salomon, J. Bioi. Chern. 122, 461 (1938).
[98] P.F. Zagalsky, Thesis (London 1964).
[99] H.J. Ceccaldi, D. F. Cheesman and P.F. Zagalsky, C. R. Soc. Bioi. 160, 587 (1966).
[100] D. F. Cheesman, Proc. Roy. Soc., Ser. B 149, 571 (1958).
[101] D. A. Norden, Thesis (London 1962).
[102] J. Ganguly, N. I. Krinsky, J. W. Mehl and H.J. Deuel, Arch. Biochern. Biophys. 38, 275
(1952).
[103] N.I. Krinsky, D. G. Cornell and J.L. Oncley, Arch. Biochern. Biophys. 73, 233 (1958).
[104] J.L. Oncley, F.R. Gurd and M. Melin, J. Amer. Chern. Soc. 72, 458 (1950).
[105] F. Leuenberger and H. Thommen, J. Insect Physiol. 16, 1855 (1970).
[106] K.H. Trautmann and H. Thommen [F. Hoffmann-La Roche & Co. Ltd., Basle], un-
published results.
[107] W.L. Lee, Cornp. Biochern. Physiol. 19, 13 (1966).
[108] W.L. Lee, Cornp. Biochern. Physiol. 18, 17 (1966).
[109] D. F. Cheesman and J. Prebble, Cornp. Biochern. Physiol. 17, 929 (1966).
[110] D. B. Gammack and J.H. Raper, Molecular Properties of Crustacyanin (1966).
[111] R. W.G. Wyckoff, Science 86, 311 (1937).
[112] P.F. Zagalsky, H.J. Ceccaldi and R. Daumas, Cornp. Biochern. Physiol. 34, 579 (1970).
[113] B. Ke, M. Green, L. P. Vernon and A. F. Garcia, Biochirn. Biophys. Acta 162, 467 (1968).
[114] L.P. Vernon and A.F. Vernon, Biochirn. Biophys. Acta 143, 144 (1967).
[115] E. Fujimori, Biochirn. Biophys. Acta 180, 360 (1969).
[116] A. Hager, Planta (Berlin) 91, 38 (1970).
[117] J. Wolken, J. Cell. Bioi. 43, 354 (1969).
[118] M.S. Bamji and N.I. Krinsky, J. Bioi. Chern. 240, 467 (1965).
[119] W.L. Lee and P.F. Zagalsky, Biochern. J. 101, 9C (1966).
[120] N.I. Krinsky and H.M. Lenhoff, Cornp. Biochern. Physiol. 16, 189 (1965).
[121] D. Thirkell and M.I.S. Hunter, J. Gen. Microbial. 62, 125 (1970).
[122] P.J. Herring, Syrnp. Zoo/. Soc. London 19, 215 (1967).
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 669

IX. Function
NORMAN I. KRINSKY

Department of Biochemistry and Pharmacology,

Tufts University School of Medicine, Boston, Massachusetts 02111, USA

A. Introduction . . . 670
B. Photofunctions . . 670
1. Photoprotection 671
a) The Stanier hypothesis 671
b) Photoprotection and chromophore length 676
c) Mechanisms of photoprotection 679
2. Photosynthesis . . . . . . . 686
a) Accessory pigments. . . . . . 686
b) 0 2 evolution and transport . . 690
c) Cofactors of photosynthetic reactions 691
d) The 515 nm effect . . . . . . . . 692
e) Association with different photosystems . 698
3. 'Blue-light' effects 698
a) Phototropism . . 699
b) Phototaxis . . . 701
c) Photoinhibitions . 702
d) Photostimulations 702
e) Photoreception . 702
f) Photoperiodism . 703
g) Protoplasmic viscosity and streaming 703
C. Non-Photofunctions . . . . . . . . 703
1. Protein and membrane stabilization . 703
2. Reproduction . . . . . 704
3. Miscellaneous functions . . . . . . 705
D. Metabolite Functions . . . . . . . . 705
1. Vitamin A and retinene (retinol/retinal) 705
2. Abscisic acid . 706
3. Trisporic acid . . . 706
4. Sporopollenin . . . 708
5. Grasshopper ketone. 709
E. Conclusions 710
References . . . . . . . 711
670 NORMAN I. KRINSKY

A. Introduction
Approximately twenty years ago, two extremely important books were
published that brought together an immense amount of knowledge concerning
the chemistry and biochemistry of carotenoid pigments. The first, Carotinoide,
by Karrer and Jucker [1] appeared in 1948, and in Braude's [2] excellent
English translation, Carotenoids, in 1950. This book dealt primarily with the
chemistry of carotenoids. In 1952, The Comparative Biochemistry of the Caro-
tenoids by Goodwin [3] appeared, dealing with biochemical aspects of caro-
tenoid pigments. The two books treated the functions of carotenoids quite
differently. Karrer and Jucker [2] dealt with the function of carotenoids in
plants very briefly and concluded that 'All these investigations are still at a
preliminary stage and further researches will be required in order to elucidate
the importance of carotenoids in plants'. In addition, they also discussed the
functions in animals as consisting of carotenoids acting as provitamins A a.nd
their role in the visual process. Goodwin on the other hand, approaching the
subject from a biochemist's point of view, was much more concerned about
function, and devoted considerable space to this area. Although he was unable
to generalize concerning the biological role of carotenoids, he felt that the
ability to absorb light seems to be the common factor. Even so, in retrospect,
we must agree with his conclusion that 'with regard to formation and function
(of carotenoids), knowledge is rudimentary'. Thus, he summarized most (if
not all!) of the available data dealing with carotenoids functioning in photo-
synthesis, phototropism, photoreception and reproduction.
Since then, numerous articles have appeared which have dealt, among
other things, with the functions of carotenoid pigments. These articles [e. g.
4-11] have served an important purpose in bringing together information
from many different areas.
This chapter will treat carotenoid functions as consisting of three separate
and distinct areas. The first can be considered the area of photofunctions and
deals with the direct and indirect effects of light as mediated or modified by
carotenoid pigments. The second deals with an area much more difficult to
define. Since it presumably does not involve light, I have called it non-photo-
functions. Lastly, some of the functions attributed to compounds formed from
carotenoids by cellular metabolism, the metabolite functions, will be discussed.
It is not my intention to treat exhaustively these functions but to survey
our present knowledge and point out areas which are either particularly well
understood or, because of the contradictory nature of available results, should
profit most from continued explorations. The reader will be referred to the
original literature and, when available, to critical reviews of specific subject
areas.

B. Photofunctions
This section deals with the multitude of responses of cells towards light
which have, rightfully or not, been attributed to direct interaction of caro-
 IX. Function 671

tenoids and light, or interaction of carotenoids and other molecules which

have either absorbed light or been formed by the action oflight. I have divided
this section into three divisions, though there is some overlap between them:
1. photoprotection, 2. photosynthesis, 3. 'blue-light' effects.

1. Photoprotection
A large number of observations can best be explained by assuming that
carotenoid pigments can protect cells and tissues against the potentially harm-
ful effects of visible light. In 1968, this extensive material was reviewed by
Krinsky [10], and many examples in bacteria, lower and higher plants, and
even animals of protection by carotenoids were described and discussed. At
that time, attempts were made to explain the mechanism of this important
function of carotenoid pigments. Since then, a new development led by Foote
(Section B.1 c) seems to give added strength to the suggestion that photo-
protection may be a wide-ranging function of carotenoid pigments. This sec-
tion will review the development of these concepts, summarize some of the
experimental work, and discuss the results from the viewpoints of mechanism
of action and potential usefulness.

a) The Stanier hypothesis

The experiments and insights of Stanier and his colleagues cannot be
underestimated in considering the development of the idea that carotenoids
function as photoprotective agents. It was first demonstrated in 1955 [12]
that a mutant strain of the non-sulphur purple bacterium, Rhoda pseudomonas
spheroides, which lacks coloured carotenoid pigments was photosensitive in
the presence of air. This mutant strain, called blue-green (B-G), 'accumulated
the colourless polyene phytoene (32), presumably as a result of a single-step
mutation in the normal pathway of carotenoid synthesis. The photosensitiza-
tion was lethal, for not only did growth cease for the B-G mutant strain in
light and air, but massive killing occurred. Under other environmental condi-
tions (light-anaerobic atmosphere or dark-aerobic atmosphere) there was no
difference in the growth of the wild-type (W-T) strain or the B-G mutant
strain of Rps. spheroides, which indicated to the authors that they were observ-
ing a photodynamic action, defined by Blum [13] as 'the sensitization of a
biological system to light by a substance which serves as a light absorber for
photochemical reactions in which molecular 0 2 takes part'. They concluded
that the light absorber sensitizing the entire process was the cells' own bacterio-
chlorophyll (BCHL).
From all of these observations, Stanier and his colleagues [12] postulated
that carotenoids are universally associated with photosynthetic tissue because
they can protect cells from photodynamic destruction catalysed by either
chlorophyll (CHL) or BCHL. Some of the consequences of this hypothesis
were: (i) only bacterial photosynthesis could proceed in the absence of caro-
tenoid pigments, inasmuch as this process does not result in 0 2 evolution;
672 NORMAN I. KRINSKY

(ii) mutation in carotenoid biosynthesis in green plants would be lethal, for

any 0 2 produced photosynthetically could be utilized in a photodynamic
reaction.
The potential danger of photodynamic destruction in cells which contain
an endogenous photosensitizer (such as metalloporphyrins, present in all cells
except strict anaerobes) was discussed by Franck [14] with respect to green
plants. These possess all the attributes of systems which can undergo photo-
sensitized oxidations, they contain high concentrations of effective photo-
sensitizers and directly evolve 0 2 in the presence oflight, yet they have utilized
their photosensitizers for reductive purposes. The hypothesis of Stanier and
his colleagues was the first directed to understanding the essential process
of the control of photooxidation which enables green plants to survive photo-
sensitizing conditions. Clayton [15] also discussed the question of preventing
harmful photooxidations in photosynthetic tissues. ,
During the four years following their original publication [12], Stanter
and his co-workers published a series of papers which amplified and extended
their original observations and raised a number of important issues, some of
which still remain to be clarified. In 1956, Sistrom et al. [16] showed the
importance of BCHL in catalysing the lethal effect in Rps. spheroides by
growing this organism for many generations in air and complete darkness.
The presence of air in such cultures had been known to inhibit pigment syn-
thesis so that under these conditions the BCHL was diluted to a very low
concentration. These bleached cells of the B-G mutant strain could then be
exposed to high light intensities in air without any change in growth rate.
Sistrom et al. [16] postulated that the carotenoid pigments, which protected
the W-T strain in the presence of BCHL, might be acting as chemical buffers
by becoming preferred substrates for the BCHL-catalysed photooxidations.
They suggested that epoxide groups might be formed across carotenoid double
bonds, and postulated an enzymic mechanism to reduce the epox.ide caro-
tenoid back to an olefinic compound. Although epoxide carotenoids are not
found in photosynthetic bacteria, this concept has been invoked by Krinsky
[17] to explain epoxide interconversions in algae as manifestations of the
carotenoid protective function.
In the first review of the hypothesis, Stanier and Cohen-Bazire [18] argued
that carotenoids must have been part of the photosynthetic apparatus of
bacterial photosynthetic systems as accessory light-absorbing pigments prior
to the emergence and development of 0 2 -evolving green plant photosynthesis,
for only those organisms which contained carotenoids could have survived
the establishment of the proper factors for lethal photosensitized oxidation,
i.e., light, a photosensitizing pigment and, of course, 0 2 . They also suggested
that carotenoids should function in non-photosynthetic organisms to protect
them against potentially lethal photosensitized oxidations. Kunisawa and
Stanier [19] were able to demonstrate that this protection did occur when
an exogenous photosensitizing pigment was used. They worked with the
chemoheterotrophic bacterium Corynebacterium poinsettiae, using both the
 IX. Function 673

coloured W-T and a white mutant strain which contained only phytoene (32)
and traces of phytofluene (30). In the presence of the photosensitizing dye
toluidine blue, the white mutant strain displayed a lethal aerobic sensitivity
whereas the coloured W-T strain was unaffected. The ecological importance of
this phenomenon was first demonstrated by Mathews and Sistrom [20] using
the yellow bacterium Sarcina lutea and a colourless mutant strain. They used
sunlight as their source of light and were able to demonstrate that the caro-
tenoidless mutant strain was killed in the presence of visible light and air whereas
theW-T strain remained viable under these conditions. They therefore proposed
that the extent of the protective function of carotenoid would be very broad.
For example, they were led to expect that all aquatic bacteria should contain
carotenoids, inasmuch as they are exposed to light and air, and the same
argument could be extended to asporogenous bacteria. Similar ecological
arguments have been advanced by Singer and Ames [21] to explain the high
percentage of guanine and cytosine in the deoxyribonucleic acid (DNA) of
bacteria exposed to sunlight.
In addition to mutations, colourless strains of normally coloured organisms
can be obtained by the use of diphenylamine (DPA), which blocks coloured-
carotenoid biosynthesis leading to the accumulation of colourless carotenoids.
Cohen-Bazire and Stanier [22] treated the non-sulphur photosynthetic bac-
terium Rhodospirillurn rubrurn with DPA and obtained cells which lacked the
more unsaturated carotenoids and accumulated phytoene (32), phytofluene
(30), (-carotene (26) and neurosporene (22). These cells showed a lethal aerobic
sensitivity, comparable to that of the carotenoid-deficient mutants of Rps. sphe-
roides. In addition, it was observed that BCHL, which is itself destroyed under
the conditions oflethal photosensitization, bleaches at a rate slower than the rate
of photokilling. Dworkin [23] studied this phenomenon in the B-G mutant
strain of Rps. spheroides and found a temperature coefficient (Q 10) of 1 for
photodynamic killing between 4 oc and 40 oc indicating that the reaction is
probably purely photochemical. In fact, he was able to demonstrate that at a
sufficiently low temperature even the W-T strains were killed. For example,
at 1 oc W-T strains of both Rps. spheroides and R. rub rum were killed upon
illumination in air, without any loss of BCHL [24]. Along similar lines,
Mathews [25] showed that even W-T cells of S. lutea were more susceptible
to toluidine blue-sensitized photokilling at 4 oc than at 34 oc. Since there
was no measurable destruction of BCHL at low temperatures, Dworkin [23]
concluded that an energy transfer must occur from BCHL to an adjacent
acceptor molecule. Since BCHL is located in the membrane-associated chro-
matoph'Ores, he proposed the membrane as the site of the lethal photooxida-
tion. This sam~ conclusion was reached by Mathews and Sistrom [26], who
found that the photodynamic killing of S. lutea, using toluidine blue as the
photosensitizer, was temperature-independent between 6.5 oc and 34 °C.
Since they had already demonstrated that the carotenoid pigments of S. lutea
are localized in the cell membrane [27] they investigated several membrane
parameters during the photosensitizing process. They found an increase in

Carotenmds 43
674 NORMAN I. KRINSKY

permeability, as measured by an increase in the amount of 260 nm-absorbing

material released into the medium and also by binding of the dye sodium
8-anilino-1-naphthalenesulphonate in a fluorescent form. They also observed
a decrease in the activity of two membrane-associated enzymes, succinic
dehydrogenase and diphosphopyridine nucleotide oxidase. They were able to
exclude the permeability change from being the primary lethal event during
photodynamic killing by comparing the effects on membrane permeability
of the antibiotic polymyxin with photosensitization; at equivalent levels of
killing, polymyxin has a much more profound effect on membrane permeability
than has the photodynamic action. Recently, Rottem et al. [28] have demon-
strated the ability of carotenoid pigments to protect the membrane enzyme
adenosine triphosphatase against a toluidine blue-sensitized photodestruction
in Mycoplasma laidlawii membranes.
By 1960, when Stanier [29] reviewed the work of his group in his Harvey
Lecture, the hypothesis that carotenoid pigments act as protective agents was.
well established. What remained to be done was the elucidation of how caro-
tenoid pigments could carry out this important function. Before discussing
the mechanisms, it should be pointed out that the hypothesis did not rest
solely on the observations made by Stanier and his colleagues. There is in the
literature a long history of reports of aerobic photosensitivity, both lethal and
non-lethal. Many of the observations have been related to alterations or
deficiencies in carotenoid pigments, and have been discussed in carotenoid
reviews [5-7, 9, 15, 30, 31]. In 1968, Krinsky [10] brought together the material
from a variety of fields to support the Stanier hypothesis.
Aerobic photosensitivity associated with carotenoid deficiencies in plants
was initiated by the observations of Koski and Smith [32], who studied
the mutant seedling, white-3, of Zea mays. This mutant strain was photo-
sensitive and contained no detectable carotenoids. Koski and Smith [32]
concluded that the CHL deficiency in this mutant was not due to an inhibition
in the biosynthesis of CHL but rather to the inability of this mutant to pre-
serve the CHL that had been formed. We would now interpret their observa-
tions to indicate that the absence of carotenoid pigments permitted the CHL
to act as a photosensitizer and led to its own bleaching. Support for this idea
came from the observations of Faludi et al. [33] and Anderson and Robert-
son [34] working with corn mutants. Wallace and his colleagues [35, 36] made
similar observations on a white mutant of the sunflower, Helianthus annuus.
These studies on mutants can be summarized as follows: in the absence of
coloured carotenoid pigments, plants are capable of converting protochloro-
phyll to CHL in dim light, but in the presence of relatively high-intensity
visible light and air, the CHL acts as a photosensitizer leading to its own
bleaching; in addition to CHL, catalase and presumably other porphyrino-
proteins can be photooxidized, as demonstrated by Anderson and Robertson
[37] and Mitchell and Anderson [38, 39].
An interesting observation has been made recently which should help to
explain the morphological change's which occur in the chloroplasts of mutant
 IX. Function 675

plants deficient in carotenoid pigments. These changes were originally de-

scribed by Walles [40, 41] and von Wettstein [42] for sunflowers and barley.
In both cases, the albino mutant formed normal early proplastids but dis-
played abnormal organization beyond this stage. Walles [41] concluded that
the photooxidation of CHL in the chloroplasts in the albino mutant led to
the disintegration of the chloroplast grana. Similar alterations in chloroplast
structure have now been reported by Bartels et al. [ 43] and Millerd et al. [ 44].
Bartels et al. [ 43] treated wheat with the herbicide 3-amino-1,2,4-triazole (AT),
which led to the development of albino leaves with abnormal chloroplasts
when the plants were germinated in the light. In the dark, the proplastids
appeared normal. The authors also did an ultracentrifugal analysis of the
ribosomes of these plants and found that the 70S ribosome, characteristic of
chloroplasts, was absent in the light-grown leaves in the presence of AT. Using
a low-temperature mutant of maize, Millerd et al. [ 44] observed a similar
phenomenon in the light. At 15 °C, abnormal chloroplast structures were
observed, and very few 70S ribosomes were present in the chloroplasts, an
effect not obtained in the dark. Another herbicide tested by Bartels and
Hyde [ 45], 4-chloro-5-(dimethylamino )-2-(a,a,a-trifluoro-m-tolyl)-3(2H)-pyrid-
azinone, had the same effect as AT described above. In addition, Bartels and
Hyde [ 45] found that this herbicide blocked carotenoid biosynthesis almost
completely. Another effect of herbicides was reported by Merkle et al. [ 46],
who observed that paraquat (1,1'-dimethyl-4,4'-bipyridinium dichloride)
caused the photooxidation of the pigment system of beans. They suggested
that this photooxidative bleaching was due to the inhibition or destruction
of a protective system which normally prevents photooxidation. Burns et al.
[47] have since proposed that the action of a number of herbicides is to inhibit
specifically carotenoid biosynthesis in plants leading to the development of
aerobic photosensitivity, bleaching, and eventual death. They propose this as
the mechanism of action of the herbicides pyriclor (2,3,5-trichloro-4-pyridinol),
dichlormate (3,4-dichlorobenzyl methylcarbamate) and AT.
An interesting variation in plastid structure has been described by Sander
et al. [ 48] in the albescent mutant of maize. In this case, they find that the
albescent corn seedlings are very deficient in coloured carotenoids, but when
grown under low-intensity light ( <0.5 ft-c) for 5 hours, the albescent plastids
can be stabilized and will lead to green tissue on subsequent illumination.
Before this stabilization period, however, additional light will lead to bleach-
ing of the seedlings. It would appear that the stabilization requires a threshold
level of carotenoids to be effective as photoprotective agents. This conclusion
is simiMr to one reached recently by Mathews-Roth and Krinsky [49] in
studying the protective action of carotenoids in pigment mutants of S. lutea.
It would appear then that the harmful effects of light in the absence of
carotenoid pigments can lead to gross morphological changes in plastids as
well as more subtle changes in terms of the absence of the characteristic 70 S
ribosomes of chloroplasts. This may be related to the observations of LefT
and Krinsky [50], who used high-intensity visible light to develop aerobic
676 NORMAN I. KRINSKY

photosensitivity in the alga Euglena gracilis. The sensitivity was expressed not
only in terms of a lethal effect on this alga, but bleached mutants were also
formed. These mutants lacked the characteristic DNA of the W-T strain
chloroplasts.
There are other examples of lethal aerobic photosensitizations described
in algae. These include the work of Claes [51] on mutant strains of the green
alga Chlorella vulgaris and of Sager and Zalokar [52] on the pale green mutant
strain of Chlamydomonas reinhardi. In both cases, severe carotenoid deficien-
cies in these mutant strains led to death when aerobic cultures were exposed
to light. An interesting observation was made on the mutant strain of C. rein-
hardi. Chance and Strehler [53] observed an absorbancy increase at 518 nm
when the organism was illuminated under anaerobic conditions, a change
reported for many photosynthetic species. However, in the pale green mutant
strain, Chance and Sager [54] could not observe the 518 nm change upon
illumination. They concluded that the change in absorbancy was due to'-a
reaction between a carotenoid and excess oxidizing equivalents produced
either by photosynthesis or by oxygenation. Furthermore, since the mutant
strain was photosensitive and did not show the 518 nm change, they concluded
that there was some relationship between the protective function of carotenoid
pigments in algae and this absorbancy change. The question of the relationship
between carotenoid pigments and absorbancy changes in photosynthetic
organisms in the 510-520 nm range will be discussed further in Section B.2d.
There are other examples of photosensitivity in algae which are not lethal.
These have been discussed by Krinsky [10].
A very interesting example of how organisms respond to the hazard of
visible light can be demonstrated in photochromogenic bacteria. These
organisms do not produce coloured carotenoid pigments when grown in the
dark, but on exposure to light, they display carotenogenesis. Simultaneously,
they begin to produce endogenous photosensitizing pigments. Fortunately,
the quantity of light needed for the lethal photodynamic action is about ten
times as much as that needed for the stimulation of carotenogenesis, as de-
scribed by Wright and Rilling [55] and by Mathews [56]. This wide field has
been recently reviewed by Batra [57].

b) Photo protection and chromophore length

From early results, it has been apparent that not all carotenoid pigments
act as photoprotective agents, and a distinction has long been drawn between
the colourless carotenoids such as phytoene (32) and phytofluene (30) and the
more unsaturated, coloured carotenoids such as /3-carotene (3). It was again
Stanier [58] who pointed out that the photoprotection of carotenoids is
related to the extent of conjugation of the double bonds in the carotenoid
molecule. This conclusion was originally based on unpublished observations
by Cohen-Bazire [58], who found a difference in the relative stability of the
pigment systems in the green and the B-G mutant strains of Rps. spheroides
in comparison to theW-T strain. 'fhe work has been pursued by observations
 IX. Function 677

both in vivo and in vitro. In her original publication on the mutant strains of
C. vulgaris, Claes [51] found that the three photosensitive mutant strains all
lacked {3-carotene (3), and accumulated hydrocarbon carotenoids with shorter
conjugated chromophores such as phytoene (32), phytofluene (30) and (-
carotene (26). In addition, mutant strain 9a also had a hydrocarbon carotene,
called P 426, which may actually be /3-zeacarotene (9), a carotenoid with 9
conjugated double bonds. This mutant strain also accumulated some xantho-
phylls, but these were different from the xanthophylls present in the W-T
strain. Somewhat similar results were obtained by Faludi et al. [33] and Lang
et al. [59] working with carotenoid mutants of corn. Both groups worked with
a mutant strain which accumulated (-carotene (26), a compound with 7 con-
jugated double bonds, and found that these strains were photosensitive. In
addition, Lang et al. [59] worked with a mutant strain which accumulated
lycopene (19), a carotenoid with 11 conjugated double bonds, and was also
photosensitive, although less so than the (-carotene (26)-accumulating strain.
Crounse et al. [60, 61] also noted that the protective action of carotenoid in
several mutant strains of Rps. spheroides was a relative phenomenon. They
concluded that neurosporene (22), a carotenoid containing 9 conjugated double
bonds, is the least unsaturated pigment capable of protecting cells against
photooxidations.
A series of in vitro experiments involving the action of carotenoid pigments
containing different numbers of conjugated double bonds was carried out by
Claes and Nakayama [62] and Claes [63, 64]. These authors studied two
CHL-sensitized light reactions in the presence of carotenoids containing either
3, 5, 7, 9 or 11 conjugated double bonds. These reactions consisted of an
irreversible photooxidation or a reversible photoreduction of CHL. Similar
experiments were carried out using {3-carotene (3) by Krasnovskii et al. [65].
Both groups found that carotenoids containing 11 conjugated double bonds
could confer some protection against the CHL-catalysed photooxidations and
photoreductions. There was however a relationship between this protection
and the polyene chain-length as demonstrated by Claes [62-64] and by
Krasnovskii and Drozdova [66]. The latter workers observed that violaxanthin
(135), a carotenoid containing 9 conjugated double bonds, was not as effective
as lutein (73) or /3-carotene (3), carotenoids containing 10 or 11 conjugated
double bonds respectively. Claes [62-64] observed a small amount of pro-
tection with (-carotene (26), a pigment containing 7 conjugated double bonds,
but phytofluene (30) and phytoene (32) containing 5 and 3 conjugated double
bonds respectively offered no protection. With respect to photoreduction of
CHL, the results were somewhat different. The carotenoid pigments were
more effective in inhibiting this reaction, for there was not only complete
inhibition with.lycopene (19) and /3-carotene (3), both containing 11 conjugated
double bonds, and neurosporene (22), containing 9 double bonds, but also (-
carotene (26). There even was some protection with phytofluene (30) but none
with phytoene (32). As will be discussed below in Section B.l c, Foote et al.
[67] have found a parallel relationship in their studies of the quenching of
678 NORMAN I. KRINSKY

Table. The relationship between the protective action of carotenoid pigments and their conjugated
chromophore length

Year Authors Conjugated chromophore length Reference

Effective Partially Ineffective

effective

Photooxidations
1959 Claes and Nakayama 9, 11 7 5 [62]
1960 Claes 9, 11 7 5 [63]
1961 Claes 9, 11 7 5 [64]

Photoreductions
1960 Claes 7, 9, 11 5 [63]
1961 Claes 7, 9, 11 5 [64]
1961 Krasnovskii and Drozdova 10, 11 9 [66]

Photoisomerizations
1959 Claes and Nakayama 9, 11 [97]
1960 Claes 9, 11 [63]
1961 Claes 7, 9, 11 [64]
1970 Foote eta!. 11 [95]

10 2quenching
1970 Foote eta!. 9, 11 [67]

Biological protection
1954 Claes 11 3, 5, 7, 9 [51]
1959 Stanier 11 9 3 [58]
1960 Faludi eta!. 7 [33]
1963 Crounse et a!. 9 7 [60]
1963 Crounse et a!. 9 [61]
1970 Mathews-Roth and Krinsky 9 [49]
1970 Mathews-Roth and Krinsky 9 8 [68]

singlet excited oxygen by carotenoid pigments. They worked with pigments

containing 5, 7, 9 or 11 conjugated double bonds and found that the effect
of the pigments containing 5 and 7 conjugated double bonds was very small
in comparison with those with 9 and 11 conjugated double bonds respectively.
Most naturally-occurring carotenoid pigments contain odd numbers of
conjugated double bonds. Recently, Mathews-Roth and Krinsky [68] have
isolated a mutant strain, mutant 7, of S. lutea, in which the major carotenoid
pigments have an absorption spectrum very similar to that of oc-zeacarotene
(12), with maxima at 448, 422 and 398 nm. Although the pigment has not
been identified, the evidence indicates that it is a conjugated octaene. Using
the toluidine blue-sensitized photo killing assay described earlier [ 49], the
authors find that the killing rate of mutant 7 is the same as that of mutant 93 a,
a colourless mutant strain. From• these observations, they concluded that a
 IX. Function 679

minimum of 9 conjugated double bonds is necessary to confer photoprotection

in vivo.

c) Mechanisms of photoprotection
Before discussing the mechanisms of photoprotection, it must be re-
emphasized that carotenoid pigments are only effective in those systems which
have a true photodynamic action as defined by Blum [13]. This requires the
simultaneous interaction of three components: visible light, a photosensitizing
dye to absorb it, and 0 2 . Unfortunately, in many cases cited in the literature
as photosensitizations it has not been documented that either visible light
(400-700 nm) or 0 2 is really required for the effect. Both Kunisawa and
Stanier [19] and Mathews and Krinsky [69] have demonstrated that the
presence or absence of coloured carotenoid pigments have no effect on the
viability of bacterial strains exposed to ultraviolet light. In addition, Mathews
and Krinsky [69] studied the effects of both X-irradiation and gamma-ray
irradiation on pigment mutants of radiation-resistant bacteria. Again, the
absence or presence of carotenoid pigments had no effect on the viability of
these organisms. Using the photosensitizer 8-methoxypsoralen, which absorbs
long-wave ultraviolet light, Mathews [70] found that, in terms of sensitiza-
tion, there was no difference between pigmented and non-pigmented cells, and
in addition, the presence of 0 2 instead of being an absolute requirement
inhibited the action of the photosensitizer. In vivo, both Forssberg et al. [71]
and Bamji [72] investigated the effect of carotenoid pigments on the survival
and physiological changes in mice and rats exposed to whole body X-irradia-
tion. In both reports, the effects were quite marginal.
Working with pigmented and colourless strains of the yeast Rhodotorula
glutinus, Maxwell et al. [73] and Chichester and Maxwell [74] presented
evidence that carotenoid pigments do not protect the cells against lethal
photosensitizations. It must be pointed out however, that the action spectrum
for this phenomenon shows a maximum in the ultraviolet with only a small
effect at 410 nm. Thus, it is not at all clear that this type of photosensitization
should be considered a photodynamic action.
Having thus limited the involvement of carotenoid pigments to protecting
reactions catalysed by visible light and 0 2 in the presence of a suitable photo-
sensitizing dye, we can look at the following mechanisms proposed by Krinsky
[10] in 1968:
1) An absorbing system in the cell envelope to filter out potentially harm-
ful light.
2) Systems to interact with and quench photosensitizer triplet states.
3) Systems to serve as preferred substrates for photosensitized oxidations.
4) Systems to stabilize membranes or repair damaged membranes.
In addition; numerous chemo-autotrophic bacteria, which lead either
obligatory anaerobic or dark existences, would not require this type of pro-
tective mechanism and, in fact, do not synthesize carotenoid pigments.
680 NORMAN I. KRINSKY

All four mechanisms were discussed in great detail by Krinsky [10]. For
example, the idea that carotenoids might serve as filter pigments was pro-
posed by Goldstrohm and Lilly [75], working with the fungus Dacryopinax
spathularia. Vail and Lilly [76] have now demonstrated that the carotenoid
pigments in this fungus are present in the cell wall fraction where they could
act as a light filter. It must be pointed out that in order for carotenoids to
serve in this way, they should be located between the source of light and the
photosensitizer. In almost all cases, however, the exact location of the endo-
genous photosensitizing pigment is not known. In addition, this concept
implies that the carotenoid pigments absorb light of the same wavelength
used to excite the photosensitizer. This seems to preclude the possibility that
carotenoids function by this mechanism in photosynthetic organisms where
the photosensitizer (CHL or BCHL) absorbs at wavelengths considerably
longer than the absorption peaks of the carotenoid pigments. In the case of
non-photosynthetic organisms, the data of Mathews [56] and Burchard et a'l·
[77] indicate that a porphyrin-type compound is the photosensitizer, and even
these compounds show absorption maxima at wavelengths beyond those of
the carotenoid pigments.
Until 1968, the interaction of carotenoid pigments with the triplet state
of CHL, first described by Fujimori and Livingston [78], had always been
considered an important means whereby carotenoid pigments could protect
photosynthetic cells from lethal photosensitization. The process has been dis-
cussed in detail by Gaffron [79] and by Clayton [15]. They argue that photo-
synthesis is normally carried out by the singlet excited state of CHL. In the
presence of excess quanta however, some of the excited CHL can be converted
to the metastable triplet excited state, which can then proceed to carry out
abnormal photoreactions, including photosensitized oxidations. The quench-
ing by carotenoids of the triplet state of CHL, first described by Fujimori and
Livingston [78] and further documented by Fujimori and Tavla [80], Chessin
et al. [81] and Mathis [82, 83], would then prevent the accumulation of the
potentially harmful CHL triplet state. As our understanding of photosensitized
oxidations has advanced, the observations cited above remain valid, although
they cannot now be considered to play as important a role as was previously
thought.
Until very recently, the following mechanism advanced by Schenck [84]
and others had been used to explain the photosensitized oxidations catalysed
by a suitable sensitizer (SENS), 1SENS being the singlet excited sensitizer and
3 SENS the triplet excited sensitizer:

SENS~ 1 SENS (1)

1 SENS-----+ 3 SENS (2)
3 SENS+0 2 -----+ SENS-0 2 (3)
SENS-0 2 ...PA----+A0 2 +SENS. (4)
 IX. Function 681

In this case the sensitizer-oxygen complex reacts with the acceptor (A) to
give a peroxide (A0 2 ), and the sensitizer is regenerated. Another mechanism
had been proposed earlier by Kautsky [85, 86] but it had been supplanted by
the Schenck hypothesis. Kautsky [85, 86] suggested that the triplet sensitizer
reacted with oxygen to generate the singlet excited form of oxygen eo2) and
regenerate the original sensitizer. He then proposed that 10 2 reacted with the
acceptor to yield the oxidized product. This is shown below, in which 3 0 2
represents the triplet ground state of oxygen:
3 SENS+ 3 0 ~SENS+ 1 0 2 (5)
2

102 +A~A0 2 . (6)

In 1964, Foote and Wexler [87] and Corey and Taylor [88] simultaneously
presented evidence that 10 2 can carry out reactions identical with photo-
sensitized oxidations. Foote and Wexler [89] therefore suggested that 10 2
was the intermediate in photosensitized oxidations, in support of the proposal
initially put forth by Kautsky [85, 86]. The work in this field has progressed
very rapidly due primarily to the work of Foote and his associates. Two
excellent reviews on the developments in this field appeared in 1968 by Foote
[90, 91], and more recently the chemical and biological aspects have been
reviewed by Wilson and Hastings [92]. The importance of singlet oxygen in
catalysing numerous chemical oxygenations has been well documented by
Foote [90]. The biological significance still remains to be assessed and docu-
mented; it appears likely, however, that 10 2 may be involved in various
oxygenase reactions in biological systems.
The historical development of the concepts involved in considering 10 2
as an intermediate in photosensitized oxidations and other reactions has been
recently reviewed by both Wilson and Hastings [92] and Khan and Kasha
[93]. In both articles, the importance of the two lowest-lying excited states
of 0 2 , the 1L1g state and the 1~:-+ g state is very well developed. We shall be
concerned here only with the involvement of carotenoid pigments with 10 2 .
In 1968, the first report of the interaction between carotenoids and 1 0 2
appeared by Foote and Denny [94]. These authors argued that the protective
action of carotenoid pigments against photosensitized oxidations could not
be adequately explained by the ability of carotenoids to quench triplet sensiti-
zers, inasmuch as this reaction is diffusion-controlled and would compete with
the similarly controlled quenching of triplet sensitizers by 0 2 • Since both
reactions occur at the same rate, the carotenoids could only protect if their
local cotJ.centration greatly exceeded that of 0 2 • The alternative possibility,
that the carotel).oid interacts directly with 10 2 was proposed and experimen-
tally verified. They showed that a low concentration (10- 4 M) of {3-carotene
(3) was effective in inhibiting the methylene blue-sensitized oxidation of 2-
methyl-2-pentene. At this concentration, 95% of the photooxidation was
inhibited. They proposed a direct transfer of energy from 10 2 to {3-carotene (3)
to yield the carotenoid triplet ecAR) plus ground-state triplet oxygen as
682 NORMAN I. KRINSKY

shown below, where 1CAR represents the singlet ground state carotenoid.
10 2 + 1CAR_. 3 CAR+ 3 0 2 • (7)
This could only occur if the triplet energy level of P-carotene (3) was near or
below that of the 1L1g state of 0 2 , 22.5 kcal. Unfortunately, the exact values
of the triplet states of P-carotene (3) or other carotenoid pigments are still
unknown. Even without an exact understanding of the mechanism of the
phenomenon, Foote and his collaborators [67, 94-96] have now adequately
demonstrated the ability of a variety of carotenoid pigments to quench 10 2
and therefore to break the series of reactions which would normally lead to
photosensitized oxidations. Foote and his collaborators have made two
important observations that should help us to understand the mechanism.
The first is that the quenching by carotenoids appears to be primarily a trans-
fer of excitation energy, and not an oxidation of the carotenoid pigment.
Foote et al. [96] have presented evidence that only one out of a thousand carg:
tenoid molecules involved in the quenching of 10 2 undergoes a chemical oxida-
tion. The dissipation of the excitation energy acquired by the carotenoids in the
quenching process is presumably related to their second observation. It has
been known for some time that carotenoid pigments can be isomerized fol-
lowing light absorption by CHL pigments. This has been well documented
by the work of Claes and Nakayama [97] and Claes [63] in which it was
found that the isomerization occurs only in the direction of cis__. trans. Foote
et al. [95] confirm this observation and report that the isomerization is sens-
itized by 10 2 , as well as by triplet sensitizers such as methylene blue, with
extremely high efficiency. In fact, they calculate that each 10 2 molecule pro-
duced in a dye-sensitized reaction isomerizes only slightly less than one mole-
cule of 15,15'-cis-P-carotene to all-trans-P-carotene. Similar results were ob-
tained when the 10 2 was generated chemically from Na0Cl-H 2 0 2 • They
propose that the product formed from the interaction of the carotene with
either the triplet sensitizer or 10 2 is 3 CAR, which then collapses largely to
the all-trans-P-carotene, whether 3 CAR is formed from the cis isomer or the
trans isomer.
The biological importance of 10 2 quenching by carotenoids is greatly
strengthened by an experiment carried out by Foote et al. [67] in which they
show a very close relationship between the ability of carotenoid pigments to
quench 10 2 in vitro and the ability of carotenoid pigments to protect CHL a
from a photosensitized oxidation (Fig. 1). The authors measured the rate of
quenching by carotenoids of dye-sensitized photooxidations and compared it
to the results ofClaes and her collaborators. As Claes [63] and Claes and Naka-
yama [97] had shown earlier, the ability of carotenoid pigments to protect CHL
a from photo bleaching was a function of the number of conjugated double bonds
in the polyene chain. Similar results were obtained by Foote et al. [67] in
determining the quenching constants for carotenoid pigments with different
numbers of conjugated double bonds. The observations can now be correlated
with those reported above in Section B.1 in which the in vivo effectiveness of
 IX. Function 683

~
% iii...
~-
50
'"g
g
40 0
(")

6·
30 ::s
e.,
n
20 :=
t"'
Q

10 'f
i,
7 9 11 0
No. conj. C=C in polyene chain
Fig. 1.
10 2 quenching rates (k0 , o-o) and protective action against photo bleaching of CHL a
(lr---A from Claes [63] and Claes and Nakayama [62]) as a function of length of conjugated
system (from Foote eta!. [67])

carotenoid pigments in protecting cells against lethal photosensitizations was

a direct function of the number of conjugated double bonds. The data brought
together in the table would indicate that the minimum number of conjugated
double bonds in carotenoid pigments able to protect against photosensitized
oxidations, and therefore quenching 10 2 , must be nine. The lack of protection
shown by a carotenoid pigment presumably containing 8 conjugated double
bonds as described by Mathews-Roth and Krinsky [68] may be related to
the triplet energy levels of the carotenoid pigments. As mentioned above, the
triplet energy level of P-carotene (3) is not known. Since P-carotene (3) quenches
CHL a fluorescence (at a triplet energy level of 29 kcal) it would seem that
the triplet energy level of P-carotene (3) must be below 29 kcal and may well
lie in the range limit for the 1LI g state of oxygen (22 kcal). In addition, the
dependence on the length of the conjugated system would be understood in
terms of the increase in triplet energy expected as the length of the conjugated
system decreases. Presumably therefore, carotenoid pigments with 8 con-
jugated double bonds would have triplet energies above 22 kcal, whereas
those containing 9 conjugated double bonds or more would have triplet
energies below this level.
The mechanisms whereby carotenoid pigments can protect cells against
photosensitized oxidations are depicted in Fig. 2, modified after Krinsky [10]
and Foote et al. [96]. In this scheme, the 3 Roman numerals indicate the fol-
lowing three mechanisms:
I. In this case 10 2 , the actual causative agent of photosensitized oxidations,
can interact with a suitable acceptor to form an oxidized product which
presumably then· goes on to lead to photodynamic action. Ifthe acceptor, A,
were a carotenoid pigment, we would form an oxidized carotenoid, and this
could then be interpreted in terms of the preferred substrate hypothesis of
684 NORMAN I. KRINSKY

Calvin [98] and Sistrom et al. [16]. Krinsky [17] has proposed that this
mechanism, accompanied by a dark regeneration of the oxidized carotenoid
to its original acceptor state, may function in algae such as E. gracilis. An
interesting corollary to this suggestion is that this reaction may be the natural
oxidative process which leads to the development of allenic carotenoids. Both
Foote and Brenner [99] and Mousseron-Canet et al. [100-102] have ob-
served the formation of allenic products following the exposure to 10 2 of
compounds closely related to carotenoids. The allenic moieties formed are
identical to the allenic portions of such widely occurring carotenoids as fuco-
xanthin (190) and neoxanthin (122). In addition, Mousseron-Canet et al. [100]
have proposed that IX-carotene (5) may be, via 10 2 oxidation, a precursor of
the naturally occurring 4-oxygenated carotenoids such as isocryptoxanthin
(40), canthaxanthin (193) and astaxanthin (203). Foote et al. [96] rule out this
mechanism as a major pathway whereby carotenoid pigments can protect
cells because as already noted they find that for every thousand molecules
of P-carotene (3) involved in quenching 10 2 (mechanism III) only one is
oxidized. This mechanism may still offer a valid explanation for the formation
of the allenic carotenoids, which are so widely distributed in nature.
Photodynamic
~Ao2-- action

rctRJC~R-02
02
1 CAR

III

Photosynthesis
Fig. 2. The three mechanisms whereby carotenoid pigments (CAR) protect cells against photo-
sensitized oxidation. Light (wavy line) excites the photosensitizer (SENS), which in the case of
photosynthetic organisms is deactivated primarily through the process of photosynthesis. Those
singlet excited sensitizers (1SENS) molecules which are not deactivated by photosynthesis can be
converted to the long-lived triplet excited sensitizer esENS). These molecules can react with
ground state oxygen eo2) to form singlet excited oxygen (102), which can react with a suitable
receptor molecule (A) to form an oxidized product (A0 2 ) capable of leading to a photodynamic
action. In mechanism I, the CAR act as preferred substrates and can be oxidized to CAR-0 2 ,
which may be regenerated by a dark, enzymic process. In mechanism II, CAR can quench 3 SENS,
presumably going to an excited state, CAR*, and then being deactivated to CAR and energy.
In mechanism III, CAR can quench 10 2 forming an excited state, CAR*, which can be again
deactivated to CAR and energy.
In addition, the accessory pigment function of carotenoid is depicted by the direct excitation
of CAR by light and the transfer of this radiant energy to the SENS.
 IX. Function 685

II. This mechanism, the quenching of the triplet sensitizer by carotenoids,

was first described by Fujimori and Livingston [78] and has been amply con-
firmed. It is difficult to evaluate its importance in photosynthetic organisms,
for the reaction between the sensitizer and the carotenoid is diffusion-limited
and would therefore compete with 10 2 formation from the triplet sensitizer.
Since most endogenous photosensitizers and the protective carotenoid pig-
ments coexist in membrane structures (CHL-carotenoids in photosynthetic
organisms; porphyrin-carotenoids in bacterial membranes), it is impossible
to discount the importance of this mechanism as a protective device. So far,
the experiments which have demonstrated the more rapid rate of quenching
by mechanism III than by mechanism II have been done in free solution, an
environment which in no way can be compared to a naturally-occurring bio-
logical membrane, where the carotenoids and photosensitizers may be present
in a manner excluding diffusion-controlled reaction rates. Evidence of this
nature has been presented by Mathis [83] in his studies on the formation of
3 CAR using CHL a as the photosensitizer. In an organic solvent, the energy

transfer was diffusion-controlled, but if he mixed his carotenoid and CHL a

in a digitonin micelle, the energy transfer was greatly accelerated. This type
of micelle may only be an approximation to the relationship that would exist
between carotenoids and photosensitizers in membranes.
III. It should be obvious by now that the quenching of 10 2 by carotenoids
must play a prominent role in explaining the protective action of carotenoid
pigments. As pointed out by Foote et al. [96] the ratio of the rate constants
for the quenching of 10 2 by carotenoids in comparison to the oxidation of
an acceptor is approximately 1.5 x 104 . This would seem to ensure that any
10 2 generated by the triplet sensitizer could not readily go on to produce a

photodynamic action in the presence of carotenoid pigments. What remains

is to gain further understanding of the fate of the energy transferred from 10 2
to carotenoid. Although the scheme presented in Fig. 2 and comparable ones
developed by Foote et al. [95, 96] and Wilson and Hastings [92] indicate
that 3 CAR is formed, there is as yet no direct evidence for this. It would seem
that the spectrophotometric observations of Chessin et al. [81] and Mathis
[82, 83] should be applicable to the experimental systems used by Foote in
observing carotenoid quenching. Using these techniques, it should be possible
to determine whether in fact 3 CAR is formed and then how the energy is
dissipated when it is reconverted to ground state carotenoid, either as heat
or as the emission of light.
Before leaving the field of photoprotection, it is imperative to discuss the
potential use of this information. As mentioned earlier [ 45, 46] it appears that
some herbicid~s may act by blocking the synthesis of carotenoids and thus
interfering with the photoprotective action. However, experience seems to
indicate that the temporary gains which can be harvested from the use of
herbicides frequently lead to long-term problems. On a more productive note
the experiments of Mathews [103] indicated that the addition of P-carotene
(3) to experimental animals supplemented with photosensitizers could lead to
686 NORMAN I. KRINSKY

a marked protection of these animals when they were exposed to light. At that
time she suggested that carotenoids might prove useful in human patients
suffering from light-sensitive diseases. This prediction has been fulfilled in an
exciting publication of Mathews-Roth et al. [104] which has just appeared.
Three patients suffering from a light-sensitive erythropoietic protoporphyria
were treated with large amounts of f:J-carotene (3) in a water-soluble beadlet
form. These patients received up to 90 mg/day and all reported the develop-
ment of tolerance to sunlight and showed increased tolerance to test lights.
Although this sample was small, it did indicate that this important function
of carotenoids, observed and studied for so long in bacteria and plants, could
indeed have direct applicability to man. We can now anticipate that these
results of Mathews-Roth et al. [104] will lead to further experimentation in
the treatment of light-sensitive diseases in man.

2. Photosynthesis
The list of possible functions of carotenoid pigments in photosynthesis is
very large, and in this section we shall attempt to concentrate on those func-
tions which appear to be well documented. The more speculative functions
will be mentioned, but space does not permit a full discussion of them.

a) Accessory pigments
The ability of carotenoid pigments to function as accessory pigments in
photosynthesis was first observed by Engelmann [105, 106] in 1883-1884.
These elegant experiments were carried out by projecting a micro-spectrum
across algal strands or filaments, and the production of 0 2 was measured by
the accumulation of motile bacteria. All of the technical improvements which
have taken place over the last 90 years have served to verify Engelmann's
original observations that the accessory pigments, such as carotenoids, can
participate in photosynthesis. Forty years later, Warburg and Negelein [107]
made similar observations when they determined the quantum yield of photo-
synthesis in Chlorella. Rabinowitch [108], in reviewing the role of carotenoids
and other compounds as accessory pigments in photosynthesis, interpreted
the results of Warburg and Negelein [107] as indicating that light absorbed
by carotenoids can be active in photosynthesis, although somewhat less so
than that absorbed by CHL a. By the early 1940's, Dutton and Manning [109]
and Emerson and Lewis [110, 111] were observing that the activities of carote-
noids in photosynthesis were not always comparable and varied from the
situation in diatoms, where Dutton and Manning [109] found that fuca-
xanthin (190) was as effective for photosynthesis as CHL whereas carotenoid
absorption in Chlorella was only partially active [111] and the carotenoids
in blue-green algae were very ineffective [110].
A similar situation was observed in studies on photosynthetic bacteria
with respect to the differences in the efficiency of carotenoids functioning as
accessory pigments. This work wa!i also initiated by Engelmann [112] and was
 IX. Function 687

revitalized by the studies of Thomas [113] and Duysens [114, 115]. The entire
field of accessory pigments has been reviewed several times [4, 11, 30, 108, 116,
117]. The reader is referred to the excellent review by Blinks [30], who summa-
rized .the available data up to 1964 including presentations of Engelmann's
[105, 106] original data and the data obtained by Blinks and others using the
polarographic method introduced by Haxo and Blinks [118].

rx) Mechanisms of energy transfer. The ability of carotenoid pigments to

elicit a photosynthetic response is presumably due to their ability to transfer
radiant energy to CHL. The most convenient means of evaluating this process
is to look at the sensitized fluorescence of CHL when carotenoids are illum-
inated. This was first carried out successfully in vivo by Dutton et al. [119],
who observed that the fluorescense intensity of the diatom N itschia closterium
was as great when the cells were illuminated with green light absorbed primarily
by fucoxanthin (190) as with red light absorbed primarily by CHL. This
efficient transfer of energy was confirmed by Wassink and Kersten [120] also
working with diatoms. It remained for Arnold and Oppenheimer [121] to
offer the physical explanation for this type of energy transfer. They concluded
that the high efficiency of transfer could not be attributed to molecular colli-
sion or to emission and reabsorption, but rather to internal conversion or
resonance transfer. This process, which involves the resonance transfer of
energy from one oscillator to another in resonance with it, was based on the
extremely high efficiency (97 %) of transfer of energy from phycocyanin to
CHL in blue-green algae. The assumption that this identical process obtained
with carotenoids has been supported by the studies of Teale [122], who
measured the carotenoid-sensitized fluorescence of CHL in detergent micelles.

f3) Energy transfer in developing chloroplasts. An early example of the ability

of carotenoids to function as accessory pigments was reported by Chen [123]
in 1952 in which he found very good agreement between the action spectrum for
the Hill reaction in isolated chloroplasts of Swiss chard and the photosynthetic
action spectrum of the intact leaf. Further investigation by Jagendorf et al.
[124] and Black et al. [125] indicated that the action spectrum for photosyn-
thetic phosphorylation and reduced triphosphopyridine nucleotide (NADPH)
formation were identical with those for the intact leaf. However, a discrepancy
was observed by Butler [126]. He was studying the action spectrum of CHL
fluorescence in bean leaves during development and observed that the reso-
nance transfer of energy between carotenoids and CHL did not occur imme-
diately tlfter the photoconversion of PCHL to CHL, but could be observed
after about one. hour. This observation has been effectively pursued by Goed-
heer leading to some extremely interesting results. In 1961, shortly after
Butler's [126] original observations, Goedheer [127] also reported that there
was no measurable energy transfer between carotenoids and CHL in the 'post-
etiolated' state of greening bean leaves. At that time, he suggested that the
transfer might require CHL b, which is not present in the 'post-etiolated'
688 NORMAN I. KRINSKY

leaves, but Losev and Gurinovich [128] have now shown, working with the
0 2 BX mutant strain of E. gracilis which lacks most of its CHL b that they
can still obtain a good transfer of energy from carotenoids to CHL a. More
recently, Goedheer [129] has studied the fluorescence action spectra of green-
ing bean leaves and algae at both room and liquid nitrogen temperatures. In
the case of blue-green algae and red algae, he found negligible transfer of
energy from carotenoids to CHL at room temperature and only barely dis-
cernible peaks in the fluorescence action spectra in the carotenoid region at
liquid nitrogen temperatures. In the case of green algae, he was able to demon-
strate energy transfer very readily. On extracting these algae with methanol
and petroleum ether, he found that most of the carotenoid from the blue-
green and red algae was hypophasic, indicating the presence of xanthophylls,
whereas the major part of the pigment from the green algae was epiphasic,
indicating the presence of carotenes. Based on this, he assumed that carotenes
can transfer to CHL a with a high efficiency, whereas the xanthophylls are
negligible with respect to transfer. This would therefore be analogous to the
situation that obtains in greening bean leaves, where the 15-minute sample,
which does not show carotenoid transfer, contains primarily xanthophylls,
whereas the 4-hour sample, at which time transfer is observed, contains
relatively large amounts of carotenes [130]. As he has refined his procedures,
Goedheer [131] has most recently presented evidence that the trace of carote-
noid absorption in the low temperature fluorescence action spectra in blue-
green and red algae can be removed by extracting the cells with petroleum
ether. This extract removes only P-carotene (3) without extracting the xantho-
phylls. The same effect can be observed in both green algae and in etiolated
bean leaves 3.5 hours after exposure to light. In both cases, transfer occurs
from carotenoid to CHL a, and this transfer can be removed by a petroleum
ether extraction which specifically removes carotenes and not xanthophylls.
He therefore concludes that the light absorbed by /3-carotene (3) can be trans-
ferred at approximately 100% efficiency to CHL a of both photosystem I or
photosystem II depending on the organism, but that light absorbed by the
xanthophylls cannot be transferred to CHL. This inability of xanthophylls to
transfer to CHL may be due either to a difference in the chemistry of these
compounds from the carotenes, or it may be due to an actual physical separa-
tion in the photosynthetic membranes. This explanation would explain the
report ofTeale [122], who found that {3-carotene (3), lutein (73) or fucoxanthinol
(189) can all transfer excitation energy to CHL a when they are in a detergent
micelle, when the pigments are at 0.1 M and at a calculated average inter-
molecular spacing of about 10 A. This is the same concentration estimated
by Losev and Gurinovich [128] of CHL in greening plants. However these
authors raise a point that it is difficult to understand the transfer of energy
from carotenoids to CHL by an inductive resonance mechanism between
singlet excited carotenoid and CHL. This is based on the fact that CHL does
not have strong absorption in the green region of the spectrum, and carotenoids
have little or no fluorescence. They suggest that the energy transfer may be
 IX. Function 689

from carotenoids to CHL when the contact between the molecules permits
an exchange-resonance interaction to develop.

y) Fluorescence quenching. A somewhat related topic deals with the ability

of carotenoid pigment to quench fluorescence in photosynthetic bacteria.
Although this appears to be another 'blue-light' effect (Section B.3) it will be
discussed here since it seems to involve a relationship with the ability to
transfer excitation energy from carotenoids to CHL. The phenomenon, the
quenching of fluorescence from BCHL, was initially described by Bril [132]
and was studied more extensively by Goedheer [133] and by Mayne [134].
Bril [132] was primarily concerned with the oxidative photobleaching of
BCHL in normal and carotenoid-depleted Chromatium and suggested that the
carotenoid pigments might protect BCHL against an oxidative photobleach-
ing by undergoing cis-trans isomerizations which could lead to structural
changes in the chromatophore and thus stabilize the BCHL. Goedheer [133],
on the other hand, was much more concerned about the mechanism of the
fluorescence decline in purple bacteria. He observed that the initial decline
in fluorescence of either R. rubrum or R. molischianum was associated with
light absorbed by carotenoid pigments. He also studied Rps. spheroides and
found a much smaller quenching effect. Inasmuch as he had demonstrated
earlier [135] that light absorbed by carotenoids was very efficiently transferred
to BCHL in Rps. spheroides (90 %) and only poorly transferred in R. rubrum
(25 %), he suggested that the quenching was caused by the fraction of quanta
not transferred to BCHL. Furthermore, he observed that the fluorescence was
also quenched by the addition of oxidants such as potassium ferricyanide and
hydrogen peroxide. He concluded that the light-induced fluorescence quench-
ing was due to a photooxidation reaction resulting from carotenoid absorp-
tion. At the same time, Mayne [134] observed very similar results on fluores-
cence quenching in photosynthetic bacteria, but arrived at a different inter-
pretation of this data. In Chromatium chromatophores he found that blue
light led to a decrease in BCHL fluorescence, and that this was a photooxidative
process. He also looked at the W-T strain and the B-G mutant strain of Rps.
spheroides and found a similar effect with blue light quenching BCHL fluores-
cence in both strains. However, in the B-G mutant strain, he found that red
light also decreased fluorescence and concluded that the carotenoids in the
W-T strain were preventing the formation of a quencher upon illumination
with red light. Mayne [134] interpreted these results to indicate that carote-
noids can act as protective pigments in photosynthetic bacteria by preventing
the formation of a fluorescence quencher. A directly opposed point of view
has now been presented by Goedheer and van der Tuin [136]. Having deter-
mined on action spectrum for the inhibition of BCHL fluorescence for both
R. molischianum and R. rubrum they find a much greater effect of 566-nm light
in R. rubrum thari in R. molischianum, which indicates that spirilloxanthin (108),
the major carotenoid pigment absorbing at 566 nm, is active in quenching
BCHL fluorescence in R. rubrum, although it has virtually no effect in trans-

Carotenmds 44
690 NORMAN I. KRINSKY

ferring energy to BCHL. They therefore suggest that there may be two carote-
noid systems operating, one which transfers light energy to BCHL with
approximately 100% efficiency, and the other system, though extremely ineffi-
cient in transfer, being capable ofleading to reactions which result in a decrease
in BCHL fluorescence. They suggest that their system II carotenoids photo-
oxidize a compound which is capable of quenching. They argue that the
fluorescence intensity is a measure of the mean life of the BCHL singlet excited
state, which can be readily photooxidized, and that the quenching of fluores-
cence is equivalent to stabilizing BCHL. They conclude that their system II
carotenoids function to protect the fluorescing species of BCHL.
Inasmuch as Goedheer [129] has already demonstrated that only carotenes
transfer their excitation energy to CHL in algae, whereas xanthophylls do not,
it might be assumed that in the photosynthetic bacteria the system II carote-
noids described may be xanthophylls, incapable of transferring radiant ener_g.y
to BCHL.

b) 0 2 evolution and transport

With the discovery, characterization and widespread distribution of epoxide
carotenoids, particularly in photosynthetic organisms, Karrer [137] suggested
that these epoxide carotenoids might function as intermediates in 0 2 trans-
port. Shortly thereafter, Sapozhnikov and Lopatkin [138] reported that inter-
conversion could be demonstrated for carotenes and xanthophylls in photo-
synthetic tissues. Such interconversion served as a stimulus for the suggestion
of Dorough and Calvin [139] that epoxide carotenoids might generate either
0 2 or H 2 0 2 during photosynthesis and in either case serve as intermediates
in the pathway of 0 2 evolution. Although the data obtained by Dorough and
Calvin [139] were inconclusive, the idea was intriguing and was pursued fur-
ther by a number of investigators. At the same time Cholnoky et al. [140], on
the basis of observations made with green leaves or ripening fruit, suggested
that the epoxide carotenoids function as intermediates in 0 2 transport. Sup-
port for the idea that epoxide carotenoids are involved in 0 2 liberation in
photosynthetic organisms comes primarily from the laboratories of Sapozhni-
kov and Saakov. Both quantitative measurements of carotenoids under dif-
ferent physiological conditions and the use of isotopes have convinced Sapozh-
nikov et al. [141] and Saakov [142, 143] that epoxide carotenoids are involved
in this process whereas very similar types of experiments carried out by Yama-
moto et al. [144, 145] and Shneour and Calvin [146] have convinced the
latter workers that carotenoids cannot function as important intermediates
in either 0 2 liberation or transport. The available evidence would seem to
indicate that carotenoid pigments do not play an important role in this pro-
cess. However, the reader is referred to a recent review by Sapozhnikov [147]
for his views on the role of carotenoid pigments in photosynthesis. Another
suggestion for the functional role of carotenoids in 0 2 evolution in photo-
synthesis has been investigated by Komissarov [148], and he has been attempt-
ing to demonstrate that a photo battery composed of alternate layers of CHL
 IX. Function 691

and carotene would be capable of decomposing water into 0 2 under the

influence of light. So far he has not substantiated this hypothesis.
c) Cofactors of photosynthetic reactions
Although there have been many suggestions that carotenoids serve as
cofactors for various photosynthetic reactions, none of them appear to be
valid. The ubiquity of carotenoids in photosynthetic tissues favoured the idea,
but difficulty was met in attempting to test the hypothesis. Such a task usually
requires much more imagination than does the formulation of the original
hypothesis. So, for example, Warburg and his collaborators [149-152] pos-
tulated the existence of a carotenoid-containing enzyme involved in enhancing
the quantum efficiency of photosynthesis when the system was illuminated
with blue light. This has not been confirmed. Kahn and his collaborators
[153-155] have suggested that a specific carotene-protein was necessary for
CHL-protein complexes to carry out various photoreductions. At one time,
they claimed a 200-300% increase in NADP reduction upon the addition of
naturally extracted carotenoids. This work has not been pursued.
On a more positive note, the original observations of Bishop [156] and
Lynch and French [157] in which it was suggested that the removal of P-
carotene (3) led to inactivation of the Hill reaction, helped to show the impor-
tance of quinones in electron transport in photosynthetic organisms [158].
Recently, Bishop [159] has demonstrated that a carotenoid-deficient strain
of Scenedesmus obliquus cannot carry out the reactions associated with photo-
system II, whereas the reactions associated with photosystems I are unaffected.
This might indicate a specific role for carotenoid pigments in photosystem II,
and the report should be followed up.
A series of publications by Lundegardh [160-166] makes a number of
claims with respect to the role of carotenoids in photosynthesis. These include
suggestions that illumination of chloroplasts results in the oxidation of carote-
noids to xanthophylls [160], that blue light leads to the reduction of xantho-
phylls to carotenoids, and red light reverses this reaction, and-in the same
publication-that a mixture of ferredoxin and P-carotene (3), when illuminated
with blue light under nitrogen, results in the reduction of ferredoxin and the
oxidation of the carotenoid [161]. In addition, he claims that a white flash
will lead to the reduction of xanthophylls to carotenes [162]. In the same
year, he repeats that he has demonstrated the reduction of ferredoxin by P-
carotene (3) and claims that blue light can both reduce xanthophylls to caro-
tenes and oxidize carotenes to xanthophylls. In this mixture of ferredoxin and
P-carotene (3), he finds that the difference spectrum following illumination
yields a band at 340 nm due to the isomerization and development of a cis
peak of the carotenoid [163]. In a subsequent publication in that year, he
shows that a mixture of ferredoxin, NADP and P-carotene (3) results in the
formation of a peak at 340 nm, which he assigns to the reduced form of NADP,
and concludes that the light absorbed by carotenoids results in the reduction
of NADP [164]. In 1967, he proposes a new photosynthetic system, photo-
692 NORMAN I. KRINSKY

system III, which involves the transfer of electrons from photostimulated

carotenoids to NADP and claims that the mixture of ferredoxin, NADP and
/3-carotene (3) results in the reduction of ferredoxin as demonstrated by the
difference spectrum obtained between 400-500 nm. This difference spectrum
shows no change for the carotenoid which might be expected to occur if it
were being oxidized in the process [165]. Most recently, he has continued to
advocate that the reductive reaction of photosynthesis is initiated by light
absorbed by /3-carotene (3), which transfers electrons to ferredoxin and then
to NADP. In addition, he now finds that light absorbed directly by cyto-
chrome f can be utilized for photosynthesis, which is the first claim for direct
photochemical involvement of cytochromes in photosynthesis [166]. It is
extremely difficult to accept the interpretations of these observations, partic-
ularly when there seems to be direct contradiction in some of the results. One
must suspect that those observations of Lundegardh which truly involvct
carotenoid changes may be a reflection of the types of observations which ha~
been coming out of the laboratory of Hager [167], indicating that carotenoid
interconversions may be a result of photosynthetic activity.
Finally, the suggestions of Strehler [168] and Platt [169] that carotenoid
pigments interact electronically with CHL in facilitating the collection of
radiant energy in photosynthetic organisms have been very stimulating to
many workers, but unfortunately no experimental evidence is available to
support adequately these hypotheses.
We must therefore conclude that, up to this time, claims that carotenoid
pigments act as cofactors in photosynthesis have not been substantiated.

d) The 515 nm effect

It would be impossible to discuss the function of carotenoid pigments
without referring to the phenomenon which I will call the 515 nm effect.
Although it is still not known how carotenoid pigments are related to this
phenomenon, even the most outspoken critics of carotenoid involvement are
now convinced that there is a relationship between the 515 nm effect and
carotenoids.
In 1954, Duysens [170] reported this effect for the first time. Upon illu-
minating Chlorella he found a change in the absorption spectrum with an
increase at 515 nm and decreases at 478 and 420 nm. He suggested that the
420 nm change was due to cytochromes and that the 515 nm and 478 nm
change would be due to a different pigment. In the following year, Chance
and Smith [171] reported similar observations on the photosynthetic bacte-
rium R. rubrum and were also able to demonstrate that a similar spectral
change occurred when an anaerobic culture was made aerobic by admitting
0 2 . They concluded that the peaks in the difference spectrum in the visible
which change with oxygenation or illumination are presumably due to pig-
ments other than cytochromes. By 1956, Strehler [168] made the suggestion
that these spectral changes may be due to a radical form of carotenoid pigment
which interacts with CHL, based on experiments of Strehler and Lynch [172].
 IX. Function 693

Then Chance and his associates [53, 54] demonstrated the 515 nm effect in
algae and, based on the absence of this effect in a mutant strain of C. reinhardi
lacking /3-carotene (3), concluded that the effect was due to changes in the
spectral properties of carotenoid pigments. By 1959, Smith and Ramirez [173]
also concluded that the spectral changes induced by either oxygenation or
illumination in photosynthetic bacteria were due to a new compound formed
from carotenoids which shows a spectral shift to longer wavelengths. Since
then, there have been many papers which have demonstrated a spectral change
on illuminating photosynthetic organisms, and some of these papers have
attempted to correlate these changes with the carotenoid pigments present in
these cells. This material has been very thoroughly reviewed in a recent
publication of Fork and Amesz [174], with particular emphasis on the rela-
tionship between photosystem I, photosystem II and the carotenoid spectral
changes.
Although this spectral change, which has a maximum between 510-520 nm,
is now generally attributed to carotenoid pigments, the exact change which
takes place is still under active investigation. There appear to be two major
concepts as to what brings about the spectral change, and the ideas behind
these concepts are not mutually exclusive. One group of investigators presents
evidence which indicates that the changes can be directly attributable to a
chemical change of the carotenoid pigments within the cell following illumina-
tion. Another group has suggested that a physical change within the photo-
synthetic membranes is reflected by a change in the spectrum of the carotenoid
pigments. In this way, the latter group suggests that the spectral changes can
serve as 'membrane signals' and indicate different membrane states. After
sixteen years of intensive work in this field, the question raised by Chance
[175] appears to be still appropriate, when he describes the spectral shift as
'a weird and wonderful phenomenon, but what is the significance of the
absorbance changes?'

rx) The chemical basis ofthe 515 nm change. There have been several expla-
nations for the spectral change which involve either a photochemical or a
chemical change in the carotenoid pigments. These have all attempted to take
into account the fact that the change looks as if there has been a shift to a
longer-wavelength absorbing species. As mentioned earlier, Strehler [168]
suggested that the 515 nm change might be due to a radical form of a carote-
noid, an explanation which has been supported by Clayton [176]. Chance
[177] suggested that the spectral change may be due to an oxidation of carote-
noids ow to a tighter binding of the pigment to a protein. The latter suggestion
was supported by Coleman and Rabinowitch [178], whereas the former sug-
gestion was supported by Nishimura and Chance [179] and Smith and Ramirez
[180]. The latter workers, however, could not understand how the oxidation of
spirilloxanthin (108) would lead to a pigment absorbing at longer wavelengths
and therefore suggested that there might be a change in the whole structure of
the bacterium, with a resultant perturbation of the carotenoid molecules,
694 NORMAN I. KRINSKY

resulting in a spectral change. This suggestion of Smith and Ramirez [180] was
the first attempt to explain the spectral changes on the basis of a structural or
conformational change in the cell itself. These topics will be discussed in the
following section (B.2 d {3).
Another suggestion, which appears to be receiving increasing support, is
that the 515 nm change is due to the formation of a metastable form of a
carotenoid pigment. This idea was first suggested by Chessin et al. [81] and
has received considerable support from the experiments of Mathis [82, 181]
and Mathis and Galmiche [182]. These workers indicate that red light absorbed
by a photosynthetic pigment such as CHL can result in spectral changes in
the blue-green region in the spectrum, which they attribute to the formation
of a metastable compound, presumably the triplet state of a carotenoid pig-
ment. Although the exact nature of the metastable pigment is not known,
Mathis [82] has presented very compelling evidence that the 515 nm change
can only be due to a new species of carotenoid, which presumably is a tripfei
compound inasmuch as its lifetime is quenched by the addition of paramagnetic
gases such as 0 2 or NO. The formation of a metastable or triplet state of
carotenoid could explain the spectral changes brought about by illumination
of photosynthetic organisms, but it would be difficult to understand how
oxygenation alone could bring about this species. Of course, the other way
to produce the metastable or triplet state of carotenoids is by reacting a
carotenoid with 10 2 , as described in Section B.1 c. Witt and his colleagues
have now suggested that this type of reaction does occur and can explain
some of the 515 changes observed by them. For many years now Witt and his
colleagues have referred to a type I spectral change which has a maximum
at 520 nm. This type I is characterized by a decay time of ~ w- 5 seconds.
Wolff and Witt [183] have now succeeded in quenching all absorption chang-
es caused by electrons, protons or electrical fields in spinach chloroplasts
by the addition of 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU). Under
these conditions, there was no change in the type I absorption or in its kinetics,
and they conclude that the type I change is caused by an excited energy trap
and not by a chemical intermediate in electron transport. Since the type I
change increases above the saturating light energy of photosynthesis, they feel
that more CHL becomes excited than can be used for photosynthesis, and the
excess energy can presumably result in the formation of 3 CHL. Under these
conditions, 10 2 can be formed and in the presence of carotenoids can be
quenched resulting in the formation of a metastable excited carotenoid mole-
cule, according to the hypothesis of Foote [91]. This process has also been
called the 'valve' reaction by Witt and Wolff [184] as it serves as the reaction
which is capable of draining off excess radiant energy.
Although the evidence for photochemical reactions involving carotenoid
pigments and the 515 nm change seems cogent, one cannot overlook the
possibility that there may be chemical changes occurring among the carotenoid
pigments that would result in a 515 nm change. For several years now, inves-
tigators appear to have been discouraged from looking for chemical changes
 IX. Function 695

by an observation of Amesz and Vredenberg [185] that approximately three

carotenoid molecules were altered per quantum absorbed in photosynthetic
bacteria, apparently obviating a chemical explanation for this phenomenon.
There are, however, well-established changes in carotenoid pigments which
occur upon illumination. In particular, the interconversion of epoxy- and non-
epoxy-carotenoids brought about by light as studied by Sapozhnikov et a!.
[141], Yamamoto eta!. [144] and Krinsky [186] would appear to be worth
investigating for a relationship to the 515 nm change. The closest approach
to the problem of measuring carotenoid changes and spectral changes directly
is that of Meister and Maslova [187]. They illuminated leaves with white light
and studied both the spectral changes and the carotenoid changes which
involved conversion ofviolaxanthin (135) to zeaxanthin (67) under the influence
of light. Both the spectral and chemical reactions were reversed in the dark.
By utilizing the data that they present for Zea mays, one can deduce that a
spectral change involving an increase in absorbancy at 515 nm brought about
by illumination is accompanied by a conversion of violaxanthin (135) to zea-
xanthin (67). Inasmuch as the 515 nm change seems to be related to a high-
energy state of photosynthetic organisms (see next section), the report of Hager
[167] seems to make an important contribution in this area. He observed the
conversion of violaxanthin (135) to zeaxanthin (67) when isolated spinach
chloroplasts were illuminated with white light. This carotenoid conversion
appeared to occur only when photosynthetic phosphorylation (PSP) was
occurring, inasmuch as PSP uncouplers were able to eliminate the light-
dependent interconversion of these carotenoids. In addition, Hager [167] was
able to observe a dark conversion of violaxanthin (135) to zeaxanthin (67)
when either adenosine triphosphate (ATP) was added or if the pH was lowered
to 5. Since illumination also causes a decrease in the pH within the chloroplast
of 1-2 units, he concluded that the xanthophyll interconversion is an indi-
cator of a pH decrease within the chloroplast and, as such, an indicator for
PSP. In a related study, Krinsky [188] has observed that the direct reduction
of epoxy carotenoids to non-epoxy carotenoids in E. gracilis results in a spec-
tral change similar to that observed in algae and plants upon illumination,
with a positive maximum at 508 nm. It is therefore possible that some chemical
interconversion of carotenoids can contribute to the overall 515 nm effect.

fJ) The structural basis of the 515 nm effect. As mentioned above, Smith
and Ramirez [180], in 1960, suggested that the 515 nm change may be due to
a perturbation of the carotenoid molecules caused by a structural change in
the assc;>ciated tissue. Since then, there have been many reports which would
support this idea that the absorbance changes observed upon illumination
are the result 6f a structural or conformational change in the membranes con-
taining the photosynthetic pigments, and furthermore this conformational
change is a reflection of some of the energy-transducing properties of these
membranes. In particular, Fleischman and Clayton [189] and Baltscheffsky
[190] have suggested that the carotenoid change is a reflection of a high-
696 NORMAN l. KRINSKY

energy state or intermediate involved in PSP in photosynthetic bacteria. The

high-energy state is still somewhat elusive, not only in PSP, but in oxidative
phosphorylation as well. It has been ascribed to a pH gradient, an electrical
potential, a conformational change or a chemical compound. The results of
Jackson and Crofts [191] strongly support the idea that the carotenoid chan-
ges are energy-linked, inasmuch as they can generate the spectral shifts in
photosynthetic bacteria in the dark by either generating electrical potentials
with ionic gradients or by generating a transmembrane pH gradient. In fact,
they find a linear relationship between the spectral shift and the electrical
potential developed across the membrane. Using this relationship, they can
utilize the magnitude of the spectral shift to evaluate the membrane potential.
Support for this concept comes from the work of Emrich et al. [192], who
were able to discharge the 515 nm signal by utilizing gramicidin D, a compound
that increases ionic fluxes and discharges the electrical field across the thylakoid
membrane. The idea that the 515 nm change is a measure of the membrant)
potential has been utilized recently by both Neumann et al. [193] and by
Hauska et al. [194] to investigate the relationship of the chemiosmotic hypo-
thesis with respect to PSP. Again, there appears to be a relationship between
the 515 nm change and the ability to carry out PSP, although the relationship
is still not known. Wolff et al. [195] have attempted to relate the 515 nm
change to the light-induced electrical field developed across the thylakoid
membrane. They observe an extremely rapid 515 nm change having an upper
limit of the half-rise time of 20 ns. They suggest that this rapid response is
caused by electron translocation across the membrane which is carried out
by carotenoids containing conjugated double bonds in which the carotenoid
serves as a 'pipe line' between an electron donor on one side of the membrane
and an electron acceptor on the other. This hypothesis is essentially identical
to that initially formulated by Platt [169] in 1959. A very brief note by Chance
et al. [196] raises some interesting questions. These authors claim that they
can observe spectral shifts in photosynthetic membranes when relatively small
hydrostatic pressures are applied to the membrane. This would indicate to
them that the energized state corresponds to a small-scale volume decrease,
possibly due to the extrusion of water. It would be interesting to see whether
agents that are capable of causing a swelling of photosynthetic membranes
would result in reversed spectral shifts.
As mentioned earlier, Amesz and Vredenberg [185] argued that the quan-
tum yield for the carotenoid spectral change was approximately 3, and that
this might be due to a chemical or structural change in the environment of
the molecule. Presumably it could not involve a direct change of the carotenoid
pigment itself. Although this was only a brief report and supporting data were
not presented, this paper has been widely accepted as the experimental evidence
for concluding that chemical reactions of the carotenoids do not occur upon
illumination. Only recently has another attempt been made to determine the
quantum yield for the spectral changes of carotenoids induced by light. This
is the work of Okada et al. [197] who attempted to measure the quantum
 IX. Function 697

yield for the light-induced absorbance change of carotenoids. By making

several assumptions, they calculate a quantum yield for the process averaging
2.5 and conclude that the phenomenon cannot be photochemical, but may
be a structural change leading to a change in the electronic state of the carote-
noids. In attempting to repeat their calculations, using the spectral data of
Jensen and Jensen [198] for the carotenoids of Rps. spheroides, I have been
unable to duplicate the calculations of Okada et al. [197]. My calculations
indicate that the quantum yield from their data is approximately 1.0. Although
this does not argue either in favour or against a structural or conformational
change as the basis of the 515 nm change, it must be borne in mind that until
evidence is presented that the quantum yield for the process is significantly
greater than 1, one cannot eliminate the possibility that a photochemical or
chemical reaction is occurring that involves the shift in the carotenoid spec-
trum to longer wavelengths. If indeed there is a conformational change which
results in a change in the carotenoid spectrum, one might be able to work
backwards from the difference spectrum to attempt to generate the actual
absorption spectrum of the carotenoids involved in the conformational change.
The only attempts in this direction have been made by Fork and Amesz [199]
and Fork [200]. These workers have investigated the spectral shifts induced
by light in a number of algae and CHL b-deficient plants and have found that
the light-induced spectrum is the mirror image of the difference spectrum
obtained on turning off the light. They suggest that the absorption change
they observe is due to a shift toward longer wavelengths of a compound
which, on the basis of integrating the difference spectrum, has 3 maxima at
504-506, 473 and 440-445 nm in both red algae and in a yellow-green alga.
Unfortunately, the carotenoid pigments of these organisms have not been
sufficiently characterized to indicate whether a pigment having those maxima
exists whithin the cells.
In summary then, carotenoid pigments appear to be related to the 515 nm
effect in several different ways. The spectral change may be a reflection of an
actual chemical change in carotenoid pigments, or it might reflect either a
conformational change in the supporting membrane or in the development
of an electrical field across the supporting membranes. The latter two pos-
sibilities may in fact be equivalent insofar as one may not be able to develop
electrical fields across membranes without conformational changes or con-
versely conformational changes might induce the development of electrical
fields. In either case, it is still necessary to explain the shift toward longer
wavelengths of the carotenoid pigments that are involved in the spectral chan-
ges. As is well known, the absorption spectrum of conjugated polyenes will
be markedly affected by the polarizability of the solvent, or by its refractive
index. This is particularly well documented in the case of the visual pigments,
where conformational changes in the proteins associated with the polyene
chromophores result in spectral shifts that are comparable to those expected
in the carotenoid pigments in photosynthetic organisms. One must therefore
conclude that the spectral changes attributed to either conformational changes
698 NORMAN I. KRINSKY

or the development of an electrical field across the membrane must reflect a

reversible change in the immediate environment of the carotenoid pigments
presumably exposing a more polarizable region of the membrane substructure
to the electron chain of the carotenoids.

e) Association with different photosystems

More and more reports are appearing which deal with the physical separa-
tion of the various photosystems in photosynthetic organisms. Many of these
reports have included analyses of the carotenoid pigments in the different
fractions and indicate that there is a differential distribution of carotenoids
in photosystem I and photosystem II. No clear pattern has yet emerged from
these studies.
As discussed throughout this section, the only clear-cut function for carote-
noids in photosynthesis is as an accessory light-absorbing pigment. In adgi-
tion, under conditions of high-light intensity, carotenoids can se~ve as photo-
protective pigments. Whether their relationship to the 515 nm effect is an
expression of a carotenoid function is still an open question. It may be that
the importance of this relationship will be a result of the ability of investiga-
tors to use carotenoid spectral shifts as membrane signals, even without clearly
understanding the mechanism of the effect.

3. 'Blue-light' effects
There have been many biological phenomena described which are initiated
or dependent upon illumination with 'blue-light'. In some cases, the 'blue-
light' has consisted of a very broad range of light passed through a simple
glass or gelatin filter, and as such, almost useless in trying to determine the
nature of the photoreceptor pigment. In other cases, quite accurate action
spectra have been obtained which, in theory, should coincide with the absorp-
tion spectrum of the photoreceptor pigment. In this way, one should be able
to identify the pigment responsible for the 'blue-light' effect. In many of the
reported 'blue-light' effects, carotenoids have been implicated as the photo-
receptor pigments. With the single exception of bacterial phototaxis, which
seems to be related to alterations in the rate of photosynthesis, the evidence
that carotenoids are involved as photoreceptor pigments is inadequate. The
most controversial area with respect to carotenoid involvement has been that
of phototropism. Although the question is still unsettled for higher plants the
evidence for fungal phototropism almost excludes carotenoids as the photo-
receptor pigment.
One of the major problems in trying to understand the possible role of
carotenoids in these processes is to try to formulate a reasonable mechanism
whereby the photoreceptor initiates the series of events which leads to the
specific photofunction. We now know that the photoprotective and photo-
synthetic functions described above (Sections B.l and B.2) are both results of
transfer of electronic energy and. are not chemical phenomena. It has been
 IX. Function 699

difficult for proponents of carotenoid involvement in other photofunctions to

propose experimentally verifiable hypotheses to explain how carotenoids
interacting with light carry out these functions. Until such experiments are
designe~, one must bear in mind that the suggestions regarding carotenoid
function are based on circumstantial evidence.

a) Phototropism
Two types of tissues are used to study phototropism, or the bending of an
organ under the influence of light. The first involves use of higher plants such
as the coleoptile of oats or barley. The other utilizes sporangiophores of
fungi such as Phycomyces or Pilobolus. Although the phenomenon is simi-
lar, i.e. an alteration in the rate of growth upon illumination, and the action
spectra are essentially identical, the mechanisms that have been postulated to
explain the effect are somewhat different, and the two systems will be treated
separately.
rx) Higher plants. Carotenoids were first suggested as the photoreceptor
pigments in phototropism in higher plants by BUnning [201] in 1937. An
alternative explanation was presented by Galston [202] in 1949 in which he
assumed that the photoreceptor pigment was a flavin, based on the observa-
tions that flavins could catalyse the photooxidation of indoleacetic acid. A
detailed action spectrum of this phenomenon was published by Thimann and
Curry [203] in their review which appeared in 1960. At that time they presented
arguments against the idea that carotenoids acted as masking pigments and
concluded that they were the photoreceptors for phototropism in higher
plants. This view was primarily based on the similarity of the action spectrum
to the absorption spectrum of the Avena coleoptile despite the fact that the
absorption peak in the ultraviolet did not coincide with any carotenoid band.
Since then, there has been very little new evidence to suggest that either a
carotenoid or a flavin is the primary photoreceptor pigment. Some work has
been done using either mutant plants or plants treated with chemicals to
decrease the carotenoid content, and phototropic sensitivity has been measured.
These results have not been conclusive. For example, Bara and Galston [204]
decreased the carotenoid content of Avena sativa to approximately 20% of
the normal level without significantly affecting phototropic curvature. Un-
fortunately, this type of experiment does not rule out the possibility that the
20% carotenoid remaining could still be involved in the process. In an attempt
to reconcile some of the divergent views, Briggs [205] suggested that both
carotenoids and flavins might function as photoreceptive pigments with a
transfer of energy from flavins absorbing in the ultraviolet to the carotenoids
absorbing in the visible region of the spectrum by means of resonance trans-
fer. This idea has not yet received support from the advocates of flavin, but
Thimann [206] ·has endorsed the suggestion of Briggs [205] that the 370 nm
band in the ultraviolet may be due to a flavin able to transfer energy to a
carotenoid.
700 NORMAN I. KRINSKY

It is unfortunate that the study of phototropism in higher plants has not

as yet revealed the nature of the photoreceptor involved in this important
process. On the basis of observations on fungal phototropism to be discussed
in the next section, it appears that even experiments with carotenoidless
mutants of higher plants would not necessarily lead to total exclusion of
carotenoids as functioning in higher plant phototropism. Presumably, only
the isolation of a light-sensitive pigment with an absorption spectrum coin-
ciding with the reported action spectrum would permit identification of the
photoreceptor pigment, and resolve the longstanding flavin-carotenoid contro-
versy. At this point, one can only conclude that although the action spectrum
of phototropism resembles the absorption spectrum of carotenoids in the
visible region, this information by itself is insufficient to assign a phototropic
role to carotenoids.

f3) Fungi. Similar arguments have been presented by both the flavin advd:-
cates and carotenoid advocates with respect to fungal phototropism. The early
qualitative observations were replaced by the detailed action spectra published
in 1959 by Curry and Gruen [207] and in 1960 by Delbrlick and Shropshire
[208]. Both groups recorded a correspondence between the action spectrum
and the absorption spectrum of carotenoids in the visible region, and also
observed in the ultraviolet region a peak in the action spectrum which could
not be accounted for by carotenoid absorption, but did coincide with a peak of
flavin absorption. Curry and Gruen [207] concluded that the photoreceptor
was a carotenoid, while Delbrlick and Shropshire [208] concluded that it
could not be. The subject has been reviewed frequently since these two pub-
lications with advocates of both flavin and carotenoid involvement presenting
their respective points of view. The reader is referred to the excellent review
of Bergman et al. [209] for a very complete study of Phycomyces and in
particular of phototropism in this organism.
Two very significant publications have appeared in the last few years
related to the action spectrum of fungal phototropism. In 1966, Page and Curry
[21 OJ presented data on the phototropic response of young sporangiophores of
Pilobolus kleinii. They obtained both an action spectrum and the absorption
spectra of very narrowly defined regions of the sporangiophore utilizing an
ultra-microspectrophotometer. In addition, they grew the fungus in DPA and
found no difference in the photoresponse of almost colourless sporangio-
phores in comparison with the normally pigmented sporangiophore. They
found that the action spectrum in P. kleinii was very similar to that obtained
for both Phycomyces and Avena with a very weak response near 500 nm. This
should be contrasted with the absorption spectrum they obtained for the
extreme tip of sporangiophore, the only region which showed an orientation
of the direction of growth when illuminated. The extreme tip showed a broad
absorption maximum centered around 440 nm, in comparison with the
densely pigmented region basal to the tip which displayed two absorption
peaks at 465 and 490 nm. On the basis of these results, Page and Curry [210]
 IX. Function 701

concluded that the photoreceptor was a flavin pigment and not a carot-
enoid.
Another series of very important experiments were reported by Meissner
and Delbrtick [211], who worked with mutant strains of Phycomyces that had
been isolated and were either deficient in coloured carotenoids or produced
lycopene (19) rather than P-carotene (3) as the major carotenoid pigment.
They found small traces of retinal, presumably present as an intermediate in
abscisic acid formation as described in Section D.2. They found that in both
the carotenoidless mutant strains and the W-T strain grown with DPA there
was no change in the photosensitivity, which would indicate that carotenoids
were not involved in the phototropic response. They then tried to rule out
quantitatively the possibility that even the traces of P-carotene (3) could be
involved as the photoreceptor pigment. They calculated that the photoreceptor
pigment would have an optical density less than 0.003 OD units/sporangio-
phore. In their mutant strain, alb-10, they found only 0.1 micrograms P-caro-
tenejgram sporangiophore. However, even this trace amount would be 10 x
as much P-carotene (3) as the lower limit of the photoreceptor pigment based
on their physiological data. On the basis of these measurements they pointed
out that P-carotene (3) still cannot be ruled out as a possible photoreceptor
pigment. In their review Bergman et al. [209] point out the possibility that
the ultraviolet and visible peak in the action spectrum may actually be due
to two separate pigments. The first indication that this may, in fact, be the
case has appeared in a very recent publication of Berns and Vaughn [212],
who have made the first successful attempt to isolate a soluble photosensitive
pigment from Phycomyces. Although these results are very preliminary, they
do present some evidence that the visible peak and ultraviolet peak behave
differently when digitonin extracts are illuminated. With this type of approach
it should be possible to resolve the nature of the photoreceptor pigment in
fungal phototropism and to identify it.
b) Phototaxis
Phototaxis in photosynthetic bacteria appears to be related to a change
in the rate of photosynthesis. The initial experiments in this field were carried
out in 1888 by Engelmann [213], who showed that the action spectrum for
phototaxis was similar to that of the action spectrum for photosynthesis and
involved light absorbed by both BCHL and carotenoids. This work was
extended and substantiated by Manten [214] and Clayton [215] and has been
reviewed recently by Clayton [216]. The question whether all the carotenoid
pigments present in the organisms are involved in phototaxis arose because
there was not. complete agreement between the action spectrum for photo-
taxis and the absorption spectrum of the organisms. The question was re-
solved by van Niel et al. [217], who showed that photosynthetic bacteria
such as R. rubrum contained large amounts of spirilloxanthin (108), the ab-
sorption peaks of which could not be observed in the action spectrum of
phototaxis. The reason was that physiologists had worked with young cui-
702 NORMAN I. KRINSKY

tures of R. rubrum, and at that stage the organism is very low in spirilloxan-
thin (108).
The study of phototaxis in algae is much more complicated. Again, the
initial experiments in this field were carried out by Engelmann [218], who
demonstrated that the photoreceptor in Euglena was in the stigma, and not
in the coloured eye spot. The work done in the following 75 years has been
reviewed by Halldal [219], who concluded that none of the major pigments
present in algae corresponded with the action spectra that he and others had
obtained. Recently, Tollin and Robinson [220] and Diehn [221] have pres-
ented evidence that algal phototaxis uses either a flavin or a flavonoid as the
photoreceptor pigment, as opposed to carotenoids. As with phototropism in
higher plants, the work on the nature of the photoreceptor pigment in algal
phototaxis is still quite incomplete and much more must be done before
carotenoids can be included in or exluded from the process.

c) Photoinhibitions
Many processes are inhibited by 'blue-light'; they include effects on PSP,
Hill reaction, germination, growth, respiration, respiratory enzymes, flowering
and circadian rhythm. Although some people have tried to implicate carote-
noids in these photoinhibitions, the evidence is very meager, and there are
specific cases where carotenoids can be excluded on the basis of mutant stu-
dies. An example is the observation that the effect of 'blue-light' on circadian
rhythm in Neurospora is the same as that observed in an albino mutant, as
described by Sargent and Briggs [222]. The remaining effects are presumably
due to flavins or cytochromes.

d) Photostimulations
This is another broad area where various biological reactions are stim-
ulated by 'blue-light'. For some time, it was felt that the stimulation of respira-
tion observed in algae upon illumination with 'blue-light' might be due to
carotenoid pigments, but the recent experiments of Pickett and French [223]
and Schmid and Schwarze [224] now indicate that the effect is due to the
absorption of light by flavins. A stimulation of 0 2 evolution in algae illuminated
with blue-green light has been observed by Adler [225], and he suggests that
this is brought about by the absorption oflight by a carotenoid pigment. From
the nature of the effect, he excludes an Emerson-enhancement phenomenon
and considers the effect to be more specific.

e) Photoreception
Although almost all photoreceptor processes in animals seem to be built
on a rhodopsin-like pigment, a few exceptions have been reported in a recent
review by Kennedy [226]. Most of the work has come from the laboratories
of Arvanitaki and Chalazonitis [227]. These workers recorded both on-
responses and off-responses in Molluscan neurons and claimed that hemo-
 IX. Function 703

proteins mediate the on-response and carotenoproteins mediate the off-

responses. Whether these results are specific for carotenoproteins or are com-
parable to the results obtained by Furchgott et al. [228] using stained nerve
preparations remains to be seen.

f) Photoperiodism
Inasmuch as the action spectrum of photoperiodism in insects has maxima
in the blue region of the spectrum, Lees [229] suggested that this effect might
be due to a carotenoid pigment. It should be possible to test this experimentally
by rearing insects on carotenoid-free diets and observing whether there is any
change in their response to visible light in setting photoperiodism.

g) Protoplasmic viscosity and streaming

Virgin [230] has reviewed the effects of light on chloroplasts and plant
protoplasm. In both cases, 'blue-light' stimulates protoplasmic streaming and
increases viscosity changes, but it is still not possible to decide whether the
effect is due to carotenoid pigments or to flavins.

C. Non-Photofunctions
In addition to the functions of carotenoids which involve their interaction,
either directly or indirectly, with light, other functions have been proposed
for this class of compounds. Although a wide variety of observations and
suggested relationships between carotenoid pigments and various biological
systems have been reported, there is little direct evidence of functional roles
for carotenoids outside their photofunctions. Proposals which have experi-
mental support are discussed below.

1. Protein and membrane stabilization

The possibility that carotenoid pigments stabilize the proteins with which
they are associated in carotenoproteins has been reviewed recently by Chees-
man et al. [231]. This article deals primarily with carotenoproteins of inverte-
brates for it is here that most of the evidence has been accumulated that carot-
enoids stabilize protein molecules. In an earlier paper, Cheesman [232] pro-
posed that in nature carotenoids stabilize polypeptide configurations, and there
have been several reports substantiating the proposal. Very recently, Zagalsky
et al. [2'33] suggested that the stabilization may arise as a result of n-orbital
interaction between the polyene molecules on the apoprotein units with the
carotenoid acting as an additional 'cement' between subunits and providing
the driving force necessary for dimerization. Much of this seems to be based on
the work of Wald and his collaborators on the ability of retinal to stabilize the
protein opsin when the two components are associated as the visual pigment
rhodopsin. The evidence for such stabilization has been reviewed by Wald and
704 NORMAN l. KRINSKY

Hubbard [234]. In a very different context, Ji et al. [235] have isolated a {3-
carotene (3}-protein from chloroplast lamellae which appears to be very similar
to the material isolated earlier by Nishimura and Takamatsu [236]. In both
cases, these groups observed a very marked red-shift of the spectrum in the
isolated carotenoprotein and found approximately one mole {3-carotene (3) per
23,000 to 28,000 grams of protein. Ji et al. [235] suggested that the carotenoid
is a specific structural component of the chloroplast lamellae and may confer
some stabilization on this structure. As discussed earlier, Walles [40] has
presented evidence that carotenoid mutants of sunflowers can develop normal
proplastids and show the initial steps of chloroplast formation, but that these
chloroplasts are unstable and do not form normal grana.
The possibility that carotenoids may be involved in the stabilization of
membranes can be inferred from the suggestions of Lucy and Dingle [237]
that vitamin A may stabilize membranes by acting as a cross-link betwee,n
lipid and protein. Conversely, the absence of carotenoids from membranes
that normally contain them could lead to a decreased stability of these mem-
branes. A suggestion that this may be occurring was seen in the data presented
by Salton and Freer [238] and Salton and Ehtisham-ud-din [239], who treated
various gram-positive bacteria with DPA and in this way prevented the forma-
tion of coloured carotenoids. They observed that certain batches of cells grown
in the presence ofDPA were more labile and had a tendency to lyse on harvest-
ing. They suggested that this membrane system might be useful in investigating
the possible role of carotenoids in stabilizing bacterial membranes. Two sub-
sequent reports have appeared which would indicate that carotenoid pigments
do not play a role in stabilizing bacterial membranes. The first is that of Razin
et al. [240] who worked with M. laidlawii and were able to demonstrate that
a 10-fold change in membrane carotenoid concentration, brought about by
varying the amount of sodium acetate in the medium, resulted in no signi-
ficant change in osmotic fragility. This is to be contrasted to the marked
changes in osmotic fragility which occurred when unsaturated fatty acids were
added to the medium or when PPLO * serum fraction was added to the
medium. They did not, however, study a carotenoid-free membrane for osmo-
tic fragility. This was done in a recent publication of Mathews-Roth and
Krinsky [241], who worked with both a colourless mutant strain of S. lutea
and a colourless culture produced by adding D PA to the medium of the W-T
strain. Protoplasts were made of all three cultures, and Mathews-Roth and
Krinsky [241] could find no difference in the osmotic fragility of the W-T
strain or the two carotenoidless strains. On this basis they concluded that
carotenoid pigments do not play a role in stabilizing cell membranes.

2. Reproduction
Although there have been many reports of the apparent relationship
between the accumulation of carotenoids and the development of reproduc-
* Pleuropneumonia-like organisms.
 IX. Function 705

tive structures in both animals and plants, there has never been an adequate
explanation for this phenomenon in terms of a functional role for carotenoids.
The subject was reviewed very extensively by Goodwin [242] in 1950, at which
time he concluded that very little was known about the direct participation
of carotenoids in reproduction. There has been little evidence in the past
20 years to change that statement. In a more recent review, Burnett [5] dis-
cusses the relationship between carotenoid pigments and reproduction, with
particular emphasis on Phycomycetes. It now appears that the relationship
is not between the carotenoid pigments themselves, but between a carotenoid
metabolite, trisporic acid, and reproduction. This is discussed below in Sec-
tion D.3. It seems that light is involved in converting various epoxy carotenoids
to trisporic acid, and the trisporic acid, in turn, stimulates carotenogenesis,
as described by Taylor and Smith [243]. Other postulated roles for carotenoids
in reproduction are quite speculative and lack experimental evidence. Even
the possibility that carotenoid pigments have an indirect role in reproduction
by serving to attract opposite sexes in animals or pollen-carriers in plants
remains to be tested experimentally.

3. Miscellaneous functions
Many other functions have been suggested for carotenoid pigments. Some
of these, such as the possibility that carotenoid pigments in the retina in the
form of oil droplets can in some species serve as filters, as discussed by Wald
[244] in one of his earliest reviews on photoreception, seems to be well founded.
The suggestion by Smith [245] that carotenoid pigments can function to
transport glucose into cells of Mycoplasma was subsequently refuted by the
work of Razin and Rottem [246], who were able to grow M. laidlawii under
conditions in which neither cholesterol or carotenoids were deposited in the
cell membrane, and the organism was still able to grow. A number of sug-
gestions of this nature seem plausible for a time, but are later shown to be
incorrect. Some may be functions not of carotenoid molecules themselves but
rather of their metabolites as will be discussed in the following section.

D. Metabolite Functions

In addition to the functions described above which pertain to the intact

carotenoid molecule, a number of important biological functions have now
been shown to be due to metabolites of carotenoids. These compounds are
discussed below.

1. Vitamin A and retinene (retinol/retinal)

The best known metabolite of carotenoids is vitamin A. The conversion
of carotenoids to vitamin A and the function of vitamin A are discussed in the
following Chapter X by Pitt.

Carotenmds 45
706 NORMAN I. KRINSKY

2. Abscisic acid
Abscisic acid (ABA) is a recently discovered growth-regulating substance
which promotes senescence and abscission of leaves and induces dormancy
in buds and seeds. The reader is referred to the recent review by Wareing and
Ryback [247] for a description of discovery, chemistry and biochemistry of
ABA. We shall only be concerned here with the experiments indicating that
ABA is formed by a photooxidation of epoxy carotenoids. The suggestion
was first made by Taylor and Smith [243], who were aware of the fact that
light stimulates the production of ABA. They illuminated the total carotenoid
mixture from dried nettle leaves (Urtica giocia) and found that, in the presence
of 0 2 , the illuminated products would promote germination of cress seeds
(Lepidium sativum L.). A more complete study was published in 1969 by Taylor
[248] in which he identified the individual carotenoids that were responsible
for producing active fractions on illumination in the presence of 0 2 . The only
two active compounds were neoxanthin (122) and violaxanthin (135), both
xanthophyll 5,6-epoxides. Support for the idea that the epoxy xanthophylls
are precursors of ABA comes from an experiment of Sondheimer et al. [249].
These workers measured the biological activity of the epidioxide isomer of
ABA, following the suggestion of Cornforth et al. [250] that this compound,
which is a chemical intermediate, might also be a biological intermediate.
Sondheimer et al. [249] could find no evidence that the epidioxide was in fact
a precursor. The ability of carotenoid 5,6-epoxides to be converted to the ring
structure of ABA was suggested by Bergmann and Meyers [251] on the basis
of the reactivity of steroid epoxides under various conditions. The appearance
of ABA in autumn leaves which seems to follow the disappearance of CHL
would indicate that the normal photoreductive process in plants has become
inoperative, and that more of the radiant energy is channelled into photo-
oxidative reactions. Since ABA antagonizes the effects of growth-promoting
hormones, Taylor and Smith [243] suggested that the formation of ABA may
explain some of the 'blue-light' effects such as phototropism in Avena. Now
that ABA has been identified, it would be important to determine whether
there is an increased production of ABA under conditions where one can
observe a phototropic bending of a plant such as Avena.

3. Trisporic acid
It is only since 1968 that the structures of a number of compounds which
function as regulators of sexual reproduction in fungi have been elucidated.
Among these is trisporic acid, a compound which controls gametogenesis in
Blakeslea trispora and in other mucoraceous genera including Phycomyces,
Mucor and Pilobolus. The chemistry and biology of these sexual hormones
have been reviewed recently by Barksdale [252]. There is an interesting dual
relationship between trisporic acid and carotenoids.
For almost fifty years, it has been known that the mating of the sexual
forms of fungi frequently resultetl in an increased synthesis of carotenoids. In
 IX. Function 707

fact, this process at one time served as a commercial source of P-carotene (3)
[253]. We now know that trisporic acid itself can stimulate carotenogenesis in
the (-) strain of B. trispora. It has been suggested by Thomas and Goodwin
[254] that trisporic acid stimulates a rate-limiting enzyme, presumably acting
before farnesyl pyrophosphate is converted to geranyl pyrophosphate, inas-
much as ergosterol synthesis is also stimulated following the administration
of trisporic acid to B. trispora.
The other relationship stems from observations of Heisenberg and Cerda-
Olmedo [255]. These workers obtained three albino mutant strains of Phyco-
myces and found that the mutant strains showed abortive zygote formation.
At first, it was not clear why a block in the synthesis of carotenoids-normally
stimulated by trisporic acid-would lead to the development of incomplete
se~ual reactions. This has now been clarified by the work of Austin, Bu'Lock
and their colleagues. Austin et al. [256] studied the biosynthesis of trisporic
acid in B. trispora and found that labelled P-carotene was a better precursor
of trisporic acid than was labelled mevalonic acid. In addition, DPA, which
blocks the conversion of phytoene (32) toP-carotene (3), also blocked the forma-
tion of trisporic acid. They therefore suggested that trisporic acids are formed
from carotenoids, and since they also promote their synthesis, this would
represent a positive feedback system. Austin et al. [257] have now shown that
tritiated retinyl acetate can be converted to both trisporic acid and the
corresponding alcohol trisporol in mated B. trispora. They concluded on the
basis of labelling experiments that the pathway for trisporic acid formation
in fungi is as shown below:
P-carotene--. retinol--. trisporol--. trisporic acid.
This would then explain the presence of retinol in Phycomyces, as observed
earlier by Meissner and Delbriick [211]. Finally, Bu'Lock and Austin [258]
have presented arguments based on the lack of label in the carboxyl carbon
at C-1 in trisporic acid following the addition of [2- 14 C]mevalonic acid to
B. trispora, that the cyclization of carotenoid precursors to P-carotene (3)
cannot occur by the selective elimination of an axial H at C-6 from a carbo-
nium ion intermediate as shown below:

If the carbonium ion described above were an intermediate, then the loss of
an axial H at C-4 would lead to the formation of an oc-ionone ring but it would
be the enantiomer of natural oc-carotene (5). It is still possible that the carbo-
nium ion intermediate is used for both oc-ionone and P-ionone synthesis, in
which case one might resort to assigning the stereospecificity to the enzyme
708 NORMAN I. KRINSKY

catalysing the two reactions and assume that it might abstract the equatorial
H from this intermediate to form oc-ionone.

4. Sporopollenin
A very interesting function has been described by Shaw and co-workers
for carotenoids. Shaw and Yeadon [259] had been studying spore coats and
pollen grains and found that the outer layer (exine) of both spores and pollen
grains consisted of a polymer called sporopollenin, which analysed as
C 90 H 134 0 31 to C 90 H 150 0 33 • This material was difficult to work with and on
oxidative degradation gave a mixture of mono- and dicarboxylic acids, with
a maximum at 16 carbons. Brooks and Shaw [260] then attempted to study
the formation of sporopollenin in the anthers of buds of Lilium henryii plants.
They found that sporopollenin formation parallelled the formation of the
carotenoid pigments in these anthers; both free and esterified carotenoid~
were present, with more than 90% of the esters having straight-chain fatty
acids. They developed polymerization methods using BF3 as a catalyst in the
presence of 0 2 and were able to form polymers from either the carotenoid
mixture of L. henryii or from {J-carotene (3) or vitamin A palmitate. These
polymers were degraded with ozone along with the naturally occurring sporo-
pollenin, and the products were separated by gas-liquid chromatography. They
observed a very similar array of products from the synthetic carotenoid poly-
mers as well as from the natural L. henryii carotenoids and suggested that the
sporopollenin in the pollen exine is formed by an oxidative polymerization
of a mixture of carotenoids and carotenoid esters contained in the anthers.
In another publication, Brooks and Shaw [261] indicated that the greater
part (95 %) of lipid material formed during anther formation consisted of
carotenoids, and that the concentration of carotenoid increased most drama-
tically when the exine was being deposited prior to the formation of the pollen
grain proper. The sporopollenin which they isolated was a highly insoluble
lipid-like macromolecule retaining a substantial degree of unsaturation. In
addition, they observed that the {J-carotene polymer could form p-hydroxy-
benzoic acid after potash fusion, indicating the ability to form aromatic com-
pounds from the carotenoid polymers.
Due to the great stability of sporopollenin, resisting both microbiological
and non-oxidative attack, Brooks and Shaw [262] suggested that sporo-
pollenin might be identical with some of the older forms ofkerogen, the organic
insoluble material known to occur in Pre-Cambrian sediments. This hypo-
thesis was also based upon a comparison of the spectrum of organic compounds
produced from sporopollenin and those from the Green River Shale kerogen,
which is a very young deposit, approximately 60 x 106 years old. Brooks and
Shaw argue that the isoprenoid nature of many degradation products of
kerogen could be understood in terms of the carotenoid-carotenoid ester
nature of sporopollenin. The paleobiological implications of this hypothesis
are quite important, for it would indicate, if true, that there were spore for-
mers in the oldest known rocks implying that chemical evolution was much
 IX. Function 709

quicker than is now thought. The hypothesis was attacked by Cooper and
Murchison [263], who based their arguments on the fact that the kerogen
from the Green River Shale had a lower 0 2 content than is normally found
in sporopollenin. In addition, they point out that the n-alkanes from Pre-
Cambrian rocks differs from the n-alkanes in the Green River Shale. They
also raise the question whether sporopollenin would be expected to serve as
the cover for the polymerized cell contents of aquatic algae and bacteria.
Brooks and Shaw [264] have replied to this criticism by pointing out that there
are many algal and fungal spore exines that contain sporopollenin. For example,
the germinated spores of Mucor mucedo contain 4% sporopollenin derived
almost entirely from {3-carotene (3), whereas the asexual form contains no
sporopollenin. They also argue that the sporopollenin polymer would contain
the long chain fatty acids from the carotenoid esters, and these would then
give rise to the n-alkanes found in kerogen. In addition, they have already
shown that the polyenoid polymer is almost equivalent to an aromatic system
due to the ability of the polyenoid structures to cyclize to aromatic compounds.
Increasing numbers of bacterial carotenoids which are aromatic are also
being discovered.
The demonstration by Brooks and Shaw [260] that the product of poly-
merizing carotenoids is similar to sporopollenin is very convincing and deser-
ves further investigation. One approach would be to use a suitable labelled
precursor to demonstrate that this can be incorporated into carotenoids and
subsequently into sporopollenin during pollen grain formation. The possible
relationship of sporopollenin to kerogen is much more difficult to test, and
it may have to await better techniques for analysing these complicated mole-
cules.

5. Grasshopper ketone
Although there is no evidence that this compound is formed directly from
carotenoids, the structural relationship between the grasshopper ketone first
described by Meinwald et al. [265] and the known structure of carotenoids
such as fucoxanthin (190) or neoxanthin (122) is unmistakable. The grasshopper
ketone, shown below, is a conjugated allenic ketone which appears in a brownish
froth that is emitted by the grasshopper from its anterior respiratory opening

N·~o
HO~······oH
when the insect is disturbed. This froth has a repellent effect on ants and other
predators. The structure was synthesized by Russell and Weedon [266], who
found that it was identical with a compound isolated from an oxidative degrada-
tion of fucoxanthin (190). Furthermore, the unusual allenic nature of this
compound is in keeping with allenic compounds formed from carotenoid
analogues by photooxidation, i.e. through the use of 1 0 2 as described by
710 NORMAN I. KRINSKY

Foote and Brenner [99] and Daile et al. [101]. It would thus appear likely
that at least in grasshoppers, carotenoids can be metabolized to yield mole-
cules which have specific and unique functions in this insect.

E. Conclusions
It should be apparent by now that there are only two functions which are
firmly established for carotenoid pigments. The first is that they act as protec-
tive agents to prevent cells from undergoing damage due to a photodynamic
action. The second is that they can act as accessory pigments in photosyn-
thetic organisms, transferring radiant energy to the actual pigments involved
in photosynthesis. Both of these functions are depicted in Fig. 2. We have
good evidence for the existence of these functions and understand the mecha-
nisms whereby carotenoid pigments take part in these functions, as described
in Sections B.1 and B.2a. In fact, the principles of the protective action of
carotenoids, elucidated in microorganisms and plants, has resulted in the suc-
cessful treatment of humans suffering from a specific type of photosensitivity
by Mathews-Roth et al. [104]. It is anticipated that other types of photo-
sensitivity will also respond to the application of these principles.
The remaining functions of carotenoids discussed in this chapter, outside
of those assigned to carotenoid metabolites, must still be considered as specula-
tive. Although many have a long history of support by various investigators,
the available evidence is insufficient to substantiate the claims made for
carotenoid function. It is hoped that as new techniques become available,
these issues can become clarified and resolved.

Acknowledgments
The work in the author's laboratory was supported in part by the United
States Public Health Service and by the National Science Foundation.

Abbreviations
ABA abscisic acid
AT 3-amino-1 ,2,4-triazo le
ATP adenosine triphosphate
BCHL bacteriochlorophyll
B-G blue-green
CAR
1 singlet ground carotenoid
3CAR triplet excited carotenoid
CAR* excited carotenoid
CHL chlorophyll
DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea
DNA deoxyribonucleic acid
DPA diphenylamine
 IX. Function 711
NADPH reduced triphosphopyridine nucleotide
302 triplet ground oxygen
102 singlet excited oxygen
PCHL protochlorophyll
PSP photophosphorylation
SENS sensitizer
1SENS singlet excited sensitizer
3 SENS triplet excited sensitizer
W-T wild-type

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 717

X. Vitamin A
G.A.J. PITT

Department of Biochemistry, University of Liverpool, Liverpool L69 3BX, England

A. Structural Specificity . . . . . . . . 718

B. Formation from Carotenoid Precursors 718
C. Metabolism . . . . . . 719
D. Function in the Eye . . . 722
E. Systemic Mode of Action 732
References . . . . . . . . 738
718 G.A.J. PITT

A. Structural Specificity
The essential physiological function of carotenoids in animals is to act as
precursors of vitamin A; one of the enzymic transformations in animals on
which the very existence of higher animals depends is the splitting of plant
carotenoids to give the active vitamin.
The structural specificity for a molecule to show vitamin A activity is very
narrow. The substance first designated vitamin A, and subsequently as vit-
amin A 1, is the primary alcohol now known as retinol [1] (1) which, as dis-
cussed later, still retains, more than does any other molecule, the right to be
called in colloquial usage simply 'vitamin A'.

11 3 ~
~o ~ 2 ~. CH20H

(I) (2)

Retinol (vitamin A1 ) 3-Dehydroretmol (vitamin A 2 j

Numerous variants on the retinol molecule have been synthesized and

tested for their ability to promote growth in animals deficient in vitamin A.
In general it can be stated [2, 3] that any change in the molecule except at the
terminal carbon atom 15 considerably reduces growth-promoting power. For
example, vitamin A activity is completely eliminated by modifications such
as the saturation of a side-chain double bond, removal of the methyl group
at C-13 or shortening of the side chain. The only change in the C-1-C-14
portion of the retinol molecule which does not reduce growth-promoting
activity to a very low level is further dehydrogenation. The only naturally
occurring compounds of this type are those of the vitamin A 2 , 3-dehydro-
retinol or simply dehydroretinol [ 4], series with an extra conjugated double
bond in the ring (2). They commonly have about half the activity of corre-
sponding A 1 compounds [5].
Earlier reports that the introduction of a 5,6-epoxy grouping had little
effect on biological activity were not confirmed by other workers [6] and have
been partially withdrawn [7]. The generalization still appears valid that any
modification to the retinol structure except at the primary alcohol end greatly
diminishes its activity. Only a limited number of naturally occurring carote-
noids will therefore be capable of acting as a precursor of vitamin A in the
animal body; of these, P-carotene (3) is quantitatively the most important.

B. Formation from Carotenoid Precursors

The conversion of P-carotene to retinol has been known for many years
and recognized as occurring mainly in the intestinal mucosa [8]; yet the
preparation of an enzymic system capable of carrying out this transformation
in vitro proved a difficult task.
 X. Vitamin A 719

The rat is a useful animal in which to study this conversion in that almost
no unchanged P-carotene passes through the intestinal wall and, after the
demonstration of the conversion of P-carotene to vitamin A in rat intestinal
sections in vitro, a soluble enzyme system capable of cleaving P-carotene was
isolated from rat intestinal mucosa. It has been studied in detail [9]; the
enzyme, P-carotene 15,15'-oxygenase, appears to catalyse an oxygenase reac-
tion in which molecular oxygen reacts across the central double bond with
the 15 and 15' carbon atoms to form a transient 15,15'-peroxide of P-carotene
which rapidly cleaves, yielding two molecules of retinaldehyde [9]. Similar
enzymes have also been found in rabbit [10] and pig [11] intestine-and in
rat liver and probably kidney too [12]. Although they differ slightly in their
properties [10, 11] the general impression is of a strong resemblance between
all these enzymes.
The specificity of the rabbit intestinal enzyme has been tested on some
P-apo-carotenals and intact carotenoids [10]. It produced retinaldehyde from
the three P-apo-x-carotenals used [x= 10' (256), 8' (248) and 4'] and (rather
surprisingly) from torularhodinaldehyde (142) faster than from P-carotene; it
split a-carotene (5) but did not attack P-carotene 5,6,5' ,6'-diepoxide (133). From
this one might have deduced that the enzyme was relatively non-specific to
the rest of the carotenoid molecule, provided that one unsubstituted P-ionone
moiety was present. An exception however was found in P-carotene 5,6-mono-
epoxide (114), which was not attacked by the enzyme, even though it can
be cleaved in vivo by the rat. On the other hand, 3,4,3',4' -tetradehydro-P-
carotene (1) was attacked at a slow but significant rate to yield 3-dehydro-
retinaldehyde [10]. Although certain aspects of this enzymic conversion remain
to be clarified, it seems probable that the enzyme is the major one involved
in the conversion of dietary carotenoids to vitamin A.
The activity of this enzyme is dependent on, among other things, the level
of protein in the diet [13]. A reduced protein intake drastically diminishes
the intestinal conversion of P-carotene to retinol [14], an effect which may
have serious consequences for the inhabitants of many of the poorer countries
of the world who rely on conversion of dietary carotenoids for their supply
of vitamin A rather than on preformed retinol or retinyl esters.
A little of the retinaldehyde formed in the intestinal wall from carotenoids
is oxidized to retinoic acid [15, 16], which passes into the portal blood [15],
but the majority is reduced to retinol [15] by an enzyme which in rat intestinal
mucosa can utilize either NADH or NADPH; although not a specific retinal-
dehyde reductase in that it will also reduce a number of other aldehydes, it
is not a. classical alcohol dehydrogenase for it does not oxidize ethanol [17].

C. Metabolism
The newly formed retinol is then handled in the same way as is dietary
retinol. Many animals, including most humans in the prosperous countries
720 G.A.J. PITT

of the world, receive their vitamin A not in the form of the provitamin carote-
noids but as retinol, which has been either formed by another animal from a
carotenoid, or synthesized commercially. Dietary retinol is usually in the
esterified form, and during absorption these esters are normally completely
hydrolysed in the small intestine, either by a powerful pancreatic hydrolase
or by a hydrolytic enzyme probably on the brush border of the intestinal
mucosal cell [14].
The free retinol is transferred across the mucosal cell membrane by an
'active transport' mechanism dependent upon oxidative phosphorylation;
metabolic inhibitors or lack of oxygen prevent its accumulation in mucosal
cells and its transport across the intestinal wall [18].
Whether obtained by hydrolysis of dietary retinyl esters or by reduction
of retinaldehyde formed in the cleavage of carotenoids, retinol is reesterified
inside the mucosal cells with long-chain fatty acids-mainly palmitate, fol-
lowed by stearate and oleate [19]. The retinyl esters are incorporated intO
chylomicrons in the mucosa and put into the lymph [19]. A few hours after
a meal the newly absorbed esters can be found in the blood, primarily with
the low-density lipoproteins [20, 21].
Retinol in excess of the body's immediate needs is stored mainly in the
liver esterified with long chain fatty acids, predominantly palmitate [22].
These reserve esters can be hydrolysed [23] and resynthesized [24], and
undergo a steady turnover [25, 26].
In the post-absorptive state the vitamin A in the blood is almost wholly
in the form of retinol, and the level remains reasonably constant provided the
liver contains adequate reserves of retinyl esters. Once these have been lost
from the liver, the blood level of retinol falls [27], the vitamin A content of
the eye [27] and of other tissues [28] declines, and eventually the overt signs
of deficiency appear [27]. The function of the liver reserve esters is therefore
to maintain a constant level of retinol in the blood to supply the vitamin to
peripheral tissues.
Retinol is carried in the plasma attached to a specific protein-retinol-
binding protein-which has a molecular weight of 21,000 and binds one mole-
cule of retinol [29]. It has been reported as being joined in plasma with pre-
albumin as a protein-protein complex [29].
Since the maintenance of a constant blood retinol level is dependent on
an adequate supply of retinol-binding protein, it is influenced by the slowing
down of protein biosynthesis brought about by inadequate protein nutrition.
Protein deficiency consequently interferes seriously with the transport of vit-
amin A by lowering the blood level of retinol and is thus a very important
factor in precipitating vitamin A deficiency in man [23, 30].
Retinol-binding protein serves as the vehicle to deliver retinol to the tis-
sues. It can there be oxidized to its aldehyde [31], which in turn can be further
oxidized irreversibly to retinoic acid, the two enzymes involved presumably
accounting for the conversion of retinol to retinoic acid observed in vivo
[33].
 X. Vitamin A 721
Retinoic acid very rapidly disappears from the animal body [34], in part
because it is readily converted to its jj-glucuronide, which is excreted in the
bile. Retinol can also form a jj-glucuronide, which is excreted in the bile,
although probably to a smaller extent than is the retinoic acid derivative.
This biliary excretion of vitamin A metabolites can be used as a 'detoxica-
tion' mechanism to deal with excessively large doses of the vitamin, but it
appears to operate also at low levels of dosing and probably in normal physio-
logical conditions. Although some of these glucuronides can be reabsorbed
from the gut in what is presumably an enterohepatic circulation, retinoic acid

Intestinal lumen
I Bile Liver Blood
I
I
RCOOG
+----------r-:-:~-t----__1
+-<_ _ _ _ _ _ _

RCH 2 0G'
[3~91 l
__,l..:.;l':..:8.:. . l- - - - h i
1

RCH 20G RCOOG+------------+------------~

ir 3 9J r 36 ~ Unidentified
I ~ metabolites
RCOOH +-------------------+-----------...
[39]
jr 4 oJ RBP i
RCHO :RCH 2 0H-RBP

1l[J 1
] 41] [43] :

RCH 2 0H----;;;- RCH,OCOR' ~

~-------Int;;t~lm-;;~s;---- ~----1-Jiymph;
l1BJ £44! J45J 1 I
R CH 2 0 H----1---'__:_----> RCH 2 0 H ;;::==>: R CH ,OCO R' -------> 1RCH20COR1
j£37] : 1------_j
RCH 2 0COR' I I
F-r-om--d-ie--,t~
.-1 : P 7J :

Provitamin A I r9 • 101 RCHO r16 l RCOOH P 5l IRCOOH

carotenoids I I, I
: f9J I
1RCOOG

r-tKiin;y-- ---
r46 l RCOOG
i

I47J
RCOOH

- [47J
t -.,;e~e;
----l
?
:1
I

I
:t
I I39J
!RCOOG~ RCOOH

Unidet~~~;ied
f-- __ .!!l~t§]l2)i~s-
: Urine 11351

RCOOG RCOOH RCH,OG I 1RCOOG Unidentified

I I metabolites

Scheme 1. Some m~;tabolic transformations of vitamin A (after Olson [32]). Abbreviations:

RCH 2 0H =retinol, RCH 2 0COR' = retinyl esters, RCH 2 0G = retinyl /3-glucuronide, RCHO =
retinaldehyde, RCOOH = retinoic acid, RCOOG = retinoyl /3-glucuronide, RBP =retinol-binding
protein

Carotenoids 46
722 G.A.J. PITT

is not extensively conserved in the rat by this means, and the faeces probably
constitute one of the major routes for the excretion of vitamin A derivatives
[32].
Metabolites are also excreted to a considerable extent in the urine, but
have not been identified further than that they contain P-glucuronides [32].
From a study of the metabolism by the rat of retinol and retinoic acid
radioactively labelled in different parts of the molecule, it was deduced [35]
that there were three pathways by which these molecules were eliminated by
the body. The first involved no shortening of the side chain, and would have
included the formation of retinoyl and retinyl glucuronide derivatives. The
second involved a decarboxylation of carbon atom 15 at the end of the chain,
and the third a much more extensive degradation of the side chain.
Retinoic acid can be decarboxylated by a peroxidase type of microsomal
enzyme [36], which would account for the second pathway postulated from
in vivo work, but no details are available of enzymic processes that might be
involved in more extensive degradation of the molecule.
The patterns of breakdown of retinoic acid and retinol appear to be roughly
similar in the rat, but retinol is disposed of much more slowly [35]. It is not
yet clear to what extent retinol is oxidized to retinoic acid for removal from
the body, for although this conversion can be demonstrated in vivo [33], it
is very difficult to measure how much retinoic acid is formed, presumably
because of the very rapid metabolism of any retinoic acid produced.
The metabolic transformations of vitamin A in the body are represented
in Scheme 1. They are influenced by many factors, particularly by the state
of protein nutrition of the animal which has severe repercussions on the
handling of vitamin A in the body [23, 30].
This is an extremely important practical problem for it is a shortage of
dietary protein that commonly accentuates or precipitates vitamin A deficiency
in man. Vitamin A deficiency is a very frequent concomitant of kwashiorkor
[23, 48], and it appears to be the unfortunate coincidence of a low dietary
level of vitamin A or carotenoid precursors with inadequate protein nutrition
that is responsible for much of the vitamin A deficiency still so distressingly
common in many parts of the world with a deplorably high mortality rate
[49, 50].

D. Function in the Eye

We therefore have a fairly detailed knowledge of what the body does to

vitamin A, but when we come to consider what vitamin A does to the body
-the physiological mode of action of vitamin A-we have only a very incom-
plete picture.
That vitamin A has more than one function was elegantly demonstrated
by using retinoic acid, the carboxylic acid derivative of retinol. If retinoic acid
is given to rats fed on a diet deficient in retinol, it maintains growth and the
animals outwardly appear healthy, but they become blind. Animals are not
 X. Vitamin A 723

able to reduce retinoic acid to retinaldehyde, which is required to form the

prosthetic group of their visual pigments [34].
It is this role in vision which is by far the best understood function of
vitamin A. Retinol is oxidized to its aldehyde, originally called retinene, but
now known as retinal or retinaldehyde [1, 4]; the retina contains more than
one enzyme capable of catalysing this oxidation, and the reverse reduction
[31, 51]. Retinaldehyde then joins with proteins of the type called opsins, to
form visual pigments.
The interactions of retinaldehyde, opsins and light in the functioning of
the visual pigments were elucidated in a brilliant series of investigations by
Wald, Hubbard and their colleagues in terms of the cis-trans isomerization
of retinaldehyde [52, 53]. The most stable form of retinaldehyde is the all-
tr~ns isomer (3), but to unite with an opsin to form a visual pigment, the 11-

11
~ ~ ~ CHO

(3) (4) HO

ali-trans- Retinaldehyde 11-cis-Retinaldehyde

cis isomer of retinaldehyde (4), which has a bent chain, is necessary; the straight
chain of all-trans-retinaldehyde will not link with opsin to form a pigment.
(The 9-cis isomer will also join with opsins in vitro but does not occur in
natural visual pigments [52].)
Visual pigments therefore consist of an 11-cis-retinaldehyde-opsin com-
plex; when struck by a photon the bent 11-cis-retinaldehyde chromophoric
group is isomerized to a steady-state mixture consisting mainly of the all-
trans form and some unchanged 11-cis chromophore [54]. The situation is
similar to what happens when one simply irradiates retinaldehyde in solu-
tion: any one isomer gives rise to a steady state mixture of isomers in which
the all-trans form predominates with, in some conditions, not inconsiderable
amounts of the 11-cis isomer. In the visual pigment, however, the straightening
of the polyene chain from the bent 11-cis form to the straight all-trans form
causes it to pull away from the opsin, initiating a train of events that results
in the transmission of an impulse up the optic nerve. In the complex series
of processes involved, the only thing that light appears to do directly to visual
pigment's is to isomerize the 11-cis-retinaldehyde portion to the all-trans form
[52, 53]. .
Retinaldehyde therefore operates with proteins of the opsin family in a
cis-trans isomerization cycle shown in its simplest form in Scheme 2 [53].
Although they are well established in outline, the processes summarized in
Scheme 2 are still under intensive investigation, for they pose many problems
still unsolved.
724 G.A.J. PITT

Visual pigment
(11-cis-retinaldeh yde-opsin complex)

/ ~Light
Retinaldehyde isomerase ~
Opsin+ 11-cis-retinaldehyde all-trans- Retinaldehyde +opsin

NADP
NADP1l
(or NAD)
J r (or NAD)

11-cis-Retinol ? all-trans- Retinol

1l Jr
11-cis-Retinyl esters all-trans- Retinyl esters

Scheme 2. Cis-trans isomerization cycle of retinaldehyde in vertebrate eyes (after Wald [53])

This basic cycle appears in a number of variant forms. In some amphibia

and fishes, usually fresh-water species, retinaldehyde is replaced by 3-dehydro-
retinaldehyde (5), the aldehyde of vitamin A 2 (once called retinene 2 ). Extensive

(5)
3-Dehydroretinaldehyde

investigations have been made into the distribution of retinaldehyde- and 3-

dehydroretinaldehyde-based pigments [55, 56]; it has been found that vitamin
A 1::;;:"::vitamin A 2 interconversions can take place [57-59] so the type of pig-
ment found is determined more by the genetic characteristics of the species
than by the dietary intake of vitamin or precursors.
The visual pigments of invertebrates differ in many respects from those
of vertebrates but still operate by the cis-trans isomerization of retinaldehyde
[60].
All naturally occurring visual pigments of which we know the chemical
make-up contain either retinaldehyde or 3-dehydroretinaldehyde [60]. In
contrast there are many opsins, as evidenced by the large number of known
visual pigments differing in their absorption maxima from 433 nm to around
565 nm in the case of retinaldehyde pigments, and from 510 to 620 nm
among those based on 3-dehydroretinaldehyde. Wald [52] has suggested
trivial names for these visual pigments, depending on the aldehyde involved
 X. Vitamin A 725

and on whether the opsin comes from rods or cones, the retinal cells concerned
with low-intensity (night) and high-intensity (day) vision respectively; the cones
are also responsible for colour vision in those species where it exists. The
trivial names of the visual pigments are rhodopsin (retinaldehyde +rod opsin),
porphyropsin (3-dehydroretinaldehyde +rod opsin), iodopsin (retinaldehyde +
cone opsin) and cyan opsin (3-dehydroretinaldehyde +cone opsin). This nomen-
clature system is not universally employed and has been criticized: the names
cover a range of pigments, and it is difficult and perhaps unnecessary to draw
a hard and fast line between rod and cone pigments in general; nevertheless
the usage is serviceable in many contexts.
Opsins are characterized by their ability to unite with a vitamin A alde-
hyde and shift its absorption spectrum to longer wavelengths. Although all
opsins have this property in common they are a large family of different pro-
teins distinguished from each other by the extent to which they induce this
bathochromic shift.
For this reason alone-apart from many others-the protein component
of the visual pigments offers a challenge. Chemical investigations on visual
pigments have concentrated on the rhodopsins and in particular on cattle
rhodopsin because of its ready availability. Cattle rhodopsin is not however
an easy protein with which to work. Because it is insoluble in water it has to
be extracted from the outer segments of rods by means of detergents, of which
the most popular since its introduction 40 years ago has been digitonin,
although recently others have been used.
Rhodopsin is a major component of the membranes of rod outer segments
[61], which, like all membranes, contain a considerable amount of phospho-
lipid [62, 63]. It is therefore not surprising that extracts contain sizeable
amounts of phospholipids intimately associated with rhodopsin [64, 65]. There
is, at the time of writing, controversy on this matter: some workers [66, 67]
consider that phospholipid is not an obligatory component of rhodopsin;
others state [65] that while the amount of phospholipid can be reduced to a
very low level (1 molecule ofphosphatidylethanolamine and 2 ofphosphatidyl-
serine per molecule of rhodopsin), it is not possible to make a completely
lipid-free preparation which retains the characteristic absorption peak of the
pigment.
All agree, however, that the polypeptide chain of opsin appears essential
for the maintenance of the rhodopsin spectrum. The polypeptide chain of
opsin has been reported as having a molecular weight of 26,000-28,000 [66,
68, 69]. Its amino acid composition has been determined in three separate
laboratories and shows a remarkably high content of non-polar amino acids
[66, 68, 69]; opsin also contains a single oligosaccharide moiety linked to the
polypeptide chain [70].
Some workers however claim that it is in the phospholipid portion of
rhodopsin that ·we must look for an explanation of a problem that has exer-
cised the minds of many: the absorption maximum of retinaldehyde shifts
from near 385 nm well into the visible region of the spectrum on union with
726 G.A.J. PITT

an opsin. Numerous ingenious ideas have been put forward to explain this
large bathochromic shift, frequently without much supporting evidence; almost
all, however, assume that the aldehyde group of retinaldehyde is joined to an
amino group of opsin in a Schiff base or related linkage. This is generally
accepted from work on the intermediates found in the photochemical break-
down of rhodopsin to all-trans-retinaldehyde and opsin, a process colloquially
known as 'bleaching' from the loss of colour as the pigment breaks down to
the almost colourless aldehyde and protein.
As mentioned, the only action that light is believed to have on rhodopsin
is to convert the 11-cis-retinaldehyde prosthetic group to the all-trans form;
this is followed by a series of thermal, i.e. non-photochemical, reactions before
the all-trans-retinaldehyde finally detaches itself from opsin. Intermediate sta-
ges in these 'dark' reactions can be revealed by irradiating rhodopsin at very
low temperatures to isolate the photochemical reaction and then allowing it
to warm up slowly. By this means it has been possible to detect a number of
intermediates, shown in Scheme 3. Investigations on the photochemical break-
Rhodopsin (498 nm)
~ Li1ht
Prelumirhodopsin (543 nm)
~>-140"C
Lumirhodopsin (497 nm)
~>-40"C
Metarhodopsin I (478 nm)
~ >-15"C
Metarhodopsin II (380 nm)
~ > -5 "C

Metarhodopsin 465 (465 nm)

or Metarhodopsin III
or Pararhodopsin
~ +15 ·c
N-Retinylidene-opsin
('indicator yellow': 440 nm in acid, 365 nm in alkali)
~ +H20
all-trans- Retinaldehyde +opsin
Scheme 3. Photochemical breakdown of rhodopsin [71]; wavelengths denote absorption maxima
of derivatives of cattle rhodopsin

down of rhodopsin have been carried on for over 30 years, becoming increas-
ingly more complex with the discovery of more intermediates. Scheme 3 rep-
resents a version which is unlikely to remain unmodified in the future; probably
many of the workers in the field even now would disagree with one or more
of the details.
 X. Vitamin A 727
For example, the exact status of N-retinylidene-opsin is far from clear, but
this molecule is of importance in that it is the only one of which we have firm
knowledge of the chemical structure. N-Retinylidene-opsin is a Schiff base
formed by the reaction of the aldehyde group of retinaldehyde with an amino
group in the opsin complex, and provides the best chemical clue to the nature
of the link between retinaldehyde and opsin.

Retinaldehyde N-Retinylidene-opsin

The compound was originally named 'indicator yellow' because of its

behaviour on changing pH. In alkaline solution it exists as an uncharged
Schiff base with an absorption maximum near 365 nm, so appearing colour-
less; in acid solution it is protonated to the yellow conjugate acid absorbing
near 440 nm [72].
Ell
C19 H 27 CH=N-Opsin +HEll ~ C19 H 27 CH=NH-Opsin

It is thought [72] that a carbon-to-nitrogen link is present in rhodopsin

and remains intact during its breakdown to N-retinylidene-opsin, a process
which in certain conditions can be reversed [73]. The hypothesis has been
widely accepted that a major link between retinaldehyde and opsin is formed
by the aldehyde group and an amino group on opsin in a Schiff base or some-
thing derived from a Schiff base.
Suggestions have not been lacking as to how this might induce the batho-
chromic shift observed when retinaldehyde unites with opsin. Since protona-
tion of the Schiff base will account for about half the required shift, a protonated
Schiff base structure has attractions as the basis of the rhodopsin chromophore
[74]. A difficulty is that retinylidene Schiff bases are normally protonated only
at unphysiologically low pH. In anhydrous conditions, however, the Schiff
base formed from retinaldehyde and the phospholipid phosphatidylethanol-
amine-retinylidene phosphatidylethanolamine-can undergo internal proto-
nation from the phosphoric acid group of phosphatidylethanolamine [75]. It
has been reported [76] that if this Schiff base is just left to stand in anhydrous
chloroform-methanol, it will complete the required bathochromic shift; its
absorption maximum moves to near 500 nm-but only if the phosphatidyl-
ethanolamine contains unsaturated fatty acids (which are known to be present
in large amounts in the lipids found in rhodopsin preparations [62, 63]; it is
suggested that the unsaturated fatty acids of the phospholipid show a n-rc
interaction with the retinaldehyde.
Some, however, would consider this extra interaction with unsaturated
fatty acids to be unspecific or physical rather than chemical. Protonated Schiff
bases show very large bathochromic shifts (up to 515 nm) in the non-polar
solvent 1,2-dichloroethane [77]; it has been proposed [77] that this pheno-
728 G.A.J. PITT

menon is involved in formation of the visual pigment chromophore, the exact

absorption maximum being simply dependent on the dielectric constant of
the environment of the chromophore and (since the shift in the analogues
depended to some extent on the acid used for protonation) on the association
between the cation and its counter-ion. The same basic hypothesis has been
presented in a rather more elaborate and refined form [78]; this suggests that
the opsin matrix provides an anhydrous microenvironment, of which the
polarizability (rather than its non-polarity [77]) is the essential property for
forming a pigment, the iminium nitrogen in the Schiff base generating induced
dipoles in this microenvironment.
Currently popular hypotheses on the chromophoric site therefore lay great
stress on the physical conditions at the active site of opsin. Workers, however,
are divided on how much onus they place on phospholipids for the formation
of the chromophore, the enthusiasts for phospholipids implying that the func-
tion of the polypeptide portion of opsin is merely to provide a hydrophobi\;'
pocket in which retinaldehyde and phospholipid can interact in an anhydrous
environment possessing the correct physical characteristics.
It is therefore essential to know whether phospholipids are present at the
active site of rhodopsin, and, in particular, to what group retinaldehyde is
linked in the intact pigment.
The latter problem has caused much difficulty in recent years. It appeared
to have been solved independently in two laboratories almost simultaneously,
by the use of sodium borohydride to reduce the Schiff base linkage of N-
retinylidene-opsin [79, 80]. Sodium borohydride has no action on rhodopsin
itself, but if the pigment is illuminated in the presence of borohydride, a
reduced Schiff base is obtained in which the original retinaldehyde portion
is present as a retinyl group believed to be still attached to opsin at the chromo-
phoric site-the compound N-retinyl-opsin. The site of attachment of the
retinyl group was identified as an ~:-amino group of a lysyl residue [81, 82].

BH£'

N-Retinylidene-opsin N-Retinyl-opsin

A few years later, however, doubt was cast upon this apparently satisfactory
conclusion. If rhodopsin is bleached in the presence of borohydride, the retinyl
group is recovered attached to lysine, but if rhodopsin is treated in the dark
with dry acid methanol, the retinyl group can be found attached instead to
the amino group of phosphatidylethanolamine [83]. There is evidence [84]
however that this is an artifact formed only in a lipophilic non-polar environ-
ment such as is believed to exist in the intact rhodopsin molecule and its
breakdown products up to and including metarhodopsin I.
The changes occurring in the conversion of metarhodopsin I to meta-
rhodopsin II are thought [84] to involve the disruption of the non-polar lipid
microenvironment of retinaldehyde in metarhodopsin I to a polar aqueous
 X. Vitamin A 729

microenvironment in metarhodopsin II. This would eliminate the possibility

of forming the phosphatidylethanolamine Schiff base as an artifact-and, in
accordance with the hypothesis described earlier, would 'bleach' the pigment
by destroying the hydrophobic pocket responsible for shifting the absorption
peak of retinaldehyde into the visible region.
The topic is controversial and with several laboratories working on it,
further developments can be expected. At present, an eclectic view of the
active site on rhodopsin is that it contains a protonated retinylidene Schiff
base with its absorption maximum shifted to longer wavelengths as a conse-
quence of the physical properties of its microenvironment. One function of
the polypeptide chain of opsin is to maintain this hydrophobic pocket which
must contain phospholipid-whether the retinylidene group is or is not nor-
mally attached to phosphatidylethanolamine, the two molecules must be in
very close proximity to allow the formation of N-retinylidene-phosphatidyl-
ethanolamine, even if only as an artifact.
Much interest in the intermediates of bleaching stems from the fact that
during these stages a nervous impulse is triggered by some mechanism as yet
unclarified [85].
A popular hypothesis involves changes in the opsin portion of the molecule
[71, 86]. There is much evidence that during the' dark' reactions opsin changes
its configuration [69], although such findings have to be interpreted with
caution since the behaviour of rhodopsin in rod outer segments is not always
the same as in detergent micelles in an aqueous medium [69, 87], the most
common experimental conditions employed for investigations of this type.
The changes in opsin are set in train by the photoisomerization of the
11-cis chromophore to the all-trans configuration, when rhodopsin is changed
to prelumirhodopsin [54] (see Scheme 3). This is presumed to release opsin
from the specific configuration in which it has been held by the 11-cis aldehyde.
11-cis-Retinaldehyde also has the property of causing the free opsin to rearrange
itself back into the specific configuration of the visual pigment [88, 89]. The
induced refolding of the protein is a normal part of the regeneration process
since only 'bleached' opsin and 11-cis-retinaldehyde are required for the
spontaneous formation of a visual pigment in vitro. For this reason it is possible
to regenerate rhodopsin at various stages of the bleaching process by irradiat-
ing the intermediates shown in Scheme 3. The retinaldehyde attached to them
in the all-trans form can be photoisomerized to give a mixture containing
some 11-cis form, which then can pull the opsin back into its original con-
figuration, i.e. it is possible to use light to reverse to some extent the bleaching
process -started by light [53]. Regeneration of visual pigments either from
retinaldehyde and opsin or from the intermediates in the breakdown process
appear to provide a spectacular analogy to the 'induced fitting' of the active
site of an enzyme to the substrate molecule [90].
Indeed in the case of the rat, retinaldehyde is required not merely to hold
opsin in the correct configuration but to maintain the very existence of opsin.
As mentioned previously, rats maintained on retinoic acid become blind for
730 G.A.J. PITT

lack of retinaldehyde in their visual pigments. The absence of retinaldehyde

from the rat's retina leads to the disappearance of opsin too and to the sub-
sequent degeneration of the visual cells [34]. This startling effect of the lack
of retinaldehyde on the retinal histology of the rat is not however a general
one, as it is not seen in guinea pigs [91] or chickens [92]; it appears to be a
consequence of the great instability of rat opsin [93].
There is considerable controversy regarding precisely what changes occur
in the dark reactions and exactly where they occur in going from prelumi-
rhodopsin to N-retinylidene-opsin [71, 85] for this is a complex and difficult
problem to tackle. But all workers would agree with the perceptive suggestion
made many years ago [94] that the effect of light is to bring about a reversible
denaturation of the protein.
The major functions of retinaldehyde in vision are clear. It joins, in the
11-cis form, with opsin, inducing configurational changes in the protein which
produce a bathochromic shift enabling the retinaldehyde prosthetic group (o
absorb, to the considerable advantage of animals, the much more plentiful
radiation in what we consequently know as the visible region of the spectrum
[60]. By being readily isomerized by light, to the all-trans form, the retinalde-
hyde releases the opsin from its fixed configuration and thus, by what is
probably an indirect triggering action, initiates events resulting in the genera-
tion of a nerve impulse.
It is far from clear just how retinaldehyde carries out these functions, but
not all the molecule appears necessary for its interactions with opsin. Although
all naturally occurring visual pigments appear to contain retinaldehyde or
3-dehydroretinaldehyde, this is not a restriction imposed by the opsins. Arti-
ficial visual pigments have been made by uniting opsin in vitro with variants
on retinaldehyde. The first made was from 4-oxoretinaldehyde which joined
with cattle, sheep or rat opsins to give a pigment showing A.max 465-470 nm
[95]. Many others have since been made from retinaldehyde 5,6-epoxide [96]
and 5,8-epoxide [96], IX-retinaldehyde [85], 5,6-dihydroretinaldehyde [97],
9-demethylretinaldehyde [98], 9,13-didemethylretinaldehyde [98] and 13-
demethylretinaldehyde [98, 99].
Attempts have been made to equip animals with an artificial visual pig-
ment by having them form one of these new pigments in vivo, but with only
scant success. A claim to have done so with retinaldehyde 5,6-epoxide has not
been confirmed: rats given the epoxide had almost no artificial pigment in
their retinas [96]. 4-0xoretinaldehyde completely failed to form any pigment
in vivo when fed to rats; one reason here for the great contrast between in vivo
and in vitro behaviour was that 4-oxoretinaldehyde was very rapidly destroyed
in the rat body [3, 95]. ·Certain structural features of vitamin A appear to
be necessary to ensure reasonable stability in animal tissues [3]. It seems
probable that many carotenoids could give rise to a half molecule capable
of joining with opsin in vitro to form a visual pigment, but it is exceedingly
unlikely that they would ever be formed in vivo: they would be either destroyed
by animal tissues, or shut out by the eye.
 X. Vitamin A 731

Although cattle rhodopsin is by far the best studied visual pigment, other
rhodopsins have been investigated, also porphyropsin and the cone pigment
iodopsin. Th!.!re are differences from cattle rhodopsin, notably among inverte-
brate visual pigments (which are commonly referred to as rhodopsins), but
cattle rhodopsin, on which most of this account has been based, exemplifies
well the role that vitamin A plays in all visual pigments.
Vision in the normal animal involves the operation of the cycle shown in
Scheme 2 at different rates and with different levels of visual pigment depend-
ing on the intensity of light to which the retina is exposed. In relatively bright
light, the level of visual pigment falls, to rise again on moving into darker
conditions-with related changes in the visual sensitivity of the animal. The
rat eye has been studied in detail [100]. Exposure to light liberates from the
bleached rhodopsin retinaldehyde, which is reduced to retinol probably in
the rod outer segments [51], esterified elsewhere in the retina [101] and trans-
ferred to the pigment epithelium lying between the retina and choroid; most
of the vitamin in the pigment epithelium is in the form of retinyl esters [100],
again mainly as the palmitate plus some stearate and a little oleate [102].
These processes are reversed when the animal moves out of strong light and
constitute part of the familiar experience of dark adaptation.
A feature of these stores of ester which has not yet been wholly explained
in a satisfactory fashion is that they contain up to 65% of 11-cis-retinyl esters
[103]. A specific enzyme, retinaldehyde isomerase (EC 5.2.1.3), exists in the
eyes of cattle and frogs [104], and it has been briefly reported that cattle rod
outer segments can isomerize retinaldehyde non-enzymically [105], but these
processes do not appear able to account adequately for the high 11-cis content
of the retinyl esters in the frog pigment epithelium and for the maintenance
of these proportions regardless of whether the eye is illuminated or not: the
percentage of 11-cis esters present is independent of the rate at which the
visual cycle operates [106]. The pigment epithelium must possess either another
enzyme capable of isomerizing retinol or retinyl esters or some mechanism
involving specific binding of the 11-cis isomer [107]. Retinol isomerases are
known to be present in other tissues [108] but the eye alone contains 11-cis
isomers, which are not detectable in other tissues [106]. The problem is
complicated by species differences: in contrast to frogs and cattle, which have
eye reserves of retinyl esters in all conditions [103, 107], the rat when fully
dark-adapted has almost all the vitamin A in its eye bound as retinaldehyde
to opsin in rhodopsin [100].
The eye is capable of running a fairly closed system for vitamin A which
can in some circumstances be isolated from the general metabolic pool of
vitamin A. This was recognized many years ago in the eyes of fish which have
a characteristic A 1/A 2 ratio different from that found in their livers [103]. A
striking demonstration was seen in rats depleted of retinol then given 3-
dehydroretinol. Although their liver stores would almost certainly have con-
sisted of 3-dehydroretinyl esters, the eye pigment contained predominantly
retinaldehyde [58].
732 G.A.J. PITT

The eye stores of vitamin A in the rat can exchange fairly rapidly with the
general metabolic pool in favourable conditions [109], but when necessary
the eye can hold tenaciously to its stores of retinaldehyde, long after other
tissues have lost all their retinol [93], perhaps because the retina lacks the
enzymes retinaldehyde oxidase and glucuronyl transferase involved in the
removal of vitamin A from the body [39]. When a rat is maintained on a
marginal intake of dietary retinol, the eye takes up retinol much more avidly
than other tissues which also have a need for it [28, 110].

E. Systemic Mode of Action

The metabolism of vitamin A in the eye can therefore be separated in

various ways from its metabolism in the body as a whole, not surprisingly,
since the role of the vitamin in vision is clearly different from its more general
function in the body.
While admittedly many aspects of vision remain to be solved, the opera-
tion of vitamin A in the eye is clear in principle, but this must be considered
a subsidiary role of the vitamin. Experimental animals, when fed on a diet
deficient in vitamin A, die, and not from a lack of visual pigments. The more
fundamental task of vitamin A in maintaining growth, health and life is often
referred to as the systemic function of the vitamin, i.e. relating to the body as
a whole rather than to any specific organ or tissue. It is this systemic function
of the vitamin which raises the greatest unsolved problem in the vitamin A field.
A well-tried approach to the mode of action of a vitamin is to observe
what goes wrong in deficiency and then track back to see what specific bio-
chemical mechanism has failed. Unfortunately vitamin A deficiency signs are
very numerous and diverse [2, 50, 111], involving the eyes, skin, nervous
system, bone, respiratory system, reproductive organs, gastro-intestinal tract,
urinary system and liver, to name some of the organs in which a variety of
lesions have been reported in vitamin A deficiency. To some extent this situa-
tion is predictable: any primary dietary deficiency inevitably leads to secondary
consequences; for example, a very characteristic lesion of vitamin A deficiency,
the keratinization of numerous epithelia [112], opens the body to infection,
which is usually the cause of death in experimental animals on an A-deficient
diet [113]. A major difficulty with vitamin A, however, is to sort out which
(if any) of the many observed deficiency signs are primary lesions, and which
rather more distant sequelae.
A minor simplification of this confusing picture was brought about by the
use of retinoic acid. Rats -on a diet deficient in retinol but containing retinoic
acid as the sole source of vitamin A grew well and outwardly appeared healthy,
but, as already mentioned, became blind [34]. Furthermore, they lost the
ability to reproduce [114].
Female rats on such a diet had normal oestrus cycles, mated and con-
ceived and the foetuses began to develop normally, but during the third week
 X. Vitamin A 733

of pregnancy the foetuses were invariably resorbed, apparently as a conse-

quence of placental failure. In male rats, spermatogenesis stopped and the
seminiferous epithelium sloughed off. The lesions can be reversed by giving
retinol [114, 115].
Retinoic acid therefore acts as a partial vitamin A: it carries out some
roles of the vitamin but it cannot replace it in vision or reproduction in the
rat or guinea pig. Because it carries out the function of the vitamin in maintain-
ing general health and life, the use of retinoic acid eliminates the effects of
general vitamin A deficiency on the reproductive organs [115].
There has been abundant evidence for many years that vitamin A is
required for reproduction in all species studied [2, 115], but this in itself is
not surprising since it is well known that vitamin A deficiency results in inani-
tion, defective epithelial tissues, infection, endocrine disturbances and other
defects likely in themselves to affect reproductive processes. These multiple
consequences of vitamin A deficiency have bedevilled the study of the defects
in reproduction, which were frequently considered non-specific consequences
of general vitamin A deficiency. The work with retinoic acid demonstrates
however that vitamin A has a more direct action on reproduction.
What this direct action is remains uncertain. What common action can
there be requiring vitamin A of which the failure results in cessation of sper-
matogenesis and placental lesions which cause foetal resorption? Although
it has been suggested that endocrine processes may be affected by lack of
retinol [116], other evidence indicates that this is not the major explanation
of the reproductive failure [117-119]. The problem is slightly complicated
by the subsequent finding that the cock can maintain normal spermatogenesis
on a diet containing no retinol, provided retinoic acid is present [92].
In the species that have been found to show these defects when maintained
on retinoic acid-rat [114], guinea pig [91], hamster [115] and pig [120]-
one can demonstrate the lesions in the reproductive system, and describe them
clearly even though the role that vitamin A plays in preventing them remains
obscure. By the use of retinoic acid one can therefore carry out a partial dis-
section of some of the effects of vitamin A deficiency-in vision and repro-
duction-from the mass of lesions usually observed in A-deficiency. A com-
parison of the reproductive system of animals maintained on retinoic acid
with that of orthodox A-deficient animals illustrates the complicating conse-
quences of the failure of the systemic function of the vitamin [114].
Many have sought to explain the numerous and diverse deficiency signs
in terms of some fundamental lesion that can manifest itself in such disparate
ways. Similarly ramifying consequences are found in deficiencies of vitamins
of the B complex-which are known to work as parts of co-enzymes. Many
workers have therefore looked to see if the fundamental lesion or lesions of
vitamin A deficiency can likewise be explained in terms of the failure of some
enzyme or related enzymes.
A number have been reported as affected by vitamin A. They include:
ATP sulphurylase (EC 2.7.7.4) involved in the activation of sulphate [121-127];
734 G.A.J. PITT

sulphotransferase (EC 2.8.2.1 ), which passes the sulphate group from adenosine
3' -phosphate 5' -sulphatophosphate ('active sulphate') to the substance being
sulphated [122, 128]; ,1 5 -3{1-hydroxysteroid dehydrogenase (EC 1.1.1.51),
an important enzyme in steroid hormone biosynthesis, involved in the con-
version of pregnenolone to progesterone and of dehydroepiandrosterone into
androstenedione [129, 130]; steroid 11{1-hydroxylase (EC 1.14.1.6), e.g. in
the formation of corticosterone from deoxycorticosterone [131]; squalene
oxidocyclase (EC 1.14.1.3) involved in the synthesis of the steroid ring
[132]; L-gulonolactone oxidase (EC 1.1.3.8) in the synthesis of ascorbic acid
from glucose [133, 134]; p-hydroxyphenylpyruvate oxidase (EC 1.14.2.2)
[133]; and codeine demethylase, a microsomal enzyme involved in the me-
tabolism of drugs [135, 136]; plus some mitochondrial oxidative enzymes
[137].
All have been reported as falling off in activity in A-deficient animals, and
being restored to normal when vitamin A is refed. It is not surprising that some
enzymes should so behave. Deficient animals usually stop eating, lose weight
and become ill. Even though devices such as pair-feeding can partially com-
pensate for such effects, one might expect the activity of some enzymes to be
affected indirectly by vitamin A deficiency [134, 138 -140].
The hypothesis that vitamin A directly affected some enzymes was how-
ever greatly strengthened by reports that some of these enzymes could be
activated by the addition of vitamin A in vitro: for example, ATP sulphurylase
[121-123, 125], sulphotransferase [122, 128], ,1 5 -3{1-hydroxysteroid dehydro-
genase [130] and steroid 11 P-hydroxylase [131].
Much controversy has arisen over the effects of vitamin A deficiency on
enzymes, especially over reactivations in vitro, for it has often proved difficult
to reproduce findings in different laboratories and even at times in the same
laboratory [125, 130, 139-145]. That competent workers in different labora-
tories can demonstrate these effects on enzymes confirms their reality. That
they cannot always be obtained suggests that one is probably concerned here
with secondary consequences of more primary lesions [134, 138-140, 142,
145]-this would account for the species differences observed [146, 147]-and
the reactivation of enzymes by adding vitamin A in vitro probably involves
some non-specific phenomenon [139, 141, 148-150].
If one looks at the great variety of enzymes involved, it is hard to think
of any simple co-factor role for vitamin A. Advances in our knowledge make
less plausible the idea that vitamin A will operate as an enzymic co-factor in
any way analagous with those of the B complex vitamins. For the co-enzymes
involving B vitamins are required in biochemical reactions essential for all
forms of life-animal, plant and microbial-whereas vitamin A seems to be
required only by intact animals or animal tissues. Many animal cells can grow
in culture in vitro without any apparent need for vitamin A [142]; the moth
Manduca sexta does not require vitamin A other than for vision [151], and
the housefly also probably requires vitamin A only in the eye and not for
growth [152].
 X. Vitamin A 735

That vitamin A is however required in some very fundamental mechanism

in higher animals was demonstrated [92] by feeding hens on a diet containing
retinoic acid as the sole source of vitamin A. The hens eggs could be fertilized
and began to develop in the usual way. After about 48 hours development
became abnormal, the embryos became disorganized and always died. Death
was due to vitamin A deficiency: if retinol was injected into the egg, the develop-
ment of the embryo continued, and some chicks were hatched. These embryos
cannot be affected by many of the traditional lesions of A-deficiency seen in
experimental animals in tissues such as epithelia, bone and the central nervous
system. The defect must be in some very fundamental mechanism.
Several lines are currently being pursued in the hope of identifying some
common primary lesion, but there is space to mention only a few. For example,
the biosynthesis of glycoproteins very characteristically falls off in vitamin A
deficiency; a possible role has been suggested for vitamin A in a sugar carrier
intermediate in the biosynthesis of glycoproteins [153].
A popular hypothesis has been that vitamin A is implicated in some way
in the structure of membranes and exerts its action by means of its effects on
membranes [23, 154-156]. Much of the experimental work in support of this
hypothesis has come from work with hypervitaminosis A. For example erythro-
cytes, when suspended in high concentrations of retinol in vitro, are haemolysed
as a consequence of the penetration of retinol into their membranes [154].
Similar conditions cause lysosomes to release hydrolytic enzymes [154, 155],
and this is believed to be part of the mechanism whereby large doses of vit-
amin A produce their characteristic damage to embryo chick limb bones in
vitro and to the bones of experimental animals in vivo [154, 155, 157].
If hypervitaminosis A is an excessive uncontrolled manifestation of the
normal mode of vitamin A, then one may deduce that the latter is concerned
in some way with membrane structure [155-157]. The relevance of hyper-
vitaminosis A to the physiological mode of action, however, now appears
more doubtful as a result of work [158] with ex-retinol (6), i.e. retinol with an
oc-ionone rather than a P-ionone ring.

(6)
IX- Retinol

oc-Retinol can induce the membrane effects of hypervitaminosis A in vitro

as well as retinol itself; in vivo it is just as effective in causing thinning and
fractures of bones, but has only 2% of the growth-promoting activity of
retinol. Since we have a compound which is very active in inducing hyper-
vitaminosis A, but has little growth-promoting activity for animals on an A-
deficient diet, the conclusion seems inescapable that hypervitaminosis A is not
necessarily related to the normal mode of action of the vitamin [158]. As the
hypothesis that vitamin A works in membranes is based to a large extent on
736 G.A.J. PITT

evidence from hypervitaminosis A, that hypothesis must have its plausibility

diminished proportionately by the findings on a-retinol.
Reports of electron microscope studies differ in whether a change can be
seen in membranes in vitamin A deficiency [136, 159], but there still remains
other evidence [155, 160] in support of the suggestion that vitamin A has a
function in the structure of membranes.
Along with others not mentioned here for lack of space, these hypotheses
based on the direct study of lesions in vitamin A deficiency or vitamin A excess
have not therefore enabled big steps to be made towards solving the problem
of what vitamin A actually does. Nevertheless if one considers the effects of
such conditions on epithelial tissues, one can discern a pattern. In vitamin A
deficiency mucous epithelia change over to keratinizing epithelia [2, 112]. On
the other hand an excess of the vitamin inhibits keratinization in epithelia
normally keratinized and can produce a mucus-secreting tissue [161]. Similar
changes can be demonstrated in a more dramatic fashion in the more con-
trollable conditions of tissue and organ culture [162].
These findings suggest that vitamin A controls the differentiation of some
cells, a field which is difficult to investigate because of the lack of detailed
biochemical knowledge of the mechanisms involved. Attention has however
been given to the DNA-+ RNA-+ protein synthesis series of reactions to see
if vitamin A can be shown to have a controlling effect on these fundamental
biochemical reactions regulating the behaviour of cells.
While vitamin A deficiency can be shown [163] to have consequences
attributable to action at the translational level (i.e. the RNA-+ protein syn-
thesis stage), again it is hard to distinguish direct from indirect effects of vit-
amin A deficiency. Referring to an earlier stage in this sequence of reactions,
there are reports that the administration of retinol or retinoic acid will stimulate
the biosynthesis of RNA in A-deficient rats. The suggestion is that vitamin A
may act at the transcription stage, i.e. on the formation of messenger RNA
[159]. In support of this hypothesis can be cited the findings that actino-
mycin D, which inhibits DNA-dependent RNA synthesis, reduces the action
of vitamin A on bone resorption in tissue culture or on the restoration of
codeine demethylase activity in the liver of previously deficient rats [136, 160].
It remains to be seen whether any of these approaches will penetrate the
cloud of ignorance that surrounds the molecular mode of action of vitamin A,
for it has to be admitted that despite the great volume of competent and
thoughtful work that has gone [23], and is still going, into investigations on
the systemic mode of action of the vitamin only a preternatural optimist would
suggest that the problem looks likely to be solved in the foreseeable future.
If vitamin A controls processes of growth and differentiation of which we do
not understand the basic mechanisms there is at present little chance of being
able to explain how it does so. Should that pessimistic view turn out to be
justified, those investigating the mode of action of vitamin A can at least
console themselves with the thought that they are really working ahead of
their time.
 X. Vitamin A 737

One consequence-or could it be the cause?-of our lack of knowledge

of the mechanism in which vitamin A is involved is that there is not even
certainty of the form in which it operates, apart from in the visual pigments.
Moore [2] has summarized some of the evidence which led workers to postulate
an active form of vitamin A other than retinol itself: notably the discrepancy
between the dose required to maintain full growth and that needed to accu-
mulate liver reserves; the delay between the disappearance of vitamin A from
storage tissues and the appearance of deficiency signs; and the inability to
detect vitamin A chemically in tissues where it is known to act. All these
fostered the idea that the function of retinol might be to act as the source of
some other substance discharging the role of vitamin A in tissues. This some-
what vague notion received a boost from the recognition that retinoic acid
could be formed in the body from retinol [33], that it had considerable growth-
promoting activity without being converted to retinol [34], and that it was
more effective than retinol in inducing hypervitaminosis A [164-166]. The
idea gained in credence that the systemically active form of the vitamin might
be retinoic acid or, in view of the very rapid disappearance of retinoic acid
from tissues, some substance formed from it [34], and a number of reports
appeared mentioning an active metabolite [ 123, 167 -170].
Some of these have subsequently been withdrawn [139] or explained away
satisfactorily in other terms [171]; none has won acceptance as the hypo-
thetical 'active vitamin A'. Although a number of metabolites of retinol and
retinoic acid have been reported [172], there is no reason to postulate any-
thing more than the normal degradative metabolism of the alcohol and acid
[36] to biologically inactive material. No conclusive evidence exists of vit-
amin A activity in any metabolite of retinol past the stage of retinoic acid (or
its p-glucuronide [ 4 7]). When very small doses of radioactive retinol were given
to rats, the major form found in tissues was retinol rather than a metabolite
[173]. Even when the animals were on the brink of deficiency and clearly in
need of all the vitamin A they could get into the active form, retinol pre-
dominated in all tissues, except in the liver where there was an approximately
equal amount of retinoic acid [6, 28].
Even though the positive evidence for an active metabolite of retinol is
almost non-existent, the concept persists. At a conference in 1968 attended by
a large number of workers in the vitamin A field a straw vote was taken [6]:
only a very small number present favoured retinol itself as the active form of
the vitamin; very slightly more considered an active metabolite to be more
likely; the overwhelming majority of those present cautiously abstained, indi-
cating the general feeling that more needs to be done on this topic before a
reliable judgem.ent can be given. Declaring a personal prejudice, I would opt
for retinol as the active form of vitamin A, on the principle of Occam's razor,
in the absence of direct evidence to the contrary.
Perhaps one ·of the most intriguing aspects of vitamin A, though alas not
one susceptible to scientific investigation is why higher animals need it at all.
In the course of evolution animals seem to have seized upon certain plant

CarotenOids 47
738 G.A.J. PITT

carotenoids to break them down to smaller molecules to utilize for their own
characteristic function of vision. The ubiquity of 11-cis-retinaldehyde in visual
pigments has been commented upon by Wald [60]. At least three times dur-
ing evolution eyes have developed independently: in arthropods, cephalopods
and vertebrates. Although the anatomy of these eyes varies greatly they have
all separately taken 11-cis-retinaldehyde as the prosthetic group of their visual
pigments, presumably because of the peculiarly suitable properties of 11-cis-
retinaldehyde as a photoreceptor molecule.
This situation, implausible prima facie, can be rationalized in terms of
evolutionary theory [60]. The basic improbability is amplified by the realiza-
tion that during the course of evolution higher animals have seized once more
upon carotenoids to obtain a breakdown product, retinol, needed for a quite
different function, without which higher organisms cannot even exist. What
process is there in higher animals which renders them wholly dependent upon
this molecule taken originally into animal bodies for one purpose and then
used in a slightly different form for a separate function? By fastening on to
it animals have tied their very existence to the procuring of a portion of certain
carotenoids either directly or indirectly through other animals. The price for
this quirk of development has been and still is being paid by multitudes of
men, women and-above all-children. For vitamin A de_ficiency is a major
nutritional disease of man responsible for many cases of irreversible blindness
in children and, although perhaps less harrowing because less conspicuous,
even more general ill-health and suffering [ 49].

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 743

XI. Use of Carotenoids

J.C.BAUERNFEIND, G.B.BRUBACHER, H.M.KLXUI and W.L.MARUSICH

F. Hoffmann-La Roche & Co. Ltd., Basle, Switzerland,

and Hoffmann-La Roche Inc., Nutley, New Jersey, USA

A. Significance as Vitamin A Precursor . 744

1. Scientific aspects . 744
2. Practical aspects . 745
B. Pigmenters in Feed . 746
1. Introduction 746
2. Stability of carotenoids in feed 746
3. P-Carotene in cattle products . 747
4. Carotenoids in poultry products 748
a) General remarks 748
b) Egg yolk . . . . . . . . . 749
c) Broiler . . . . . . . . . . 752
d) Other applications in the poultry industry . 753
5. Carotenoids for pigmenting zoo-birds and fishes 754
C. Pigmenters in Food . . . . . . . 754
1. Introduction . . . . . . . . 754
2. Natural occurrence and stability 755
3. Food colours . . . . . . . . 758
a) General . . . . . . . . . 758
b) Natural carotenoid extracts available as food colours . 758
c) Products prepared from pure chemical compounds . 759
D. Other Carotenoid Uses 762
1. Pharmaceuticals 762
2. Cosmetics . . 762
E. Assay Problems . 762
1. Assay in feeds . 762
2. Assay in foods . 764
References . . . . . 764
744 J. C. BAUERNFEIND, G. B. BRUBACHER, H. M. KLAUI and W. L. MARUSICH

A. Significance as Vitamin A Precursor

Apart from synthetic vitamin A and pre-formed vitamin A contained in

milk, eggs, liver and fish liver oils and their derivatives, the main source of
supply of this vitamin for man and the farm animal is the so-called provitamin A
present in plants and belonging to the group of carotenes and related carot-
enoids. In fact, /3-carotene is the most important vitamin A precursor in human
nutrition and in animal rations, since its presence in food and feed ingredients,
particularly ofleafy origin, greatly exceeds in concentration the other vitamin A
active compounds. Cryptoxanthin is present in yellow corn (maize) and
contributes significant vitamin A activity when high levels of yellow corn are
used in animal rations.
Because of the general importance of /3-carotene, other carotenes and
carotenoids as a main source of vitamin A supply in human and animal
nutrition, many efforts have been made to evaluate the exact vitamin A
potencies of individual carotenes and carotenoids. For a detailed discussion
of results found by different experimental techniques see [1-7]. When com-
paring the conclusions of the various authors, many discrepancies are found.
In general, these discrepancies arise from the different points of view, since the
problem has not only a scientific but also a practical aspect. For solving the
scientific problem, pure carotenes and carotenoids are investigated under
laboratory conditions, whereas the practical problem consists in determining
the vitamin A potency of naturally occurring mixtures under practical field
conditions.

1. Scientific aspects
The vitamin A activities of pure compounds have been measured in bio-
logical trials .under well-defined laboratory conditions. Table 1 is a compila-
tion of results obtained by different authors in the so-called rat growth curative
assay.
Table 1. Vitamin A activities of carotenoids (rat growth curative assay data)

Carotenoid Vitamin A activity*

i.u./g

P-Carotene (3) 1,667,000

()(-Carotene (S) 880,000
y-Carotene (8) 750,000
Echinenone (148) 890,000
Cryptoxanthin (39) 950,000
Torularhodin (211) active
P-Apo-8'-carotenal (248) 1,100,000
P-Apo-8'-carotenoic acid (250) ethyl ester 420,000
P-Apo-12'-carotenal (262) 2,000,000
P-Zeacarotene (P 1 -Zeacarotene) (9) 420,000 (210,000)
Citranaxanthin (237) active

* 1 i. u. vitamin A corresponds to the biblogical activity of 0.344j.ig all-trans-vitamin A acetate.

 XI. Use of Carotenoids 745

A detailed discussion of the vitamin A activity of carotenes and carotenoids

can be found elsewhere [8]. Most of the carotenoids not mentioned in Table 1
have no, or very weak, vitamin A activity. Since most of the carotenoids
mentioned in Table 1 occur only in the natural mixture contained in food and
feed ingredients, the information given in Table 1 has, from a practical point
of view, only limited value. The three carotenoids P-apo-8'-carotenal, the
ethyl ester of P-apo-8' -carotenoic acid and citranaxanthin are applied in
avian feeding as pure stabilized preparations, and the vitamin A potencies of
these substances determined by this kind of exact laboratory trials have also
to be evaluated. Being measured in different animals and with different methods,
the vitamin A potencies of these carotenoids may vary to a certain degree. For
instance, in the chicken-growth assay for P-apo-8'-carotenal, Haslach [9] has
observed a biological potency of only 658,000 i.u.jg. Besides these three caro-
tenoids, synthetic P-carotene is used for human nutrition. Only if it is admin-
istered in diets at low levels of intake [10, 11], such as margarine and similar
products, is the theoretical value valid as given in Table 1. Otherwise, due to
limited absorption or transformation to vitamin A, the true value for vitamin A
potency is lower. In the rat curative growth assay water-soluble P-carotene
preparations may have the same biological activity as P-carotene indicated
in Table 1 [11, 12]. Therefore, in food coloured by this type of preparation the
full biological activity can be considered.
2. Practical aspects
In food and feed ingredients of natural origin, all-trans-{J-carotene is always
accompanied by numerous other carotenes and carotenoids. Apart from all-
trans-{J-carotene of theoretical vitamin A activity 1,667,000 i. u./g, we have to
consider (a) the geometric isomers of all-trans-{J-carotene, e.g. neo-U-P-
carotene and neo-B-{J-carotene, each having a certain vitamin A activity,
which is, however, in all cases lower than that of all-trans-{J-carotene, (b) struc-
tural isomers, such as ex- (5) and y-carotene (8) etc., with vitamin A activities
also lower than the activity of P-carotene (3), (c) certain related carotenoids with
vitamin A activity, e.g. echinenone (148), and (d) degradation products of the
compounds mentioned above, which may still have some vitamin A activity,
or have lost it all.
In most of the analytical methods applied to food and feed ingredients,
these congener compounds are extracted with all-trans-{J-carotene and are
not completely separated from it. In general, therefore, P-carotene values
measured in food and feed ingredients are too high (see p. 763).
Under practical conditions, many external and internal factors may in-
fluence the conversion of vitamin A-active carotenes and carotenoids to
vitamin A by farm animals so that the actual vitamin A potency will be signif-
icantly lower than the theoretical value [ 4, 13].
The vitamin A potencies of natural sources of P-carotene shown in Table 2
are compiled from data found in literature which can be used under most
practical conditions where the conversion of P-carotene is not greatly impaired.
746 J.C. BAUERNFEIND, G.B. BRUBACHER, H.M. KLAUI and W.L. MARUSICH

Table 2. Biological potency of P-carotene in various animals {for details see [3])

Animal I. u. vitamin A formed per mg P-carotene

Poultry 536-1667
Dairy cattle 333-476
Beef cattle 400-476
Sheep 400-578
Swine 476-533

The same problems arise regarding the activity of naturally occurring

provitamin A in food. For practical purposes, if no further information on the
availability and isomer-content of fJ-carotene in such food-stuffs is available,
IUPAC [1] has proposed a vitamin A potency of fJ-carotene which puts
1 J.lg P-carotene equivalent to 1 i. u. vitamin A, whereas a group of experts
from F AO/WHO [2] decided to use the relation putting 1 J.lg fJ-carote'ae
equivalent to 0.167 J.lg retinol or 0.56 i. u. vitamin A.
In most food tables where the vitamin A activity of raw, processed and
prepared food is given [14, 15], other conversion factors are used. Such tables
can be of service only when the conversion factor used is known. It is therefore
more reasonable to tabulate the P-carotene content of food in weight units
than in vitamin A potency.

B. Pigmenters in Feed
1. Introduction
Carotenes and carotenoid pigments are extremely abundant in the animal
kingdom (see [8, 16-18] and Chapter II). They are, however, not synthesized
by the animal itself and have to be ingested with feed, as such, or in the form
of precursors. From a practical point of view, many carotenes and carotenoids
are responsible for colour in animals or animal products used as human food.
Therefore, the content of carotenes or carotenoids of such products is con-
sidered to be a sign of quality. In the following, the term 'pigmenters' is used
to refer to carotenoids that are present in the animal's ration and can lead to
the colouring of body tissues such as skin, fat and shanks, or animal products
such as eggs, butter and cheese. The addition of these pigmenters to animal
rations may be used as an indirect method of colouring food. The addition of
a pigmenter to the ration can enhance either the visual impression of colour
or the actual concentration of a pigment in the animal product; it is important
to note that one of these "effects does not necessarily imply the other.

2. Stability of carotenoids in feed

During storage of the hay crop the carotene content decreases at a sub-
stantial rate. The rate of decrease depends on temperature, amount of exposure
 XI. Use of Carotenoids 747

to air and sunshine, original content and other factors. Under average con-
ditions the carotene content of hay can be expected to decrease by about
6-7% per month. When the temperature is above 19 °C the loss may be over
18% per month [19]. Somewhat higher losses were found by Fassler et al. [20]
in chicken rations stored under normal conditions. Tiews and Zucker [21]
observed rather high losses in green meals of different origin during storage.
Losses of carotenoids from dehydrated alfalfa hays and calf starters were
investigated by Dolge et al. [22]. Losses were found to occur essentially at a
linear rate. Reviewing the literature, Orth and Koch [23] pointed out that
during preparation and storage of conserved green crops (hay, silage and dried
grass) a considerable proportion of the carotene content is often destroyed.
While most of the carotene is preserved in silage, the losses in hay are ex-
tremely high.
Halverson and Hendrick [24] reviewed the literature on factors affecting
the stability of carotene in mixed feeds, and reported that losses increased
markedly in high trace-mineral diets. Other studies have also been made [87].
The stability of the xanthophyll carotenoids depends upon the source and
storage conditions [20, 21, 25-28]. While isomerization of carotenoids occurs
during alfalfa meal processing, their rate of decrease during storage has been
shown to be influenced by oxygen and storage temperatures. The stability of
the xanthophylls of alfalfa meal has not been examined as thoroughly as that
of carotenes. Thompson and Maclay [29], Livingston et al. [30] and Guglie-
melli and Mitchell [31] report that during storage the xanthophylls of alfalfa
meal and certain xanthophyll pigmenting concentrates are destroyed to a
lesser extent than the carotenes. Also the concentration of xanthophyll in
corn does not decrease as rapidly during storage as in alfalfa meal.
Since all carotenes and carotenoids are susceptible to oxidative destruction,
the addition of an antioxidant to carotenes or carotenoid-containing mixtures
has a stabilizing effect [24, 32]. The best antioxidant for addition to feed
mixtures for the stabilization of carotenoids is ethoxyquin [21, 33].
Stabilized synthetic carotenoids are much less sensitive to deleterious
influences than the carotenoids naturally occurring in feed ingredients.

3. [3-Carotene in cattle products

In addition to its role as the most potent vitamin A precursor, [3-carotene
is also a pigmenter for the dairy cow. Absorbed [3-carotene not converted to
vitamin A is stored in the fatty tissue deposits in the body as well as in the
butterfat. A great breed and species specificity exists in the metabolism of
{3-carotene [8].
Since the dairy cow has a carotene intake variable over the year, influenced
by the alternate feeding of fresh pasture and dry roughage and feed, the [3-caro-
tene and vitamin A content of the milk fluctuates accordingly [34, 35]. How-
ever, the feeding of synthetic [3-carotene is not of practical significance for
maintaining a standard colour of butter [36]. Addition of [3-carotene in butter
processing would be a more convenient way (see p. 760).
748 J. C. BAUERNFEIND, G. B. BRUBACHER, H. M. KLAUI and W. L. MARUSICH

4. Carotenoids in poultry products

a) General remarks
{3-Carotene and other carotenes have little pigmenting value for the avian
family but certain hydroxy- or oxo-carotenoids are excellent in that respect.
While the avian family converts carotene to vitamin A like mammals, it differs
from mammals in preferentially storing xanthophylls and other carotenoids
in the liver, eggs, body fat, skin, feathers and shanks. In the poultry field
pigmentation is concerned both with the skin colour of meat birds and the
degree of yolk colour. In most parts of the world the use of materials to
improve yolk colours is encouraged because consumers prefer deeply coloured
yolks [61, 261]. In addition, yolks with a high pigment content are demanded for
the commercial production of bakery products, noodles, mayonnaise, prepared
cake mixes and other egg products. Also breeder hens should be provided
with adequate pigment so that the chicks would have an acceptable colotir.
Chicks with pale shanks and down are less acceptable to the poultry breeder,
even though they may be produced from eggs laid by hens fed a nutritionally
complete ration.
In many parts of the world a well-pigmented broiler is preferred [62], and
it is, therefore, of economic importance for the broiler grower to be able to
produce birds with the degree of pigmentation desired in the market. In other
areas of the world the very opposite tendency is observed, and the consumer
demands broilers with absolutely white meat. In parts of the USA a high
degree of broiler skin and shank pigmentation is desired since the consumers
consider a bright yellow colour in skin, shanks and body fat an indication of
good quality [262]. The skin of guinea-fowl should also be well coloured [260].
When the barnyard flock was the principal method of raising poultry, the
problem of skin and yolk pigmentation did not arise because a lower physio-
logical response (slower growth and fewer eggs) and ample feeding of green
grass and yellow corn resulted in adequately pigmented eggs and birds. Today,
however, without grass feeding and with modern methods of intensive poultry
raising, such as the use of low-fiber, high-energy feeds, and birds with high
physiological capacity, etc., the production of well-pigmented birds and highly
pigmented eggs has become difficult to achieve [264].
The advent of high-energy, low-fiber feed adds to the pigmentation problem
because this concept restricts the use of high-fiber but xanthophyll-rich
ingredients such as alfalfa leaf meal. High-energy rations need more xantho-
phyll than low-energy rations to assure an adequate intake of pigment, since
faster growth due to improved strains and a more favourable calorie/protein
ratio brings the bird to market weight sooner and with less feed. This lessened
feed intake and more rapid growth necessitates much higher levels of xantho-
phylls for attaining adequate pigmentation.
In modern commercial feeds the pigmenting burden falls on yellow corn
(maize), which does not produce adequate results without the aid of some
alfalfa meal and/or corn gluten meal. Within the past few years, milo, wheat,
 XI. Use of Carotenoids 749
rice and barley have become recognized as high-energy and economical cereal
grains for poultry. When these grains replace yellow com the natural carot-
enoids of the ration are further depressed [37, 38].
In order to overcome all the difficulties described, the poultry industry
now uses concentrates or extracts of natural sources of carotenoids or dry,
stabilized synthetic carotenoids which are readily transferred to the poultry
product and can be mixed into the feed without major alteration of the feed
formula [73, 89].
The most important of the first type of pigmenters are lucerne (or alfalfa)
meal and maize gluten meal [39]. Extracts of lucerne, rich in lutein (73), are
sold under various brand names, as are also those of the marigold plant
(Tagetes species), which confer a pure yellow colour. Reddish concentrates are
prepared from extracts of red pepper [27, 40], whereas the use of annatto-seed
extracts is of doubtful value [41].
The most important of the second type of pigmenters are the ethyl ester of
P-apo-8'-carotenoic acid (250)* and canthaxanthin (193)*, which have a wide
application in poultry layer and fattener rations, whereas two other carotenoids,
which are also available in stabilized form as a commercial commodity, namely
P-apo-8'-carotenal (248) [73, 89, 263] and citranaxanthin (237), contribute to
the colour of eggs but are not efficiently deposited in the skin of broilers.
b) Egg yolk
a) Significance of the egg yolk colour and of the carotenoid content of egg
yolk. A clear distinction has to be made between eggs for immediate con-
sumption (table eggs) and eggs for industrial purposes (manufacture of noodles,
bakery products). The table egg is examined by the consumer's eye and its
visual appearance is the decisive factor in assessing of quality. In most parts
of the world consumers prefer a golden-yellow yolk without noticing its
actual pigment content. Orange yolks can be produced even with small
quantities of yellow components, such as naturally occurring xanthophylls or
P--apo-8'-carotenoic acid ethyl ester, and the addition of relatively small
amounts of red ones, such as canthaxanthin. The resulting eggs have a relatively
low pigment content but, nevertheless, appear to be well coloured because of
the orange hue produced by the pigmenter combination. The pigment content
of eggs produced in this way may not be sufficient for the colouring of noodles
and bakery products. The use of red pigments alone for industrial eggs is not
recommended. It has been observed that very dark pigmented reddish yolks
produce an unattractive colour in noodles, particularly if too little yellow
pigment is present. Hence, a visual evaluation of yolks for industrial purposes
may be misleading. Reddish yolks may appear dark despite lower pigment
contents, while bright yellow yolks may have a much higher pigment content
though they may appear paler. Consequently, the pigmenting power of such
* P-Apo-8'-carotenoic acid ethyl ester and canthaxanthin are sold in stabilized form as
pigmenters under the trade mark 'carophyll yellow' and 'carophyll red' by F. Hoffmann-
La Roche & Co. Ltd., Basle, Switzerland.
750 J.C. BAUERNFEIND, G.B. BRUBACHER, H.M. KLAUI and W.L. MARUSICH

yolks has to be judged by a quantitative analysis of their pigment content, and

by practical trials for their intended usage.

P) Measurement of the egg yolk colour and pigment content of egg yolk.
Many devices are described in the literature for scoring egg yolk colour. For
a review see [ 42-44]. Of these devices the 'Roche Colour Fan' developed by
co-workers at F. Hoffmann-La Roche & Co. Ltd. is the most commonly used
instrument. The colour fan consists of 15 blades varying in colour from a pale
yellow to a deep orange red and encompassing the whole range of egg yolk
colours found under practical conditions. Each colour is defined by three
co-ordinates according to the internationally recognized tri-stimulus method
of the CIE (Commission lnternationale de l'Eclairage). For reasons of con-
venience, the blades are numbered from 1 to 15, starting with the palest yellow
shade [ 43] *.
The measurement of the pigment content of egg yolks can be carried OJ!t
by the quantitative determination of every individual carotene or carotenoid
occurring in the egg yolk, a task which is very difficult, since many different
carotenoids may be found in egg yolks, separable only by sophisticated analyt-
ical methods. Wildfeuer et al. [ 41] have recently discussed suitable methods.
For practical purposes, the optical density of egg yolk extracts is compared
with the optical density of standard solutions, and the results are expressed in
equivalents of these standard solutions. A procedure widely used in the USA
for measuring yolk pigments is the NEPA (National Egg Products Association)
method [45], in which an acetone extract is compared with aqueous potassium
dichromate standards of arbitrary numbers of 1 to 10, indicating increasing
colour from light yellow to deep orange.
In a revision of this method, now referred to as the AOAC method [ 46-48],
a P-carotene solution is used as the standard, though the yolk colour results
are still expressed as NEPA numbers [ 49: p. 224]. More recently, P-carotene
is being used as the standard, and the results are expressed as P-carotene
extinction equivalents in JLg/g [50: p. 708, 41]. This procedure has been used
in the ANRC (Animal Nutrition Research Council) studies [51]. Scott et al. [52]
have compared the results gained in the different ways described above.

y) Relationship between quality and quantity ofcarotenoids present in feed and

egg yolk colour. Before 1956 practically all research on avian carotenoids
concerned 'total oxycarotenoids' or 'total xanthophylls' as assayed or esti-
mated in the natural ingredients or in crude carotenoid extracts and concen-
trates. When synthetic carotenoids became available, pure compounds were
investigated. Steinegger et al. [53], Steinegger and Zanetti [54, 55] and Geisen-
dorff and Streiff [56] determined the influence of feeding pure zeaxanthin (67),
canthaxanthin (193), isozeaxanthin (71), physalien (68), P-apo-8'-carotenoic
acid (250) ethyl ester, P-apo-8'-carotenal (248), other P-apo-carotenals, iso-
* The 'Roche Colour Fan' is available on request from F. Hoffmann-La Roche & Co. Ltd.,
Basle, Switzerland.
 XI. Use of Carotenoids 751

zeaxanthin dimethyl ether and isozeaxanthin diacetate on pigmentation of

egg yolk. Further research on these and other carotenoids has been reported
[42, 57-59] or reviewed [60-67, 264]. Of more recent papers in which rela-
tionships between the quality and quantity of carotenoids present in feed and
egg yolk colour are investigated, the following should be mentioned [52,
68-73].
From a practical point of view, if no pigmenter is added to the feed, only
the xanthophylls of yellow corn, grass and lucerne meal are of interest. In the
Figure the relationship between the xanthophyll content of the ration under
practical conditions and the egg yolk colour is given [74]. In actual trials the
egg yolk colour may deviate considerably from the ideal curve. This may be

14 Roche colour fan value

12

/;;:~~:===::=:
10

6
//• /-" of all observations
4 • "'.··"' - - Probable values
/ ..·
2

5 10 15 20 25
mg xanthophylls/kg feed

Fig. Relationship between xanthophyll content of the feed and egg yolk colour

due to different carotenoid patterns, the presence of factors such as toco-

pherols, or the effect of dietary free fatty acids [75]. Marusich [51] found a
similar relationship, using only dehydrated alfalfa meal as pigmenting agent.
From the same paper [51] the relationship between the xanthophyll content
of feed and the carotenoid content of egg yolk can be deduced. Quacken-
bush et al. [76] and others [257] show similar relationships. .
Concerning pigmenters added to the ration (see p. 749) we can submit
only general rules. Yellow pigmenters, such as extracts from grass or lucerne
meal, corn gluten meal, extracts from tagetes, P-apo-8' -carotenal and P-apo-
8'-carotenoic acid ethyl ester, can be used to produce table eggs as well as
eggs forindustrial purposes. The colour achieved has a bright or deep yellow
hue. The pigmt:nting potency per milligram of the pure carotenoids is similar
to or higher than that of the lutein and zeaxanthin fraction of yellow corn or
alfalfa. Paprika powder or paprika extracts, canthaxanthin and citranaxanthin
should only be used if the basic ration contains enough yellow pigmenters such
as xanthophylls or P-apo-8' -carotenoic acid ethyl ester, otherwise an un-
attractive reddish off-colour of the egg yolk is obtained.
752 J.C. BAUERNFEIND, G.B. BRUBACHER, H.M. KLAUI and W.L. MARUSICH

(J) Factors influencing the deposition of carotenoids in the egg yolk. Apart
from genetic differences [52] many internal and external factors are responsible
for an increase or decrease of yolk pigmentation. Among the factors which
improve the egg yolk pigmentation are other feed ingredients such as vitamin E
and certain antioxidants, and the fat content of the ration [77, 78]. The main
factors decreasing pigmentation seem to be pro-oxidants such as trace minerals,
unsaturated fatty acids such as those present in fish liver oils, and certain
animal protein sources [59]. Excessive levels of vitamin A in the feed have also
been reported to depress the amount of carotenoids absorbed and deposited
in the yolk. Normal vitamin A levels in the feed have little or no effect on the
yolk pigmentation. At increased levels of vitamin A applied temporarily during
stress a certain drop in the yolk colour may be expected. Finally, conditions
such as coccidiosis [265] and chronic respiratory disease [266] will result in
low pigmentation. Environmental temperature is also a factor, since Couch [79]
and Farr et al. [80] report a decrease in egg yolk colour amounting to ab6ut
33% during hot summer weather on an unchanged diet.

c) Broiler
r:x) Measurement of broiler pigmentation. There is no generally accepted
method for scoring the quality of broilers according to their pigmentation.
Acceptability by the consumer has therefore to be taken as a measure. Other
attempts in which, for example, the colour of the skin and shank is estimated
by the Roche egg yolk colour fan, or the xanthophyll content of certain tissues
such as the toe web is determined by chemical means [81] give results which
may be related to the subjective assessment of skin pigmentation and are
useful tools for the scientific evaluation of the pigmenting potencies of different
carotenoids, though these should always be checked.

f3) Relationship between the quality and quantity of carotenoids present in

feed and broiler pigmentation. From experiments in which different methods
of judging broiler pigmentation were used, certain general rules can be deduced.
In practical rations without added pigmenters the so-called xanthophyll
fraction of cereals, grass and alfalfa meal is responsible for broiler pigmentation.
This fraction contains mainly lutein (73) and zeaxanthin (67). It is difficult by
simple analytical procedures to separate this fraction from carotenoids of
minor importance. For example, in the (WRRL) analytical procedure [82]
the carotenoids lutein (73), zeaxanthin (67), cryptoxanthin (39), neoxanthin (122)
and violaxanthin (135), carotenoid oxidation products and various cis iso-
mers of these, are determined together. The pigmentation potency of these
mixtures is grosso modo equal, whether it is derived from cereals, grass meal
or from alfalfa meal [81] but it seems that carotenoids from grass and alfalfa
meal, which contain a high amount of lutein (73), produce a bright lemon-
yellow colour, whereas carotenoids from corn, which contains a relatively
higher proportion of zeaxanthin (67), lead to a golden-yellow colour [83].
The polyoxy carotenoids contaifted in this mixture are probably poor skin
 XI. Use of Carotenoids 753

pigmenters [81, 84]. Neoxanthin (122) has only 8% of the pigmenting capacity
of lutein (73), and violaxanthin (135) is essentially ineffective [85]. Lutein
(73) and zeaxanthin (67) are deposited in the tissues, as such, or as their
esters [86].
Quackenbush et al. [76] propose a technique for the determination of a
'dihydroxy pigment' (DHP) equivalent as a measure of pigmenting efficacy for
avian skin. Of the pigmenters available on the market, P-apo-8' -carotenoic
acid ethyl ester has the highest pigmenting activity [88]. P-Apo-8' -carotenal
and citranaxanthin have little skin pigmenting potency (own observations),
and canthaxanthin, which is even better deposited in the avian body than
P-apo-8'-carotenoic acid ethyl ester, gives rise to reddish-hue colours which
may be softened if the ration contains enough yellow pigments. Recently
Marusich and Bauernfeind [89] studied the potency of stabilized P-apo-
8' -carotenoic acid ethyl ester and canthaxanthin in comparison with naturally
occurring oxycarotenoids. An evaluation of stabilized P-apo-8' -carotenoic acid
ethyl ester as a potential ANRC (Animal Nutrition Research Council) reference
standard for broiler pigmentation was made in a collaborative study by eight
investigators [90]. (For guinea-fowl see [260].)

y) Factors influencing the deposition of carotenoids in the tissue. Mainly

the same internal and external factors are responsible for the increase or
decrease of pigmentation which we discussed in the chapter on egg yolk
pigmentation.
The colour-enhancing effect of fat in broiler rations is due not so much to
its influence on absorption as to the fact that the finished broiler has a higher
amount of fat tissue in which carotenoids are deposited and that these fat
tissues hide the bluish colour of muscle tissue.
The total amount of carotenoids in the finished broiler does not depend so
much on the carotenoid content of the ration as on the total amount of caro-
tenoids ingested. This total may be spread over its whole life time or fed only
during the last three or five weeks when it leads to the same degree of pigmen-
tation. Experiments showed that a strong skin pigmentation can be produced
even within one week. Depending on the consumer's wishes, the total amount
of oxycarotenoids fed to broilers during the fattening period should be between
100-150 mgjkg of body weight of the finished broiler.
Factors decreasing the deposition of carotenoids are mainly of interest
in regions where white meat is desired. Methods for decreasing pigmentation
consist of feeding charcoal or inorganic adsorbents. Other interesting sub-
stances which may solve the problem are vitamin A as well as retinoic acid,
mentioned by Dua et al. [91].

d) Other applications in the poultry industry

The opinion that broiler or replacement breeder pullets should have a
well-coloured beak and shanks is justified to the extent that animals suffering
from chronic disorders, especially of the alimentary tract, have, in general,

Carotenoids 48
754 J.C. BAUERNFEIND, G.B. BRUBACHER, H.M. KLAUI and W.L. MARUSICH

impaired ability to absorb carotenoids from the intestine. Here again, an

abundant supply of carotenoids in the feed may help to attain the desired effect
by which healthy animals can be distinguished from sick ones. Bauernfeind [92]
has shown that canthaxanthin in stabilized form added to the feed in amounts
of 3 to 6 g per ton provides sufficient shank pigmentation of a deep orange-
yellow tone with a 60% yellow corn ration, giving a colour score of 7-8 meas-
ured by the old Roche egg yolk colour fan (1-12), equivalent to a score of
10-12 on the new Roche fan (1-15).
In the commercially produced day-old chick, similar views lead to the
preference for animals with fine, yellow to creamy colour of the feathering,
which is, in turn, dependent on the carotenoid reserve of the yolks of the eggs
from which the chicks are hatched. By means of a high amount of pigmenting
carotenoids given in the feed of the breeder hens the colour of the hatching
chick may be adjusted [93].

5. Carotenoids for pigmenting zoo-birds and fishes

The main source of the red and yellow colouration of birds exhibited in
zoos or kept for pleasure are the carotenoids contained in their food [16].
Since, however, the feed which they receive in captivity in general differs
fundamentally from that which they find in their natural environment, the
birds often tend to lose their natural colours. The feed of zoo- and cage-birds
should, therefore, be reinforced with carotenoids of natural or synthetic ori-
gin [94].
The natural food of fish contains carotenes and carotenoids which can be
deposited in the flesh, as such, or metabolized, giving the flesh an attractive
pink colour. Kanemitsu and Aoe [95] reported the presence of astaxanthin (203)
in the flesh of five species of Pacific salmon. In addition to this pigment, Thom-
men and Gloor [96] found canthaxanthin (193) and /3-carotene (3) in the flesh
of the lake trout (Salmo trutta). Since the most notable differences between
wild types of fish and those bred in ponds lies in the colouration of the flesh,
many attempts are made to correct this sign of inferior quality by feeding
carotenoid-rich feed. One of the most promising ways consists of feeding
synthetic canthaxanthin (193) in stabilized form [97-99].

C. Pigmenters in Food
1. Introduction
The colour of food is a significant factor in determining its acceptability.
We expect to see food in its 'natural' colour; a 'natural' appearance is appetiz-
ing, and we get cautious when a food shows an unexpected colour, interpreting
it as a possible sign of spoilage, poor processing or as an indication of adultera-
tion. The association of colour and acceptability of food is universal, although
there may be significant differences depending on geographical, ethnical,
 XI. Use of Carotenoids 755

historical and social factors. What may be attractive to one group may be
unappetizing or even repulsive to another (see for instance [100]).
The main groups of natural colouring substances in food are carotenoids,
anthocyanins, porphyrins and chlorophylls. The carotenoids are responsible
for many of the brilliant red, orange and yellow colours of edible fruits and
berries, vegetables and mushrooms, flowers, and also of some animals. These
compounds occur very widely in nature (see for instance Chapter II}, and it
has been estimated that the annual natural production of these pigments
amounts to about 100 million tons [101]. Though oxygenated carotenoids
occur in the largest quantities, the hydrocarbon P-carotene is of special
interest because of its provitamin A activity.
Since the colour of food is one of the criteria used for commercial standards,
it is important to know the carotenoid content of the product in question.
Many factors affect, however, colour such as variety of fruit, maturity, place
of origin, seasonal and climatic changes, processing methods. A review of the
seasonal colour fluctuations and carotenoid changes during maturation was
published by Bauernfeind [102].
Natural extracts containing carotenoids have been used for colouring food
for centuries: annatto with bixin (265) as the main colouring component,
saffron with derivatives of crocetin (269) and other carotenoids, paprika
containing the two pigments capsanthin (170) and capsorubin (205), xantho-
phyll extracts from leaves, carrot extracts of varying purity, and red palm oil.
The latter two contain considerable amounts of carotenes and have been
widely used as colouring agents mainly for fats and margarine.
P-Carotene (3} was the first synthetic carotenoid to be marketed in 1954.
Other carotenoids which have since become commercially available for food
colouring are P-apo-8'-carotenal (248) in 1962 and canthaxanthin (193) in
1964.
In a review Bauernfeind et al. [103] gave a survey of the development of
preparations based on the pure carotenoids and their application in colouring
food and pharmaceuticals.

2. Natural occurrence and stability

A review of carotenoids occurring naturally in foods is given in an excellent
survey by Borenstein and Bunnell [104]. Benk [105] has analysed various
European fruits, particularly wild fruits from Germany. These analyses illus-
trate the highly variable ratio between P-carotene (3) and total carotenoids in
these fruits.
Large amounts of carotenoids are present in nature very finely dispersed
and, in that fopn, are capable of colouring aqueous media. The carotenoids
in oranges, tomatoes and carrots serve as well-known examples. The out-
standing stability of the colour of these natural water-dispersible carotenoid-
protein or -lipoprotein complexes is probably due to their ultrastructure.
But remarkably little is known of the nature of protein-carotenoid link-
ages. This applies also to the carotenoproteins, which contain carotenoids
756 J. C. BAUERNFEIND, G. B. BRUBACHER, H. M. KLAUI and W. L. MARUSICH

and protein in stoichiometric proportions, and which are of extraordinary

interest in view of the green and blue and other colours of these particular
combinations.
In higher plants, carotenoids are present in the chromoplasts; as in all
photosynthetic organisms, the xanthophylls are not esterified. In autumn
leaves the xanthophylls liberated from the disintegrating chloroplasts are
esterified and thereby become more lipophilic [106-108].
Fruit carotenoids are attached to proteins. In the few roots that contain
significant amounts of carotenoids, carotenes generally predominate, e.g.
carrots and sweet potatoes [8]. Carotene in carrots is located in lipidic droplets
-or 'globuli'-in filaments and crystals. The crystalline chromatoplastids
contain, in addition to protein, RNA, DNA and phospholipid [109]. Frey-
Wyssling and Schwegler [110] showed by electron microscopic and polariza-
tion microscopic studies that the 'crystalline' chromoplasts contain aniso-
tropic sheets of lipoprotein, in which the surplus carotene is stored. A stu(iy
of the 'links' of carotenoids to lipoproteins of chloroplasts in green leaves
indicates a heterogeneity of strength [111] which is associated with a hetero-
geneous metabolism.
In animal tissues carotenoids may be present in fats in dissolved form.
Proteins in which carotenoids are present in stoichiometric proportions as
prosthetic groups constitute a very interesting set of compounds, since the
combination of a carotenoid with protein can extend the range of colours to
green, purple, blue and black. Despite these interesting properties, remarkably
little is known of these compounds. Cheesman et al. [112] recently prepared
a review in which they compiled a list of papers reporting the presence of
carotenoproteins in invertebrates. The table includes over 120 species of
animals. According to Cheesman et al. [112] the protein-carotenoid com-
plexes may be subdivided into (i) true carotenoproteins, with a strictly stoichio-
metric relationship between carotenoid and protein, and (ii) lipoproteins, in
which carotenoids are associated with the lipid component, and in which a
stoichiometric relationship has not been proved. A characteristic property of
both types of complexes [112] is shown in an important modification of the
absorption spectrum of the carotenoid. The majority of carotenoprotein com-
plexes are blue to green with their main absorption maxima lying between
560 and 680 nm; the others are red to purple with their maxima between 490
and 532 nm. Rather exceptionally, their maxima are lower than those of the
free carotenoid; and some green complexes appear to result from a blue
stoichiometric complex in combination with a lipid-dissolved free carotenoid.
The carotenoids found in carotenoproteins all commonly have reactive keto
groups in the 4- and 4'~positions of the ionone ring (e.g. astaxanthin (203),
astaxanthin esters and canthaxanthin (193)). Attempts at making artificial
carotenoproteins with various carotenoids have succeeded only when a keto
group was present in at least one of these positions [113].
The facts discussed above make clear that the characteristics of the colour,
its stability and chemical behaviour depend not only on the chemical nature
 XI. Use of Carotenoids 757

of the colouring carotenoids present in food but in a high degree on the

physico-chemical distribution.
The main cause of damage to carotenoids during food processing and
storage is oxidation. The naturally-occurring carotenoprotein complexes and
the protein-adsorbed carotenoids are more stable than the carotenoids as
such. Destroying the natural complexes by heating, or treating with water-
soluble solvents such as alcohol and acetone, usually leads to a decrease in
stability. Enzymatic destruction of carotene and carotenoids may also be
involved [114]. Losses of carotenoids occur during food processing [115],
during storage and processing of milk and milk products [116] and during
storage and processing of citrus fruits [117].
The stability of P-carotene in dehydrated vegetables was investigated by
Schillinger and Zimmermann [118], Spiess et al. [119] and by Kieckebusch
and Lang [120]. Dehydration and increase of surface, e. g. in powdered or
lyophilized fruit and vegetable products, generally leads to very poor stability
unless the products are protected from air by storing in an inert atmosphere
or in vacuum [121]. Hoover [122, 123] reported that sodium acid pyrophos-
phate improved natural colour retention in pre-cooked frozen sweet-potato
products and in sweet-potato flakes.
Borenstein and Bunnell [104] have reviewed earlier literature on the effect
of canning on carotenoids. The type of pretreatment used and the storage
temperature are significant canning variables controlling the stability of
carotenoids in the finished product [124, 125]. In general, little or practically
no losses occurred during canning and subsequent storage at ordinary tem-
peratures. Addition of BHA and BHT and mixtures with citric acid or sodium
acid pyrophosphate improves stability during storage of canned, pre-cooked
and dehydrated sweet-potato flakes [126].
The carotenoids in macaroni products undergo oxidative changes during
manufacture, and Menger [127] studied the influence of various factors on
the destruction of pigments. The factors affecting oxidative stability of caro-
tenoid pigments of milled durum products (semolina and flours) were investi-
gated by Dahle [128].
Irradiation may also lead to losses of P-carotene [129-132]. Microwave
and normal cooking have little or no effect on carotenoids. When vegetables
are cooked and kept hot [118] or cooking times are prolonged [134] appreciable
losses may occur.
The prevention of losses of carotenoids in citrus fruit juices and corre-
sponding beverages is still a technical problem. The degree of destruction of
colour depends on the amount of oxygen and the duration of exposure to
light [133]. Natural [135] or added antioxidants may counteract this effect.
The most suitable antioxidant is ascorbic acid [133, 136, 137, 204] since this
compound naturally functions this way [135, 138]. Since the mechanism of
destruction depends on many other parameters [139], e.g. traces of heavy
metals [140], the maximum stability and efficacy of ascorbic acid can be
achieved [141] only if all precautions are strictly followed during processing.
758 J.C. BAUERNFEIND, G.B. BRUBACHER, H.M. KLAUI and W.L. MARUSICH

3. Food colours
a) General
Two main groups are distinguished:
1) Extracts of plants, animals or microorganisms which contain carotenoids
as colouring agents. These extracts are often processed before use with the
result that the native carotenoids are transformed in chemical structure with
concomitant changes in physico-chemical behaviour.
2) Well defined preparations prepared from pure chemical compounds.
The legal regulations for the use of these two different groups as food col-
ourants may vary from country to country. According to the joint F AOjWHO
Expert Committee on Food Additives [142] natural colours have been used
in food over a long period of time and have been accepted for such use without
supporting toxicological evidence in much the same manner as vegetables a~
cereal products. Lack of published information related to adequate identifica-
tion and chemical composition creates serious problems; it is hoped that
future studies will provide much more detailed information which will allow
the toxicological evaluation of natural colouring matters.
On the other hand, preparations containing pure chemical compounds can
be evaluated toxicologically on the same lines as those acquired for other food
additives. This creates no problems. Based on the chronic toxicity data and
available specifications, the joint FAOjWHO Expert Commitee on Food
Additives has classified the four carotenoids P-carotene (EEC number*:
E 160a), P-apo-carotenal(C 30) (EEC number*: E 160e), canthaxanthin (EEC
number*: E 161a) and ethyl ester of P-apo-carotenoic acid(C 30 ) (EEC num-
ber*: E 160 f) in Class A, and they are, therefore, 'found acceptable for use in
foods'. Their legal status in different countries has been summarized as of
1968 [259] **.

b) Natural carotenoid extracts available as food colours

Annatto. The term 'annatto' includes a whole series of colouring prepara-
tions, all based on extracts of the seed of the annatto tree, Bixa ore Ilana. The
pigments are present in the thin, resinous coating of the seeds, and the major
component consists of bixin (265). Bixin is the main pigment of oil-soluble
annatto preparations, and norbixin, the saponification product of bixin, the
main colouring matter of the water-soluble products. The composition of
annatto preparations has been reported by McKeown and Mark [143].
Annatto preparations are used for colouring butter, bakery products and
salad oil; mixtures with paprika oleoresin yield redder colours and may be
used in processed cheese; mixtures with curcumin are more yellow in colour
and are claimed to be more stable [145]. Water-soluble preparations consisting
* EEC number= number of the European Economic Community; see for instance [258].
** Information about the current legal status of carotenoid food colours in different countries
is available on request from F. Hoffmann-La Roche & Co. Ltd., Basle, Switzerland.
 XI. Use of Carotenoids 759

mainly ofnorbixin at about 0.1 to 3% in aqueous potassium hydroxide, or of

water-soluble powders or of tablets containing potassium carbonate, are
applied in colouring cheese, cereals and ice cream. An insoluble powder is
available for colouring spices and meat.
Since the main components of annatto preparations are carboxylic acids,
all these products are pH-sensitive. This sensitivity is of practical significance
only in low-pH foods, where the colour may turn pink. By a special method
the skin of citrus fruits may be coloured by annatto preparations [146]. Assay
methods are available [147-149].
The stability of commercial butter-colouring annatto preparations was
studied by Rajan et al. [150].
Oleoresin of paprika is the oil extract of paprika, Capsicum annuum. The
colour imparted to a food product can range from a deep crimson red to a pale
pinkish-yellow, depending on the concentrations used. The material can be
used in salad dressings, sauces and particularly in meat products, including
sausages, where it is allowed as a spice. It is often used in combination with
annatto to colour processed cheese.
This oleoresin consists of about 37 to 54 pigments depending on the mode
of preparation (extract of unbleached or bleached paprika), of which only 21
and 33, respectively, could be completely or even tentatively identified [104,
151]. The main pigments are in general esters of capsanthin (170) and capso-
rubin (205). Most assay methods therefore are based on the determination of
these two carotenoids.
Xanthophylls. Several xanthophyll preparations originating from different
sources ofleafy material are described in the literature [152, 153], but it seems
that this kind of food colour is, for the time being, of no importance.
Tomato extracts may be used in certain cases as a food colour. They contain
lycopene (19) as the main pigmenting carotenoid [152]. The isolation and
identification of tomato colour in meat is described by Giinther and Grau [154].
Saffron consists of the dried stigmas of Crocus sativus. It contains
crocin (271). Since it is a spice it has the advantage of being widely accepted
in soups, meat products and cheese. Its pure yellow colour is attractive in
beverages and special foods, such as risotto and curry. Montag [155] has
described the identification of saffron in bakery products and in sausages.

c) Products prepared from pure chemical compounds

KHiui et al. [156] and KHiui [157] gave an excellent survey of the problems
involved in transforming the pure chemical compounds P-carotene, cantha-
xanthin, P-apo-8' -carotenal and the ester of P-apocarotenoic acid into market-
able forms suitable for colouring food. According to these authors we have to
distinguish between products for colouring fat-based foods, and products for
colouring water-based foods.
oc) Products for colouring fat-based foods. The main pure compound used
for colouring fat-based foods on an industrial scale is P-carotene (3). Since this
760 J.C. BAUERNFEIND, G.B. BRUBACHER, H.M. KLAUI and W.L. MARUSICH

compound is available from synthesis, today only a few preparations originating

from carrots or other natural sources are on the market [159].
The main products for colouring fat-based foods contain the compounds
in an oily suspension corresponding to a concentration of 20 to 30%. The
particle size has to be very small to overcome the slow rate of dissolution ofthe
crystalline pigment when the oil suspension is added to the food to be col-
oured [158]. Other preparations have been proposed [160-167, 181]. The
main fields of application for these products are margarine, oils, fats and
shortenings. Synthetic {3-carotene was first used to colour margarine and butter,
and successful industrial trials were carried out by Schuchardt [168]. The
technology of colouring fat-based foods has been described in detail by Bauern-
feind and co-workers [169].
The stability of the added {3-carotene in margarine, butter and shortenings
is very good. Only in rare cases may /3-carotene be destroyed and give rise
to a green discolouration [170]. According to McWeeny [171] this pherr'o-
menon is caused by oxidation of {3-carotene by peroxy acids. Stabilizing
additives may retard appearance of the green discolouration [172]. Of these
citric acid and ascorbyl palmitate are the most potent ones. Furthermore, it
was shown that the ethyl ester of {J-apo-8' -carotenoic acid did not undergo
colour changes.
For maintaining a standard colour it is desirable to colour butter with
{3-carotene, since large differences may occur in its natural content (seep. 747)
during the seasons. The use of {3-carotene for colouring butter is described by
Parman [173]. However, a universal method of application of {3-carotene has
not yet been adopted, since commercial butter production is still in rapid
development. Present marketable forms of {3-carotene may not be suitable for
all methods under production.
fJ) Products for colouring water-based foods and processes for solubilizing
carotenoids. The conversion of crystalline carotenoids with their highly un-
satisfactory solubility characteristics into water-dispersible preparations is
quite a problem. One approach, therefore, omits the pure crystalline compounds
and instead uses natural carotenoid sources [174-178].
Using pure crystalline carotenoids it is, however, possible to prepare water-
dispersible carotenoids by 1) formation of colloidal suspensions, 2) emulsifica-
tion of oily solutions, and 3) dispersion in suitable colloids particularly with the
addition of surface-active agents.
1) Colloidal suspensions are usually formed by first dissolving carotenoids in
a water-miscible solvent, such as alcohol or acetone, pouring this solution
into water, and finally removing the organic solvent by evaporation [156,
179]. However, concentrations in these suspensions are very low, and they
show, in general, an unnatural pinkish tinge.
2) Emulsions of oily solutions of carotenoids in aqueous media can be prepared,
but the poor solubility of carotenoids in oil limits such emulsions to low
levels of potency [180]. Higher concentrations are achieved by heating the
oil to above 100 oc [181]. The emulsion obtained can be used either as
 XI. Use of Carotenoids 761
liquid, or it can be converted into dry beadlets, according to one of the
many methods described for fat-soluble vitamins [156]. Besides the applica-
tion of heat, it is also possible to increase the concentration by using a better
solvent. Such solvents include: orange oil [182, 183], diterpene acid esters,
e.g. abietic acid esters [184, 200], acetoglycerides (acetylated partial fatty
acid glycerides) [185], aromatic alcohols or their derivatives [186]. Par-
ticularly interesting solvents are vitamin A and vitamin E [187].
3) Very fine dispersions of carotenoids are obtained by using solvents in com-
bination with surface-active agents, such as polyhydric alcohol esters of
fatty acids (e.g. sucrose fatty acid esters) and emulsifying these solutions
into water or aqueous dispersions of special colloids. Klaui et al. [156]
discuss the various possibilities which are described in many patents
[188-191].
The emulsions obtained following the above procedures are quite stable,
but as in aqueous dilutions the carotene can be further stabilized by the
addition of ascorbic acid [157, 192].
A congealing process is described in a Japanese patent [193]. Carotenoid
pigments based on protein complexes are obtained according to [194].
Stable, water-soluble, transparent compositions are obtained by heating
carotenoids with hydrogenated castor oil-polyoxyethylene ether under
nitrogen [195].
Easily water-dispersible carotenoid preparations are obtained by emulsifying
a solution of a carotenoid in a volatile solvent into an aqueous solution of a
hydrophilic colloid, evaporating the solvent and optionally converting the
dispersion into a dry product* [157, 196-198]. The product obtained is
described by Klaui et al. [156, 199].
The main fields of application of water-soluble carotenoids are: juices and
beverages, dry soups, canned soups and gravies, dairy products such as cheese,
yoghurt, ice cream and ice cream mixes, dairy substitutes such as coffee
whiteners, desserts, jellies and puddings, preserves and syrups, confectionery
products and candies, bakery products, dressings, meat products and meat
substitutes, macaroni and pasta products and eggs and egg products.
Bauernfeind et al. [103] have given a few examples. Applications to juices
and beverages and stability problems are discussed by [12, 102, 133, 138,
201-204]. Bunnell and Borenstein [205] studied the behaviour of cantha-
xanthin 10% water-soluble in canned tomato soup and in spaghetti sauce.
The performance of P-carotene and of P-apo-8' -carotenal was studied by
Bauernfeind and Bunnell [206] in both natural and processed cheese. Bunnell
et al. [204] described the stability of P-carotene in ice cream. The inclusion of
canthaxanthin -in gelatine dessert powder is described by Bunnell and Boren-
stein [205].
• Suitable dry stabilized water-soluble powders of P-carotene, P-apo-8' -carotenal, P-apo-
8'-carotenoic acid ethyl ester and canthaxanthin containing 10% of the pure chemical compound
were developed by F. Hoffmann-La Roche and Co. Ltd., Basle, Switzerland, and are commercially
available.
762 J.C. BAUERNFEIND, G.B. BRUBACHER, H.M. KLAUI and W.L. MARUSICH

Klliui et al. [207] described the use of carotenoids for colouring sucrose
syrup and sugar-coated tablets, and Jager [208] for colouring confectionery
products and candies. Bunnell et al. [204] studied the problem of colouring
bakery products with carotenoids. French-type dressings can be prepared by
using canthaxanthin [205] or P-carotene and P-apo-carotenal [206]. Cantha-
xanthin is best suited for colouring meat products [205], whereas a water-
soluble form of P-carotene is the easiest to apply in maintaining a standard
colour of industrially used egg yolks in the mixing or churning process prior
to freezing, drying or further processing [204].

D. Other Carotenoid Uses

1. Pharmaceuticals
Carotenoids are useful in colouring sugar-coated tablets [209-21f:J,
Coloured sugar coatings for tablets containing carotenoids may be prepared
according to [212]. Depending on the amount of carotenoid used, the following
colours can be obtained:
P-carotene: yellow to deep orange; P-apo-8'-carotenoic acid ethyl ester:
yellow to orange; P-apo-8'-carotenal: peach to red-brown; canthaxanthin:
peach to red.
Since the matrix of the water-soluble carotenoid preparation contains
gelatine, these products are fully compatible with encapsulation gelatine for
gelatine capsules; on the other hand, fat-based suppositories can be coloured
by using oily suspensions of micronized carotenoids [213].

2. Cosmetics
Carotenoids are potentially useful in cosmetic products, such as suspensions,
emulsions, lotions, lipsticks and powder bases [214, 215].

E. Assay Problems

The exact chemical determination of every single compound of food or

feed ingredients with provitamin A or pigmenting capacity is a very difficult
task and is only performed for research purposes. As an example, the deter-
mination of the carotene and carotenoid pattern of dehydrated alfalfa meal
by Bickoff et al. [216] should be mentioned. Fiasson et al. [217] have given a
general scheme for qualitative and quantitative assay of naturally occurring
carotenoids.
1. Assay in feeds
Depending on the problem, simplified methods are used by which certain
fractions containing a certain number of various carotenes or carotenoids are
isolated. The optical density of these fractions gives a measure for the quantity
 XI. Use of Carotenoids 763

of the main carotenes or carotenoids present in every fraction. With regard to

carotenes and carotenoids possessing provitamin A activity, Bickoff et al. [218]
point out that most of the older analytical methods for P-carotene (3) measured
only total carotene, and no allowance was made for the difference in activity
of the various provitamin A carotenoids-oc- (5), P- (3), y-carotene (8) and
cryptoxanthin (39)-that might be present. The problem is further complicated
by the fact that each of the carotenoids can exist in a number of stereoisomeric
forms, some with widely different biological properties. The various carotenoids
described can be separated by chromatographic procedure and the individual
carotenoids determined photometrically. However, the widely used chromato-
graphic procedure for the determination of the P-carotene (3) in feed ingredients
such as the AOAC method [219, 220] and the method of Papendick [221] fail
to separate the isomers of lower biological potency from all-trans-P-carotene
so that, in fact, the biological activity may be overestimated by as much as
30%. Workers who have compared the biological response of a feed containing
a mixture of carotenoids and their isomers point out that while it is possible
to work out a factor for each feed-stuff converting the chemical value to that
obtained biologically, the biological evaluation is the only method which
indicates the actual vitamin A activity of such feed ingredients.
However, with the availability of stabilized pure synthetic vitamin A, from
a practical point of view the need to solve the analytical problem exists only for
fresh forage and silage as feed of ruminants [222], since vitamin A is added to
most rations independently of their real provitamin A content.
The problem of determining the pigmenting carotenoids is much more
complicated because of the plurality of the compounds involved. Brubacher
and Vuilleumier [223] gave a review of the available methods. One of the first
papers on practical methods of xanthophyll determination was that of Bickoff
et al. [218], in which two alternative methods applicable to alfalfa meal were
described. These methods have been widely used over the past decade. Various
workers have adapted the procedure to the sample on hand or the type of
xanthophylls encountered but only a few cases need be mentioned. Quacken-
bush et al. [224] determined the carotenoids in corn. Blessin [225] reported
on the carotenoid composition of corn and sorghum, Kohler et al. [82] gave
a practical procedure for determining the so-called total xanthophylls and
carotene in fresh and dehydrated alfalfa and grass meal. Livingston et al [227]
recorded the xanthophyll and carotene content of lucerne, clovers and grasses
found by the method of Kohler et al. [82]. A practical approach was given by
Quackenbush et al. [76]; the method was checked in a comparative trial by
12 different collaborators [226], and this technique was recommended in
place of the offi~ial AOAC method. A technique was also proposed by the same
authors [76] for the determination of a 'dihydroxy pigment' (DHP) equivalent
which may be used as a measure of pigmenting efficacy for avian skins and
yolk. Another collaborative trial with broilers has shown good correlation
between the DHP equivalents and shank pigmentation [144]. With the
availability of stabilized pure synthetic carotenoids, analytical methods were
764 J.C. BAUERNFEIND, G.B. BRUBACHER, H.M. KLAUI and W.L. MARUSICH

developed to assay rations enriched with these pigmenters. Brubacher and

Vuilleumier [223] gave an outline of the possibilities in this field. Assay
methods for [3-apo-carotenal, the ethyl ester of [3-apo-8'-carotenoic acid,
canthaxanthin and the major xanthophyll components in feeding stuffs are
available at request from F. Hoffmann-LaRoche & Co. Ltd., Basle, Switzer-
land [228- 230].

2. Assay in foods
Similar problems arise in the assay of carotenoids in food as were discussed
in the previous section. However, a considerable number of publications deal
with the differentiation between carotenes and other carotenoids that are
normally present and those that have been added. This aspect is of particular
significance for those countries where colouration of certain products is not
legally allowed.
Several conventional methods exist for analysing certain fractions of caro-
tenoids. The most important ones are described by Vuilleumier et a!. [50],
Booth [231], the Association of Official Agricultural Chemists [220], the
'Schweizerisches Lebensmittelbuch' [232], the British Margarine Order [233],
and by Usher et a!. [234].
The problem of analysis of [3-carotene (3) is treated by Booth [231] in a
very thorough and critical manner (see also [235]). For conversion of [3-caro-
tene values to vitamin A activity see p. 745.
Procedures for the assay of naturally occurring as well as added carotenoids
in food are given by Vuilleumier et a!. [50] and in fats by Brubacher [236].
Osadca eta!. [237] described assay methods for apo-carotenal (248) and cantha-
xanthin (193) in food.
It is possible to detect [3-carotene (3) added to citrus fruit juices and citrus
fruit-based products due to the experimentally based observation that the
amount of natural [3-carotene (3) does not exceed a certain proportion of the
total carotenoids [238-252].
Using similar methods, added [3-carotene can be detected in vegetable
oils [253], in egg yolks and egg powders [254, 255] and in macaroni products
[256].

References
[1] IUPAC, The Vitamin A Potency of {3-Carotene (Butterworth, London 1959).
[2] FAO/WHO, FAD Nutr. Meet. Rep. Ser. No.41/World Health Organ. Tech. Rep. Ser. No.362
(FAO, Rome 1967).
[3] Canada Department of Agriculture, Biological Values of Provitamins A for Man, Domestic
Animals and Birds (Can. Dep. Agr. Pub/. No.1238) (Roger Duhamel, F.R.S. C., Ottawa 1966).
[4] J. Nadai and G. Brubacher, in Fat-Soluble Vitamins (International Encyclopaedia of Food
and Nutrition, Vol. 9 ), ed. by R.A. Morton (Pergamon Press, Oxford 1970), p. 449.
[5] R. Ferrando, Cah. Nutr. Diet., in the press (1971).
[6] H. Simmonet, Wiss. Veroff. Deut. Ges. Erniihr. 9, 205 (1963).
[7] J. Tiews, Wiss. Veroff. Deut. Ges. Ernii~r. 9, 235 (1963).
 XI. Use of Carotenoids 765

[8] T. W. Goodwin, The Comparative Biochemistry of the Carotenoids (Chapman and Hall,
London 1952).
[9] H. Haslach, Die biologische Aktivitiit von 13-mono-cis-Vitamin A und {J-Apo-8' -carotinal im
Wachstumstest am Kuken, Diss. (Munich 1964).
[10] W.L. Marusich, E. De Ritter and J.C. Bauernfeind, J. Amer. Oil. Chem. Soc. 34, 217 (1951).
[11] W.L. Marusich and J.C. Bauernfeind, Poultry Sci. 42, 949 (1961).
[12] J.C. Bauernfeind, M. Osadca and R.H. Bunnell, Food. Techno!. 16, No.8, 101 (1962).
[13] J. Nadai, The Vitamin A Requirement of Cattle and its Satisfaction, Proc. World Congr. Anim.
Feed. 1966, Vol.1 (Gimenez, Madrid 1966), p. 600.
[14] B.K.Watt and A.L. Merrill, U.S., Dep. Agr., Agr. Handb. No.8 (1950), Rev. Ed. (1963).
[15] R.A. McCance and E.M. Widdowson, Med. Res. Counc. (Gt. Brit.), Spec. Rep. Ser. No. 297
(1960).
[16] 0. Volker, Wiss. Veroff. Deut. Ges. Erniihr. 9, 282 (1963).
[17] H. Thommen and H. Wackernagel, Biochim. Biophys. Acta 69,387 (1963).
[18] H. Thommen and H. Wackernagel, Naturwiss. 51, 87 (1964).
[)9] P.N. Davis and F. H. Kratzer, Poultry Sci. 37, 851 (1958).
[20] C. Fassler, J.P. Vuilleumier and G.B. Brubacher, Internat. Z. Vitami'!forsch. 32, 454 (1962).
[21] J. Tiews and H. Zucker, Tieriirztl. Umsch. 18, 590 (1963).
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[253] E. Benk and L. Brixius, Fette, Seifen, Anstrichm. 67, 65 (1965).
[254] E. Benk, L. Brixius and I. Wolff, Deut. Lebensm.-Rundsch. 62, 139 (1966).
[255] E. Benk, R. Dietl and L. Brixius, Deut. Lebensm.-Rundsch. 63, 110 (1967).
[256] E. Benk and H. Seibold, Gordian 68, 275 (1968).
[257] H. Hartel, Arch. Gefliigelk. 34, 109 (1970).
[258] Verordnung tiber farbende Stoffe (Farbstoff-Verordnung, Bundesrepublik Deutschlarlp)
vom 19. Dezember 1959, Bundesgesetzblatt I, 756; geandert durch VO. vom 22. Dezember
1960, Bundesgesetzblatt I, 1073.
[259] H. K!aui, Proc. Inst. Food Sci. Techno/. (U.K.), Vol.1, No.5 (Dec. 1968).
[260] M. Bougon, Bull. Inform. Sta. Exp. Avicult. Ploufragan (Cotes-du-Nord) 9, No.4 (1969).
[261] F. Hoffmann-LaRoche &Co. Ltd., Egg quality and yolk colour (Roche News and Reviews)
(Basle 1966).
[262] H.V. Courtnay and R. E. Branson, Texas, Agr. Exp. Sta., Bull. No. 989 (1962).
[263] G.O. Kohler, Proc. 9th Alfalfa Conf, Lincoln, Nebraska 1965, p. 50.
[264] R.H. Bunnell and J.C. Bauernfeind, Proc. 11th World's Poultry Congr., Mexico City 1958.
[265] J. K. Bletner, R. P. Mitchell, Jr., and R. L. Tugwell, Poultry Sci. 45, 698 (1966).
[266] F. H. Bird, Proc. Texas Nutr. Conf. 1952.
 771

XII. Lists of Natural Carotenoids

O.STRAUB
F. Hoffmann-La Roche & Co. Ltd., Basle, Switzerland

A. Carotenoids of Known Structure 772

1. Hydrocarbons . . . . . . . 772
2. C 40 -Xanthophylls . . . . . 779
a) Monohydroxy compounds 779
b) Dihydroxy compounds . . 784
c) Polyhydroxy compounds . 789
d) Alkoxy compounds 791
e) Epoxy compounds . 795
f) Aldehydes . . 800
g) Monoketones . 803
h) Diketones . 813
i) Polyketones . 818
j) Acids . . . . 818
k) Seco compounds 818
3. Homo-carotenoids . 819
4. Apo-carotenoids . . 823
5. Nor-carotenoids . . 830
B. Carotenoids of Unknown Structure 830
C. Old Names 833
References . . . . . . . . . . . . 833
 772 0. STRAUB

A. Carotenoids of Known Structure

Each carotenoid is designated by a prefix number in bold type, which is

used for reference throughout this book. Above the structural formula, the
commonly used trivial name and additional designations are given, with
obsolete or erroneous names in double quotation marks. Shown last is the
semi-systematic name (cf. nomenclature rules in the Appendix of this mono-
graph). Uncertainty in a structural formula is indicated by a question mark
and uncertain natural occurrence by the term 'natural?'. A query after a trivial
name leaves open the question whether the structural formula shown is correct
for the pigment concerned.
Below the formulae are found literature references for proton magnetic
resonance (PMR), mass (MS) and infrared (IR) spectra, for optical rotatory
dispersion (ORD) and circular dichroism (CD). Crystal structural studies are
denoted 'X-ray' and chemical structural studies 'Chern.'. Literature on partial
and total syntheses (excluding patent literature) is indicated by 'Synth.', other
miscellaneous literature by 'Misc.'. Occasionally indications on occurrence
are found under 'Occur.'. The literature references are in chronological order,
beginning with the latest publications.
Within the subsections the Greek alphabetical order is followed, for
example: 1-0H-p,p-, 2-0H-p,p, ... , 1-0H-p,e-, ... , 1-0H-e,t/J-, ... ,20'-0H-t/J,t/J-
carotene. Hydroxy alkoxy compounds are found under ethers, alkoxy ketones
under the ketones, etc. Glycosides are arranged according to the related
aglycones, acyl compounds according to the related carotenols and esters under
the corresponding acids.
The list in its present form displays the fully established characteristics of the
individual carotenoids. It also indicates where there are gaps or uncertainties.

1. Hydrocarbons

1 3,4,3',4'-Bisdehydro-P-carotene, "Dehydrocarotene III" [338]; 3,4,3',4'-Tetradehydro-P,P-carotene

Chern. [812, 338] Synth. [312, 307, 304] Occur. [260, 204]

2 3,4-Dehydro-P-carotene; 3,4-Didehydro-p,p-carotene

PMR [36] Synth. [307, 338] Occur. [260,.204]

 XII. Lists of Natural Carotenoids 773

3 P-Carotene; p,p-Carotene

PMR [737, 324, 643, 638, 318, 36] MS [643, 638] IR [638, 111, 681, 315, 306, 305, 525] X-ray [657]
Chern. [794,352,348,345, 791] Synth.[584,688,687,685,336,88,45,619,682,603,312,315,310,309, 306,
221,190,300,298,297,296,549,408,406] Misc. [77,226,653]

3a "p,y-Carotene" [26], P 444 [22]; 5,18-Didehydro-5,6-dihydro-p,p-carotene

PMR, MS, IR [26] Occur. [26, 518, 22]

4 17-Carotene, Tetrahydro-p-carotene, "Octahydrolycopene" [333]; 7,8, 7',8'-Tetrahydro-p,p-carotene

PMR [753, 750] Misc. [645, 333, 610, 236]

5 0(-Carotene; (6' R)-p,s-Carotene

PMR, MS [643, 41, 638] IR [638, 667, 666, 525] ORD [41] CD [69a] Chern. [359, 351]
Synth. [194,41,619, 720, 191,302,411] Misc. [194,653]

6 P- Isorenieratene; p,rp-Carotene

PMR, IR [110] Synth. [110, 423] Occur. [510]

 774 0. STRAUB

7 Torulene, 3',4'-Dehydro-y-carotene; 3',4'-Didehydro-P,I/f-carotene

IR, Synth. [620] Misc. [767, 482, 471]

8 y-Carotene, "Sphaerobolin" [636, 216]; p,l/f-Carotene

PMR [643, 537,638,619] MS [638] IR [537,638,619, 525] Chern. [456, 766, 765] Synth. [537,619,221]

9 P-Zeacarotene, P1-Zeacarotene [649, 598], 7',8'-Dihydro-y-carotene, "Carotene X" [89, 254, 252, i05],
"Pigment X"? [764, 679,256,677,150,334,738,106,105,241, 238]; 7',8'-Dihydro-P,I/f-carotene

PMR [753] MS [76] IR [620] Chern. [598] Synth. [751, 752, 620] Misc. [612, 649, 599, 544]

10 s-Carotene; (6R,6'R)-s,s-Carotene

PMR [643, 537, 638] MS [643, 638] IR [537, 638] CD [69a] Synth. [537, 721, 410] Occur. [82, 660]
Misc. [194]

11 .5-Carotene; (6R)-s,l/f-Carotene

PMR [643, 537, 638] MS [638] IR [537, 638, 337] CD [69a] Chern. [337, 606] Synth. [537]
Occur. [766]

12 a-Zeacarotene, 7',8'-Dihydro-.5-carotene; 7',8'-Dihydro-s,l/f-carotene

PMR [753] Chern. [598] Synth. [751, 752] Misc. [544, 599]
 XII. Lists of Natural Carotenoids 775

13 Isorenieratene, Leprotene; rp,rp-Carotene

PMR [426, 110, 36] MS [426] IR [426, 588, 110,774, 772] Synth. [426, 110,423, 775, 774]
Misc. [507, 509, 770, 701, 699, 266]

14 Renieratene; rp,x-Carotene

IR [110, 771] Chern. [773, 771] Synth. [110, 775] Misc. [770]

15 Chlorobactene; rp,ljJ-Carotene
C40Hs2

""":::: """:::: """:::: """:::: """:::: """:::: """:::: """:::: """:::: """::::

PMR [59, 508] MS [188, 187] IR [59, 508] Synth. [59]

16 Renierapurpurin; x,x-Carotene
C40H4s

""":::: """:::: """:::: """:::: """:::: """:::: """:::: """:::: """::::

PMR [426, 750, 59] MS [426, 189] IR, Synth. [426, 110, 775] Misc. [770]

17 Bisdehydrolycopene, "Dehydrolycopene"; 3,4,3',4'-Tetradehydro-1/1,1/J-carotene

PMR [36] MS [188] Synth. [683, 394] Misc. [768]

18 3,4-Dehydrolycopene, Monodehydrolycopene; 3,4-Didehydro-1/J,I/J-carotene

Occur. [734, 512, 768]

 776 0. STRAUB

19 Lycopene, Rhodopurpurin? [500, 493, 245, 370, 368]; 1/J,I/f-Carotene

PMR [426, 643, 638, 36] MS [426, 737,643, 758, 188, 187, 638] IR [426, 424, 537, 638, 500, 667, 528, 310,
525] Chern. [454, 350, 348] Synth. [426, 537, 336, 310, 220, 409] Misc. [226, 693, 653]

20 1,2-Dihydro-3,4-dehydrolycopene; 3,4-Didehydro-1,2-dihydro-1/1,1/J-carotene
C40Hs6

"""'-:: """'-:: """'-:: """'-:: """'-:: """'-:: """'-:: """'-:: """'-:: """'-:: """'-:: """'-::

MS, Misc. [533, 530]

21 1,2-Dihydrolycopene; 1,2-Dihydro-1/1,1/J-carotene
C40Hss

"""'-:: """'-:: """'-:: """'-:: """'-:: """'-:: """'-:: """'-:: """'-:: """'-:: """'-::

PMR, MS [425, 533, 530] IR, Synth. [ 425]

22 Neurosporene, 7,8-Dihydrolycopene, Flavorhodin? [513, 368], "Poly-cis-1/f-carotene" [527, 605, 526],

"Tetrahydrolycopene" [719, 605, 604]; 7,8-Dihydro-1/J,I/f-carotene

PMR [159, 158] MS [155, 758] IR, Synth. [159, 158] Misc. [500, 491, 527, 269]

23 1,2-Dihydroneurosporene; 1,2,7,8-Tetrahydro-1/J,I/J-carotene

PMR, MS, Misc. [533, 530]

24 1,2,1',2'-Tetrahydrolycopene; 1,2,1',2'-Tetrahydro-1/J,I/J-carotene

PMR [425] MS [425, 533] IR, Synth. [425]

25 7,8,11,12-Tetrahydrolycopene, Asym. (-carotene; 7,8,11,12-Tetrahydro-1/J,I/J-carotene

MS [154] Synth. [159, 158] Misc. [156, liS, 152]

 XII. Lists of Natural Carotenoids 777

26 (-Carotene, 7,8, 7',8'-Tetrahydrolycopene, "5,6, 7,8,5',6', 7',8' -Octahydrolycopene" [556],

"~-Carotene"? [332 a, 736a]; 7,8, 7',8'-Tetrahydro-1/J,I/f-carotene

PMR [159, 158] MS [155, 154, 758] IR [159, 335, 158] Chern. [556] Synth. [159, 158]
Misc. [599, 271, 556, 604, 660]

27 1,2, 7,8,11, 12-Hexahydrolycopene; 1,2,7,8,11, 12-Hexahydro-1/1 ,1/f-carotene

'-'::::::

MS, Misc. [533]

28 1,2,1',2'-Tetrahydroneurosporene; 1,2, 7,8,1',2' -Hexahydro-1/J,I/f-carotene

'-':::::: '-':::::: '-':::::: '-':::::: '-':::::: '-':::::: '-':::::: '-':::::: '-':::::: '-'::::::

MS, Misc. [533]

29 7,8, 1',2', 7',8'-Hexahydrolycopene; 1,2, 7,8, 7' ,8'-Hexahydro-1/J ,1/J-carotene

MS, Misc. [533]

30 Phytofluene, 7,8,11,12,7',8'-Hexahydrolycopene, "Dodecahydrolycopene" [605];

15-cis-7 ,8, 11 ,12, 7',8'-Hexahydro-1/1 ,1/f-carotene

PMR [159, !58] MS [155, 758] IR [159, 335, 158] Chern. [814, 593, 743] Synth. [159, 158]
Misc. [811, 809, 437, 807, 604, 806, 805]

31 "1 ',2'- Dihydrophytofluene" [533]; 1,2, 7,8, 11, 12, 7',8' -Octahydro-1/J,I/f-carotene

'-'::::::
MS, Misc. [533]
 778 O.STRAUB

32 Phytoene, 7,8,11,12,7',8',11',12'-0ctahydrolycopene, 15,15'-Dehydrolycopersene,

"Hexadecahydrolycopene" [ 605]; 15-cis-7 ,8,11,12, 7',8',11 ',12' -Octahydro-r/1 ,rjl-carotene

PMR [758, 159, 335, 158] MS [155, 758] IR [692, 159, 335, 158] Chern. [609, 811] Synth. [159, 158]
Misc. [148, 608, 604]

33 1,2-Dihydrophytoene; 1,2,7,8,11,12,7',8',11',12'-Decahydro-rjl,rjl-carotene

MS, Misc. [533]

34 Lycopersene, Dihydrophytoene, Decahydrolycopene;

7,8,11,12,15, 7',8',11',12',15' -Decahydro-rfr,rjl-carotene
natural?

MS[188] Synth.[319,386] Occur.[626,644,87,718,257,255,585,760,43,149,548,15,263,590,147,262]

35 Anhydroeschscholtzxanthin, Dianhydroeschscholtzxanthin;
2,3,2',3',4',5'-Hexadehydro-4,5' -retro-P,P-carotene

IR [112, 561] Chern. [413, 659] Synth. [413] Occur.[112] Misc. [338, 810]

36 Isocarotene, Dehydroretrocarotene, Retrodehydrocarotene, Retrodehydro-P-carotene,

"Dehydro-P-carotene" [81 0], 4,4'-Didehydro-P-carotene;
4',5'-Didehydro-4,5' -retro-p,p-carotene

natural?

PMR [36] IR [561, 265] Synth. [689, 702,.309, 810, 449] Occur. [27] Misc. [814, 379]
 XII. Lists of Natural Carotenoids 779

37 7, 7'- Dihydro-P-carotene, P- Dihydrocarotene [380]; 7,8-Dihydro-8,7'-retro-P.P-carotene

natural?

Synth. [309,299,380] ()ccur. [414]

2. C40 -X anthophylls

a) Monohydroxy compounds

38 Anhydrolutein, 3'-Hydroxy-3,4-dehydro-p-carotene, "Deoxylutein I" [813];

3',4'-Didehydro-P.P-caroten-3-ol

artifact!
HO

()ccur. [166, 75, 74] Misc. [813]

39 P-Cryptoxanthin, Cryptoxanthin, Physoxanthin = Neo-P-cryptoxanthin A? [99, 49, 96, 48, 95, 47, 46;
see also 43], "Caricaxanthin" [12, 361, 776]; (3R)-P,P-Caroten-3-ol

PMR [523] MS [523, 737] IR [523, 697, 51, 667, 313] C>RD [41] Chern. [235, 415, 462]
Synth. [523, 313] Misc. [162, 653]

40 Isocryptoxanthin, 4-Hydroxy-p-carotene; p,p-Caroten-4-ol

OH

MS [76] IR [265, 51] Synth. [689, 184, 596, 744] C>ccur. [275]
 780 0. STRAUB

41 Crocoxanthin; (3R,6' R)-7,8-Didehydro-p,e-caroten-3-ol

:><Y'*'
HO~
PMR [753] MS [534] ORO [41] Synth. [755, 754] Misc. [84]

42 IX-Cryptoxanthin (see also 43), Zeinoxanthin, 3-Hydroxy-tX-carotene; (3R,6'R)-P,e-Caroten-3-ol

PMR [523, 697, 41] MS [523, 41] IR [523, 697] ORO [233, 756, 41] CD [233] Synth. [523]
Misc. [162, 600, 94]

43 IX-Cryptoxanthin? (see 42), Physoxanthin? (see 39), 3'-Hydroxy-tX-carotene; p,e-Caroten-3'-ol

OH

44 Celaxanthin, 3-Hydroxytorulene, 3',4'-Dehydrorubixanthin; 3',4'-Didehydro-P,I/J-caroten-3-ol

Chern. [488] Synth. [2, 55]

45 Rubixanthin, 3-Hydroxy-y-carotene; (3R)-P,if!-Caroten-3-ol

PMR [23, 66] MS [23, 66, 187] IR [23, 508] ORO [23, 41] CD [23] Chern. [23, 463]
Misc. [162, 66, 271]
 XII. Lists of Natural Carotenoids 781

46 Gazaniaxanthin; (3 R)-5' -cis-{J,r/J-Caroten-3-ol

PMR, MS [23, 66] IR [23] ORO [23, 41] CD [23] Misc. [162, 804, 637]

47 Myxobactin, 1',2'-Dihydro-1'-hydroxy-3,4-dehydrotorulene glucoside;

1'-Glucosyloxy-3,4,3',4' -tetradehydro-1 ',2' -dihydro-{J,r/J-carotene

MS, IR [ 435] Occurrence as fatty acid esters [ 435, 434]

47 a I ',2'-Dihydro-1' -OH-torulene; 3',4'-Didehydro-1 ',2' -dihydro-{J,r/J-caroten-1'-ol

OH

Chern. [613a] Misc. [57]

48 1',2'-Dihydro-1 '-hydroxy-y-carotene; 1',2'-Dihydro-{J,r/J-caroten-1 '-ol

OH

PMR [59] Chern. [276] Synth. [59]

49 Aleuriaxanthin; 1',16'-Didehydro-1',2'-dihydro-{J,r/J-caroten-2'-ol
OH

PMR, MS, IR [519] Chern. [519, 512] Misc. [211a]

50 "3',4'-Didehydro-18' -hydroxy-y-carotene "; 3',4' -Didehydro-{J,r/J-caroten-16' -ol

CH 2 0H

Synth. [618] Occur. [58]

 782 O.STRAUB

51 3-Hydroxy-.5-carotene; e,t/J-Caroten-3-ol

Misc. [230, 128]

52 3-Hydroxyisorenieratene; q>,q>-Caroten-3-ol

HO
PMR [18] MS [586, 18] IR, Chern. [586, 18, 17] Synth. [18, 17]

53 OH-Chlorobactene, 1',2'-Dihydro-1' -hydroxychlorobactene; 1',2' -Dihydro-q>,t/J-caroten-1 '-ol

PMR, IR [59] Chern. [508] Synth. [59]

54 OH-Chlorobactene glucoside; 1'-(p-o-Glucopyranosyloxy)-1 ',2' -dihydro-q>,t/J-carotene

Misc. [607]

55 3,4-Dehydrorhodopin, 1,2-Dihydro-3,4-dehydro-1-0H -lycopene;

3,4-Didehydro-1,2-dihydro-t/J,t/J-caroten-1-ol

PMR [635, 320] MS, IR, Synth. [635] Misc. [613a]

56 Rhodopin, 1-Hydroxy-1,2-dihydrolycopene, 1,2-Dihydro-1-0H -lycopene,

OH-Lycopene, "Bacterioerythrin {J" [245]; 1,2-Dihydro-t/J,t/J-caroten-1-ol

PMR [7, 59, 622] MS [188, 187, 547] IR [59, 622, 500, 493] Chern. [500, 493, 376] Synth. [684, 59]
Misc. [491, 245, 370, 368]
 XII. Lists of Natural Carotenoids 783

57 Rhodopin glucoside; 1-(p-o-Glucopyranosyloxy)-1,2-dihydro-t/f,t/1-carotene

PMR, MS, IR, Chern. [634]

57 a Dernethylated spheroidene; 3,4-Didehydro-1,2, 7',8' -tetrahydro-t/1,t/f-caroten-1-ol

H
natural?

Misc. [64a, 258, 177, 506]

58 Chloroxanthin, 1-Hydroxy-l ,2-dihydroneurosporene, 0 H-Neurosporene,

1-Hydroxy-1,2, 7',8' -tetrahydrolycopene; 1,2,7',8'-Tetrahydro-t/1 ,t/1-caroten-1-ol

PMR [39, 536, 158] MS [39] IR [39, 536, 503, 500, 493] Chern. [553] Synth. [39, 536]
Misc. [633, 491, 246, 240]

59 1-Hydroxy-1,2, 7',8',11 ',12' -hexahydrolycopene; 1,2,7',8' ,11 ',12'-Hexahydro-t/l,t/f-caroten-1-ol

Misc. [156]

60 1-Hydroxy-1,2-dihydrophytofluene, Hydroxyphytofluene? [508, 491], Phytofluenol? [131, 808];

1,2,7,8,11,12, 7',8' -Octahydro-t/l,t/f-caroten-1-ol C 40 H 640

MS, Chern. [531] Misc. [156]

61 1-Hydroxy-1,2-dihydrophytoene, Hydroxyphytoene? [436], Phytoenol? [131];

1,2,7,8,11, 12,7',8', 11 ',12'-Decahydro-t/f,t/f-caroten-1-ol

MS, Chern. [531]

 784 O.STRAUB

62 Lycoxanthin, Lycopen-16-ol, 16-Hydroxylycopene; 1/1,1/1-Caroten-16-ol

HOH 2C ~

PMR [426a, 543, 103] MS [426a, 188, 103, 187, 543] IR [426a, 543, 500, 494]
Chern. [421, 103,543,500, 799] Synth. [426a,518] Misc. [622,245,250]

63 Anhydrowarrningol, 13-cis-Lycopen-20-ol; 13-cis-1/f,I/1-Caroten-20-ol

Misc. [601]

b) Dihydroxy compounds

64 3,4-Dihydroxy-p-carotene; p,p-Carotene-3,4-diol

HO
OH

Synth. [53] Occur. [557]

65 Alloxanthin, Cynthiaxanthin, Pectenoxanthin, Cryptornonaxanthin [41],

7,8, 7',8'-Tetradehydrozeaxanthin; (3R,3' R)-7,8,7',8'-Tetradehydro-P,fl-carotene-3,3'-diol

;><y~

HOAA

PMR [756, 753, 79, 534] MS [79, 534] IR [643, 79, 534] ORO [41] Chern. [79]
Synth. [755, 754, 751] Misc. [162,616,84,8~,272,481,476,475,474]
 XII. Lists of Natural Carotenoids 785

66 Diatoxanthin, 7,8-Didehydrozeaxanthin; (3 R,3' R)-7,8-Didehydro-/),/)-carotene-3,3' -diol

OH C•oHs402

;><y'*
HO~
PMR [672, 534] MS [672] IR [672, 674] ORD [41] Chern. [577, 674] Synth. [579, 577]
Misc. [162, 173, 661]

67 Zeaxanthin, Anchovyxanthin? [287]; (3R,3' R)-{J,fi-Carotene-3,3' -diol

PMR [523, 41, 104, 643, 36] MS [523, 76, 41, 104, 188, 187] IR [523, 675, 51, 667, 493, 311, 525]
ORD [41] CD [41] Chern. [674, 552, 375,457,453, 354,443, 347]
Synth. [523, 104, 312, 313, 311, 366] Misc. [162, 653, 415, 343]

68 Physalien, Zeaxanthin dipalrnitate; (3R,3'R)-/),/)-Carotene-3,3'-diol dipalrnitate C 72 Htt604

OCO(CH 2 )J.CH3

IR [305] Chern. [443] Synth. [312, 311, 464] Misc. [442]

69 Caloxanthin; 6, 7-Didehydro-5,6-dihydro-/),p-carotene-3,3'-diol
OH

N·~
HO~
IR [675, 267] Chern. [267] Misc. [284b, 289]

70 Nostoxanthin; 6,7,6',7'-Tetradehydro-5,6,5',6' -tetrahydro-p,p-carotene-3,3' -diol

YYOH

~-~
N·~
HO~
IR, Chern. [675, 267] Misc. [284b, 289]

Carotenoids 50
 786 0. STRAUB

71 Isozeaxanthin, 4,4'-Dihydroxy-P-carotene; p,p-Carotene-4,4'-diol

OH
PMR [750] MS [737, 76, 187] IR [265, 51, 264] Chern. [595] Synth. [183, 308, 595]
Misc. [145, 483]

72 Monadoxanthin; 7,8-Didehydro-p,e-carotene-3,3'-diol

~'*
HOAA
PMR [753] MS [534] ORD [41] Misc. [84, 83]

73 Lutein, "Xanthophyll", "Cucurbitaxanthin" [796, 680], 3,3'-Dihydroxy-IX-carotene;

(3 R,3'S,6' R)-p,e-Carotene-3,3' -diol*
H

PMR [41, 36] MS [76, 41, 188, 187] IR [41, 51, 176, 500,666, 667] ORD [41]
Chern. [401a,360,347, 792] Misc. [233, 162,745, 104,653,415,447,346]

74 Helenien, Lutein dipalmitate, Xanthophyll dipalmitate;

(3R,3' S,6' R)-P,e-Carotene-3,3' -diol dipalmitate

Synth. [349] Misc. [447, 444]

75 Saproxanthin; 3',4'-Didehydro-1',2'-dihydro-P,t/t-carotene-3,1'-diol

OH
HO
IR, Chern. [2]

* Added in proof: For C-3' the opposite (R) configuration has been claimed [69b].
 XII. Lists of Natural Carotenoids 787

76 Plectaniaxanthin, 1',2'-Dihydroxy-1 ',2' -dihydrotorulene;

3',4'-Didehydro-1 ',2' -dihydro-P,ifi-carotene-1 ',2' -diol

OH

PMR [28a, 283, 641] MS [28a, 188] IR, Chern. [28a, 21] Synth. [641]

77 Phleixanthophyll; 1'-(p-o-Giucopyranosyloxy)-3',4' -didehydro-1',2' -dihydro-P,ifi-caroten-2'-ol

PMR, MS, IR, Chern. [279] Misc. [276, 630]

77a 1'-[(x-0- Palmitoyl-P-o-glucopyranosyl)oxy]-3',4'-didehydro-1 ',2' -dihydro-P,ifi-caroten-2'-ol

Misc. [733]

78 Tunaxanthin, 3,3'-Dihydroxy-e-carotene; e,e-Carotene-3,3'-diol

OH

Misc. [288, 114, 287, 113, 727, 286]

79 3,3'-Dihydroxyisorenieratene; tp,tp-Carotene-3,3' -diol

OH

HO

PMR [18, 17] MS [586, 18] IR, Chern. [586, 18, 17] Synth. [18, 17]
 788 O.STRAUB

80 Rhodopinol, Warrningol; 13-cis-1,2-Dihydro-Y,,Y,-carotene-1,20-diol

PMR [7] MS [213] IR, Chern. [7] Misc. [601, 505]

81 "Dihydroxylycopene ", 1,1 '-Dihydroxy-1,2, 1',2' -tetrahydrolycopene, OH-Rhodopin;

1,2,1 ',2'-Tetrahydro-Y,,Y,-carotene-1 ,1 '-diol

OH
MS [188, 187, 547] Chern. [622] Synth. [683] Misc. [431, 543, 500, 491]

82 1,1'-Dihydroxy-1,2, 1',2' -tetrahydro-(-carotene; 1,2,7,8,1',2', 7',8'-Octahydro-Y,,Y,-carotene-1,1 '-diol

?
Chern. [202] Misc. see also [757, 491]

83 Lycophyll, Lycopene-16,16'-diol; Y,,Y,-Carotene-16,16'-diol

~ CH 2 0H
HOH 2C ~

PMR [103, 543] MS [188, 103, 543] IR [543] Chern. [103, 543, 799] Synth. [426a]
Misc. [421, 399]

84 Eschscholtzxanthin, "3,3'-Dihydroxyretro-P-carotene ", "3,3'-Dihydroxydehydro-P-carotene";

4',5' -Didehydro-4,5' -retro-P.P-carotene-3,3'-diol C40 H 54 0 2
OH

HO

IR [561, 667] Chern. [323, 413] Synth. [18~] Misc. [659]

 XII. Lists of Natural Carotenoids 789

c) Polyhydroxy compounds

8S 3,4,4'-Trihydroxy-p-carotene; p,p-Carotene-3,4,4' -trio!

?
HO
OH

Misc. [146, 731]

86 Deepoxyneoxanthin, Trollein?; 6, 7-Didehydro-5,6-dihydro-p,p-carotene-3,5,3' -trio!

OH

Chern. [577, 575] Misc. [628, 439, 117, 116]

87 5-Hydroxy-5,6-dihydrozeaxanthin; 5,6-Dihydro-p,p-carotene-3,5,3'-trio!
OH

IR, Misc. [142]

88 Loroxanthin, 19-Hydroxylutein; p,s-Carotene-3,19,3'-triol

OH

HO
PMR [11] MS [746, 11] IR, Chern. [11] Misc. [433]

89 Pyrenoxanthin; p,s-Carotene-3,20,3' -trio!

H 2 0H

HO
PMR, MS, IR, Chern. [777]
 790 0. STRAUB

90 Myxoxanthophyll, Myxol 2' -rhamnoside; 2' -(/J-L- Rhamnopyranosyloxy)-3',4' -didehydro-1',2' -dihydro-

fJ,I/t-carotene-3,1 '-diol

OH
HO
PMR [283, 641] MS, IR [283] Chern. [283, 278, 389] Misc. [212, 276, 251, 274]

91 Myxol 2' -0-methyl-methylpentoside; 2' -(0-Methyl-5-C-methylpentosyloxy)-3' ,4' -didehydro-

1',2'-dihydro-fJ,I/t-carotene-3,1' -diol C47 H 68 0 7
C 7 H 13 0 4

OH
HO
PMR, MS, Chern. [212]

92 Myxol 2' -glucoside; 2' -(fJ-o-Giucopyranosyloxy)-3',4' -didehydro-1',2' -dihydro-fJ,I/t-carotene-3,1' -diol

HO
Lit. see 90

93 Crustaxanthin, 3,4,3',4'-Tetrahydroxy-fJ-carotene; fJ,fJ-Carotene-3,4,3',4' -tetrol

HO
HO
IR [54, 53] ORD [41] Chern. [562, 54] Synth. [485, 562] Misc. [145, 557]

94 Heteroxanthin; 7' ,8'-Didehydro-5,6-dihydro-fJ,fJ-carotene-3,5,6,3' -tetrol

(or "3,3',5'-Trihydroxy-6' -hydro-7,8-dehydro-fJ-carotene"; 7',8'-Didehydro-
5,6-dihydro-fJ,fJ-carotene-3,5,3'-triol? [674]) see also 187
I("'OH

'*/X
HO
PMR [672, 671] MS [672, 671, 669] IR [6?1, 674, 669] Chern. [671, 674]
 XII. Lists of Natural Carotenoids 791

94a Aphanizophyll, 4-Hydroxyrnyxoxanthophyll;

2' -(/3-L-Rharnnopyranosyloxy)-3',4'-didehydro-1 ',2' -dihydro-{3,1/t-carotene-3,4, 1'-trio!

OH
HO
OH

MS, Chern. [284a] Misc. [278, 713, 711]

95 Oscillaxanthin, Oscillol 2,2' -dirharnnoside, "1,1'-Dihydroxy-2,2' -di-P-L-rharnnosyl-1,2,1 ',2' -tetrahydro-

3,4,3',4' -tetradehydrolycopene"; 2,2'-Bis(/3-L-rharnnopyranosyloxy)-3,4,3' ,4' -tetradehydro-
1,2,1',2' -tetrahydro-1/t,ift-carotene-1,1' -diol C 52 H 7 6 0 12

OH

PMR, MS, IR, Chern. [284] Misc. [212, 389]

96 Oscillol2,2'-di(O-rnethyl-rnethylpentoside); 2,2'-Bis(O-rnethyl-5-C-rnethylpentosyloxy)-
3,4,3',4'-tetradehydro-1,2,1 ',2' -tetrahydro-ift,ift-carotene-1,1 '-diol

OH

OH
MS, Chern. [212]

d) Alkoxy compounds

96 a 1'-Methoxy-1 ',2' -dihydro-3',4' -dehydro-y-carotene; 1'-Methoxy-3' ,4' -didehydro-1',2' -dihydro-{3,1/t-carotene

CH 3 0

[44]

97 Anhydrorh.odovibrin, "P 481 "; 1-Methoxy-3,4-didehydro-1,2-dihydro-ift ,ift-carotene

PMR [39] MS [188, 187] IR [500, 492, 490] Chern. [500] Synth. [39, 684]
Misc. [499, 498,491,245, 244]
 792 0. STRAUB

98 3,4-Dihydroanhydrorhodovibrin, 1-Methoxy-1 ,2-dihydrolycopene;

1-Methoxy-1,2-dihydro-t/J,t/J-carotene

OCH 3

PMR [426] MS [426, 532] IR, Synth. [426] Misc. [156, 500]

99 Spheroidene, "Pigment Y" [648, 554, 243, 571], "P 450" [156, 154, 153, 529, 491];
1-Methoxy-3,4-didehydro-1 ,2, 7',8'-tetrahydro-t/J ,ljt-carotene

CH 3

PMR [39, 538, 158] MS [156, 154, 529, 39] IR [39, 538, 503, 492, 554, 250] Synth. [39, 538]

100 3,4-Dihydrospheroidene, "1-Methoxy-1,2-dihydroneurosporene" [532];

1-Methoxy-1 ,2, 7',8' -tetrahydro-ljt,ljt-carotene

OCH 3

MS [156, 532]

101 11',12'-Dihydrospheroidene, "P412"? [491, 240];

1-Methoxy-3,4-didehydro-1 ,2, 7',8', 11 ', 12'-hexahydro-1/t ,ljt-carotene

MS [154] Misc. [156, 153]

102 3,4,11',12'-Tetrahydrospheroidene; 1-Methoxy-1,2, 7',8',11',12' -hexahydro-1/t,l/t-carotene

OCH 3

MS [532, 154] Misc. [156]

103 1-Methoxy-1,2-dihydrophytofluene, "Methoxyphytofluene ";

1-Methoxy-1,2, 7,8, 7' ,8', 11 ', 12'-octahydro-1/t,ljt-carotene

OCH 3

?
Misc. [532, 531]
 XII. Lists of Natural Carotenoids 793

104 1-Methoxy-1,2-dihydrophytoene, "Methoxyphytoene ";

1-Methoxy-1,2,7,8,11,12, 7',8',11',12' -decahydro-1/f,l/f-carotene

Misc. [532, 531]

105 OH-Spirilloxanthin, Monodemethylated spirilloxanthin, Dehydrorhodovibrin,

"ac-Bacterioruberin monomethyl ether" [ 500]; 1'-Methoxy-3,4,3',4' -tetradehydro-1,2,1 ',2' -tetrahydro-
1/1,1/f-caroten-1-ol

CH 3 0

PMR [635, 33] MS [188, 187, 635] IR [635, 503] Chern. [500, 497] Synth. [635]
Misc. [253,492,491, 572,240,239]

106 Rhodovibrin, OH-P 481; 1'-Methoxy-3',4'-didehydro-1,2,1',2'-tetrahydro-1/f,l/l-caroten-1-ol

CH 3 0

MS [188, 187] IR [503, 500, 495, 490] Chern. [500, 495] Synth. [684]
Misc. [33,491,245,246,572,244,376,370]

107 OH-Spheroidene, "Hydroxyspheroidene", "1'-Hydroxy-1',2'-dihydrospheroidene" [156],

"7',8'-Dihydrorhodovibrin" [503], OR-Pigment Y, OH-Y [500, 499,246, 243];
1'-Methoxy-3' ,4' -didehydro-1 ,2, 7,8,1 ',2' -hexahydro-1/1 ,1/f-caroten-1-ol

CH 30

PMR [321] MS [529] IR [503] Chern. [321, 503] Misc. [177]

108 Spirilloxanthin, Rhodoviolascin, Bacteriopurpin ac? [555, 42, 654, 570], Bacterioerythrin a? [500, 570];
1,1 '-Dimethoxy-3,4,3',4' -tetradehydro-1,2,1 ',2' -tetrahydro-1/1,1/f-carotene

CH 3 0

PMR [39, 683, 36, 33] MS [188, 187, 635] IR [39, 622, 503, 501, 500, 33, 492]
Chern. [39,500,33,492,378,370,369] Synth. [635,683,497] Misc. [653,602,569,368]
 794 O.STRAUB

109 3,4-Dihydrospirilloxanthin; 1,1 '-Dimethoxy-3,4-didehydro-1,2,1',2' -tetrahydro-t/f,t/1-carotene

CH 3 0

MS [532] Misc. [500]

110 3,4,3',4'-Tetrahydrospirilloxanthin, 1,1'-Dimethoxy-1,2,1',2' -tetrahydrolycopene;

1,1 '-Dimethoxy-1,2,1 ',2' -tetrahydro-1/t,t/f-carotene

CH 3 0

PMR [426, 5] MS [426, 188, 187] IR [426, 5] Synth. [426, 5, 683] Misc. [532, 601, 178]

Ill 3,4,7,8-Tetrahydrospirilloxanthin, 1'-Methoxy-1 ',2' -dihydrospheroidene;

1,1'-Dimethoxy-3,4-didehydro-1,2,1',2',7',8'-hexahydro-t/t,t/t-carotene

CH 3 0

MS, Misc. [532]

112 3,4,3',4',7',8'-Hexahydrospirilloxanthin, 1,1'-Dimethoxy-1,1',2,2'-tetrahydroneurosporene;

1,1 '-Dimethoxy-1,2, 7,8,1',2' -hexahydro-1/t,1/t-carotene

?
CH 3 0
MS, Misc. [532]

113 3,4,3',4',7' ,8',11',12'-Octahydrospirilloxanthin; 1,1'-Dimethoxy-1,2,7,8,11,12, 1',2' -octahydro-t/f,t/f-carotene

?
CH 3 0
Misc. [532]
 XII. Lists of Natural Carotenoids 795

e) Epoxy compounds

114 P-Carotene 5,6-epoxide, P-Carotene rnonoepoxide; 5,6-Epoxy-5,6-dihydro-p,p-carotene

MS [76, 29] Synth. [392] Misc. [180]

115 IX-Carotene epoxide, "5,6-Monoepoxy-<X-carotene"; 5,6-Epoxy-5,6-dihydro-{J,e-carotene

Synth. [ 423 a, 393] Occur. [735, 695, 223, 397]

116 Phytoene 1,2-oxide; 1,2-Epoxy-1,2,7,8,11,12,7',8',11',12'-decahydro-1/J,I/J-carotene

MS, Chern. [64]

117 Cryptoxanthin epoxide; 5,6-Epoxy-5,6-dihydro-p,p-caroten-3-ol

or perhaps 5',6' -epoxide?

HO

Chern. [674, 196, 399] Synth. [399] Misc. [783, 131, 129]

118 Diadinoxanthin; 5,6-Epoxy-7' ,8' -didehydro-5,6-dihydro-p,p-carotene-3,3'-diol

HO

PMR [672, 10] MS [76, 10] IR, Chern. [674, 10] Misc. [267, 173, 661]
 796 0. STRAUB

119 Antheraxanthin, "5,6-Epoxyzeaxanthin ", 3,3' -Oihydroxy-5,6-epoxy-/J-carotene;

5,6-Epoxy-5,6-dihydro-/J,/J-carotene-3,3' -diol
OH

HO

PMR [41] MS [76, 41] IR [723] ORO [41] Chern. [629] Synth. [391] Misc. [267, 574, 367]

120 Taraxanthin, Lutein epoxide, Xanthophyll epoxide, Isolutein [422, 658], Eloxanthin [398, 391, 285],
Tareoxanthin (cis isomer) [698, 665, 661]; (3S,5R,6S,3'S,6'R)-5,6-Epoxy-5,6-dihydro-P,e-carotene-3,3'-diol

MS [717] IR [717] ORO [41] Chern. [322, 192,391, 448] Synth. [391]
Misc. [233, 104, 574, 171, 170, 63, 662, 405, 448]

121 Taraxien, Taraxanthin dipalmitate; (3S,5R,6S,3'S,6' R)-5,6-Epoxy-5,6-dihydro-/J,e-carotene-3,3' -diol

dipalmitate

Misc. [513, 63]

122 Neoxanthin, F oliaxanthin, Trollixanthin (=stereoisomer [175]; see also 123);

(3S,5 R,6R,3' S,5' R,6' S)-5' ,6'-Epoxy-6, 7-didehydro-5,6,5',6' -tetrahydro-/J,/J-carotene-3,5,3' -trio I

HO•••••.g~OH OH

••••• """•~-······""<:::::
. ""<::::: ""<::::: ""<::::: ""<::::: ""<::::: ""<::::: ""<:::::

PMR [104, 725, 724, 535, 101, 164] MS [76, 104, 725, 724, 535, 101, 29] IR [104, 725, 724, 535, 101, 164]
ORO [104, 41, 535] Chern. [577, 175, 104, 535, 101, 629]
Misc. [162, 621, 726, 723, 61, 628, 136, 232, q1, 91, 90, 658]
 XII. Lists of Natural Carotenoids 797

123 Trollixanthin (stereoisomer of Neoxanthin ?, see 122); 5,6-Epoxy-5,6-dihydro-IJ,e-carotene-3,3',6'-triol

HO

Chern. [175, 521,404, 401] Misc. [755, 62]

124 Vaucheriaxanthin; 5' ,6'-Epoxy-6, 7-didehydro-5,6,5',6'-tetrahydro-/1,/1-carotene-3,5,19,3'-tetrol

OH
H 2 0H

;xy-~

HO~OH
MS [669] IR [674, 669] Chern. [674, 576] Misc. [201, 429]
(see [674]: 4'-0H instead of 19-0H!)

125 Mutatochrome, Citroxanthin, Flavacin (stereoisomer? [280, 711]), "5,8-Epoxy-tJ-carotene";

5,8-Epoxy-5,8-dihydro-p,p-carotene C40 H 56 0

PMR [36] MS [737, 76, 29] IR [51, 50] Synth. [180, 403, 392] Misc. [280, 390]

126 Flavochrome, "5,8-Epoxy-cx-carotene "; 5,8-Epoxy-5,8-dihydro-tJ,e-carotene

Synth., Misc. [393]

127 Cryptoflavin, Cryptoxanthin 5,8-epoxide; 5,8-Epoxy-5,8-dihydro-tJ,tJ-caroten-3-ol

natural?

or perhaps 5',8'-epoxide?
HO

IR [51] Synth. [399] Misc. [133, 131, 196]

 798 0. STRAUB

128 Rubichrome, "5,8-Epoxy-3-hydroxy-y-carotene"; 5,8-Epoxy-5,8-dihydro-P,l/t-caroten-3-ol

natural?

Synth., Misc. [ 402]

129 Mutatoxanthin, Zeaxanthin furanoxide; 5,8-Epoxy-5,8-dihydro-p,p-carotene-3,3'-diol

OH

IR [51] Synth. [391] Misc. [125, 124, 123, 116, 388]

130 Flavoxanthin, "Carcinoxanthin" [224, 487, 486], Chrysanthemaxanthin ( = epimer), "5,8-Epoxylutein ",
"5,8-Epoxy-3,3'-dihydroxy-IX-carotene"; 5,8-Epoxy-5,8-dihydro-p,e-carotene-3,3'-diol

HO

Flavoxanthin: MS [29] ORD [41] Chern. [381, 455] Synth. [391] Misc. [658]
Chrysanthemaxanthin: PMR [753] ORD [41] Chern. [387] Synth. [391] Misc. [383]

131 Neochrome, Foliachrome, Trollichrome (stereoisomer [62]; see also 132);

5',8' -Epoxy-6, 7-didehydro-5,6,5',8' -tetrahydro-p,p-carotene-3,5,3'-trio!
OH

(><Y•¥
HO~OH
MS [76, 104, 29] IR [104] Synth. [104] Misc. [535, 668, 101, 126, 125]

132 Trollichrome (see 131); 5,8-Epoxy-5,8-dihydro-p,e-carotene-3,3',6'-triol

OH
natural?
?

HO

Chern. [521, 401] Misc. [62, 404]

 XII. Lists of Natural Carotenoids 799

133 P-Carotene 5,6,5' ,6'-diepoxide, "Diepoxy-P-carotene"; 5,6,5',6'-Diepoxy-5,6,5',6' -tetrahydro-P,P-carotene

PMR [36] MS [29] Chern. [674, 392] Synth. [180, 722, 392] Misc. [735]

134 Cryptoxanthin diepoxide; 5,6,5',6'-Diepoxy-5,6,5',6' -tetrahydro-p,p-caroten-3-ol

HO
Chern., Synth. [399] Occur. [133, 131, 129]

135 Violaxanthin, Zeaxanthin diepoxide, Violeoxanthin=9-cis-Violaxanthin?;

(3S,5R,6S,3' S,5' R,6' S)-5,6,5',6'-Diepoxy-5,6,5',6' -tetrahydro-P,P-carotene-3,3'-diol

Violaxanthin: PMR [41] MS [76, 41] IR [493] ORD [41] Chern. [674, 102,388, 370a, 360,445, 353]
Synth. [41, 391] Misc. [104, 162, 658]
Violeoxanthin: IR, ORD, Chern. [698] Misc. [665, 661]

136 Luteochrorne; 5,6,5',8'-Diepoxy-5,6,5',8' -tetrahydro-p,p-carotene

natural?

Synth., Misc. [392]

137 Cryptoxanthin 5,6,5',8'-diepoxide; 5,6,5',8'-Diepoxy-5,6,5',8'-tetrahydro-p,p-caroten-3-ol

Chern. [674] Misc. [133, 131, 129]

 800 0. STRAUB

138 Luteoxanthin, 3,3'-Dihydroxyluteochrome; 5,6,5',8'-Diepoxy-5,6,5',8' -tetrahydro-P,P-carotene-3,3'-diol

Synth. [322] Misc. [668, 130, 128, 123, 116]

139 A urochrome, "~-Carotene" [68]; 5,8,5',8' -Diepoxy-5,8,5',8' -tetrahydro-p,p-carotene

PMR [36] MS [76, 29] Synth. [180, 392]

140 Cryptochrome, Cryptoxanthin 5,8,5',8"-diepoxide, "3-Hydroxy-5,8,5',8'-diepoxy-P-carotene";

5,8,5',8'-Diepoxy-5,8,5',8' -tetrahydro-p,p-caroten-3-ol

Synth. [399] Occur. [131]

141 A uroxanthin, "5,8,5',8'-Diepoxyzeaxanthin ", "3,3' -Dihydroxy-5,8,5',8' -diepoxy-P-carotene";

5,8,5' ,8'-Diepoxy-5,8,5',8' -tetrahydro-P,P-carotene-3,3'-diol

HO

MS [29] ORD [41] Synth. [391] Misc. [668, 783, 388, 385, 382]

f) Aldehydes

142 Torularhodinaldehyde, "3',4'-Dehydro-17' -oxo-y-carotene"; 3',4'- Didehydro-P,I/f-caroten-16'-al

CHO

Synth. [639, 618] Misc. [58, 653, 56, 768]

 XII. Lists of Natural Carotenoids 801

143 13-cis-Lycopen-20-al, Lycopenal, Anhydrowarrningone; 13-cis-t/f,t/f-Caroten-20-al

CHO

MS [213] Chern. [7] Misc. [633, 505]

144 13-cis-Rhodopin-20-al, Rhodopinal, Warmingone; 13-cis-1-Hydroxy-l ,2-dihydro-t/f,t/f-caroten-20-al

PMR [7] MS [213, 7] IR, Chern. [7] Misc. [601, 505]

145 Rhodopinal D-glucoside; 13-cis-1-(P-o-Giucopyranosyloxy)-1,2-dihydro-t/f,t/f-caroten-20-al

CHO

MS, Misc. [634]

Carotenoids 51
 802 0. STRAUB

145 a 1-Methoxy-1,2-dihydro-3,4-didehydrolycopen-20-al;
13-cis-1-Methoxy-3,4-didehydro-1,2-dihydro-1/J,I/f-caroten-20-al

CHO

MS, Misc. [634]

146 Methoxylycopenal; 13-cis-1-Methoxy-1,2-dihydro-1/f,1/f-caroten-20-al

CHO

MS [213] Misc. [601]

147 3,4,3',4'-Tetrahydrospirilloxanthin-20-al, P 500 [178, 5];

13-cis-1,1'-Dimethoxy-1,2,1',2'-tetrahydro-1/f,I/J-caroten-20-al

CHO

MS [213, 7] IR, Chern. [7] Misc. [178, 5]

 XII. Lists of Natural Carotenoids 803

g) Monoketones

148 Echinenone, Aphanin, Myxoxanthin; p,p-Caroten-4-one

PMR [737, 753] MS [76, 30, 188, 187] IR [140, 278, 265, 13, 748] Chern. [218, 249, 234, 389, 274]
Synth. [689, 13, 748] Misc. [653, 715, 251, 595, 235, 481, 479]

149 Phoenicopterone, 4-Keto-a-carotene; p,e-Caroten-4-one

Synth.[78, 184] ()ccur.[209,207,206]

150 4-Ketotorulene; 3',4'-Didehydro-P,t/1-caroten-4-one

Chern. [613a] Misc. [431, 3]

151 4-Keto-y-carotene; p,tj!-Caroten-4-one

PMR [484] IR [703, 484, 276] Chern. [276] Synth. [484, 78]

152 3-C>xoechinenone, Dehydrohydroxyechinenone, "Euglenanone" (see 193), 3,4-Diketo-P-carotene,

3-Hydroxy-4-oxo-2,3-dehydro-P-carotene;
3-Hydroxy-2,3-didehydro-P.P-caroten-4-one +t p,p-Carotene-3,4-dione c40 H 52 () 2
artifact?

HO
0
MS [30] Chern. [144, 163] Synth. [750, 485] Misc. [432, 169, 168,293, 167]
 804 0. STRAUB

1S3 Hydroxyechinenone, 3-Hydroxy-4-keto-P-carotene; 3-Hydroxy-p,p-caroten-4-one

HO

PMR [485] MS [30, 485] IR [485] Chern. [53] Synth. [485] Misc. [225, 169, 167, 438]

154 Asteroidenone ?, 3'-Hydroxyechinenone, 3'-Hydroxy-4-oxo-P-carotene; 3'-Hydroxy-p,p-caroten-4-one

OH

Chern. [277, 228] Misc. [225, 229, 568, 565, see also 80]

ISS 4' -Hydroxyechinenone, 4-Hydroxy-4' -keto-P-carotene; 4'-Hydroxy-p,p-caroten-4-one

H

IR [675, 265, 511, 51, 595] Synth. [511, 595] Misc. [432, 483,440, see also 80]

156 3,4-Diketo-IX-carotene; 3-Hydroxy-2,3-didehydro-p,e-caroten-4-one ;:::!: p,e-Carotene-3,4-dione

C4oHs202

[731]

1S6a Philosarniaxanthin [89c], 3'-Dehydrolutein, 3-Hydroxy-3'-keto-IX-carotene;

3-Hydroxy-p,e-caroten-3'-one
~0

HO
IR, Chern. [513a] Occur. [268a, 89a]
 XII. Lists of Natural Carotenoids 805

157 Cryptocapsin; (3'S,5' R)-3' -Hydroxy-p,K-caroten-6' -one

~ ~ ~ ~ ~ ~ ~ ~
-4~0

PMR [753, 100] IR [100] ORD [41] Chern., Synth. [100] Misc. [162, 36, 94, 91]

158 Deoxyflexixanthin, 4-Keto-1 ',2' -dihydro-1 '-hydroxytorulene;

1'-Hydroxy-3',4' -didehydro-1 ',2' -dihydro-P,I{t-caroten-4-one

OH

MS, IR, Chern. [3]

159 Myxobactone, 1',2' -Dihydro-1' -hydroxy-4-ketotorulene glucoside;

1'-G lucosyloxy-3',4'-didehydro-1 ',2' -dihydro-p,ift-caroten-4-one

MS, IR [435] Occurrence as fatty acid esters [434, 435]

160 1',2'-Dihydro-1 '-hydroxy-4-keto-y-carotene; 1'-Hydroxy-l ',2' -dihydro-P,I{t-caroten-4-one

~ ~ ~ ~ ~ ~ ~ ~ ~ ~
I
OH

PMR, IR [484] Chern. [276] Synth. [484]

161 1',2'-Dihydro-2' -hydroxy-3',4' -dehydro-4-keto-y-carotene;

2'-Hydroxy-3',4' -didehydro-1 ',2' -dihydro-P,ift-caroten-4-one C4oHs402
OH

~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~
I

Chern. [703]
 806 0. STRAUB

162 2' -Dehydroplectaniaxanthin, 1'-Hydroxy-2-keto-1 ',2' -dihydrotorulene;

1' -Hydroxy-3',4' -didehydro-1 ',2' -dihydro-{3,1/J-caroten-2' -one
0

OH

PMR [24] MS [24, 188, 732] IR [21] Chern. [21, 19] Synth. [641]

163 Rubixanthone, 4' -Ketorubixanthin, 4' -Keto-3-hydroxy-y-carotene; 3-Hydroxy-{3,1/J-caroten-4' -one

HO

IR, Chern., Misc. [558]

164 OH -Okenone, Dernethylated okenone; 1'-Hydroxy-l ',2' -dihydro-x,l/J-caroten-4' -one

Chern., Synth., Misc. [601]

165 4-Keto-3' -hydroxylycopene; 3'-Hydroxy-1/J,l/J-caroten-4-one

OH
0

Misc. [559]

165a {3- Doradecin, Dehydroadonixanthin, 3' -Hydroxy-3,4-diketo-{3-carotene;

3,3'-Dihydroxy-2,3-didehydro-{3,{3-caroten-4-one <=! 3'-Hydroxy-{3,{3-carotene-3,4-dione

OH artifact?

HO
0

Misc. [416, 432, 167]

 XII. Lists of Natural Carotenoids 807

166 Pectenolone, Glycyrnerin?, Hydroxyasteroidenone? (see 167);

3,3'-Dihydroxy-7',8' -didehydro-fl,fl-caroten-4-one
YYOH

'*~
HO

PMR, MS, IR, Chern. [79]

167 'Adonixanthin, Hydroxyasteroidenone? [416, 79, 228, 565], 3,3'-Dihydroxyechinenone,

fl-Doradexanthin [418], 3,3'-Dihydroxy-4-keto-fl-carotene; 3,3'-Dihydroxy-fl,fl-caroten-4-one

Chern. [167, 228] Misc. [432, 168, 427, 227, 568]

168 4'-Hydroxy-3-oxoechinenone, 4-Hydroxy-3',4'-dioxo-fl-carotene;

3,4'-Dihydroxy-2,3-didehydro-fl,fl-caroten-4-one ;:::t 4' -Hydroxy-fl,fl-carotene-3,4-dione

natural?

HO

MS [30] Misc. [ 432]

168a IX-Doradecin, "3-Hydroxy-3',4'-diketo-IX-carotene";

3,3'-Dihydroxy-2,3-didehydro-fl,e-caroten-4-one ;:::t 3'-Hydroxy-fl,e-carotene-3,4-dione

artifact?
OH

HO

PMR, MS, IR, Chern. [418] Misc. [416]

 808 0. STRAUB

169 ex-Doradexanthin, "3'-Dihydro-ex-doradecin ", 4-Ketolutein, "3,3'-Dihydroxy-4' -keto-ex-carotene";

3,3'-Dihydroxy-p,e-caroten-4-one
OH

HO
0

PMR, MS, IR [418] Misc. [417, 416]

170 Capsanthin; (3R,3' S,5' R)-3,3' -Dihydroxy-{J,K-caroten-6' -one OH

--6-i i

PMR [200, 38, 34] MS [76, 30, 211] IR [38, 34, 186, 747] ORD [41] Chern. [552, 200, 696, 38, 199,
198,34,186; 93,92, 181,798,797,795,793, 790] Synth.[752,109] Misc.[162, 751,653, 789]

171 Flexixanthin; 3,1 '-Dihydroxy-3' ,4' -didehydro-1 ',2' -dihydro-P,t/J-caroten-4-one

OH
HO

PMR, MS, IR, Chern. [3]

171a 1',2'-Dihydro-1 ',2' -dihydroxy-4-ketotorulene;

1',2' -Dihydroxy-3',4' -didehydro-1',2' -dihydro-P,t/J-caroten-4-one
OH

OH

Occurrence as fatty acid esters at 2' [613 a]

172 4-Ketophleixanthophyll;
1'-(P-o-Glucopyranosyloxy)-2'-hydroxy-3' ,4' -didehydro-1 ',2' -dihydro-P,t/J-caroten-4-one
H

0
PMR, MS, Chern., Synth. [279]
 XII. Lists of Natural Carotenoids 809

173 Idoxanthin, 3,3',4'-Trihydroxy-4-keto-P-carotene; 3,3',4'-Trihydroxy-p,p-caroten-4-one

OH
OH

HO
0

IR, Synth. [53] Occur. [275]

174 Siphonaxanthin, 3,3',19-Trihydroxy-7,8-dihydro-8-oxo-et-carotene;

3,19,3'-Trihydroxy-7,8-dihydro-P,E-caroten-8-one

H 2 0H

0
HO
PMR, MS [673a, 746] IR [433] Chern. [746, 433, 428] Misc. [670, 664]

175 Siphonein; 3,19,3'-Trihydroxy-7,8-dihydro-p,e-caroten-8-one 19-laurate (?)

CH,OCO(CH 2 ) 10 CH, OH

0
HO
PMR [673a] MS [673a, 746] Chern. [746, 433, 428] Misc. [664]

176 4-Ketornyxol2'-(rnethylpentoside);
3, 1'-Dihydroxy-2' -( 5-C-rnethylpentosyloxy)-3',4' -didehydro-1',2' -dihydro-P,!/J-caroten-4-one

OH
HO
0
MS, Chern. [212].

177 2'-Dihydrophillipsiaxanthin;
1,1 ',2'-Trihyaroxy-3,4,3',4' -tetradehydro-1,2, 1',2' -tetrahydro-1/J ,!/J-caroten-2-one

?
I OH
0
Misc. [19]
 810 0. STRAUB

178 Isofucoxanthinol (artifact?), Pentaxanthin? [217, 481,479,475, 474];

3,5,3',5'-Tetrahydroxy-6', 7' -didehydro-5,8,5',6'-tetrahydro-P,fl-caroten-8-one

HOJ;)OH

ec:P'
-:::-

HO

PMR, [62] MS [76, 62] IR [62] Chern. [62, 61] Misc. [41]

179 lsofucoxanthin; 3,5,3',5'-Tetrahydroxy-6', 7'-didehydro-5,8,5',6' -tetrahydro-p,p-caroten-8-one 3'-acetate

HO

PMR, MS, IR [62, 332, 331] Chern. [61, 331] Misc. [41, 551]

180 Thiothece-478; 1'-Methoxy-1 ',2'-dihydro-q>,I/J-caroten-4'-one

182 Spheroidenone, "Pigment R" [246, 243, 571], "1-Methoxy-2-keto-7',8'-dihydro-3,4-dehydrolycopene";

1-Methoxy-3,4-didehydro-1,2, 7',8'-tetrahydro-1/1,1/J-caroten-2-one

PMR [39, 538, 158] MS [188, 39] IR [39,.538, 502, 501, 554, 250] Chern. [39, 502] Synth. [538, 39]
 XII. Lists of Natural Carotenoids 811

183 3, I ',2'-Trirnethoxy-3',4' -didehydro-1 ',2'-dihydro-fl,Y,-caroten-4-one

?
CH 3 0
CH 3 0

IR, Chern. [31]

184 2-Keto-OH-spirilloxanthin, "2-Ketorhodovibrin" [19, 513, 177, 504];

I'-Hydroxy-1-rnethoxy-3,4,3',4' -tetradehydro-1,2,1 ',2' -tetrahydro-Y,,Y,-caroten-2-one

OH

Misc. [19]

185 OH-Spheroidenone, "Hydroxyspheroidenone", "OH-R";

1'-Hydroxy-1-rnethoxy-3,4-did.ehydro-1,2,1 ',2', 7',8' -hexahydro-Y,,Y,-caroten-2-one

PMR [321] IR [502] Chern. [321, 502] Misc. [177, 500,499,250,246, 243]

186 2'-Hydroxy-3,1 '-dirnethoxy-3',4'-didehydro-1 ',2'-dihydro-fl,Y,-caroten-4-one

OH

IR, Chern. [31]

187 Heteroxanthin (see 94); 5,6-Epoxy-3,3'-dihydroxy-5,6,7,8-tetrahydro-fl,p-caroten-8-one

OH

?
HO
Chern., Misc. [578]
 812 0. STRAUB

188 Capsanthin rnonoepoxide; 5,6-Epoxy-3,3' -dihydroxy-5,6-dihydro-p,K-caroten-6' -one

OH

HO

Chern., Synth. [396] Misc. [560, 130]

189 Fucoxanthinol;
(3S ,5 R,6S,3'S,5 'R,6' R)- 5,6-Epoxy-3,3',5'-trihydroxy -6', 7'-didehydro-5,6, 7,8,5 ',6'-hexah ydro-p,p-caroten -8-one

'<::::,.····~.~
0
H O + J... ~'OH
PMR [62] MS [76, 62] IR, Chern. [62] Misc. [41, 104, 217]

190 Fucoxanthin; (3S,5R,6S,3'S,5' R,6' R)-5,6-Epoxy-3,3',5' -trihydroxy-6', 7' -didehydro-5,6, 7,8,5',6'-

hexahydro-p,p-caroten-8-one 3' -acetate C 42 H 58 0 6

'<::::,.·······~.~

HO
Ho~·····ococH,
PMR [737, 62, 61, 332, 330, 329] MS [76, 62, 60, 61] IR [62, 751,61, 332,329,327,489, 716] ORD [16]
X-ray [162] Chern. [579, 62, 61, 60, 330, 329, 328, 327, 489] Misc. [41, 104, 621]

191 Capsochrorne; 5,8-Epoxy-3,3' -dihydroxy-5,8-dihydro-P,K-caroten-6' -one

OH

natural?

HO
Synth. [396] Misc. [130]

192 Eschscholtzxanthone; 3'-Hydroxy-4' ,5'-didehydro-4,5' -retro-p,p-caroten-3-one

OH

IR [561, 52] Chern., Synth. [52] Misc. [563]

 XII. Lists of Natural Carotenoids 813

h) Dike tones

193 Canthaxanthin, Aphanicin [278, 713, 711], Chlorellaxanthin [139, 161],

Euglenanone? [169, 752, 750, 151, 167, 438], 4,4'-Diketo-/J-carotene; p,p-Carotene-4,4'-dione

PMR [643, 751, 750, 36] MS [76, 30, 188, 187] IR [675, 108, 278, 51, 13, 748, 219] X-ray [40]
Chern. [625] Synth. [689, 13, 815, 183, 748, 595] Misc. [653, 594, 270]

194 Diketopirardixanthin, 5,6,5',6'-Tetrahydrocanthaxanthin; 5,6,5',6'-Tetrahydro-/J,/J-carotene-4,4' -dione

Misc. [440]

194a 1',2' -Dihydro-4,2' -diketotorulene; 3',4'-Didehydro-1',2' -dihydro-/J,I/J-carotene-4,2' -dione

Chern., Misc. [613a]

194 b 4,4' -Diketo-y-carotene; p,I/J-Carotene-4,4' -dione

0

Chern., Misc. [613a]

 814 O.STRAUB

194c 4,4'-Diketolycopene; t{l,t{I-Carotene-4,4'-dione

Chern., Misc. [613a]

195 Phoeniconone, Dehydroadonirubin, 3-0xocanthaxanthin, 3,4,4'-Triketo-P-carotene;

3-Hydroxy-2,3-didehydro-P,P-carotene-4,4' -dione ~ p,p-Carotene-3,4,4' -trione

artifact?

MS [30] Chern. [206] Synth. [751] Misc. [432, 145, 143, 169, 168, 167]

196 Adonirubin, Phoenicoxanthin, 3-Hydroxycanthaxanthin, 3-Hydroxy-4,4' -diketo-P-carotene;

3-Hydroxy-p,p-carotene-4,4' -dione

Chern. [427, 430, 168, 206, 167] Misc. [143, 169]

197 Capsanthinone, Capsanthone, Ketocapsanthin; (3R,5'R)-3-Hydroxy-P,K-carotene-3',6'-dione

A . :

PMR [38, 34] MS [76, 30] IR [34, 186] ORD [41] Chern., Synth. [186]
Misc. [560, 160, see also 798]
 XII. Lists of Natural Carotenoids 815

198 Astacene, "Euglenarhodon" [714, 469, 709], "Salmon acid" [ 480], 3,4,3',4'-Tetraketo-P-carotene;
3,3'-Dihydroxy-2,3,2' ,3' -tetraaehydro-p,p-carotene-4,4'-dione .,z p,p-Carotene-3,4,3' ,4'-tetrone

OH

artifact?

HO
0
PMR [3, 157] MS [30, 282] IR [3, 51, 157, 489] Chern. [365, 363] Synth. [157]
Misc. [138, 141, 467, 458]

199 Astacein, Astacene dipalrnitate;

3,3'-Dihydroxy-2,3,2',3' -tetradehydro-p,p-carotene-4,4' -dione 3,3'-dipalrnitate

OCO(CH 2 ) 1.CH3

CH3(CHz)14COO

Chern. [365] Synth. [ 469]

200 7,8,7',8'-Tetradehydroastaxanthin (in asterinic acid; see also 202);

3,3' -Dihydroxy-7,8, 7',8' -tetradehydro-p,p-carotene-4,4' -dione
~OH
*~
(<Y*
HOY
0

PMR [214] MS, IR, Chern. [214, 652] Misc. [195]

201 3-Hydroxy-3',4,4' -triketo-{J-carotene;

3,3'-Dihydroxy-2,3-didehydro-p,p-carotene-4,4' -dione .,z 3'-Hydroxy-p,p-carotene-3,4,4' -trione
0
OH
artifact?

HO
0
Misc. [432]
 816 0. STRAUB

202 7,8-Didehydroastaxanthin (in asterinic acid, see 200);

3,3'-Dihydroxy-7,8-didehydro-Jl,Jl-carotene-4,4'-dione
OH

(<y~

HOY
0
Chern. [214, 652] Misc. [195]

203 Astaxanthin, 3,3'-Dihydroxycanthaxanthin, 3,3'-Dihydroxy-4,4' -diketo-Jl-carotene;

3,3'-Dihydroxy-Jl,Jl-carotene-4,4'-dione

OH

HO
0
PMR [79, 485, 686] MS [30, 188,79, 485] IR [79, 485, 3, 489] ORD [41. 72] CD [73] Chern. [467]
Synth. [ 485, 686]

204 Guaraxanthin, 7,8-Dihydroastaxanthin; 3,3'-Dihydroxy-7,8-dihydro-Jl,Jl-carotene-4,4'-dione

0
OH
?

n
HO
0

Chern., Misc. [207]

105 Capoxubm; (3S~R,3'S,5'R)- 3,3'-D;hy-y-<,<-carot~o-6,6'-di~

-7
o '"""" '"""" '"""" '-":::: '-":::: '-":::: '-":::: '-":::: '-":::: '<:::o

OH
PMR [200, 109, 38, 34] MS [76, 30, 211] IR [109, 38, 747] ORD [41]
Chern. [200,696,38,98, 186,92,93, 797] Sxnth. [109] Misc. [162, 199,198,181, 795]
 XII. Lists of Natural Carotenoids 817

206 7,8, 7',8'-Tetrahydrocapsorubin; 3,3'-Dihydroxy-7,8, 7',8' -tetrahydro-K,K-carotene-6,6'-dione

OH

Chern., Misc. [130]

207 Phillipsiaxanthin, 1,1 '-Dihydroxy-2,2' -diketo-1,1 ',2,2' -tetrahydro-3,3',4,4' -tetradehydrolycopene,

"2,2'-Diketobacterioruberin" [640];
1,1 '-Dihydroxy-3,4,3',4' -tetradehydro-1,2,1 ',2' -tetrahydro-ljl,ljl-carotene-2,2'-dione C40H szO 4

OH

MS [640] IR, Chern. [19] Synth. [640]

208 2,2'-Diketospirilloxanthin, P 518 [513, 506], "2-Ketospirilloxanthin" [501, 321];

1,1 '-Dirnethoxy-3,4,3',4' -tetradehydro-1 ,2,1 ',2' -tetrahydro-1/1,ljl-carotene-2,2'-dione

PMR [538, 321] MS [188, 640] IR [538, 501] Chern. [321, 501] Synth. [538, 640] Misc. [177]

209 Rhodoxanthin, 3,3'-Diketoretrodehydro-P-carotene, "3,3'-Diketoretro-P-carotene ";

4',5'-Didehydro-4,5' -retro-P,P-carotene-3,3'-dione

PMR [545] MS [545, 188, 187] IR [689, 545,561,51, 489] Chern. [366, 459]
Synth. [689,545,323, 185]

Caroten01ds 52
 818 0. STRAUB

i) Polyketones

210 Capsorubone, Capsorubindione, Ketocapsorubin; ~e,~e-Carotene-3,6,3',6'-tetrone

MS [30] ORD [41] Chern. [97] Synth. [186] Occur. [160]

j) Acids

211 Torularhodin, "Torulenecarboxylic( 16') acid", "16' -Carboxyl-3',4'-dehydro-y-carotene ",

"Lusomycin "? [ 580]; 3',4'-Didehydro-P,I/J-caroten-16'-oic acid

Chern. [395, 384] Synth. [317] Misc. [653, 400,482, 471]

212 Torularhodin methyl ester; Methyl 3',4'-didehydro-P,I/J-caroten-16'-oate

PMR [36] MS see also [737] Chern. [20] Synth. [317]

k) Seco compounds

213 Semi-P-carotenone; 5,6-Seco-p,p-carotene-5,6-dione

PMR [784] MS [211, 784] IR [784] Ohern. [784, 466, 461]

 XII. Lists of Natural Carotenoids 819

214 Serni-a-carotenone; (6'R)-5,6-Seco-p,e-carotene-5,6-dione

PMR, MS, IR [786] CD [69] Chern. [786, 364, 362] Synth. [786]

215 Triphasiaxanthin, 3-Hydroxyserni-P-carotenone; 3'-Hydroxy-5,6-seco-P,P-carotene-5,6-dione

C4oHs603
OH

PMR, MS, IR, Chern. [785]

216 P-Carotenone; 5,6,5',6'-Diseco-p,p-carotene-5,6,5',6' -tetrone

PMR [784] MS [737, 211, 784] IR [784] Chern. [466, 465, 461, 451] Synth. [784]

3. Homo-carotenoids

216a 7',8',11 ',12' -Dehydrononaprenoxanthin, P 452 [757];

2-(4-Hydroxy-3-rnethyl-2-butenyl)-e,I/J-carotene

HOH 2 C ~

Misc. [258]

217 11 ',12'-Dehydrononaprenoxanthin, Dehydrogenans-P 422 [758, 757];

2-(4- Hydroxy~ 3-rnethy1-2-buteny1)-7',8' -dihydro-e,I/J -carotene

HOH 2C ~

MS [758]
 820 0. STRAUB

218 Nonaprenoxanthin, Dehydrogenans-P 373 [758, 757];

2-(4-Hydroxy-3-rnethyl-2-butenyl)-7',8', 11 ', 12'-tetrahydro-e,t/1-carotene

HOH2C "=::::

MS [758]

219 2-Isopentenyl-3,4-dehydrorhodopin; 2-(3-Methyl-2-butenyl)-3,4-didehydro-1,2-dihydro-t/l,t/1-caroten-1-ol

PMR [583, 581] MS [583] IR [583, 581]

220 2-(Dihydroxyisopentenyl)-2'-isopentenyl-P-carotene, C.p. 450;

2-(4-Hydroxy-3-hydroxyrnethyl-2-butenyl)-2'-(3-rnethyl-2-butenyi)-P.P-carotene

HOH,C "=::::

PMR, MS, IR, Chern. [583]

221 1'-Hydroxy-1',2' -dihydro-2-isopentenyl-2' -(hydroxyisopentenyl)torulene, C. p. 473;

2' -(4-Hydroxy-3-rnethyl-2-butenyl)-2-(3-rnethyl-2-butenyl)-3',4' -didehydro-1 ',2' -dihyaro-P,t/1-caroten-1 '-ol

CH 20H

"=:::: "=:::: "=:::: "=:::: "=:::: "=:::: "=:::: "=:::: "=:::: "=::::
OH

PMR, MS, IR, Chern. [583] Misc. [518]

222 Sarcinene; 2,2'-Bis(3-rnethyl-2-butenyl)-e,e-carotene

CsoHn

"=:::: "=:::: "=:::: "=:::: "=:::: "=:::: "=:::: "=:::: ?

Misc. [517, 704, 86, 85]

 XII. Lists of Natural Carotenoids 821

223 Deshydroxydecaprenoxanthin;
2-(4-Hydroxy-3-rnethyl-2-butenyl)-2'-(3-rnethyl-2-butenyl)-E,E-carotene

HOH 2C "":::

MS [758] Misc. [757]

224 Sarcinaxanthin; 2-(4-H ydroxy-3-rnethyl-2-butenyl)-2' -(3-rnethyl-2-butenyl)-E,E-caroten-18-ol

HOH 2C "'::::
?

PMR, MS, IR [25] Chern. [25, 518] Misc. [517, 516, 700]

225 Sarcinaxanthin rnono-o-glucoside;

2'-[ 4-(o-Glucopyranosyloxy)-3-rnethyl-2-butenyl]-2-(3-rnethyl-2-butenyl)-E,E-caroten-18-ol

MS, Chern. [582]

226 Decaprenoxanthin, Dehydrogenans-P 439; 2,2' -Bis(4-hydroxy-3-rnethyl-2-butenyl)-E,E-carotene

""" CH,OH
HOH,C """

PMR [642, 517, 515] MS [517, 516, 704] IR [517, 704] ORD [41] Chern. [517] Misc. [758]

227 Corynexanthin, Decaprenoxanthin rnonoglucoside;

2-[4-(fJ-o-Glucopyranosyloxy)-3-rnethyl-2-butenyl]-2' -(4-hydroxy-3-rnethyl-2-butenyl)-E,E-carotene

""" CH 2 0H

MS [759] Misc. [291]

 822 0. STRAUB

228 Bisanhydrobacterioruberin;
2,2'-Bis(3-rnethy1-2-butenyl)-3,4,3',4'-tetradehydro-1,2,1 ',2'-tetrahydro-t/1,t/J-carotene-1,1 '-diol

PMR, MS, IR [583] Chern. [583, 420] Misc. [512, 655]

229 3,4,3',4'-Tetrahydrobisanhydrobacterioruberin;
2,2'-Bis(3-rnethyl-2-butenyl)-1,2,1 ',2' -tetrahydro-t/l,t/J-carotene-1,1'-diol

OH

MS, Chern. [583]

230 Monoanhydrobacterioruberin; 2-(3-Hydroxy-3-rnethylbutyl)-2'-(3-rnethyl-2-butenyl)-

3,4,3',4' -tetradehydro-1,2,1 ',2' -tetrahydro-t/1,t/J-carotene-1,1 '-diol

OH

OH
PMR, MS, Chern. [420] Misc. [518]

231 Bacterioruberin, IX-Bacterioruberin;

2,2'-Bis(3-hydroxy-3-rnethylbutyl)-3,4,3',4'-tetradehydro-1,2,1 ',2' -tetrahydro-t/1,t/J-carotene-1,1 '-diol

MS [420, 419] IR [500, 496], Chern. [420, 419, 500, 496] Misc. [635, 497,42, 482, 597]
 XII. Lists of Natural Carotenoids 823

4. Apo-carotenoids

232 P-Apo-2' -carotenol; 3',4'-Didehydro-2'-apo-p-caroten-2'-ol

Synth. [618] Misc. [58]

233 P-Apo-2'-carotenal; 3',4'-Didehydro-2' -apo-P-caroten-2'-al

MS [188] Synth. [639, 618] Misc. [768]

234 Neurosporaxanthin; 4'-Apo-P-caroten-4'-oic acid

MS [188] IR, Chern. [1] Synth. [317] Misc. [788, 269]

235 Neurosporaxanthin methyl ester; Methyl4'-apo-P-caroten-4'-oate

Chern. [203] Synth. [317]

236 Tangeraxanthin; 3-Hydroxy-4,5'-retro-5' -apo-P-caroten-5'-one

0
~ ?

HO
Chern. [132]

237 Citranaxanthin; 5',6'-Dihydro-5' -apo-18'-nor-P-caroten-6'-one

PMR [778] MS [211] IR, Chern., Synth. [778] Misc. [782]

 824 0. STRAUB

238 Reticulataxanthin, 3-Hydroxycitranaxanthin;

(3R)-3-Hydroxy-5',6' -di~ydro-5' -apo-18' -nor-{J-caroten-6'-one

PMR [779] MS [211] IR [779] ORD [41] Chern. [779, 132] Misc. [162, 129]

239 8'-Hydroxy-7',8' -dihydrocitranaxanthin;

8'-Hydroxy-5',6', 7',8' -tetrahydro-5'-apo-18'-nor-fl-caroten-6'-one

PMR, IR, Chern., Synth. [781] Misc. [782]

240 8'-Hydroxy-7' ,8' -dihydroreticulataxanthin;

3,8'-Dihydroxy-5',6', 7',8' -tetrahydro-5' -apo-18' -nor-fl-caroten-6'-one

?
HO
Chern. [782]

241 15-Hydroxy-7',8',9',10',11',12',13',14'-octahydro-6'-apo-fl-caroten-7'-one

?
0

Misc. [295]

242 Apo-6' -lycopenal, "Apo-2-lycopenal"; 6' -Apo-Y,-caroten-6' -al

Chern. [ 452] Misc. [768, 377]

243 Methyl apo-6'-lycopenoate; Methyl 6' -apo-Y,-caroten-6'-oate

PMR, MS, IR, Chern., Synth. [ 424]

 XII. Lists of Natural Carotenoids 825

244 Sintaxanthin; 7',8' -Dihydro-7' -apo-P-caroten-8'-one

PMR, IR, Chern., Synth. [780]

244a Triophaxanthin; 3-Hydroxy-7,8-didehydro-7',8'-dihydro-7' -apo-P-caroten-8'-one

li~
HO

MS, IR, Chern. [ 546]

245 3-Hydroxysintaxanthin; 3-Hydroxy-7',8'-dihydro-7' -apo-P-caroten-8'-one

HO

Chern. [782]

246 Paracentrone; (3S,5R,6R)-3,5-Dihydroxy-6, 7-didehydro-5,6, 7',8' -tetrahydro-7'-apo-P-caroten-8'-one

PMR, MS, IR [294, 217] Misc. [41]

247 Hopkinsiaxanthin; 3-Hydroxy-7,8-didehydro-7',8'-dihydro-7' -apo-P-carotene-4,8'-dione

(Y~

HOY
0

IR, MS, Chern. [564] Misc. [663]

 826 0. STRAUB

248 P-Apo-8'-carotenal, "{3-Apo-2-carotenal" [372], "{3-Carotenal" [372, 371]; 8'-Apo-{3-caroten-8'-al

CHO
""'-::: ""'-:::
I I

PMR [643, 318, 36] IR [737, 708] Chern. [371] Synth. [639, 613, 618] Misc. [172, 653, 782, 707, 768]

249 {3-Citraurin, "Apo-2-zeaxanthinal" [372, 371]; (3R)-3-Hydroxy-8'-apo-{3-caroten-8'-al

ORD [41] Chern. [371, 802, 803, 801] Misc. [162, 375, 374, 372]

250 {3-Apo-8'-carotenoic acid; 8'-Apo-{3-caroten-8'-oic acid

PMR [643, 36] Synth. [317] Misc. [653, 32, 706]

251 Methyl {3-apo-8'-carotenoate; Methyl8'-apo-fl-caroten-8'-oate

C0 2CH 3
""'-::: ""'-:::
I I

PMR [643] Synth. [317] Occur. [67]

252 see244a

253 "1-Hexosyl-1,2-dihydro-3,4-didehydroapo-8'-lycopenol";
1-Mannosyloxy-3,4-didehydro-1,2-dihydro-8'-apo-1/1-caroten-8'-ol

""'-::: ""'-::: ""'-::: ""'-::: ""'-::: ""'-::: ""'-::: ""'-::: ""'-::: ""'-::: CH 2 0H

Chern. [8]

254 Apo-8' -lycopenal, "Apo-3-lycopenal" [684, 377]; 8'-Apo-1/1-caroten-8'-al

PMR, IR [59] Synth. [684, 59] Misc. [768]

 XII. Lists of Natural Carotenoids 827

255 "Methyl 1-hexosyl-1,2-dihydro-3,4-didehydroapo-8'-lycopenoate";

Methyl 1-rnannosyloxy-3,4-didehydro-1,2-dihydro-8'-apo-lj!-caroten-8'-oate

C0 2 CH 3
"""" I """" I
PMR, MS, IR, Chern. [8] Misc. [518]

256 P-Apo-10' -carotenal, "P-Apo-3-carotenal" [316, 372]; 10'-Apo-P-caroten-10' -al

CHO
"""" I
PMR [643] IR [782, 708] Synth. [618] Occur. [782, 707, 768, 231]

257 3-Hydroxy-p-apo-10'-carotenal? [782], "Apo-3-zeaxanthinal" [372];

3-Hydroxy-1 0' -apo-P-caroten-1 0' -al

CHO
"""" I
HO
Misc. [782]

258 Azafrinaldehyde; 5,6-Dihydroxy-5,6-dihydro-1 0' -apo-P-caroten-1 0' -al

CHO
"""" I
Misc. [513, 768]

259 Apo-10' -violaxanthal; 5,6-Epoxy-3-hydroxy-5,6-dihydro-10' -apo-P-caroten-10'-al

CHO
?
"""" I
HO
Chern. [137]

260 P-Apo-10'-carotenoic acid; 10'-Apo-p-caroten-10'-oic acid

C0 2 H

PMR [36] Synth. [317] Misc. [763]

 828 O.STRAUB

261 Azafrin; 5,6-Dihydroxy-5,6-dihydro-10' -apo-fl-caroten-10'-oic acid

PMR [650, 36] MS [188, 547] IR [552, 750, 650] ORD [41] Chern. [552, 460]

262 fl-Apo-12' -carotenal, "fl-Apo-4-carotenal" [372]; 12'-Apo-fl-caroten-12'-a!

PMR [737, 643, 36] Chern. [372] Synth. [613, 618, 617] Misc. [231]

263 Apo-12' -violaxanthal; 5,6-Epoxy-3-hydroxy-5,6-dihydro-12'-apo-fl-caroten-12' -a!

?
HO
Chern. [135]

263a fl-Apo-12'-carotenoic acid; 12'-Apo-fl-caroten-12'-oic acid

[231a]

264 Carotenonaldehyde; 5,6-Dioxo-10'-apo-5,6-seco-fl-caroten-10'-a!

artifact?

Chern. [ 466, 465] Misc. [769]

265 Bixin, Isobixin (isomer) [259, 373, 342]; Methyl hydrogen 9'-cis-6,6'-diapocarotene-6,6'-dioate

C2sH3o04

C02 CH 3

PMR [37, 35] Chern. [373, 357, 356, 450, 4<\1] Synth. [603, 70, 314, 305, 9] Misc. [653, 407, 358, 355]
 XII. Lists of Natural Carotenoids 829

266 trans- Methylbixin; Dimethyl 6,6' -diapocarotene-6,6'-dioate

C02CH 3
CH 302C "':::: natural?

PMR [37, 36, 35] IR [314, 525] Synth. [751, 603, 70, 314, 303, 9]

266a Methyl-cis- (natural) bixin; Dimethyl9-cis-6,6'-diapocarotene-6,6'-dioate

natural?

CH302
PMR [589, 37, 35] IR, Synth. [589]

267 Crocetindial, Crocetindialdehyde; 8,8'-Diapocarotene-8,8'-dial

CHO
OHC "'::::

PMR [193, 639, 36] MS [193, 188, 639] IR [193] Synth. [603, 310] Misc. [653]

268 Crocetinsemialdehyde; 8'-0xo-8,8'-diapocaroten-8-oic acid

CHO
H0 2C "'::::

PMR, MS, IR [193]

269 Crocetin, "IX-Crocetin "; 8,8'-Diapocarotene-8,8'-dioic acid

C02H
H02

Chern. [356, 355, 341, 340] Misc. [754, 339]

270 Dimethylcrocetin, Crocetin dimethyl ester, "y-Crocetin "; Dimethyl 8,8'-diapocarotene-8,8'-dioate

C0 2CH3
CH 302C "'::::

PMR [36]_ IR [314, 525, 470] Chem. [446] Synth. [754, 71, 603, 314, 301] Misc. [259, 340, 339]

271 Crocin; Digentiobiosyl 8,8'-diapocarotene-8,8'-dioate

co2-c,2H2,o,o

Chern. [344, 340] Synth. [468, 446]

 830 O.STRAUB

5. N or-carotenoids

272 Actinioerythrin; 3,3'-Dihydroxy-2,2' -dinor-P.P-carotene-4,4'-dione 3,3'-diacylate

c3sH4so4
(as diol)

PMR [282] MS [282, 281] IR [282] Chern. [282, 281] Misc. [292, 197, 473]

273 Peridinin, Sulcatoxanthin [661, 273];

5',6'-Epoxy-3,5,3' -trihydroxy-6, 7-didehydro-5,6,5',6' -tetrahydro-1 0,11 ,20-trinor-P.P-caroten-19',11 '-olide
3-acetate

N·~
CH 3C 0 0 4 0 H

PMR, MS, Chern. [673] Misc. [670, 524, 325, 661]

B. Carotenoids of Unknown Structure

This is an alphabetical list of carotenoids the structures of which are quite

unknown or not known with sufficient certainty for inclusion in list A. Some
of these carotenoids have prematurely been given a systematic designation;
others are known only by a trivial name.
However, the list omits ill-defined carotenoid fractions reported in bewilder-
ing profusion in the literature and mostly characterized only by absorption
bands or vague chromatogram locations; also excluded, with very few ex-
ceptions, are carotenoid fractions designated as similar to a specific carotenoid,
as, for example, 'zeaxanthin-like', 'phytofluene-like', etc.
Although some new carotenoids may be found among the still unresolved
carotenoid fractions excluded from the list, a large proportion of these ill-
defined materials is likely to be made up of known structures or artifacts.
The literature references to each carotenoid are listed chronologically from
the latest to the oldest publications.

Anchovyxanthin (Zeaxanthin (67)?) [287]

Asperxanthin [787]
"Bacterial phytoene" see: Compound X
Bacteriopurpurin P(Rhodopin ?, Spirilloxanthin ?, Lycopene?) [500, 42, 654, 237]
P-Bacterioruberin [500, 597]
Bisdihydrocanthaxanthin [207]
C 50 -Carotenoid C 50 H 70 0 [704: MS, IR]
 XII. Lists of Natural Carotenoids 831
Canaryxanthophyll [741, 739, 65]
Capsolutein (P,K-Carotene-3,3' -diol ?) [ 130]
Capsolutein 5,6-epoxide (5,6-Epoxy-5,6-dihydro-P,K-carotene-3,3'-diol?) [130]
Capsolutein 5,8-epoxide (5,8-Epoxy-5,8-dihydro-P,K-carotene-3,3'-diol ?) [130]
Carangoxanthin (Isomer of tunaxanthin (78)?) [287]
'cCarotene [599]
.9-Carotene [149, 554, 271]
K-Carotene, K-Carotene (Mixture of '-carotene (26) and P-zeacarotene (9)?) [611, 165, 598,242,
237,624,623,761, 762,215]
K,K-Carotene-3,3' -diol (?), P 482 [130]
p,p-Carotene-3,4,3'-triol (?)(or p,p-Carotene-3,4,4'-triol (85)?) [146]
p,E-Carotene-2,3,2'-triol (?)(or p,E-Carotene-3,4,3'-triol?) [749, 268]
p,E-Carotene-3,4,3' -trio I(?) [268]
Chrysophlein [290, 520, 248, 247, 28, 730, 729, 591, 728]
Compound X, C 30 H 48 , "Bacterial phytoene" [692, 690,691: MS, IR]
C. p. 435 (C. p. =Corynebacterium poinsettiae) [583]
C.p. 450-monool [583]
C.p. 473-monool [583]
1/1-Cryptoflavin [129]
Dehydrogenans-,-carotene-mono-OH [757]
Dehydrogenans-,-carotene-di-OH [757]
3',4'-Didehydro-1',2' -dihydro-p,ljf-caroten-2'-ol (?) [28 a]
3' ,4'-Didehydro-7 ,8,1 ',2' -tetrahydro-P,I/I-carotene-3,4,1 ',2' -tetrol (?) [613 a]
Dihydrocrocetin [179]
1,2-Dihydro-3',4' -dehydrolycopene (?) (or 1,2,1 ',2'-Tetrahydro-3,4,3' ,4' -dehydrolycopene ?) [533]
1',2'-Dihydrotrihydroxy-4-ketotorulene [613 a]
Dihydroxy-,-carotene (see also (82)) [491, 757]
Dihydroxy-carotenoid with triple bond, C 40 H 54 0 2 [669]
x,y- Dihydroxy-3,4-dehydro-a-carotene [731]
Dihydroxyneurosporene [246]
Dihydroxypirardixanthin [ 440]
Dinoxanthin [326, 524, 678, 661]
Dodecahydro-P-carotene [333, 236]
"Eicosahydrolycopene" see: Tetrahydrophytoene
5',6'-Epoxy-5',6'-dihydro-p,ljf-caroten-4-one(?) (or x-Methoxy-3,4-didehydro-7',8'-dihydro-
p,ljl-caroten-2-one?) [703]
5,6-Epoxymonohydroxy-a-carotene see: Hydroxy-a-carotene 5,6-epoxide
Euglenanone (Canthaxanthin (193)?) [169, 752, 750, 151, 167, 438]
Eugorgiaenoic acid (Dihydrobixin ?) [210]
"Fenicotterina" see: Phoenicopterin
Galloxanthin [14, 676, 742]
Glycymerin (Pectenolone (166)?) [79, 481, 197, 472]
Haematoxanthin [712, 710]
Hydroxyactinioerythrin (artifact?) [282, 567]
x-Hydroxy-p-apo-8'-carotenoic acid [706]
Hydroxycapsanthin-like compound [130]
Hydroxycapsolutein [130]
Hydroxycapsolutein 5,6-epoxide [130]
Hydroxy-a-carotene (Zeinoxanthin (42)?) [783, 550, 695, 694, 131, 124, 261, 123, 118, 115]
see also: [134, 130, 122, 120, 119, 761]
Hydroxy-,-carotene [131, 124,42, 246] see also: [757]
Hydroxy-a-carotene' 5,6-epoxide [615, 614, 131, 129]
Hydroxy-a-carotene 5,8-epoxide [783, 129, 122, 119]
x-Hydroxy-p,p-caroten-y-one [80, 225]
x-Hydroxy-p,ljf-caroten-4-one (see also (160)) [703]
832 0. STRAUB

2'-Hydroxy-1',2'-dihydrotorulene (?) [28a]

Hydroxyflavoxanthin [568]
Hydroxyphytoene, Phytoenol (see also (61)) [436, 513, 131]
Hydroxyphytofluene, Phytofluenol (see also (60)) [508, 131, 491, 808]
Hydroxypirardixanthin [ 440]
Ketohydroxypirardixanthin [ 440]
Laetiporxanthin [736]
Lycopene epoxide [64, 768]
Lycoxanthin epoxide [768]
x-Methoxy-P,t/J-caroten-4-one [703]
x-Methoxy-3,4-didehydro-7',8'-dihydro-P,t/J-caroten-2-one (or 5',6'-Epoxy-5',6'-dihydro-
p,t/J-caroten-4-one?) [703]
Micronone (Hydroxy(?)-5,8-epoxy-P-carotene-x-one?) [614]
Microxanthin (Reduction product of micronone) [614]
Monohydroxy-4-keto-y-carotene see: x- Hydroxy-p,t/J-caroten-4-one
Monomethoxy-4-keto-y-carotene see: x-Methoxy-P,t/J-caroten-4-one
'Mycoplasma laidlawii B' carotenol (pigment B, B', C) [651]
Mycoxanthin [290,223,222,592,591]
Mytiloxanthin [756, 755, 81, 646, 627]
Papilioerythrins (px-1, px-2, pp-1, pp-2) [587]
Paracentrotin A [566, 564]
Paracentrotin B [566, 564]
Persicachrome [123]
Persicaxanthin [127, 124, 123]
Petaloxanthin (Mixture of zeaxanthin (67), lutein (73) and possibly antheraxanthin (119)?)
[698a, 237, 800]
Phoenicopterin, 'Fenicotterina ', "Phoenicoxanthin" (Astaxanthin (203) ?)
[542,541,539]
Phytoenol see: Hydroxyphytoene
Phytofluenol see: Hydroxyphytofluene
Polytomaxanthin [705, 520]
Pyrroxanthin [524]
Renieraxanthin, C 40 H 56 0 2 [770]
Rosaxanthili [735]
S.g. 434 (S.g. = Saprospira grandis) [2]
S.g. 460 [2]
S.g. 500 [2]
Sinensiachrome [126, 119, 116]
Sinensiaxanthin [134, 129, 127, 122, 119, 116]
S. t. 483 (S. t. = Saprospira thermalis) [ 4]
1,2,1 ',2'-Tetrahydro-3,4,3' ,4' -dehydrolycopene (?) see: 1,2-Dihydro-3' ,4' -dehydrolycopene
Tetrahydrophytoene ("Eicosahydrolycopene") [436, 647,239, 605]
Thiothece-484 [601]
Thiothece-polar-484 [601]
2,3,2'-Trihydroxy-IX-carotene see: p,E-Carotene-2,3,2' -trio!
3,4,3'-Trihydroxy-IX-carotene see: p,E-Carotene-3,4,3' -trio!
Trihydroxy-3',4' -didehydro-1 ',2' -dihydro-P,t/J-caroten-4-one
see: 1',2'-Dihydrotrihydroxy-4-ketotorulene
Triticoxanthin [540]
Trolliflavin-like compound [62, 126, 522]
Trolliflor [62, 133, 130, 128, 522]
Tugali-carotenoid X [573]
Valenciachrome [129, 120, 119, 116]
Valenciaxanthin [134, 129, 122, 119, 242, 116]
Xanthomonas-carotenoid [656] '
 XII. Lists of Natural Carotenoids 833

C. Old Names

Apart from the trivial names mentioned in lists A and B the older literature
includes a number of trivial designations no longer in current use, which are
only of historical interest. Many such names cannot now be definitely ascribed
to a specific carotenoid. These obsolete names are simply listed here alpha-
betically; the current names are added, where possible, in brackets. Literature
references are not given; they can be traced in the older monographs and
reviews, for example [10, 107, 205, 237, 412, 477, 478, 664, 740].

Araroth, Astrogrisein, Astroviolettine, Arumin, Calendulin, Calorhodin IX (Echinenone),

Calorhodin f3 (Canthaxanthin), Capsumin, Chloriosulfurin or Coriosulfurin, Chlorophan. Chryso-
xanthophyll. Coralin, Crustaceorubin (Astaxanthin), Cucurbitene (/3-Carotene), Diatomin (mix-
ture), Dicarotene (Lycopene), Eucarotene, Fucoxanthophyll (Fucoxanthin), Gardenidin (Crocetin),
red Haematochrome (Astacene), yellow Haematochrome (/3-Carotene), Hepatocarotene, Hepato-
xanthophyll (Lutein?), Lacertofulvin, Leucocyanin (Fucoxanthin ?), Lichnoxanthin, Linacarotene
(mixture), Lipochrin, Metridene (see [208]), Metridioxanthin (see [208]), Myxorhodin IX, Myxo-
rhodin /3, Nyctanthin (Crocetin), Orangin, Paradiseofulvin, Phycoxanthin (Fucoxanthin, Myxo-
xanthophyll), Phyllorhodin (Taraxanthin ?), Phylloxanthin (Violaxanthin ?, Mutatochrome ?),
Physalin, Picofulvin, Psittacofulvin, Rhodophan (Astacene?), Solanorubin (Lycopene), Sorbusin,
Taxorhodin, Tetronerythrin. Thujarhodin (Rhodoxanthin), Vitellolutein, Vitellorubin (Asta-
xanthin), /3-Xanthophyll (mixture), Xanthophan (Lutein?. Zeaxanthin?). Zoo(n)erythrin (Asta-
xanthin ?), Zoochlorin, Zoofulvin (Lutein?), Zooxanthin.

References
[1] Aasen A.J. and Liaaen-Jensen S., Acta Chern. Scand. 19, 1843 (1965).
[2] Aasen A.J. and Liaaen-Jensen S., Acta Chern. Scand. 20, 811 (1966).
[3] Aasen A.J. and Liaaen-Jensen S., Acta Chern. Scand. 20, 1970 (1966).
[4] Aasen A.J. and Liaaen-Jensen S., Acta Chern. Scand. 20, 2322 (1966).
[5] Aasen A.J. and Liaaen-Jensen S., Acta Chern. Scand. 21, 371 (1967).
[6] Aasen A.J. and Liaaen-Jensen S., Acta Chern. Scand. 21, 970 (1967).
[7] Aasen A.J. and Liaaen-Jensen S., Acta Chern. Scand. 21, 2185 (1967).
[8] Aasen A.J., Francis G. W. and Liaaen-Jensen S., Acta Chern. Scand. 23, 2605 (1969).
[9] Ahmad R. and Weedon B.C.L., J. Chern. Soc. 1953, 3286.
[10] AitzetmiillerK., SvecW.A., KatzJ.J. and StrainH.H., Chern. Cornrnun.1968, 32.
[11] AitzetmiillerK., StrainH.H., SvecW.A., GrandolfoM. and KatzJ.J., Phytochern. 8,1761
(1969).
[12] Ajisaka M., J. Biochern. (Tokyo) 34, 421 (1941).
[13] Akhtar M. and Weedon B.C.L., J. Chern. Soc. 1959, 4058.
[14] Akiya S., J. Japan. Ophthalrnol. Soc. 67. 1168 (1963); Chern. Abstr. 62, 8045.
[15] Anderson D.G. and Porter J. W., Arch. Biochern. Biophys. 97, 509 (1962).
[16] Antia N.J., Can. J. Chern. 43, 302 (1965).
[17] Aroamone F., Camerino B., Cotta E., Franceschi G., Grein A., Penco S. and Spalla C.,
Experientia 25, 241 (1969).
[18] Arcamone P., Camerino B., Franceschi G. and Penco S., Gazz. Chirn. Ita/. 100, 581 (1970).
[19] Arpin N. and Liaaen-Jensen S., Bull. Soc. Chirn. Bioi. 49, 527 (1967).
[20] Arpin N. and Liaaen-Jensen S., C. R. Acad. Sci., Ser. D 265, 1083 (1967).
[21] Arpin N. and ·Liaaen-Jensen S., Phytochern. 6, 995 (1967).
[22] Arpin N., These (Lyon 1968).
[23] Arpin N. and Liaaen-Jensen S., Phytochern. 8, 185 (1969).
[24] Arpin N., Fiasson J. L. and Lebreton P., Prod. Probl. Pharrn. 25, 21 (1970).

Carotenmds 53
834 0. STRAUB

[25] Arpin N., Norgard S., Francis G. W. and Liaaen-Jensen S., Acta Chern. Scand., to be
published.
[26] Arpin N., Fiasson J.L., Bouchez-Dangye-Caye M.P., Francis G. W. and Liaaen-Jensen S.,
Phytochem. 10, 1595 (1971).
[27] Ashmawi H., Hussein A.A. and Shaker M.H., Bull. Fac. Sci. Cairo Univ. No. 61 (1955);
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[280] Hertzberg S. and Liaaen-Jensen S., Phytochem. 6, 1119 (1967).
[281] Hertzberg S. and Liaaen-Jensen S., Acta Chern. Scand. 22, 1714 (1968).
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[283] Hertzberg S. and Liaaen-Jensen S., Phytochem. 8, 1259 (1969).
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[284a] Hertzberg S. and Liaaen-Jensen S., Phytochem., to be published.
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[293] Hopkins T. S., Diss. Abstr. B 28, 1603 (1967); Chern. Abstr. 68, 47,383.
[294] Hora J., Toube T. P. and Weedon B. C. L., J. Chern. Soc. C 1970, 241.
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9510.
[296] lnhoffen H. H., Pommer H. and Bohlmann F., Ann. Chern. 569, 237 (1950).
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[298] Inhoffen H. H., Pommer H. and Westphal F., Ann. Chern. 570, 69 (1950).
[299] Inhoffen H. H., Pommer H. and Meth E.-G., Ann. Chern. 572, 151 (1951).
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[301] Inhoffen H. H., Isler 0., von der Bey G., Raspe G., Zeller P. and Ahrens R., Ann. Chern.
580, 7 (1953).
[302] Inhoffen H. H., Schwieter U. and Raspe G., Ann. Chern. 588, 117 (1954).
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[305] Isler 0., Angew. Chern. 68, 547 (1956).
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456 (1957).
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[322] Jaeger L. and Karrer P., Helv. Chim. Acta 46, 683 (1963).
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[340] Karrer P. and Salomon H., Helv. Chim. Acta 11, 513 (1928).
[341] Karrer P. and Salomon H., Heir. Chim. Acta II, 711 (1928).
[342] Karrer P., Helfenstein A., Widmer R. and van ltallie T. B., Helv. Chim. Acta I2, 741 (1929).
[343] Karrer P., Salomon H. and Wehrli H., Helv. Chim. Acta 12, 790 (1929).
 XII. Lists of Natural Carotenoids 841

[344] Karrer P. and Miki K., Helv. Chim. Acta 12,985 (1929).
[345] Karrer P. and Helfenstein A., Helv. Chim. Acta 12, 1142 (1929).
[346] Karrer P., Helfenstein A. and Wehrli H., Helv. Chim. Acta 13, 87 (1930).
[347] Karrer P., Wehrli H. and Helfenstein A., Helv. Chim. Acta 13,268 (1930).
[348] Karrer P., Helfenstein A., Wehrli H. and Wettstein A., Helv. Chim. Acta 13, 1084 (1930).
[349] Karrer P. and Ishikawa S., Helv. Chim. Acta 13, 1099 (1930).
[350] Karrer P., Helfenstein A., Pieper B. and Wettstein A., Helv. Chim. Acta 14, 435 (1931).
[351] Karrer P. and MorfR., Helv. Chim. Acta 14, 833 (1931).
[352] Karrer P. and MorfR., Helv. Chim. Acta 14, 1033 (1931).
[353] Karrer P. and MorfR., Helv. Chim. Acta 14, 1044 (1931).
[354] Karrer P., MorfR., Krauss E.-V. and Zubrys A., Helv. Chim. Acta 15, 490 (1932).
[355] Karrer P., Benz P., MorfR., Raudnitz H., Stoll M. and Takahashi T., Helv. Chim. Acta 15,
1218 (1932).
[356] Karrer P., Benz P., MorfR., Raudnitz H., Stoll M. and Takahashi T., Helv. Chim. Acta 15,
1399 (1932).
[357] Karrer P. and Takahashi T., Helv. Chim. Acta 16, 287 (1933).
[358] Karrer P. and Benz F., Helv. Chim. Acta 16, 337 (1933).
[359] Karrer P., MorfR. and Walker 0., Heh'. Chim. Acta 16, 975 (1933).
[360] Karrer P., Zubrys A. and MorfR., Heft'. Chim. Acta 16, 977 (1933).
[361] Karrer P. and Schlientz W., Helv. Chim. Acta 17, 55 (1934).
[362] Karrer P., Solmssen U. and Walker 0., Helv. Chim. Acta 17, 417 (1934).
[363] Karrer P. and Loewe L., Helv. Chim. Acta 17, 745 (1934).
[364] Karrer P., v. Euler H. and Solmssen U., Heh'. Chim. Acta 17, 1169 (1934).
[365] Karrer P., Loewe L. and Hiibner H., Heh'. Chim. Acta 18, 96 (1935).
[366] Karrer P. and Solmssen U., Helv. Chim. Acta 18,477 (1935).
[367] Karrer P. and Oswald A., Helv. Chim. Acta 18, 1303 (1935).
[368] Karrer P. and Solmssen U., Heh'. Chim. Acta 18, 1306 (1935).
[369] Karrer P. and Solmssen U., Helv. Chim. Acta 19, 1 (1936).
[370] Karrer P. and Solmssen U., Helv. Chim. Acta 19, 1019 (1936).
[370a] Karrer P. and Solmssen U., Heh'. Chim. Acta 19, 1024 (1936).
[371] Karrer P. and Solmssen U., He/v. Chim. Acta 20, 682 (1937).
[372] Karrer P., Solmssen U. and Gugelmann W., Helv. Chim. Acta 20, 1020 (1937).
[373] Karrer P. and Solmssen U., Helv. Chim. Acta 20, 1396 (1937).
[374] Karrer P., Koenig H. and Solmssen U., Heh'. Chim. Acta 21,445 (1938).
[375] Karrer P., Riiegger A. and Solmssen U., Helv. Chim. Acta 21,448 (1938).
[376] Karrer P., Solmssen U. and Koenig H., Helv. Chim. Acta 21, 454 (1938).
[377] Karrer P. and Jaffe W., Helv. Chim. Acta 22, 69 (1939).
[378] Karrer P. and Koenig H., Helv. Chim. Acta 23, 460 (1940).
[379] Karrer P. and Schwab G., Heh-. Chzm. Acta 23, 578 (1940).
[380] Karrer P. and Riiegger A., Helv. Chim. Acta 23,955 (1940).
[381] Karrer P. and Rutschmann J., Helv. Chim. Acta 25, 1144 (1942).
[382] Karrer P. and Rutschmann J., Helv. Chim. Acta 25, 1624 (1942).
[383] Karrer P. and Jucker E., Helv. Chim. Acta 26, 626 (1943).
[384] Karrer P. and Rutschmann J., Helv. Chim. Acta 26, 2109 (1943).
[385] Karrer P. and Rutschmann J., He h-. Chim. Acta 27, 320 (1944).
[386] Karrer P. and Kramer H., Helv. Chim. Acta 27, 1301 (1944).
[387] Ka,rer P. and Jucker E., Helv. Chim. Acta 27, 1585 (1944).
[388] Karrer P. and Rutschmann J., Helv. Chim. Acta 27, 1684 (1944).
[389] Karrer P. and Rutschmann J., Heh'. Chim. Acta 27, 1691 (1944).
[390] Karrer P. and Jucker E., Helv. Chim. Acta 27, 1695 (1944).
[391] Karrer P. and Jucker E., Helv. Chim. Acta 28, 300 (1945).
[392] Karrer P. and Jucker E., Helv. Chim. Acta 28, 427 (1945).
[393] Karrer P. and Jucker E., Helv. Chim. Acta 28,471 (1945).
[394] Karrer P. and Rutschmann J., Hell'. Chim. Acta 28, 793 (1945).
[395] Karrer P. and Rutschmann J., Helv. Chim. Acta 28, 795 (1945).
842 0. STRAUB

[396] Karrer P. and Jucker E., Helv. Chirn. Acta 28, 1143 (1945).
[397] Karrer P., Jucker E., Rutschmann J. and Steinlin K., Helv. Chirn. Acta 28, 1146 (1945).
[398] Karrer P. and Rutschmann J., Helv. Chirn. Acta 28, 1526 (1945).
[399] Karrer P. and Jucker E., Helv. Chirn. Acta 29, 229 (1946).
[400] Karrer P. and Rutschmann J., Helv. Chirn. Acta 29, 355 (1946).
[401] Karrer P. and Jucker E., Helv. Chirn. Acta 29, 1539 (1946).
[401a] Karrer P. and Jucker E., Helv. Chirn. Acta 30, 266 (1947).
[402] Karrer P., Jucker E. and Steinlin K., Helv. Chirn. Acta 30, 531 (1947).
[403] Karrer P. and Jucker E., Helv. Chirn. Acta 30, 536 (1947).
[404] Karrer P. and Krause-Voith E., Helv. Chirn. Acta 30, 1772 (1947).
[405] Karrer P., Krause-Voith E. and Steinlin K., Helv. Chirn. Acta 31, 113 (1948).
[406] Karrer P. and Schwyzer R., Helv. Chirn. Acta 31, 1055 (1948).
[407] Karrer P., Schwyzer R. and Neuwirth A., Helv. Chirn. Acta 31, 1210 (1948).
[408] Karrer P. and Eugster C. H., Helv. Chirn. Acta 33, 1172 (1950).
[409] Karrer P., Eugster C. H. and Tobler E., Helv. Chirn. Acta 33, 1349 (1950).
[410] Karrer P. and Eugster C. H., Helv. Chirn. Acta 33, 1433 (1950).
[411] Karrer P. and Eugster C. H., Helv. Chirn. Acta 33, 1952 (1950).
[ 412] Karrer P. and Jucker E., Carotenoids (Elsevier, Amsterdam 1950).
[413] Karrer P. and Leumann E., Helv. Chirn. Acta 34, 445 (1951).
[414] Karrer P. and Leumann E., Helv. Chirn. Acta 34, 1412 (1951).
[415] Karrer P., Helv. Chirn. Acta 34, 2160 (1951).
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[418] Katayama T., Yokoyama H. and Chichester C. 0., lnternat. J. Biochern. 1, 438 (1970).
[419] Kelly M. and Liaaen-Jensen S., Acta Chern. Scand. 21, 2578 (1967).
[420] Kelly M., NorgArd S. and Liaaen-Jensen S., Acta Chern. Scand. 24, 2169 (1970).
[421] Kelly M .. Authen Andresen S. and L1aaen-Jensen S.• Acta Chern. Scand. 25. in the press
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[422] Ketskhoveli E. N., Kinkladze D. C. and Gigineishvili M. N., Soobshch. Akad. Nauk Gruz.
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[423a] Kishore G. S., Subbarayan C. and Cama H. R., Indian J. Biochern. 7, 98 (1970).
[424] Kj~sen H. and Liaaen-Jensen S., Phytochern. 8, 483 (1969).
[425] Kj~sen H. and Liaaen-Jensen S., Acta Chern. Scand. 24, 2259 (1970).
[426] Kj~sen H .. Liaaen-Jensen S. and Enzell C.R., Acta Chern. Scand. 25. 85 (1971).
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[427] Kleinig H. and Egger K., Phytochern. 6, 611 (1967).
[428] Kleinig H. and Egger K., Phytochern. 6, 1681 (1967).
[429] Kleinig H. and Egger K., Z. Naturforsch. B 22, 868 (1967).
[430] Kleinig H., Z. Naturforsch. B 22, 977 (1967).
[431] Kleinig H. and Reichenbach H., Arch. Mikrobiol. 68,210 (1969).
[432] Kleinig H. and Czygan F.-C., Z. Naturforsch. B 24, 927 (1969).
[433] Kleinig H., Nitsche H. and Egger K., Tetrahedron Letters 1969, 5139.
[434] Kleinig H. and Reichenbach H., Naturwiss. 57, 92 (1970).
[435] Kleinig H., Reichenbach H. and Achenbach H., Arch. Mikrobiol. 74, 223 (1970).
[436] KleinkaufH., Arch. Mikrobiol. 53, 154 (1966).
[ 437] Koe B. K. and Zechmeister L., Arch. Biochern. Biophys. 46, 100 (1953).
[438] KrinskyN.I. and Goldsmith T.H., Arch. Biochern. Biophys. 91,271 (1960).
[439] Krinsky N.I. and LevineR. P., Plant Physio/. 39, 680 (1964).
[440] KrinskyN.I. and LenhoffH.M., Cornp. Biochern. Physio/.16, 189 (1965).
[441] Kuhn R. and Winterstein A., Helv. Chirn. Acta 11, 427 (1928).
[442] Kuhn R. and Wiegand W., Helv. qirn. Acta 12, 499 (1929).
[443] Kuhn R., Winterstein A. and Kaufmann W., Ber. Deut. Chern. Ges. 63, 1489 (1930).
 XII. Lists of Natural Carotenoids 843
[444] Kuhn R. and Winterstein A., Naturwiss. I8, 754 (1930).
[445] Kuhn R. and Winterstein A., Ber. Deut. Chern. Ges. 64, 326 (1931).
[446] Kuhn R. and L'Orsa F., Ber. Deut. Chern. Ges. 64, 1732 (1931).
[447] Kuhn R., Winterstein A. and Lederer E., Hoppe-Seyler's Z. Physiol. Chern. I97, 141 (1931).
[448] Kuhn R. and Lederer E., Hoppe-Seyler's Z. Physiol. Chern. 200, 108 (1931).
[449] Kuhn R. and Lederer E., Ber. Deut. Chern. Ges. 65, 637 (1932).
[450] Kuhn R. and Winterstein A., Ber. Deut. Chern. Ges. 65, 646 (1932).
[451] Kuhn R. and Brockmann H., Ber. Deut. Chern. Ges. 65, 894 (1932).
[452] Kuhn R. and Grundmann C., Ber. Deut. Chern. Ges. 65, 898 (1932).
[453] Kuhn R. and Winterstein A., Ber. Deut. Chern. Ges. 65, 1873 (1932).
[454] Kuhn R. and Grundmann C., Ber. Deut. Chern. Ges. 65, 1880 (1932).
[455] Kuhn R. and Brockmann H., Hoppe-Seyler's Z. Physiol. Chern. 213, 192 (1932).
[456] Kuhn R. and Brockmann H., Ber. Deut. Chern. Ges. 66, 407 (1933).
[457] Kuhn R. and Winterstein A., Ber. Deut. Chern. Ges. 66, 429 (1933).
[458] Kuhn R. and Lederer E., Ber. Deut. Chern. Ges. 66, 488 (1933).
[459] Kuhn R. and Brockmann H., Ber. Deut. Chern. Ges. 66, 828 (1933).
[460] Kuhn R. and Deutsch A., Ber. Deut. Chern. Ges. 66, 883 (1933).
[461] Kuhn R. and Brockmann H., Ber. Deut. Chern. Ges. 66, 1319 (1933).
[462] Kuhn R. and Grundmann C., Ber. Deut. Chern. Ges. 66, 1746 (1933).
[463] Kuhn R. and Grundmann C., Ber. Deut. Chern. Ges. 67, 339, 1133 (1934).
[464] Kuhn R. and Grundmann C., Ber. Deut. Chern. Ges. 67, 596 (1934).
[465] Kuhn R. and Brockmann H., Ber. Deut. Chern. Ges. 67, 885 (1934).
[466] Kuhn R. and Brockmann H., Ann. Chern. 5I6, 95 (1935).
[467] Kuhn R. and Sorensen N.A., Ber. Deut. Chern. Ges. 7I, 1879 (1938).
[468] Kuhn R. and Wang Y., Ber. Deut. Chern. Ges. 72, 871 (1939).
[469] Kuhn R., SteneJ. and Sorensen N.A., Ber. Deut. Chern. Ges. 72, 1688 (1939).
[470] Kuhn R., Inhoffen H.H., Staab H.A. and Otting W., Chern. Ber. 86, 965 (1953).
[471] Lederer E., C. R. Acad. Sci. I97, 1694 (1933).
[472] Lederer E., C. R. Soc. Bioi. 113, 1015 (1933).
[473] Lederer E. and Fabre R., C. R. Soc. Bioi. I13, 1391 (1933).
[474] Lederer E., C. R. Soc. Bioi. 116, 150 (1934).
[475] Lederer E., C. R. Soc. Bioi. 117, 411 (1934).
[476] Lederer E., C. R. Soc. Bioi. 117, 1086 (1934).
[477] Lederer E., Les Carotenoides des Plantes (Hermann, Paris 1934).
[478] Lederer E., Les Carotenoides des Anirnaux (Hermann, Paris 1935).
[479] Lederer E., C. R. Acad. Sci. 20I, 300 (1935).
[ 480] Lederer E., Bull. Soc. Chirn. Bioi. 20, 554 (1938).
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[490] Liaaen-Jensen S., Acta Chern. Scand. I2, 1698 (1958).
[491] Liaaen-Jensen S., Cohen-Bazire G., Nakayama T.O. M. and Stanier R. Y., Biochirn. Biophys.
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[492] Liaaen-Jensen S., Acta Chern. Scand. I3, 381 (1959).
[493] Liaaen-Jensen·S., Acta Chern. Scand. I3, 842 (1959).
[494] Liaaen-Jensen S., Acta Chern. Scand. 13, 2142 (1959).
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[497] Liaaen-Jensen S., Acta Chem. Scand. 14, 953 (1960).

[498] Liaaen-Jensen S., Acta Chem. Scand. 15, 1182 (1961).
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Carotenoids 54
850 O.STRAUB

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 851

Appendix. Tentative Rules

for the Nomenclature of Carotenoids

IUPAC Commission on the Nomenclature of Organic Chemistry and

IUPAC-IUB Commission on Biochemical Nomenclature

1. Class of compound . . . . . . . . . . 852

2. The stem name . . . . . . . . . . . 852
3. Specific names; end-group designations . 853
4. Numbering of carotenoid hydrocarbons . 854
5. Nor-carotenoids and seco-carotenoids 855
6. Changes in hydrogenation level . . . 856
7. Oxygenated derivatives . . . . . . . 857
8. Numbering of oxygenated derivatives . 858
9. Retro nomenclature 860
10. Apo nomenclature . 860
11. Higher carotenoids . 861
12. Stereochemistry 862
13. Trivial names . . . 864
852 Appendix. Tentative Rules for the Nomenclature of Carotenoids

Rule Carotenoid 1. Class of compound

Carotenoids are a class of hydrocarbons (carotenes) and their oxygenated
derivatives (xanthophylls) consisting of eight isoprenoid units joined in such
a manner that the arrangement of isoprenoid units is reversed at the centre
of the molecule so that the two central methyl groups are in a 1,6 positional
relationship and the remaining non-terminal methyl groups are in a 1,5
positional relationship. All carotenoids may be formally derived from the
acyclic C 40 H 56 structure (I), having a long central chain of conjugated double
bonds, by (i) hydrogenation, (ii) dehydrogenation, (iii) cyclization, or (iv)
oxidation, or any combination of these processes.

(I)

The class also includes certain compounds that arise from certain rear-
rangements or degradations of the carbon skeleton (I) provided that the two
central methyl groups are retained. This excludes retinol (vitamin A) and
related C20 -compounds.
For convenience carotenoid formulae are often written, in a shorthand
form, as:

(lA)

(General formula; broken lines indicate formal division into isoprenoid units)

Rule Carotenoid 2. The stem name

All specific names are based on the stem name 'carotene', which corre-
sponds to the structure and numbering shown in (II), where the broken lines
at the two terminations are intended only to represent two 'double-bond
equivalents'. Individual compounds may have C 9 acyclic end groups with
two double bonds at positions 1,2 and 5,6 (e.g. III) or cyclic end groups (such
as IV-VIII).

9 11 13 IS 14' 12' 10' ..

"'<:::::10 "'<:::::12 "'<:::::14 "'<:::::, •• "'<::::: 13' "'<:::::...
20'

4
(II)
 Appendix. Tentative Rules for the Nomenclature of Carotenoids 853

Rule Carotenoid 3. Specific names; end-group designations

3.1. The name of a specific carotenoid hydrocarbon is constructed by adding
two greek letters as prefixes to the stem name 'carotene' (defined in Rule
Carotenoid 2), these prefixes being characteristic of the two C 9 end groups.
3.2. The prefixes are:

Type Prefix Formula Structure

Acyclic 1/1 C 9H 15 (III)

Cyclohexene {3, e C 9H 15 (IV), (V)
Cyclopentane K C9H!7 (VI)
Aryl qJ,X CgHu (VII), (VIII)

and correspond to the following end-group modifications:

1d116R 1a11•R
1.
2 6
I. " 1
R : 16
~ 2 4 6 4
5 18
3
4
..<?, 18

tf
(III) 1/1 (IV) {3 (V) e

2R
17 18
1 R
17n-;?' R
6
2 3 1
4 :::::,.... 5 18

(VI) K (VII) qJ (VIII) X

R= ...
(6)
-'~'·· ...
8 10 12 14

Note: The choice of locants 16 and 17 for the two methyl groups at C-1
is considered in connection with stereochemistry in Rule Carotenoid 12.4.
3.3. The greek-letter prefixes are cited in alphabetical order; the first is
separated from the second by a comma, and the second is connected to the
stem name by a hyphen.
Note: The greek-letter alphabetical order is
J3 (beta), e (epsilon), K (kappa), cp (phi), x (chi), t/1 (psi).
Examples:

e,e-Carotene
854 Appendix. Tentative Rules for the Nomenclature of Carotenoids

e,x-Carotene

Notes: (i) The greek-letter prefixes are derived:

f3 and e from the symmetrical carotenoids with the trivial names
'{3-carotene' and 'e-carotene'
" from the symmetrical capsorubin
t/1 from t/J-ionone
q> for phenyl, and
x the next greek letter after q>
(ii) a, y and b, from the trivial names a-, y- and b-carotene, are not
used.
3.4. When, in a modified or degraded carotenoid, an end group can be
derived from more than one specific end group, the end group chosen as the
basis of the name is that occurring earliest in the alphabetical order (see Rule
Carotenoid 3.3).
Example:
is a derivative of the {3-end group not the
e-end group.

Rule Carotenoid 4. Numbering of carotenoid hydrocarbons

The basic system of numbering is that shown in structure (II} of Rule
Carotenoid 2; the end groups are numbered as indicated in Rule Carotenoid 3.
If the two end groups are dissimilar, lower (unprimed) numbers are given to
that end of the molecule which is associated with the greek-letter prefix cited
first in the name. (All unprimed locants are cited before primed locants *.)
Examples: 4'

9 11 13 15 14' 12' 10'

'<::::,0 '<::::,2 '<::::,4 '<::::15' '<:::: 13' '<::::11,

201 19'

e,e-Carotene

• IUPAC Nomenclature of Organic Chemistry, Section C, 1970 (published by Butterworths,

London), indicates that, in ranking locants for priority, a primed numeral ranks immediately
after the same numeral unprimed. In general organic nomenclature the common practice is
therefore to cite locants in the order
x, x', (x+n), (x+n)'.
The established sequence in the caroq:noid field is, however, to cite all unprimed numerals
before any primed numerals, and this practice is followed in these rules.
 Appendix. Tentative Rules for the Nomenclature of Carotenoids 855
4'

19
9 11 13 15 14' 12' 10' 8'

""" """ """ """ """

10 12 14 15' 13'

20'
~11'
""" """7'
9'
19' 16'

e,x-Carotene

Note: It is recommended that formulae be drawn so that unprimed

numbers are on the left hand side.

Rule Carotenoid 5. Nor-carotenoids and seco-carotenoids

a) N or-carotenoids
Elimination of a CH 3 , CH 2 or a CH group from a carotenoid is indicated
by the prefix 'nor', which in all cases is preceded by the locant of the carbon
atom that has been eliminated. The prefix is non-detachable. When alternatives
are possible, the locant attached to nor is the lowest possible*. The basic
numbering of the carotenoid is retained in the nor-carotenoid.
Examples:

""" """ """ """ """ """ """ """ """

3'

5 18

2,2'-Dinor-j:l,f:l-carotene

4'
18'
3'

""" """ """ """ """ """ """ """ """

2'

1'

16,17,16',17' -Tetranor-e,e-carotene

b) Seco-carotenoids
Fission of the bond between two adjacent carbon atoms (other than
carbon atoms J and 6) of a cyclic end group, with addition of one or more
hydrogen atoms at each terminal group thus created, is indicated by the
prefix 'seco ', the original carotenoid numbering being retained.
*The contrast with steroid usage (IUPAC-IUB 1967 Revised Rules for Nomenclature of
Steroids, IUPAC Information Bulletin No. 33, Rule 2S-6, page 52), where the prefix 'nor' is
associated with the highest permissible number, is to be noted.
856 Appendix. Tentative Rules for the Nomenclature of Carotenoids

Example:

2,3-Seco-e,e-carotene

Rule Carotenoid 6. Changes in hydrogenation level

Carotenoid hydrocarbons, including carotenoid acetylenes and allenes,
that differ in hydrogenation level from the corresponding carotenoid hydro-
carbon defined by Rule Carotenoid 3 are named from the latter by use of
the prefixes 'hydro' and 'dehydro' together with the locants specifying the
carbon atoms at which hydrogen atoms have been added or removed.
These prefixes are non-detachable* and immediately precede the gree~­
letter prefixes denoting the end groups, and, if both occur in one name, are
cited in the order: dehydro before hydro (multipliers do not affect the order).
Example:
tetradehydrodihydro.
Note: Since, to maintain valency requirements, hydrogen atoms are,
formally, always added or removed in pairs these prefixes will always be used
with an even-number multiplier, e.g., te~rahydro, didehydro.
Examples: 4'

..
7'

4
5,6,7,8,1',2',3',4',5',6',7',8'-Dodecahydro-P,x-carotene

7,8-Didehydro-e,e-carotene

6, 7-Didehydro-e,e-carotene
* IUPAC Nomenclature of Organic Chemistry, Section C, 1970 (published by Butterworths,
London), Rule C-16.1, allows the prefix 'hydro' to be detachable or non-detachable, and the
former has become the established usage in general organic chemistry. However, the common
practice in carotenoid names is now to use this prefix as non-detachable, a practice that is followed
in this set of rules. '
 Appendix. Tentative Rules for the Nomenclature of Carotenoids 857

Rule Carotenoid 7. Oxygenated derivatives

7.1. Oxygenated (and other) derivatives of carotenoid hydrocarbons are
named by use of suffixes and prefixes, according to the rules of general organic
chemical nomenclature*.
Of the oxygen-containing characteristic groups present, that which occurs
earliest in the sequence:
Carboxylic acid, ester of carotenoid acid, aldehyde, ketone, alcohol, ester
of carotenoid alcohol
is chosen as principal group (following IUPAC Rule C-10.3) and is cited by
use of a suffix; all other groups are cited as prefixes.
Examples:

HO
3-Hydroxy-p,e-caroten-3' -one

3-Hydroxy-3' -oxo-p,e-caroten-16-oic acid

7.2. A non-bridging ether group is named by use of the appropriate alkoxy

or aryloxy prefix (Rule C-211.2).
Example:

8'

1-Ethoxy-l ,2, 7',8' -tetrahydro-1/1,1/J-carotene

7.3. Oxygen bridges are indicated by use of the prefix 'epoxy'; this prefix is
preceded by the locants of the two carbon atoms that form the bridgeheads
of the oxygen bridge.
Note: The prefix 'epoxy' denotes replacement, by an oxygen bridge, of a
hydrogen atom at each of two carbon atoms already otherwise connected
to one another. An epoxide, notionally formed by adding an oxygen atom
to a double bond, is therefore an epoxy-dihydro derivative of the original
compound.
* IUPAC Nomenclature of Organic Chemistry, Section C, 1970, Subsections C-2 to C-4
(published by Butterworths, London).
858 Appendix. Tentative Rules for the Nomenclature of Carotenoids

Examples:
OH

HO

5,6: 5' ,6'-Diepoxy-5,6,5',6' -tetrahydro-p,p-carotene-3,3'-diol

5,8: 5',8' -Diepoxy-5,8,5',8' -tetrahydro-p,p-carotene

7.4. Compounds which may be formally derived from a carotenoid hydro-

carbon by the addition of the elements of water (H, OH) or of methanol
(H, OCH 3 ) to a double bond are named as 'hydroxy-dihydro' or 'methoxy-
dihydro' derivatives.
Examples:

HO

3,7'-Dihydroxy-7' ,8' -dihydro-p,e-caroten-3' -one

7, 7'-Dimethoxy-7,8, 7' ,8' -tetrahydro-e,e-carotene-3,3'-dione

Rule Carotenoid 8. Numbering of oxygenated derivatives

8.1. If the two C 9 end groups of the parent carotenoid hydrocarbon are
dissimilar their oxygenated derivatives are numbered according to Rule
Carotenoid 4, i.e., the end group designated by the greek letter occurring
earlier in the greek alphabet receives unprimed locants.
 Appendix. Tentative Rules for the Nomenclature of Carotenoids 859

Examples:

HO

2,4-Dihydroxy-p,e-caroten-3' -one

e
p
2,2' ,3'-Trimethoxy-p,e-carotene

8.2. a) If the two C 9 end groups of the parent carotenoid hydrocarbon are
identical then the lowest* locant possible is assigned to the principal group,
cited as suffix.
b) If more than one of the group chosen to be cited as suffix is present
the numbering is determined by the principle of lowest locants ** applied to
the suffixes.
c) If no group qualifies to be cited as suffix then the numbering is deter-
mined by the principle of lowest locants ** for all groups cited as prefixes.
Examples:

OCH 3
a)

2' -Methoxy-e,e-caroten-3-one
(not 2-Methoxy-e,e-caroten-3'-one)

b)

2'-Methoxy-e,e-carotene-2,3' -dione
(not 2-Methoxy-e,e-carotene-3,2'-dione)

c)
CH 3
6' 2'
,,

3,2'-Dimethoxy-1 ,2,5' ,6' -tetrahydro-1/1,.p-carotene

(not 2,3'-Dimethoxy-5,6,1',2'-tetrahydro-1/1,1/1-carotene; 1,2,3,2',5',6' having priority over 2,5,6,1',2',3')
860 Appendix. Tentative Rules for the Nomenclature of Carotenoids

Notes: * In the carotenoid series all unprimed numbers are cited before
primed numbers and the former are therefore considered as 'lower' than
primed numbers, e.g. 2, 6, 6, 1', 2', 6' etc.
** When series of locants containing the same number of terms are
compared term by term, that series is 'lowest' which contains the lowest
number on the occasion of the first difference [IUPAC Rule C-13.1l(e),
footnote].

Rule Carotenoid 9. Retro nomenclature

9.1. The prefix 'retro' (printed in italics) and a pair of locants are used to
indicate a shift, by one position, of all single and double bonds of the conjugated
polyene system delineated by the pair of locants.
9.2. The pair of locants precedes the prefix retro. The first locant is that 9f
the carbon atom that has lost a proton, the second that of the carbon atom
that has gained one.
9.3. The prefix and its accompanying locants are placed immediately before,
and hyphenated to, the greek-letter prefixes of the name defined according
to Rule Carotenoid 3.
9.4. The prefix and its associated locants are not detachable from the names
defined according to Rule Carotenoid 3.
Examples:

fJ

3-Hydroxy-6', 7-retro-f3,e-caroten-3' -one

6', 7-retro-B,K-Caroten-3-one

0
I
"""' c:::?'

8', 11-retro-f3,1/f-Caroten-11-one

Rule Carotenoid 10. Apo nomenclature

It is often necessary to designate derivatives in which the carbon skeleton
has been shortened by the formal removal of fragments from one or both
ends of a carotenoid.
 Appendix. Tentative Rules for the Nomenclature of Carotenoids 861

10.1. The unitalicized prefix 'apo ', preceded by a locant, is used to indicate
that all of the molecule beyond the carbon atom corresponding to that locant
has been replaced by hydrogen atoms. A side-chain methyl group is not
considered to be 'beyond' the carbon atom to which it is attached.
10.2. The prefix and its locant immediately precede the specific name (Rule
Carotenoid 3) unless the locant associated with the prefix 'apo' is greater
than 5, in which case there is no need to give a greek-letter end-group desig-
nation for that end of the molecule.
10.3. For purposes of numbering etc., an end that has been shortened by 5
or less skeletal carbon atoms is considered a t/1 (acyclic) end group.
10.4. The prefix diapo, preceded by two locants, is used to indicate removal
of fragments from both ends of the molecule.
10.5. If, in a diapo compound, the two ends of the carotenoid skeleton have
been shortened unequally the lower locant associated with the prefix 'diapo'
is unprimed.
Examples:

2'
CHO

2' -Apo-P,I/f-caroten-2' -al

.,
HOOC '<:::::: '<:::::: '<:::::: '<:::::: '<:::::: '<:::::: '<:::::: '<:::::: '<:::::: COOCH.

Methyl hydrogen 6,6'-diapocarotene-6,6'-dioate (trans-Bixin)

Rule Carotenoid 11. Higher carotenoids

The higher carotenoids are a class of hydrocarbons and their oxygenated
derivatives consisting of more than eight isoprenoid units joined in a manner
similar to that of the C40 -carotenoids.
They are named as mono- or di-substituted C 40 -carotenoids.
The numbering of the normal carotenoid is retained.
Examples:

2-(3-Methyl-2-butenyl)-e,l/f-carotene
862 Appendix. Tentative Rules for the Nomenclature of Carotenoids

OH

OH

2,2' -Bis(3-methylbutyl)-P.P-carotene-3",3"' -diol

or
2,2'-Bis(3-hydroxy-3-methylbutyl)-p,p-carotene

Rule Carotenoid 12. Stereochemistry

12.1. Absolute cmifiguration at chiral centers.
The absolute configuration at chiral centers is designated by use of the RS
convention*, the symbols being placed, with the corresponding locants, before
the carotenoid name.
Example:

_6_ I
I
I
I
~ ~ ""'o

(3S,5R,3'S,5' R)-3,3'-Dihydroxy-K,K-carotene-6,6' -dione (Capsorubin)

12.2. Absolute corifiguration of allenic compounds.

The absolute configuration around allene groups will be similarly
designated, when known.
Example:

H O ;4 ,q · ·OCOCH 3
5' 3'
6' 2'
-9" 1'
H ··. -9"• .. ·
~~~~~~~

HO

(3'S,5'R,6' R)-3' -Acetoxy-5,6-epoxy-3,5'-dihydroxy-6', 7'-

didehydro-5,6, 7,8,5',6' -hexahydro-P,P-caroten-8-one (Fucoxanthin)

* For a discussion on the RS convention and the use of thickened lines, broken lines, wavy
lines and wedges in displayed formulae, see IUPAC Tentative Rules for the Nomenclature of
Organic Chemistry, Section E, Fundamental Stereochemistry, IUPAC Information Bulletin
No. 35, page 68.
 Appendix. Tentative Rules for the Nomenclature of Carotenoids 863

12.3. Geometrical cmifiguration around double bonds.

The stem name 'carotene' implies trans configuration about all double
bonds unless the contrary is indicated. Following the designation of absolute
configuration (if any) geometrical configuration is indicated by citing the
double bond or bonds with a cis configuration.
Example:

Natural phytoene = 15-cis-7,8,11,12,7',8',11',12'-octahydro-t/J,t/1-carotene

At trisubstituted double bonds the term cis refers to the relative position
of the two substituents forming parts of the main chain of carbon atoms.
Example:
Natural bixin= Methyl hydrogen 9' -cis-6,6' -diapocarotene-6,6'-dioate
In the absence of definite information on geometrical configuration,
cis isomers may be distinguished by prefixes such as neo A, neo U etc. (cf.
Zechmeister, Cis-trans Isomeric Carotenoids, Vitamins A and Arylpolyenes,
Springer-Verlag, Vienna, 1962).
The stereochemical prefixes E and Z [see footnote to Rule 12.1 and
J. Amer. Chern. Soc. 90, 509 (1968)] may be used, especially when the prefixes
cis and trans might lead to ambiguity.
12.4. Numbering of gem-dimethyl groups at C-1.
In an end group of /3- or e-type, the two methyl groups attached to C-1
are distinguished as follows:
When the potential chirality is as shown in formula (A), i.e. with the polyene
chain (R) to the right of C-1, the methyl groups below and above the plane of
the paper are numbered 16 and 17, respectively.
If the end group is as shown in (B), with the polyene chain (R) to the left,
these designations are reversed.

R~•

H3C ~
(A) (B)
864 Appendix. Tentative Rules for the Nomenclature of Carotenoids

In an acyclic end group, the methyl group which is trans to the main
skeletal chain is numbered 16, and the methyl group which is cis is numbered 17,
as shown in (C).
3
17 HC CH····
c=t
"
3 " / 2

to H 3C / H
(C)

Rule Carotenoid 13. Trivial names

The preceding rules are designed to define precisely the structure of a given
carotenoid by its name. Use of the semi-systematic names derived from these
rules will greatly assist communication between scientists and enable work
to be more readily retrieved from the literature.
The appendix contains a list* of the trivial names, currently in use, for
naturally occurring carotenoids; it also gives their semi-systematic names
and structures. These trivial names will be of value in natural-product and
biochemical work but their use in organic or systematic work should be
restricted. If trivial names are used in a paper, the semi-systematic name
should always be given, in parentheses or in a footnote at the first mention.
Whilst the need to coin a new trivial name must occasionally arise (e.g.,
because the structure of the compound is unknown) the list* in the appendix
should not be unnecessarily enlarged, nor should simple derivatives of known
carotenoids be named by use of the trivial name or given a new one.
* In this book replaced by Chapter XII, Section A.

Author Index
Numbers in parentheses are reference numbers and indicate that an author's work is referred to
although his name may not be cited in the text. The complete references are listed at the end of
each chapter.
A Allen, M. B., 44 [174, 176]
Aasen, A. 1., 34 [ 42], 37 [80], 38 [97], 46 [209],
48 [226, 230], 53 [226], 63 [16], 65 [17],
69 [105], 77 [16, 105], 79 [105, 184], 82 [195],
83 [195], 85 [105, 184, 212], 87 [17, 184],
88 [105, 212], 91 [212], 98 [284], 123 [378],
126 [184, 284], 129 [184], 134 [105],
150[195], 153[184, 195], 164[212], 166[17],
174[16], 177[16], 270[30], 414[326],
416 [326], 452 [369a], 500 [326], 512 [369a],
538 [369 a], 556 [369 a], 557 [326],
559 [369 a], 626 [335]
Abrahamson, E. W., 725 [65], 726 [71],
728 [83], 729 [71, 87], 730 [71]
Achenbach, H., 38 [95], 40 [95], 52 [95],
117[413]
Acker, L., 749[41], 750[41], 751 [65]
Ackermann, W., 760 [176]
Adachi, M., 355 [145]
Adams, R. G., 725 [64], 727 [76]
Addicott, F. T., 527 [430, 432, 435-437]
Adler, K., 702 [225]
Agranoff, B. W., 583 [66, 69], 584 [73]
Ahluwalia, B., 733 [119]
Ahmad, B., 606 [225]
Ahmad, R., 167 [502], 403 [315], 420[333,
334], 429 [344], 430 [344], 431 [333],
439[334], 466[315, 384], 467[315],
469[315], 471 [315], 542[315], 544[315]
Ahrens, R., 168 [508], 281 [74], 433 [350],
470[350],543[350],544[350]
Aitzetmiiller, K., 44[179, 181, 182, 184],
46 [204], 53 [283], 83 [201], 91 [248],
106 [248, 325], 107 [325], 113 [537],
157[201], 161 [549], 163 [549], 251 [48]
Akhtar, M., 402 [310], 463 [310], 464 [310],
469[310],504[310], 517[310], 519[310],
558[310], 560[310], 728[80, 82, 84]
Aksnes, G., 393 [264J
Aldag, H.-J., 430[347], 482 [406], 520[406],
529[406],531[406],545[406]
Aldrich, P. E., 579 [11]
Alkazov, D. G., 600 [168]
Allais, A., 397 [293]
Allen, F. H., 295[130]
Carotenmds 55
865
Althausen, T. L., 720 [18], 721 [18]
Ames, B. N., 673 [21]
Amesz, 1., 687 [117], 693 [174], 695 [185],
696[185], 697[199]
Andersen, K., 34[49a], 37 [49a], 53 [49a],
91 [253], 92 [253], 119 [253], 120 [253],
121 [253], 647 [ 49]
Anderson, D. G., 31 [12], 580[20], 581 [20],
583 [20], 584 [20], 591 [124]
Anderson, I. C., 605[199], 674[34, 37-39]
Anderson, J. M., 67 [51]
Anderson, 0. R., 735 [155], 736 [155]
Anderson, R.F., 35[55, 56], 615[289]
Anderson, W. A., 239 [26]
Andre, D., 43 [159], 111 [330], 162 [330],
300[141]
Andree, F., 400 [299, 306, 307], 428 [343],
438 [343], 470 [299], 471 [306, 307],
543[299], 544[306, 307]
Andresen, S., 79[190], 86[190],
148 [ 455]
Andrewes, A. G., 32 [15], 33 [15], 46 [212,
213], 53 [213], 174 [524, 529], 257 [54],
259 [54], 268 [11], 618 [293, 296, 298],
619 [298]
Andrews, J. S., 721 [ 41], 731 [101, 102]
Antia, N.J., 289[113]
Aoe, H., 754 [95]
Arcamone, F., 41 [140], 75 [144], 78 [144],
152[144], 157[144], 336[67, 68], 499[67,
68], 552 [67, 68], 555 [67, 68]
Archer, B. L., 581 [35], 587 [105]
Arend, W., 369 [203], 395 [290]
Arens, 1. F., 16, 340 [93], 355 [93], 366 [189],
369[189,202],376[212],387[189,242 ,
243], 405 [212]
Arigoni, D., 295 [129], 580[13]
Arnaki, M., 583 [63]
Arnold, M., 35 [55, 56], 615 [289]
Arnold, W., 687 [121]
Arpin, N., 34[47], 40[124, 125], 46[199a],
52[47, 124, 125], 75[151], 79[207],
84[204], 85[207], 104[206, 206a],
117[546], 126[208], 128[394], 130[207,
208, 452], 149 [204], 151 [206a, 464],
866 Author Index
153[423], 155[207], 174[532], 176[532],
273 [ 42], 289 [ 42], 762 [217]
Arvanitaki, A., 702 [227]
Ascarelli, 1., 750[52], 751 [52], 752 [52]
Askar, A., 757 [119]
Askar, C., 757 [119]
Asmundson, C. M., 534 [ 443]
Attenburrow, J., 79 [185], 332 [48], 337 [48],
368 [ 48], 374 [ 48], 375 [ 48], 408 [ 48]
Audley, B. G., 581 [35]
Augustine, R. L., 85 [210]
Aust, H., 747 [34], 757 [116]
Austin, D. G., 707 [256]
Austin, D.J., 50[245], 270[35], 312[35, 178],
598 [157], 615 [283], 616 [283], 707 [257,
258]
Avron, M., 687[124]
Ayers, J. E., 51 [261]
Ayyoub, N.l., 721 [42]
Azuma, M., 727 [73]
B 545 [206]
Babtist, J. N., 751 [71]
Bacharach, A. D. E., 725 [67]
Bader, F., 74 [136], 78 [136], 372 [208],
393 [208], 482 [208], 483 [208], 504 [208],
554 [208], 560 [208]
Baer,H.H., 78[169]
Bagdon, R. E., 641 [27]
Bailey, G. F., 44[170], 66[19, 22-26, 31],
89[237], 111 [22, 24], 112[24], 114[347],
269 [27], 762 [216], 763 [218]
Bailey, W. C., 51 [269]
Baillarge, M., 362 [ 170]
Baker, E.G., 754[98]
Balasubramaniyan, P., 730[97, 98]
Balasubramaniyan, V., 730 [97, 98]
Baldas,J., 251 [49], 262[49], 263[49],
264[65]
Ball, S., 380 [224]
Baltscheffsky, M., 695 [190]
Bamji, M.S., 71 [112], 602 [182], 603 [182],
663 [118], 664 [118], 679 [72]
Bancher, E., 757 [130-132]
Bannister, T. T., 691 [154]
Bara, M., 699 [204]
Barber, M.S., 34[39], 42[147], 74[132],
78 [174], 121 [371], 122 [373], 123 [373],
124[386], 137[132], 204[16], 209[16],
275 [47], 278 [62], 279 [62], 280[62],
292 [118], 293 [118], 337 [70], 400 [309],
401 [309], 416 [309], 423 [309], 455 [309],
456 [309], 465 [309], 468 [309], 469 [386]:
501 [70, 309], 509 [309], 514 [309],
515[309], 516[70], 539[309], 540[309],
542 [309], 543 [309], 553 [309], 556 [309],
558 [70], 560 [309], 561 [70, 309]
Barbier, M., 43 [159], 44[191], 45 [191],
106 [323], 107 [323], 111 [330], 162 [330],
300 [141]
Barksdale, A. W., 706 [252]
Barnard, D., 587 [105]
Barnett, H. L., 35 [54], 579 [8], 614 [272]
Barnett, H. M., 760[158]
Bart, J.C.J., 285[89, 91], 286[89, 91], 287[91]
Bartels, P.O., 675[43, 45], 685[45]
Bartlett, L., 91 [247], 95 [247], 289 [114],
290 [114], 291 [114], 292 [114], 300 [114],
301 [114], 302 [114], 305 [114], 306 [114],
307[114], 308[114], 309[114], 310[114],
311 [114], 312 [114], 341 [106], 360 [106],
508 [106], 546 [106]
Bartov, 1., 749 [37, 38], 751 [75]
Bartram, K., 286 [93], 328 [ 4], 366 [191],
369 [191], 371 [206], 402 [206], 480 [206],
Bass, J. L., 276 [51]
Bassham, J. A., 604 [ 195]
Bates, R.B., 275 [48], 276[52]
Batra, P.P., 596[151], 620[301], 676[57]
Battaile, J ., 581 [38]
Battersby,A.R., 584[81]
Bauernfeind, J. C., 641 [28], 745 [10-12],
748 [264], 749 [73, 89], 751 [57-59, 73, 264],
752 [59], 753 [89], 754 [92], 755 [102, 103],
757[204], 760[160, 169, 181], 761 [12, 102,
103,204,206], 762[204,206]
Baxter, J. G., 328 [32], 380 [225]
Baxter, R. M., 63 [15]
Bazhanova, N. V., 600[168], 602[176]
Beall, G., 747 [22]
Beckman, L.D., 623 [316]
Beeler, D. A., 584 [82, 83], 591 [122, 128],
596[147]
Beevers, H., 604[190]
Belie, 1., 328 [6], 484 [6], 486 [6], 545 [6]
Bell, W.D., 675[48]
Bellin, S.A., 533 [441]
Ben-Aziz, A., 602 [172], 610[254], 627 [342],
628 [342]
Benedict, C. R., 623 [316], 627 [340]
Benk, E., 755 [105], 759 [152], 764 [239-247,
253-256]
Ben-Shaul, Y., 606 [224], 607 [224]
Benson, A. A., 704 [235]
Bentley, K. W., 21 [39]
Bentley, R., 91 [254], 92 [254]
Benton, C. H., 386 [238, 238 a]
 Author Index 867
Benton, F. L., 44 [184], 69 [93-95], 106 [325], [213],95[272-279,282,545], 101[272,278,
107 [325] 279, 282], 114[279], 133 [409], 146[116, 187,
Benz, F., 167 [505] 446, 448], 149[275], 150[462], 151 [272,
Benz, W., 243 [30], 247 [30], 262 [63] 467], 157[40, 490], 204[14], 312[177]
Berg, M. H., 376 [212], 405 [212] Boettger, H., 46 [206], 48 [223], 133 [ 406]
Bergan,J.G., 737[168] Bohlmann, F., 286 [93], 328 [3, 4, 14],
Berger, R., 750[53] 366 [191], 369 [191, 201], 371 [206],
Berges, 0., 400 [304] 387 [201 ], 402 [206], 480 [206], 482 [ 406],
Bergman, K., 700 [209], 701 [209] 483[3], 484[3], 520[406, 421], 529[406],
Bergmann, R., 759[152], 764[247] 531[406,424], 545[3,206,406]
Bergmann, W., 706 [251] Bohlmann, M., 369 [201], 387 [201]
Bern, H. A., 736[161] Boling, J. A., 720 [26]
Berns, D. S., 701 [212] Bolliger, H. R., 67 [54, 57, 58], 68 [57, 77],
Berthod, H., 287 [94] 205 [18], 209 [18], 235 [18], 248 [18],
Bertin, D., 397 [293] 252[18], 260[18], 263 [18]
Berzelius, J.J., 13 [7], 606 [218] Bonaly, R., 46 [200], 129 [396], 612 [270],
Besson, G., 578 [3, 4] 614[270]
Bestmann, H.J., 389 [250], 474 [394], 545 [394] Bonner, J., 580 [25], 581 [37], 584 [37],
Bey, G. von der, 168 [508], 281 [74], 433 [350, 591 [25]
351],470[350],543[350], 544[350] Bonnett, R., 43[151-153], 44[153], 49[242],
Beytia, E., 581 [39], 584 [39] 75[146], 95[146], 104[317], 107[146],
Bianchi, M., 614[275] 110[146], 111[146], 148[317], 151[317],
Bickoff, E. M., 66 [20], 747 [30], 762 [216], 168 [317], 251 [50], 260 [50], 285 [82],
763 [218] 289[111], 295[111], 296[111], 297[111],
Biedermann, R., 752 [77] 299[111], 300[111], 415[327], 452[327],
Bielig, H.-J., 21 [40] 456 [327], 511 [327], 512 [327], 513 [327],
Biemann, K., 243 [29], 265 [29] 514 [327], 538 [327], 539 [327], 548 [327],
Bieri,J.G., 732[113], 733[119], 734[148] 549 [327], 552 [327], 553 [327]
Biffi, G., 614 [275] Bonting, S. L., 725 [62], 727 [62, 75]
Bijvoet, J. M., 299 [140] Booth, V.H., 31[1], 63[7, 10], 69[99],
Bilz, R., 755 [100] 116[356], 764[231]
Bird, F. H., 752 [266] Borchgrevink, N.C., 757 [134]
Bishop, N. I., 691 [156, 158, 159] B~rdalen, B., 45[196], 72[124], 107[124],
Black, C. C., 687 [125] 251 [51]
Black, D. M., 337 [72] Borenstein, B., 755 [104], 757 [104], 759 [104],
Blain,J.A., 757[114] 760[162], 761 [198, 205], 762[205]
Blass, U., 67 [51] Borggreven, J. M.P. M., 725 [62], 727 [62]
Blatz, P. E., 190 [1], 727 [77], 728 [77], Bork, S., 370 [205], 431 [349], 482 [ 406],
730[97, 98] 520[406], 529[406], 531[406], 545[406]
Blessin, C. W., 763 [225] Bornstein, S., 749 [37, 38], 751 [75]
Bletner, J. K., 752 [265], 754 [93] Boschetti, A., 31 [8], 584 [87]
Blinks, L. R., 670 [ 4], 674 [30], 687 [ 4, 30, 116, Bose, A. K., 51 [269]
118] Bosshard, A., 333 [53]
Bloch, E., 306 [165], 341 [105] Bouchez, M.P., 104[206a], 151 [206a],
Bloch, K., 581 [32, 33, 45], 584 [72], 586 [103] 166[541]
Blomstrand, R., 720[19] Bougon, M., 748 [260], 753 [260]
Blosse, P. T., 728 [80, 82] Bovey, R. W., 675 [ 46], 685 [ 46]
Bluhm, H.( 341 [103] Bowden, K., 430 [346]
Blum, H. F., 671 [13], 679 [13] Bowden, R., 295 [126]
Blum, W.P., 380[225] Bownds, D., 725 [61], 728 [79, 81], 729 [86]
Bob bit, J. M., 67 [56], 68 [56] Braconnot, H., 13 [5]
Boch, J., 351 [125], 44,3 [367], 448 [367], Braekkan, 0. R., 724 [59]
536[367],537[367] Braithwaite, G. D., 579 [6], 580 [6], 582 [6],
Bodea, C., 37 [84], 38 [108], 42 [150], 66 [ 40], 590[6], 623[315]
71 [115, 116], 76[156], 79[187], 81 [40], 85 Brandsma, L., 376 [212], 405 [212]
868 Author Index
Brannock, K. C., 393 [279]
Branson, R. E., 748 [262]
Braude, E. A., 284 [79]
Breckoff, W.E., 332[51], 350[51]
Bredt, J., 295 [128]
Breidenbach, R. W., 67 [65], 604 [190]
Brenner, M., 303 [149], 684 [99], 710 [99]
Bricout, J ., 51 [265, 266], 52 [265]
Bridges, C. D. B., 732 [109]
Briggs, W. R., 699 [205], 702 [222]
Bril, C., 689 [ 132]
Britton, G., 32[32], 34[30a, 41, 50], 46[205],
89 [230], 103 [295], 116 [230], 123 [544],
139[422], 148[449], 157[422], 311[176],
588 [107], 589 [107, 108], 592 [107, 108,
130, 131], 593 [107], 596 [152], 597 [155],
599 [158], 600 [152, 161], 601 [170],
602[170-172], 603[155, 161], 605[198],
608 [253], 610 [253, 254], 624 [324],
625 [324], 626 [326, 327, 332, 333],
627[342],628[342]
Brixius, L., 764 [253-255]
Brockmann, H., 37 [83], 49 [233], 66 [ 4 7],
77[158], 78[178, 179], 86[179], 87[158],
116 [357], 130 [ 400-404], 133 [ 404],
146 [158], 167 [500], 606 [217]
Brodie, J ., 580 [ 16]
Brooks, J., 604 [197], 708 [260-262],
709[260, 264]
Brown, B. 0., 32 [15], 33 [15], 36 [78],
46 [212], 149 [ 460], 174 [524], 257 [54],
259 [54], 268 [11], 269 [22], 273 [22],
274 [22], 276 [55], 277 [22], 618 [293]
Brubacher, G., 744 [ 4], 747 [20], 748 [61, 62],
750[50], 751 [61, 62], 763 [223], 764[50,
223,235,236]
Brunnmiiller, F., 394 [282]
Brzezinka, H., 248 [39], 251 [39], 259 [39],
260 [39], 264 [39]
Buchanan, G. A., 675 [ 47]
Buchecker, R., 306 [167], 308 [167 a],
341 [ 107], 364 [ 107], 484 [ 107], 486 [ 107],
487 [107], 489 [107], 546 [107], 547 [107],
548 [ 107], 597 [ 156]
Biichi, G., 394[275]
Buchta, E., 400[299, 306, 307], 428 [343],
435 [356], 438 [343], 470[299], 471 [306,
307], 543[299], 544[306, 307]
Buchwald, H.-E., 696[195]
Buchwald, M., 39 [112], 290 [116], 291 [116]
Budick, E. M., 759 [153]
Budzikiewicz, H., 243 [31], 248 [39], 251 [39],
252[31], 259[39], 260[31, 39]
Buggy, M.J., 584[85a], 589[108], 592[108],
605 [198]
Bu'Lock, J.D., 50[245], 270[35], 312 [35,
178], 598[157], 615[283], 616[283],
707 [256-258]
Bunnell, R. H., 745 [12], 748 [264], 751 [59,
264], 752 [59], 755 [104], 757 [104, 204],
759[104], 760[160, 169, 181], 761 [12, 198,
204-206], 762[204-206], 764[237]
Bunning, E., 699 [201]
Bunt, J. S., 67 [63]
Burchard, R. P., 680 [77]
Burden, R.S., 50[248a], 305[142a]
Burger, B. V., 49 [242]
Burke, P. V., 700[209], 701 [209]
Burness, D. M., 362 [174], 376 [214], 385 [174],
393 [174]
Burnett, J. H., 670 [5], 674 [5], 705 [5]
Burns, E. R., 675 [ 47]
Burrell, J. W. K., 276 [53]
Bush, W. V., 95 [283], 101 [283]
Businger, A., 362 [ 173]
Buss, C. D., 35 [57], 615 [284, 286]
Buten, B., 72 [123], 657 [94]
Biitler, R., 578 [5], 579 [5]
Butler, W. L., 687 [126]
Byers, G. W., 728 [78]
c
Cabell, C. A., 747 [26]
Caglioti, L., 615 [278, 279]
Cahn, R.S., 312[181-184], 313[181-184],
316[181, 182]
Cahnmann, H., 306[165], 341[105]
Cainelli, G., 615 [278, 279]
Calaustro, E. Q., 733 [ 118]
Calvarono, M., 764 [249]
Calvin, M., 67[51], 684[98], 690[139, 146]
Cama, H. R., 718 [7], 733 [126]
Camerino, B., 41 [140], 75 [144], 78 [144],
152 [144], 157 [144], 336 [67, 68],
499 [67, 68], 552 [67, 68], 555 [67, 68],
615 [278, 279]
Cameron, A. F. B., 79 [185], 332 [48], 337[48],
368 [ 48], 374 [ 48], 375 [ 48], 408 [ 48]
Campbell, S.A., 44[191], 45[191], 106[323],
107 [323]
Campbell, T. W., 428 [342]
Capeder, A., 762 [207, 209]
Carington, T. R., 340 [92]
Carlson, S.D., 734[151]
Carnahan, H. L., 763 [227]
Carns, H. R., 527 [ 430, 432]
Carroll, J., 733 [122], 734[122, 150]
Carroll, M. F., 339 [80, 81]
 Author Index
Carter, M. C., 675 [ 47]
Cawley, J.D., 338 [74], 352 [128], 359 [128],
380[128, 225]
Ceccaldi, H. J., 39 [115-117], 657 [95, 99],
659[95, 99, 112], 660[112], 663[112],
703 [233]
Cederberg, E., 35 [59]
Cerda-Olmedo, E., 700 [209], 701 [209],
707[255]
Chalazonitis, N., 702[227]
Chance, B., 68[78], 676[53, 54], 692[171],
693 [53, 54, 175, 177, 179], 696 [196]
Chandler, B. V., 580[18]
Chang, J. L., 89 [235]
Chang, M. L., 737 [167]
Chang, Y. C., 677 [67], 678 [67, 95], 682 [67,
95,96],683[67,96],684[96],685[95,96]
Chanley, J.D., 332 [ 47], 337 [ 47]
Chaplin, C. E., 51 [263]
Chapman, D.J., 35[63], 44[171, 172],
104[303], 106[320, 321]
Chapman, J. H., 79 [185], 332 [ 48], 337 [ 48],
368[48], 374[48], 375[48],408[48]
Chapon, L., 757 [140]
Charley, H., 757 [134]
Charlton, J. M., 585 [93], 588 [107], 589 [107],
592[107], 593[107],605[93]
Chase, G. 0., 338 [77, 78], 339 [77, 78]
Chatterjee, G. C., 734 [134]
Chatterjee, 1., 734 [134]
Chaykin, S., 581 [32, 33]
Chechak, A.J., 337 [73], 338 [75], 352 [128],
370 [75], 380 [128], 388 [248], 425 [339],
454 [371], 457 [376], 462 [75], 464 [75],
474 [248], 476 [396], 477 [339], 478 [339,
396],480[339,403],520[403], 522[403],
529 [73, 75], 531 [75], 538 [371], 540 [376],
545 [248, 396], 549 [339], 554 [339, 403],
560 [339, 403]
Cheeseman, G. W. H., 364 [184], 408 [318]
Cheesman, D. F., 14[24], 21 [24], 39[110, 113,
115, 117, 118], 53 [110], 72 [118, 119],
73 [125], 657 [92, 95, 99, 100], 658 [92, 100],
659 [95, 99, 109], 660 [92], 661 [92],
663 [92], 703 [231, 232], 756 [112]
Chen, S. L., 687 [123]
Chessin, ~-, 680[81], 685[81], 694[81]
Chiba, N., 736 [159]
Chichester, C. 0., 32 [17], 35 [57, 58], 37 [88],
38 [88], 43 [164], 44 [164], 89 [234],
111 [332, 336], 123[336], 143[441],
506[418], 536[41.8], 545[418], 579[9, 10],
580[9, 17], 581 [47], 582[52-59],
583 [52-59, 68], 584 [68, 80, 85], 592 [85],
594[137], 600[165-167], 602[181],
869
603[181], 606[213], 612[266, 269],
613[266],615[284-286,288],616[290],
628 [345, 346], 629 [346], 649 [58, 59],
690[144, 145],695[144]
Cho, S., 111 [339], 277 [56], 304 [154]
Cholnoky, L., 36 [72], 42 [144, 149], 43 [155,
163, 164], 44[144, 164], 46[201, 203],
75 [138], 78 [175], 79 [261], 89 [242, 243],
91 [242, 243, 246], 111 [243, 246, 331, 332],
114 [243], 116 [243], 125 [261], 137 [261,
413a, 415], 139[175], 148[414, 454],
151 [466], 153[414, 454], 264[65], 270[29],
273 [29], 277 [29], 288 [95, 97], 289 [97],
293 [122], 295 [29], 296 [132], 300 [132],
302[29], 516[419], 559[419], 606[229],
607[229],690[140]
Chopard-dit-Jean, L. H., 68 [77], 205 [18],
209 [18], 235 [18], 248 [18], 252 [18],
260 [18], 263 [18], 276 [50], 400 [305],
403 [312], 436 [305], 494 [305], 545 [305],
546 [305]
Ciegler, A., 35 [55, 56], 614 [273], 615 [289],
707 [253]
Claes, H., 32 [25], 269 [14, 18], 585 [96],
589 [96, 113-115], 592 [ 113-115], 593 [96,
115, 134], 600[159], 676[51], 677[51,
62-64], 678[51, 62-64, 97], 682[63, 97],
683 [62, 63]
Clausen, J., 733 [127]
Clayton, R. K., 672 [15], 674[15], 677 [60],
678[60], 680[15], 693[176], 695[189],
701 [215, 216]
Cleland, W. W., 583 [67]
Cockbain, E. G., 581 [35], 587 [105]
Coggins, C. W., 596 [150]
Cohen-Bazire, G., 32 [14], 33 [14], 123 [374],
623 [313, 321], 671 [12], 672 [12, 18],
673 [22]
Coleman, J. W., 693 [178]
Colman, A.D., 731 [107]
Colman, B., 67 [64]
Colombi, L., 333 [53]
Conradie, W.J., 398 [295]
Conti, S. F., 627 [340]
Conway, W. G., 649 [73, 74]
Coon, M.J., 582[50]
Cooper, B.S., 709 [263]
Cooper, R.D.G., 41 [126], 78[175], 104[313],
137 [ 416], 139 [175], 285 [83], 293 [124],
295 [124], 299 [124], 336 [66], 337 [69],
497 [69], 499 [66], 501 [69], 516 [419],
522 [66], 523 [66], 546 [66], 547 [66],
548 [66], 549 [66], 559 [419], 562 [69]
Coover, H. W., 394 [273]
Corey, E.J., 681 [88]
870 Author Index
Cori, 0., 581 [39], 584 [39]
Cornell, D. G., 658 [103]
Cornforth, J. W., 304[156], 305[156, 157],
527 [ 430, 433, 434, 438-440], 584 [86],
585 [86, 101, 102], 586 [101, 102], 587 [86,
10~ 105], 588[1011 589[101]
Cornubert, R., 332[45, 46]
Cornwell, D. G., 720 [21]
Costes, C., 36 [67], 89 [241], 591 [127],
600 [164], 756 [111]
Cotta, E., 41 [140], 75 [144], 78 [144],
152 [144], 157 [144], 336 [68], 499 [68],
552 [68], 555 [68]
Couch, J. R., 747 [27, 28], 749 [79], 752 [79, 80]
Coulson, C. A., 286 [92]
Courtnay, H. V., 748 [262]
Coward, W. A., 732 [110], 733 [117], 737 [173]
Crain, F. D., 720 [24]
Creff, R., 757 [140]
Creger, C. R., 752 [80]
Crescitelli, F., 724 [58], 731 [58]
Criegee, R., 167 [501]
Critchley, J.P., 163 [497]
Crofts, A. R., 696 [ 191]
Crounse, J. B., 677 [60, 61], 678 [60, 61]
Crozier, G. F., 37 [87], 155 [ 471]
Curl, A. L., 36 [74], 42 [145], 44[170],
49 [238], 66 [18-34], 78 [171], 85 [32],
89 [237], 111 [22, 24, 28, 30, 335, 341],
112[24], 114[34, 347], 116[33], 155[171],
163[496, 498], 169[32, 498], 269[27],
639 [15]
Curry, G. M., 699 [203], 700 [207, 210]
Cymerman, J., 403 [311]
Czeczuga, B., 649 [63]
Czerpak, R., 649 [63]
Czygan, F.C., 38 [107], 39 [107], 143 [440],
151 [ 470], 156 [470], 157 [ 470, 492], 585 [971
603 [183], 607 [232], 612 [264], 628 [347,
348],645[33-36,38,39,43],649[60]
D Decker, K., 596[145]
Dabbagh, A. G., 44 [156a, 169], 76 [154, 155],
111 [154], 112 [155, 534]
Daemen, F.J. M., 725 [62], 727 [62, 75]
Dahle, L., 757 [128]
Dahmen, A., 249[41]
Dale, J., 201 [10], 202 [10]
Dales, R. P., 37 [89]
Dallacker, F., 412 [324]
Daile, J.-P., 303 [148], 335 [65], 337 [65],
341 [65], 343 [65], 344 [113], 360 [168],
527 [65], 684 [100-102], 710 [101]
Dangye-Caye, M.P., see Bouchez, M.P.
Daniel, A. F., see Faludi-Dimiel, A.
Daoud, H. N., 757 [125]
Dartnall, H. J. A., 724 [55]
Das, B. C., 49 [243]
Daumas, R., 39 [116], 659 [112], 660 [112],
663 [112], 703 [233]
David, C. N., 700 [209], 701 [209]
David, J. S. K., 721 [3 7]
Davies, A.J., 305 [164], 306 [164]
Davies, B. H., 31 [10, 11], 32 [28-30], 34 [28,
30], 35 [53], 37 [88], 38 [88], 42 [146],
43[146], 63[4], 65[4], 66[4], 67[4], 68[4,
72, 73], 69 [100], 70 [ 4], 103 [292],
124[388], 260[61], 261 [61], 584[89, 90],
585 [89], 594 [138], 596 [144], 609 [248],
618 [299], 622 [317], 623 [317], 624 [325],
625 [325], 626 [325], 628 [346], 629 [346],
649 [58, 59]
Davies, R. E., 747 [27, 28]
Davis, E. A., 754[93]
Davis, J. B., 14 [22], 31 [13], 41 [126],
89 [240], 101 [286], 103 [290], 104 [313],
113 [240], 124 [385], 137 [240], 143 [240],
146[240], 148[385], 155[240], 204[16],
209 [16], 268 [10], 269 [10], 275 [10, 47],
276 [10], 285 [83], 328 [29], 336 [66],
400 [296], 401 [296], 421 [296, 335],
423 [296], 426 [296], 428 [296, 335],
454 [296, 335], 477 [296, 335], 478 [296,
335], 489 [296, 335], 490 [296, 335],
499 [66], 509 [296, 335], 517 [296],
521 [296], 522 [66, 296], 523 [66], 529 [296],
538 [296, 335], 546 [66], 547 [66], 548 [66],
549 [66, 296, 335], 550 [296, 335], 589 [109],
623 [320]
Davis, P., 328 [6], 484 [6], 486 [6], 545 [6]
Davis, P. N., 747 [19]
Davydova, L. P., 361 [169], 388 [245 a]
Day, E. J., 749 [39], 753 [91]
Day, W. C., 244 [35]
deBruijn, H., 681 [85]
DeJongh, S., 751 [64]
Dekker, E. E., 582 [50]
DeLaMar, R.R., 759[151]
Delbri.ick, M., 596[148], 700[208, 209],
701 [209, 211], 707 [211]
del Campillo, A., 582 [50]
DeLuca, H. F., 720 [33], 721 [35, 36],
722 [33, 35, 36], 734 [143], 737 [33, 36, 170,
171]
DeLuca, L., 735 [153], 736 [163]
de Man, J. M., 759 [150]
Dembiczak, C. M., 747 [22]
 Author Index
Demole, E., 51 [262, 268]
De Nicola, M.G., see Giudici de Nicola, M.
Denney, D. B., 393 [265]
Dennison, D. S., 701 [209]
Denny, R. W., 677 [67], 678 [67, 95], 681 [94],
682 [67, 94-96], 683 [96], 684 [96],
685 [95, 96]
Denticedi Accadia, F., 645 [41, 42]
Deobald, H.J., 757 [126]
de Riel, J. K., 730 [99]
De Ritter, E., 641 [26, 28], 745 [10], 751 [57],
764[237]
Deroche, M.E., 756[111]
Dershowitz, S., 394 [274]
Deschreider, A. R., 757 [129]
Desplanques, D., 734 [130]
Deuel, H.J., 658 [102]
Deufel, J., 754 [97]
Deutsch, A., 49 [232], 78 [165], 87 [227],
146 [165], 167 [227], 288 [105], 308 [105]
DeVille, T. E., 43 [157], 45 [157], 95 [280],
109 [280], 297 [138], 302 [138], 303 [139,
151], 304[139], 305 [139], 307 [139],
309 [139], 310[139], 311 [139], 600[163]
de Waard, A., 581 [32, 45], 586 [103]
Dewhurst, P. B., 728 [80, 82], 730 [97, 98]
Dialer, K., 281 [72], 379 [217], 403 [217]
Dickey, J. B., 394 [273]
Diehn, B., 702 [221]
Diemair, W., 757 [117]
Dieterle, J. M., 342 [108], 380 [225]
Dietl, R., 764 [255]
DiGiacomo, A., 757 [135]
Dilley, R.A., 696[193]
Diment, J., 342 [109], 360 [109], 489 [109],
490[109], 510[109], 551 [109], 552[109],
554[109]
Diner, B., 46[205], 139 [422], 157 [422]
Dingle,J.T., 704[237], 735[154], 736[160]
Dinovo, E. C., 725 [ 68]
Djerassi, C., 243 [31 ], 252 [31 ], 260 [31]
Doering, W. v. E., 312 [179]
Dolge, K. L., 747 [22]
Donninger, C., 585 [101, 102], 586 [101, 102],
587 [102], 588 [101], 589 [101]
Donohue,H.V., 111[336], 114[336], 123[336],
602 [181], 603 [181]
Dornand)., 363[177], 377[177], 405[177]
Dorough, G. D., 69,0[139]
Dorsey, J. A., 583 [70]
Dorsey, J. K., 583 [70]
Douglas, C. R., 754 [93]
Dowling, J. E., 16, 720 [27], 721 [34], 723 [34],
730 [34], 731 [100, 106], 732 [34], 737 [34]
Draber, W., 304[156], 305[156], 527[439]
871
Drake, D., 50[245], 312 [178], 615 [283],
616 [283], 707 [257]
Drischel, W., 332[51], 350[51]
Driscoll, W., 757 [204], 761 [204], 762 [204]
Drozdova, N.N., 677[65, 66], 678[66]
Drumm, P.J., 86[215, 216]
Dua, P. N., 753 [91]
Dubal, V. L., 753 [91]
Duden, R., 757 [137]
Duggar, B. M., 687 [119]
Dunagin, P. E., 721 [ 46]
Durr, I. F., 580 [14]
Dutton, H.J., 686[109], 687[119]
Duysens, L. N. M., 687 [114, 115], 692 [170]
Dvolaitzky, M., 295 [129]
Dworkin, M., 673 [23, 24], 680 [77]
Dyer, M.A., 751 [76], 753 [76], 763 [76]
E
Eaton, H. D., 747 [22]
Eberhard, D., 620 [305]
Eberle, M., 580 [13]
Eddinger, C. C., 352 [128], 380 [128]
Edisbury, J. R., 49 [241]
Edmunds, F. S., 244 [33], 249 [33]
Egger, K., 38 [109], 41 [143], 43 [158],
44 [156a, 158, 169, 183, 185], 46 [158, 207],
67 [59-62, 66], 68 [61, 66-70], 76 [154, 155],
107[535], 111 [154], 112[155, 534],
113[535, 536], 116[354], 139[420, 421],
141 [433, 434], 142[433], 143 [433, 439],
269 [28], 273 [28], 756 [108]
Egger, K. W., 249 [ 43]
Eggerer, H., 581 [34], 583 [66, 69], 584 [73, 74]
Egli, R. H., 51 [265, 266], 52 [265]
Ehrenberg, L., 583 [60]
Ehrreich, S. J ., 703 [228]
Ehtisham-ud-din, A. F. M., 704 [239]
Eichenberger, W., 606 [216], 755 [106, 107]
Eichhorn, E. L., 285 [85] ·
Eichmann, G. G., 338 [78], 339 [78]
Eidel'man, Z. M., 600[168], 602[175]
Eimhjellen, K. E., 33 [37], 124 [387], 623 [323],
624[323],625[323],626[330,331,337]
Eisner, T., 50[254], 303[145], 535[448],
709[265]
Eiter, K., 328 [35], 345 [117], 351 [35],
352 [35, 130], 357 [35], 362 [35, 175, 176],
363[175, 178], 364[183], 366[35],
379 [183, 220], 380 [35], 384 [230], 386 [175],
394 [276], 424 [338], 425 [338, 340],
430 [338], 483 [338, 408], 484 [338],
487[338],545[338,408],549[338]
872 Author Index
Ellis, N. R., 747 [26]
El Ridi, M. S., 268 [ 4, 5]
Emerick, R. J., 720 [33], 722 [33], 737 [33, 171]
Emerson, R., 686[110, 111]
Emmons, W. D., 393 [270]
Emrich, H. M., 696 [192]
Engel, B. G., 615 [287]
Engelhard, H., 393 [269]
Engelmann, T. W., 686 [105, 106, 112],
687 [105, 106], 701 [213], 702 [218]
Enggist, P., 51 [262, 268]
Englert, G., 68 [77], 205 [18, 19], 209 [18, 19],
210 [19], 213 [19], 226 [19], 235 [18],
239 [19], 245 [19], 248 [18, 19], 249 [19],
250[19], 251 [19], 252 [18], 259 [19],
260[18], 263 [18], 280 [65], 531 [ 425],
535[425], 536[425]
Entschel, R., 42[148], 69[101], 78[176],
79 [192], 95 [288], 98 [192], 137 [176],
146[288], 151 [192], 156[192, 488a],
292 [119], 531 [ 428]
Enzell, C. R., 39 [121], 40[121], 45 [196],
66[41], 69[41], 72[124], 79[41], 83[203],
85[41], 87[41], 88[41], 107[124], 161 [41],
171 [41], 173[41], 246[37], 248[40],
250[40, 45, 46], 251 [37, 47, 51], 252[37],
253 [37], 257 [37, 53], 259 [37, 53], 260 [37],
261 [37], 262 [37], 285 [84]
Erdman, J. G., 244 [35]
Erickson, K., 50 [254], 303 [145], 535 [ 448],
709 [265]
Erikson, J. 0., 727 [77], 728 [77]
Ernst, R., 239 [26]
Ernster, L., 679 [71]
Escapini, H., 722 [ 49], 738 [ 49]
Escue, R. B., 271 [38], 278 [60], 279 [60]
Eugster, C. H., 44 [168], 49 [239], 69 [101],
103 [294], 104 [304, 305], 111 [342],
116 [353, 355], 168 [507, 543], 269 [23],
306 [166, 167], 308 [167 a], 328 [2], 341 [107],
364[107, 179, 179a], 391 [258, 259],
392 [261], 424 [337], 425 [341], 429 [345],
430 [345], 462 [261], 464 [261], 483 [2, 407],
484[107, 179, 179a, 337, 341], 486[107,
179, 179a, 337], 487 [107, 337, 341,
409-411], 489 [107, 409-411], 522 [258, 259,
421 a], 523 [421 a], 524 [258, 259], 525 [261],
526 [261], 531 [ 427, 428], 545 [2], 546 [107,
407, 409, 410), 547 [107, 179, 179a, 337, 411],
548[107, 179a], 549[337, 341], 564[258,
259, 261], 597 [156]
Euler, H. von, 16, 72[121], 107[121], 114[348],
133[408], 169[512], 171 [512]
Evans, M. B., 687 [124]
Evans, R. H., 79 [185]
Evans, R. J., 747 [25]
Evans, R.M., 332[48], 337[48], 368[48, 194],
374 [ 48], 375 [ 48], 408 [ 48], 420 [332]
F
Fabre, R., 171 [522]
Faigle, H., 93 [270], 137 [ 415 a], 293 [120, 121]
F aigle, J. W., 294 [ 125]
Faltis, F., 167 [504], 268 [1]
Faludi, B., 674 [33], 677 [33], 678 [33]
Faludi-Dimiel, A., 583 [60], 605 [202],
674 [33], 677 [33], 678 [33]
Farr, F. M., 752 [80]
Farrar, K. R., 369 [200]
Fassler, C., 747 [20, 35]
Faust, M., 269 [23], 391 [259], 564 [259]
Favell, D.J., 764[234]
Feichtmayr, F., 199 [7]
Feldbau, E., 306 [165], 341 [105]
Feldbruegge, D. H., 69 [103], 584 [84],
585 [84]
Feldman, R. P., 677 [60], 678 [60]
Fell, H. B., 735 [157], 736 [162]
Fenical, W., 304[152]
Ferguson, T. M., 747 [28]
Fernandez, H.R., 734[152]
Ferrando, R., 641 [29, 30], 744[5], 750[44],
753 [88]
Fetizon, M., 79 [189], 157 [189]
Feuser, B., 620 [308]
Fiasson, J.L., 104[206a], 151 [206a],
153[423], 166[541], 762[217]
Fidge, N.H., 719 [11, 15, 17], 721 [15, 17]
Fields, C. L., 720 [26]
Firch, J. G., 763 [224]
Fischer, D. R., 747 [32]
Fischer, R., 764[248]
Fish wick, M. J., 51 [261], 606 [210]
Fisscher, L. G. M., 352 [133, 134], 354 [134]
Fitzpatrick, T. B., 686[104], 710[104]
Fleischer, S., 725 [63], 727 [63]
Fleischman, D.E., 695 [189]
Fles, D., 328 [6], 438 [360], 484 [6], 486 [6],
494 [360], 545 [6]
Fletcher, G. L., 393 [278]
Folkers, K., 579 [11]
Foote, C. S., 303 [149], 671, 677 [67], 678 [67,
95],681[87, 89-91,94],682[67,94-96],
683 [96], 684 [96, 99], 685 [95, 96], 694 [91 ],
710 [99]
Foppen, F. H., 36 [66], 37 [85], 104 [307],
589 [118]
Fork, D.C., 687[117], 693[174], 697[199,200]
 Author Index
Fornasir, V., 339 [83]
Forssberg, A., 679 [71]
Forsythe, R. H., 750 [ 46-48]
Foster, K. W., 700[209], 701 [209]
Fox, D. L, 38 [100, 101, 106], 39 [100, 101],
63 [13], 141 [ 427], 143 [ 427], 146 [ 442],
638[4, 5, 7, 8], 641, 645[40], 647[50],
649[64, 69, 70, 81, 83], 650[83], 652[81],
654 [81], 655 [83], 656 [86]
Fox, H. M., 21 [38]
Fradkin, L I., 605 [202]
Frame, H. D., 66[45]
Franceschi, G., 41 [140], 75 [144], 78 [144],
152 [144], 157 [144], 336 [67, 68], 499 [67,
68], 552 [67, 68], 555 [67, 68]
Francis, F.J., 759[151]
Francis, G. W., 34[49a], 37 [49a], 39 [121],
40[121], 45[196], 48[226], 53[49a, 226],
66[41], 69[41], 72[124], 75[151], 79[41],
83[202], 85[41, 212], 87[41], 88[41, 212],
91 [212, 253], 92 [253, 262], 93 [271],
104 [206a], 107 [124, 326], 119 [253],
120 [253], 121 [253, 365], 126 [202],
127 [202, 365], 129 [202], 151 [206a, 464],
153 [202], 161 [ 41], 164 [212], 169 [517],
171 [ 41, 365], 173 [ 41, 365], 174 [532],
176[262, 532], 177[262], 246[37], 248[40],
250 [ 40], 251 [37, 51], 252 [37, 40], 253 [37],
257 [37, 53], 259 [37, 53], 260 [37], 261 [37],
262[37],285[84],645[49]
Franck, J., 672 [14]
Fredriksen, G., 82 [199]
Freer, J. H., 704 [238]
French, C. S., 691 [157], 702 [223]
Freudenberg, K., 293 [123]
Freyschlag, H., 341 [99, 100], 357 [100],
394 [284-286], 441 [364, 365], 443 [366],
444 [364, 366], 446 [364], 448 [366],
536[364-366],537[364,366]
Frey-Wyssling, A., 606 [221], 756 [110]
Friedrich, G., 400[304]
Friend, J., 163 [ 497]
Froning, J. F., 338 [76]
Fiihrer, J., 50 [256]
Fujimori, E., 662 [115], 680 [78, 80], 685 [78]
Fujita, E., 761 [184]
Fujiwara, K., 393 [271], 394 [271]
Fukami, H., 51 [264]
Fukui, H., 50 [249].
Fukui, S., 134[411]
Fiiller, W., 762 [213]
Furchgott, R. F., 703 [228]
Furia, T. E., 761 [201]
Futterman, S., 721 [41], 723[51], 731 [51, 101,
102, 105]
873
G
Gaffron, H., 680 [79]
Gaide- Huguenin, A. C., 725 [ 61]
Galasko, G., 43 [155, 159], 91 [246, 247],
95 [247], 111 [246, 330], 116 [246], 162 [330],
270 [29], 273 [29], 277 [29], 289 [114],
290 [114], 291 [114], 292 [114], 295 [29],
300[114, 141], 301 [114], 302[29, 114],
305[114], 306[114], 307[114], 308[114],
309 [114], 310[114], 311 [114], 312 [114],
341[106], 360[106],508[106], 546[106]
Gale, D. M., 275 [ 48], 276 [52]
Galliard, T., 604[197]
Galmiche, J. M., 694[182]
Galston, A. W., 699 [202, 204]
Gammack, D. B., 659[110]
Ganguly,J., 36[64], 140[426], 658[102],
719[14,15], 720[14, 30], 721 [15, 37, 40, 44],
722 [30], 733 [116, 125], 734 [125, 128, 129,
133]
Garbers, C. F., 49 [242], 385 [233, 235],
387 [244], 398 [295], 483 [ 407], 487 [ 411],
489 [ 411], 522 [ 421 a], 523 [ 421 a], 536 [244],
546[407], 547[411], 720[20]
Garcia, A. F., 662[113]
Garner, R.J., 46 [210], 618 [292], 619 [292],
620[292]
Garwood, R.F., 276[53]
Gay, E. A., 355 [149], 386 [149]
Gaylord, N. G., 85 [209]
Geisendorff, A., 750 [56]
Geison, R.L, 734[139], 737[139]
Geller, S., 734 [138]
Gerhardt, W., 177 [533]
Gewitz, H.-S., 691 [152]
Ghosh, N.C., 734[134]
Giannone, L, 757 [124]
Giannotti, C., 49 [243]
Gibas, J., 123 [337], 333 [62], 342 [62],
360 [167], 372 [167], 389 [252], 390 [167],
408 [167], 456 [374], 477 [167], 479 [167],
480 [167], 489 [167], 492 [167], 504 [167],
507 [167], 513 [374], 514 [374], 523 [167],
524 [167], 539 [374], 545 [252], 550 [167],
551 [167, 252], 553 [374], 556 [374],
557 [167, 374], 558 [167, 252], 560 [167],
561 [167], 563 [167], 564 [167]
Gibbs, M., 687 [125]
Gibbs, M. H., 579 [12]
Giesberger, G., 626 [329]
Gilchrist, B. M., 37 [89, 90, 92], 629 [350],
649 [56], 661 [124]
Gillam, A. E., 268 [ 4, 5]
Gillingham, J. T., 763 [222]
874 Author Index
Girshin, A.B., 600[168]
Giudici de Nicola, M., 38 [98], 45 [98],
142[437], 143[437], 647[55]
Glick, D., 734[144]
Gloor, U., 754[96]
Glover, J., 606 [212], 638 [1, 2], 640[2],
641 [2], 718 [8]
Go, G., 89 [235]
Goad, L.J., 593 [132], 609[245]
Godnev, T.N., 600[160]
Goedheer, J.C., 687 [127], 688 [129, 131],
689[133, 135, 136],690[129]
Goff, 0. E., 754 [93]
Goldsmith, T. H., 43 [166], 44 [166, 180],
111 [334], 141 [435], 734[152]
Goldstein, W.l., 720[18], 721 [18]
Goldstrohm, D. D., 680 [75]
Golfier, M., 79 [189], 157 [189]
Goodchild, D.J., 675 [44]
Goodell, E. W., 700[209], 701 [209]
Goodfellow, D., 289 [115], 311 [176a]
Goodman, D.S., 584[75], 640[17, 19],
719[9, 11, 15, 17], 720[19, 22, 29], 721 [9,
15, 17, 29, 45]
Goodwin, T. W., 12 [2], 15 [2], 21 [37, 42],
31 [4, 5, 10, 11], 32[4, 27, 32], 33[4, 5],
34[30a, 40, 41, 50], 35 [4, 5, 51, 53], 36[4,
5], 38[4, 5], 39[4], 41[4, 5], 42[4, 5],
43 [5], 44 [172, 174], 46 [205], 52 [5], 53 [5],
63 [3], 66 [3], 68 [72, 73], 69 [3, 100],
79 [193], 89 [230], 102 [289], 103 [295],
104[301], 106 [321], 116 [230], 123 [375,
379, 544], 124 [193, 375], 126 [375],
139[422, 424], 140[424], 148[375, 379,
449], 157 [ 422], 174 [528], 276 [55], 280 [66,
67], 311 [176], 380[224], 579[6], 580[6,
23], 581 [29-31, 40], 582 [6, 51], 583 [51,
61], 584 [89, 90], 585 [89, 93], 586 [104],
588[107], 589[107, 108, 116, 117], 590[6,
116, 117], 591 [117], 592 [107, 108, 116, 130,
131], 593 [107, 131 a], 594 [138, 139],
596[152-154], 597[155], 599[158],
600[152, 161], 601 [170], 602[170-172],
603 [153-155, 161], 605 [40, 200, 207],
606 [211], 607 [230], 608 [253], 609 [245],
610[254, 256,257,260, 261], 612[31, 262,
263], 614[271, 277], 615 [277, 282],
618 [295], 622 [311, 312], 623 [314, 315,
319],624[324],625[324],626[326-328,
332, 333], 627 [342], 628 [342], 670 [3, 6-8],
674 [6, 7], 688 [130], 701 [217], 705 [242],
707 [254], 745 [8], 746 [8], 747 [8], 756 [8]
Gordon, A., 603 [186]
Gordon, S. A., 680 [77], 687 [125]
Gore, T.S., 21 [41]
Gottfried, L., 674 [28]
Gould, R.G., 355[155]
Graebe, J. E., 585 [94]
Graham, W., 387 [243]
Grandolfo, M., 44[182, 184], 46[204],
53 [283], 66 [49], 83 [201], 106 [325],
107 [325], 113 [537], 157 [201], 161 [549],
163[549],251[48]
Grangaud, R., 734[130]
Grant, D. M., 242 [27]
Grassner, H., 341 [100], 357 [100]
Grau, R., 759 [154]
Gray, J.C., 581 [41], 605 [41]
Green, A., 168 [543]
Green, J., 649 [56, 61]
Green, M., 662[113]
Greenblatt, E., 703 [228]
Grein, A., 41 [140], 75 [144], 78 [144],
152 [144], 157 [144], 336 [68], 499 [68],
552 [68], 555 [68]
Gribanovski-Sassu, 0., 36 [66], 37 [85],
104[307], 589 [118], 645 [41, 42]
Griffiths, A. E., 594 [143], 607 [235]
Griffiths, M., 585 [100], 671 [12], 672 [12, 16],
684[16]
Grigg, R. W., 63 [13]
Grimwood, B. E., 91 [250]
Grob, E. C., 31 [7, 8], 91 [244], 114[244],
141 [431], 578[1-5], 579[5], 580[19],
584 [77, 87], 606 [216], 756 [106, 107]
Grogan, C. 0., 749 [39]
Gronowska-Senger, A., 719 [13]
Grossi, F. X., 344 [ 115], 364 [ 115]
Gruen, H. E., 700 [207]
Grundmann, C., 36 [75], 41 [133], 86 [219]
104[314], 149[456, 457], 151 [468],
168 [509], 288 [96]
Gruner, B.J., 276[52]
Grutzner, J. B., 242 [28]
Guerrero, C., 48 [222, 223], 133 [ 406]
Guex, W., 46 [199], 128 [393], 166 [393],
281 [72], 379 [217], 400[298], 403 [217],
405 [298], 411 [298], 457 [375, 378],
458 [298, 375], 459 [378], 460[378],
461 [298, 378], 472 [389], 511 [298],
517 [378], 519 [378], 540[375, 378],
541 [298, 375, 378], 542 [298, 378],
544[389], 563[298,378], 762[214]
Gugelmann, W., 87 [225], 137 [225], 169 [514]
Gugliemelli, L.A., 747 [31]
Guilluy, R., 40 [125], 52 [125], 130 [ 452]
Giinther, H., 759 [154]
Gurd, F. R., 658 [104]
Gurin, S., 579 [7], 580 [7], 581 [7], 583 [7]
Gurinovich, G. P., 688 [128]
 Author Index
Gutmann, H., 261 [62], 281 [77], 333 [60],
388 [247], 393 [262], 394 [247, 262],
405 [247], 406 [247], 410 [247, 319, 320],
411 [247], 412[60], 420[247], 423 [247],
446 [247], 447 [247], 450 [247], 458 [247],
460 [247], 467 [60, 247], 468 [247], 470 [247,
262], 471 [247, 262], 472 [247], 489 [247],
490 [247], 497 [60], 499 [60], 500 [247],
511 [247], 537 [247], 538 [247], 541 [247],
542[60, 247], 543[247, 262], 544[247, 262],
545 [60], 549 [60], 557 [247], 563 [247],
564[247]
Gyllenberg, H. G., 754[99]
Gyorgyfy, K., 42[144], 43[155, 163, 164],
44[144, 164], 89 [242], 91 [242, 246],
111 [246, 331, 332], 116[246], 270[29],
273 [29], 277 [29], 295 [29], 302 [29]
H Hayano, M., 600[162]
Haag, W., 393 [263]
Haagen-Smit, A.J., 581 [37], 584[37]
Habermann, H. M., 605 [204, 205], 674 [36]
Haeck, H. H., 366 [192], 440[192, 363],
444 [368], 457 [368], 536 [192, 363],
540[368]
Hageman, R.H., 604[192, 193]
Hagenbach, R., 50[256]
Hager, A., 63 [11], 68 [11], 107 [548], 199 [8],
603 [184, 185, 187], 662 [116], 692 [167],
695 [167]
Hall, M. 0., 725 [67], 733 [124]
Halldal, P., 702[219]
Haller, A., 332 [ 45]
Halverson, A. W., 747 [24]
Hama, T., 155[472]
Hamilton, R. H., 675 [ 48]
Hamlet, J. C., 369 [200]
Hamprecht, G., 395 [291]
Hanaoka, M., 52 [278], 332 [50]
Hanser, E., 749 [ 40]
Hara, Y., 296[135]
Harber, L.C., 686[104], 710[104]
Hardin, G., 44[173], 106[324], 163[324],
269 [26]
Hardisson, f..., 278 [62], 279 [62], 280 [62],
469 [386]
Harrington, T. M., 369 [198]
Harris, P. L., 737 [165]
Harris, R. C., 615 [282]
Harris, R. S., 18 [30] .
Hiirtel, H., 751 [257]
Hartmann, H., 352 [131]
Hartmann, M. L., 760 [ 158]
875
Hartshorn, M., 50 [254], 303 [ 145], 535 [ 448],
709[265]
Haslach, H., 641 [31], 745 [9]
Hastings, J. W., 681 [92], 685 [92]
Hata, Masahiro, 649 [57]
Hata, Mitsuo, 649 [57]
Haug, A., 45 [196], 63 [9], 72 [124], 107 [124],
251 [51]
Hausheer, W., 759 [156], 760 [156], 761 [156]
Hauska, G. A., 696 [194]
Havivi, E., 734[146]
Hawke, J. C., 604 [197]
Hawks, 0. D., 386 [239]
Hawrylyshyn, M., 67 [55]
Haxo, F., 32[16], 35[63], 38[102], 44[172],
53 [280, 283], 103 [293], 104 [303],
106[321], 141 [428], 161 [549], 163[549],
165 [539], 585 [98, 99], 644, 687 [118]
Hayaishi, 0., 640 [18], 719 [12]
Hayes, B. W., 720 [25]
Hayes, E., 760[170]
Heckman, R.A., 533 [441]
Heftmann, E., 66 [ 43]
Hege, B.P., 533[441]
Hegedus, B., 68 [74]
Hegge, E., 41 [137], 82[197], 83 [197],
104[197], 149[197], 151[197],285[81],
627 [343]
Heilbron, I., 13, 40[122], 53 [282], 87 [221],
119 [221], 139 [221], 171 [523], 281 [69],
288 [98], 328 [34], 344 [116], 345 [116],
355 [146], 364 [184, 185], 366 [190],
370[190], 403[311], 408[318], 430[346]
Heilbronner, E., 199[7]
Heisenberg, M., 700[209], 701 [209], 707 [255]
Helfenstein, A., 155 [477, 478], 268 [2],
288 [99]
Heller, J., 725 [66, 70]
Hellstrom, H., 72 [121], 107 [121]
Hemmer, E., 91 [255]
Hemming, F. W., 584[91], 587 [106]
Hems, B. A., 79 [185], 332 [ 48], 337 [ 48],
368[48],374[48], 375[48],408[48]
Hen best, H. B., 341 [101 ], 344 [101], 369 [200]
Hendrick, C. M., 747 [24]
Hendricks, S. B., 687 [124]
Hendry, L., 303 [146], 304 [155], 332 [52]
Henecka, H., 352 [130]
Henning, G.L., 596[150]
Henning, U., 581 [34], 583 [66, 69], 584 [73,
74]
Bennion, G. F., 338 [76]
Henriksen, R. A., 725 [68]
Herrick, R. B., 734 [147]
876 Author Index
Herring, P.J., 629 [349], 663 [122]
Hertzberg, S., 34 [ 48, 49 a], 37 [ 49 a, 81 ],
38[93, 96, 99, 104], 39[120, 121], 40[93, 96,
120, 121], 47[211], 53[48, 49a, 96], 66[41,
46], 69[41], 75[142, 150, 151], 77[46, 142,
150], 78[142, 172], 79[41,46, 142, 172],
82 [200], 83 [200], 85 [ 41], 87 [ 41, 142],
88[41], 91[46, 150,200,252, 253], 92[46,
150, 252, 253], 104 [ 46], 114 [200],
117 [252], 119 [ 46, 252, 253], 120 [253],
121 [253], 134[410], 139 [425], 141 [425],
142[436], 151 [410], 155[172], 161 [41,
493], 171 [41, 493], 173[41], 174[142],
233 [24], 257 [53, 59], 259 [52, 53], 260 [59,
60], 263 [52], 285 [84], 647 [ 44-49]
Herzig, J., 167 [504], 268 [1]
Hess, J. L., 704 [235]
Heumann, W., 157[492]
Higby, W. K., 764 [238]
Hill, H. M., 596 [146], 607 [236]
Hili,J. E., 749 [39], 753 [91]
Hindley, N.C., 281 [72], 379 [217],
403 [217]
Hirao, S., 155 [472]
Hirayama, K., 199[9]
Hirtenstein, M.D., 728 [84]
Hiyama, T., 68 [78]
Hodges, R., 51 [271]
Hodgkiss, W., 174[528], 618[295]
Hoffer, M., 86 [216], 333 [56]
Hoffman, C. H., 579 [11]
Hoffmann, R., 249 [ 42]
Holloway, P. W., 583 [71]
Holmes, E. A., 32 [28], 34 [28], 103 [292],
124[388], 260[61], 261 [61], 618[299]
Holyer, N. F., 69 [102], 75 [102]
Holzel, R., 40 [123], 101 [287], 171 [287],
251[49],262[49],263[49]
Hoover, M. W., 757 [122, 123]
Hoover, T., 753 [84]
Hopkins, T. S., 38 [100], 39 [100], 141 [ 427],
143[427], 146[427], 638[4], 645[40],
647 [50], 649 [83], 650[83], 655 [83]
Hora, J., 43 [159, 160], 49 [160], 75 [152],
111 [330], 162[152, 330], 300[141,
142], 303 [151], 305 [142]
Horii, Z., 52[278], 332[50]
Houser, A. R., 69 [103], 584 [84], 585 [84]
Hove, E.L., 734[137]
Howell, J. McC., 730 [91-93], 732 [93, 110,
114], 733 [91, 92, 114, 115, 117], 735 [92]
Howes, C. D., 596[151]
Hruban, L., 270 [35], 312 [35], 598 [157],
707 [258]
HSatava, M., 751 [67]
Hsu, W.-J., 37 [88], 38 [88], 628 [345, 346],
629[346],649[58, 59]
Huang, H. S., 640[17], 720[19, 22], 721 [45]
Hubbard, R., 704[234], 723[52], 724[52],
727 [74], 729 [86], 731 [104, 106, 107]
Huber, H., 249 [ 41]
Huber, W., 281 [72], 328 [1], 362 [173],
379 [1, 217], 403 [1, 217, 313, 314]
Hiibner, H., 85 [214]
Huff, J. W., 579 [12]
Huisgen, R., 249[41]
Huisman, H.O., 276[51], 352[126, 133, 134],
354[134], 369[126], 386[241], 387[126]
Hull, J. S., 393 [278]
Hummel, G., 395 [291]
Humphlett, W.J., 362[174], 385[174],
393 [174]
Hunter, M.I.S., 663[121]
Hiirlimann, H., 49[239], 168 [507]
Hursthouse, M. B., 43 [157], 45 [157], 77 [161],
95 [280], 109 [280], 297 [138], 299 [138,
139], 302[138], 303[139, 151], 304[139],
305 [139], 307 [139], 308 [139], 310 [139],
311 [139], 600[163]
Huschke, G., 759 [156], 760 [156], 761 [156]
Hyde, A., 675 [ 45], 685 [ 45]
Hyeon, S.B., 51 [267, 270], 303 [147]
I
Ichikawa, H., 51 [267], 303 [147]
Ikeda, R. M., 51 [269]
Illyes, G., 38 [108], 42 [150], 66 [ 40], 71 [115],
81 [40], 85 [213], 133 [409], 157 [40, 490],
312[177]
Impellizzeri, C., 641 [27]
Ina, K., 51 [264]
Ingersoll, R., 534 [ 443]
Ingold,C.K., 312[181-183], 313[181-183],
316 [181, 182]
Inhoffen, H. H., 14, 168 [508], 281 [74, 76],
282 [76], 286 [93], 328 [3-5, 14, 15],
345 [118], 366 [191], 369 [191, 201, 203],
370 [118, 205], 371 [206], 387 [201],
391 [118], 400[304], 402[206], 430[347],
431 [349], 433 [350, 351], 436[357, 358],
470[350, 387], 471 [387], 480[206],
482 [ 404, 406], 483 [3, 5], 486 [3], 494 [ 414],
496[414], 506[118, 418], 520[406, 420,
421], 529[406], 531 [406, 424], 536[418],
543 [350, 387], 544 [350, 387], 545 [3, 5,
206,404,406, 418], 546[118], 551[414]
Inui, M., 50[249]
Irmscher, K., 400[304]
 Author Index
Irreverre, F., 725 [69], 729 [69]
Irving, C. S., 728 [78]
Ishii, H., 51 [272]
Ishii, R., 583 [64]
Ishikawa, S., 155 [475, 476], 344[114]
Ishikawa, Y., 354 [143], 355 [150]
Ishizaka, 0., 760 [174]
Isler, 0., 18 [31], 19 [35], 36 [65], 46 [199],
49 [240], 68 [77, 79], 74 [135, 136],
78 [136], 79 [186], 87 [135], 103 [296],
104[296, 306], 128[393], 129[397],
130[399, 405], 146[444], 151[186, 469],
155[405, 483], 156[186], 166[393],
168[508], 169[397], 191[4], 205[18],
206 [20], 208 [20], 209 [18], 235 [18],
246 [38], 248 [18], 251 [20], 252 [18],
260 [18], 261 [20, 38, 62], 263 [18], 270 [33],
276[50], 281 [68, 71, 72, 74, 77], 289[109,
110], 291 [109, 110], 297[71], 328[1, 7-10,
13, 16-19, 22-24, 31, 37], 329 [24, 38],
332 [41, 44], 333 [60], 340[89, 94], 344 [1],
345[41, 44,119, 120, 122], 347[44],
355 [153], 357 [153, 164, 165], 360 [153],
362[172], 364[41, 119, 122, 186-188],
365[119], 372[41, 119, 186,187, 208],
377[216a], 379[1, 164,217, 221], 380[164],
385 [165], 388 [245, 247], 391 [186, 260],
393 [208, 262, 280], 394[247, 262], 398 [165,
294],400[298,303,305], 403[1,217,280,
312-314], 405[247, 298], 406[247],
410 [247, 321], 411 [247, 298], 412 [60, 294],
420[247], 421 [303], 422 [153], 423 [247],
431 [280], 433 [350], 435 [354], 436 [305],
437 [ 44, 359], 438 [245], 442 [280], 445 [188,
369], 446 [247], 447 [247], 448 [188],
450[188, 247], 457 [375, 378], 458 [247,
298, 375], 459[13, 378], 460[247, 378],
461 [298, 378], 462 [13, 382], 463 [13, 382],
467[60,247],468[247,38 5],470[247,262,
350], 471 [247, 262], 472 [247, 389],
474 [245], 477 [23], 479 [23], 480 [186],
482[41, 187,208,245,260, 405], 483 [41,
187, 208], 489 [247], 490[247], 494[305,
359], 496[44], 497 [60], 499[60, 416],
500 [247], 503 [385], 504 [153, 208],
506[186, 187,245, 260], 507[245, 260],
508 [153], 511 [23, 153, 247, 298], 512 [153],
516 [321],'517 [188, 378], 518 [303, 321],
519 [188, 378], 525 [321, 359], 526 [321,
359], 527 [164], 536 [188, 280], 537 [188,
247, 369], 538 [188, 247], 540 [375, 378],
541[13,247,298,375,378 ],542[60,247,
298,378],543[247,262,3 50,385],544[247,
262, 350, 389], 545 [60, 153, 186, 245, 260,
305, 359], 546 [153, 305], 547 [153, 303],
877
549 [60, 416], 550 [23, 245], 551 [187, 245],
554[41, 187,208,359, 405], 555[321],
557 [188, 247], 559 [321], 560[208],
562 [385], 563 [23, 44, 247, 298, 378, 385],
564 [247, 321, 359], 639 [9], 641 [22, 23],
718 [5], 755 [101]
Isoe, S., 51 [267, 270], 303 [147]
Ito, M., 52[278], 332[50]
J
Jackman, L. M., 31 [13], 34 [39, 46], 41 [137],
42 [147], 74 [132], 78 [174], 79 [217],
82 [197], 83 [197], 86 [217], 101 [286],
103 [290], 104 [197], 121 [371], 122 [373],
123 [373], 124 [217, 385, 386], 125 [217],
126 [217], 137 [132, 416], 148 [510],
149 [197], 151 [197], 204 [16], 209 [16],
213 [23], 268 [10], 269 [10], 275 [10, 47],
276 [10, 53], 278 [62, 63], 279 [62], 280 [62],
285 [81], 292 [118], 293 [118, 124],
295 [124], 299 [124], 337 [69, 70], 400 [296,
309], 401 [296, 309], 416 [309], 421 [296,
335], 423 [296, 309], 426 [296], 428 [296,
335], 454 [296, 335], 455 [309], 456 [309],
465 [309], 468 [309], 469 [386], 477 [296,
335], 478 [296, 335], 489 [296, 335],
490 [296, 335], 497 [69], 501 [69, 70, 309],
509 [296, 309, 335], 514 [309], 515 [309],
516 [70], 517 [296], 521 [296], 522 [296],
529 [296], 538 [296, 335], 539 [309],
540 [309], 542 [309], 543 [309], 549 [296,
335], 550 [296, 335], 553 [309], 556 [309],
558 [70], 560[309], 561 [70, 309], 562 [69],
589 [109], 623 [320], 627 [343]
Jackson, H., 40[122], 53 [282], 171 [523]
Jackson, J. B., 696 [ 191]
Jacobs, H. A.M., 376[212], 405 [212]
Jacobsberg, B., 757 [139]
Jaeger, L., 91 [249], 146[443]
Jaffe, H. H., 273 [45], 274[45]
Jagendorf, A. T., 687 [124]
Jager, A., 762 [208]
Jager, H., 614 [276], 615 [276]
James, T.L., 249[43]
Jamikorn, M., 174 [528], 591 [120], 594 [139],
607 [230], 618 [295]
Jansen, A.B.A., 79[185], 332[48], 337[48],
368 [ 48], 374 [ 48], 375 [ 48], 408 [ 48]
Jautelat, M., 242[28]
Jeffrey, S. W., 44 [175, 176], 53 [280], 68 [92]
Jencks, W. P., 39 [112], 72 [123], 290[116],
291[116],657[93,94]
Jenkin, P., 649 [75]
878 Author Index
Jenkins, J. A., 593 [133], 608 [238, 244],
609 [242, 244]
Jennings, W. H., 727 [76]
Jensen, A., 14 [18], 43 [154, 162], 45 [195],
63 [5, 6], 65 [6], 66 [6, 39], 68 [5, 6, 87,
89-91], 77[160], 84[39], 87[160], 92[262],
103 [5], 107 [39], 110 [39], 111 [328],
137 [5], 176 [262], 177 [262], 295 [131],
328 [25], 697 [198]
Jensen, S. L., see Liaaen-Jensen, S.
Ji, T. H., 704 [235]
Johannes, B., 248 [39], 251 [39], 259 [39],
260 [39], 264 [39]
John, J., 718 [7]
John, K. V., 718 [7]
Johnson, A. W., 344 [116], 345 [116], 364 [185]
Johnson, B. C., 536 [ 450], 733 [121, 131],
734[121, 131, 135, 139], 736[159], 737[139,
167]
Johnson, F. H., 50 [253]
Johnson, K. W., 610 [252]
Johnstone, R. A. W., 244 [33], 249 [33]
Jones, D., 31 [10], 68 [73], 584 [89], 585 [89]
Jones, E. R. H., 284 [79], 344 [116], 345 [116],
355 [146], 364 [184, 185], 366 [190],
369[200], 370[190], 403[311], 408[318],
420 [332], 430 [346]
Jones, R.N., 40 [122], 53 [282], 171 [523]
Joshi, B.S., 21 [41]
Jucker, E., 12 [1], 14, 31 [3], 33 [3], 35 [3],
36 [3], 39 [3], 41 [3], 42 [3], 49 [3], 53 [3],
63 [2], 66 [2], 69 [2], 74 [2], 75 [2, 148],
76 [2], 77 [2], 84 [2], 87 [2], 89 [2, 236],
95[2], 102[2], 103[2], 104[2], 111 [360],
113 [2, 343, 344], 114 [236, 345, 346, 348,
349], 116 [350, 352, 359], 137 [2], 143 [2],
146 [2], 155 [2, 148], 167 [2], 168 [2],
278 [61], 284 [61], 288 [61, 104], 300 [144],
328 [20], 370 [204], 387 [204], 670 [1, 2]
Julia, M., 355 [146], 362 [170]
Julietti, F.J., 91 [250]
Juneja, H.S., 733 [116], 734[129]
Jungalwala, F. B., 268 [12], 584[79], 589 [110],
718 [7]
Junge, W., 696[192]
Juniper, B. E., 606 [222]
K 526[261], 531 [427, 428], 545 [2, 399],
Kabbe, H.-J., 364 [183], 379 [183], 424 [338],
425 [338, 340], 430 [338], 483 [338, 408],
484 [338], 487 [338], 545 [338, 408],
549 [338]
Kadin, H., 751 [58]
Kahlenberg, O.J., 750[45]
Kahn, A., 604[190]
Kahn, J. S., 691 [153-155]
Kahn, S.G., 536[450]
Kaiser, S., 338 [78], 339 [78]
Kakutani, Y., 591 [125, 126]
Kampe, D., 400 [304]
Kanai, M., 720[29], 721 [29]
Kandutsch, A. A., 584 [72]
Kanemitsu, T., 754[95]
Kargl, T. E., 75 [149], 103 [149], 594 [141,
142], 607 [141, 231], 608 [142, 243],
609[243], 763[224]
Karmakar, G., 93 [268], 104 [268]
Karmas, G., 355 [156], 377 [216], 403 [216],
404 [216]
Karrer, P., 12[1], 13, 14[16], 16,31 [3],
33 [3], 35 [3], 36 [3, 68], 39 [3], 41 [3, 141]~
42[3, 141, 148], 44[168], 45[193], 46[1971,
49 [3], 53 [3], 63 [2], 66 [2], 69 [2, 101],
74 [2], 75 [2, 137, 148], 76 [2], 77 [2],
78 [167, 176], 79 [191, 192], 84 [2], 85 [214],
87 [2, 214, 223-225], 88 [229], 89 [2, 231,
236], 91 [249], 93 [266, 270, 271], 95 [2,
231, 288], 98 [192], 102 [2], 103 [2, 266,
294], 104[2, 304,305, 312], 111 [342, 360,
361], 113[2, 231, 344], 114[236, 345,346,
348, 349], 116 [350-353, 358, 359, 362],
119[364], 121[229, 366-369], 122
[229], 123 [229, 369], 128 [224, 392],
133 [ 407, 408], 137 [2, 176, 225, 415 a],
143 [2], 146 [2, 223, 288, 443, 445],
148 [366], 151 [192], 155 [2, 148, 474-480,
484], 156[167, 192,488, 488a], 157[191],
160[506], 167[2, 505], 168 [2, 362, 543],
169 [512-514], 171 [512], 206[21], 208 [21],
268[2], 269[23], 278[59, 61], 284[61],
288 [61, 99, 102, 104], 289 [112], 292 [119],
293 [120, 121], 294 [125], 300 [143, 144],
306 [166], 308 [175], 328 [2], 364 [179,
179a, 180], 370[204], 387[204], 391[258,
259], 392[261], 424[337], 425 [341],
429 [345], 430 [345], 462 [261], 464 [261],
477 [399, 400], 479 [ 400], 483 [2, 407],
484[179, 179a, 180,337, 341], 486[179,
180, 337], 487 [337, 341, 409-411],
489[409-411], 522[258, 259, 421a],
523 [ 421 a], 524 [258, 259], 525 [261],
546[407, 409, 410], 547[179, 179a, 337,
411], 548[179a, 180], 549[337, 341],
550[400], 564[258, 259, 261], 606[214],
670[1,2],690[137]
Karrer, W., 21 [36], 117 [363]
Karunakaran, A., 591 [123]
 Author Index
Karunakaran, M.E., 591 [123]
Kasha, M., 681 [93]
Kass, E. H., 686 [104], 710 [104]
Katayama, T., 43 [164], 44 [164], 111 [332],
143[441]
Kato, M., 760[161], 761 [182, 192, 200]
Katsumura, S., 51 [267], 303 [147]
Katz, J.J., 44 [179, 181, 182, 184], 46 [204],
53 [283], 69 [93-95], 83 [201], 91 [248],
106 [248, 325], 107 [325], 113 [537],
157 [201], 161 [549], 163 [549], 251 [48],
602 [169]
Kaufman, S., 394 [277]
Kaufmann, W., 156[487], 289[108]
Kautsky, H., 681 [85, 86]
Kay, R.E., 605 [206], 610 [255]
Kayser, F., 612 [267]
Ke, B., 662[113], 696[193]
Kean,E.L., 734[145]
Kearns,D.R., 304[152, 153]
Kekwick, R. G. 0., 581 [36, 41], 605 [41]
Kelemen, G., 674 [33], 677 [33],
678 [33]
Keller, H. J., 759 [146]
Keller, V., 578 [3]
Kelly, M., 47[219], 68[85], 70[85], 71 [85],
75 [85], 77 [85, 163], 78 [85], 79 [190],
82 [85], 83 [85], 86 [190], 123 [85, 163],
148 [ 455], 174 [85, 163]
Kemp, T. R., 51 [263]
Kennedy, D., 702 [226]
Kennedy, M., 736 [159]
Kenyon, R. L., 340 [86]
Kersten, J. A. H., 687 [120]
Kessel, I., 581 [34], 584 [74]
Kessler, E., 143 [ 440], 585 [97]
Khan, A. U., 681 [93]
Kharasch, M.S., 331 [ 40]
Khare, A., 45 [191 a]
Khodzhaev, A., 602 [175]
Khorlina, I. M., 361 [169], 388 [245a]
Khosla, M.C., 104[312], 364[180], 484[180],
486 [ 180], 548 [ 180]
Kieckebusch, W., 757 [120]
Kikuchi, R., 155 [ 472]
Kimbel, R. L., 725 [68], 728 [83]
Kimel, W., 338 [77, 78], 339 [77, 78]
Kirk,J.T.O., 42[146], 43[146], 604[196],
606[222],609[246,248],615[282]
Kirschner, K., 31 [7], 584[77]
Kisaki, T., 604[192-194]
Kitamura, S., 379 [22.2], 384 [229]
Kito, Y., 727 [73], 729 [88]
Kj¢sen, H., 48 [225], 53 [283], 103 [538],
149[455a], 161 [549], 163[549], 167[499],
879
250[45], 416 [329], 465 [383], 515 [330a],
542[383], 553 [330a], 556[330a]
K!aui, H., 328 [7], 757 [141], 758 [259],
759[156, 157], 760[156, 161, 165, 166, 179],
761 [156, 157, 197, 199, 203], 762 [207, 209,
210]
Kleinig, H., 38 [94, 95], 40 [95], 44 [183, 185],
46[207], 52[95], 67[66], 68[66, 69, 71],
107 [535], 113 [535], 117 [413], 139
[420,421], 141[434],645[43]
Klyne, W., 91 [247], 95 [247], 289 [114, 115],
290 [114], 291 [114], 292 [114], 300 [114],
301 [114], 302 [114], 305 [114], 306 [114],
307[114], 308[114], 309[114], 310[114],
311 [114], 312[114], 341 [106], 360[106],
508 [106], 546 [106]
Knappe, J., 580[15]
Knowles, R. E., 36 [71], 747 [33, 87], 752 [81,
82], 753 [81, 85, 86], 763 [82, 227]
Knypl, J. S., 596 [149]
Kobayashi, A., 379 [222]
Kobrich, G., 332[51], 350[51]
Koch, B., 285 [86], 286 [86]
Koch, G., 747 [23]
Koch, H., 760 [177]
Koch, H. P., 284 [79]
Kochhar, D. M., 737 [166]
Koe, B. K., 93 [264, 265], 269 [13]
Koen,A.L., 720[31], 721[31],
723[31]
Koenig, H., 121 [368, 369], 123 [369],
155 [ 479]
Kofler, M., 68 [77], 205 [18], 209 [18],
235 [18], 248 [18], 252 [18], 260 [18],
263 [18], 281 [72], 328 [1], 344[1], 379 [1,
217], 403[1, 217, 312-314]
Kohi,F.G., 13[8]
Kohler, G. 0., 747 [33, 87], 749 [263], 752 [81,
82], 753 [81, 82, 85, 86], 763 [227]
Komissarov, G. G., 690 [148]
Kon, S. K., 268 [5]
Konaka, R., 79[188]
Kondo, K., 395 [289]
Konig, A., 67 [57, 58], 68 [57, 77], 205 [18],
209 [18], 235 [18], 248 [18], 252 [18],
260 [18], 263 [18]
Konig, H., 380 [226]
Kornfeld, F., 412 [324]
Korver, P.K., 276[51]
Koshimizu, K., 50 [249]
Koshland, D. E., 729 [90]
Koski, V. M., 674 [32]
Kosolapoff, G. M., 394 [272]
Koster, H., 333 [54]
Kovats, E., 51 [262]
880 Author Index
Krait, T., 366[192], 440[192, 363], 444[368],
457[368], 536[192,363],540[368]
Kramer, H., 477[400], 479[400], 550[400]
Krasnovskii, A.A., 677 [59, 65, 66], 678[66]
Krasovskaya, T.A., 602[173, 174], 690[141],
695 [141]
Kratzer, F. H., 747 [19]
Kratzer, 0., 389 [250], 474[394],
545 [394]
Krause, H.-J., 431 [349]
Krause, R.F., 35 [54], 579 [8], 614[272],
719[16], 720[24], 721 [16]
Krause-Voith, E., 111 [361]
Krauss, E. von, 289 [112]
Kreutzer, E., 606[221]
Krinsky, N.l., 36 [64], 43 [164, 166], 44 [164,
166, 180], 71 [110-112], 85 [111], 91 [245],
111 [332-334], 140[426], 141 [432, 435],
142[111], 602[179, 180,182], 603[182,
186], 658 [102, 103], 663 [118, 120],
664 [118], 665 [120], 670[10], 671 [10],
672 [17], 674 [10], 675 [ 49, 50], 676 [10],
678[49, 68], 679[10, 69], 680[10], 683[10,
68], 684 [17], 695 [186, 188], 704 [241],
731 [103]
Krippahl, G., 691 [149-152]
Krisammer, R., 749[40]
Krishna, H.J. V., 340[90]
Krishna Mallia, A., 718 [7]
Krogmann, D. W., 687 [125]
Kropf, A., 727 [74], 730 [99]
Kruger, F. A., 720[21]
Kruk, C., 276[51]
Krukar, R., 641 [26]
Krzeminski, L.F., 580[21]
Kucherov, V. F., 333 [57]
Kuhlmann, K. F., 242 [27]
Kiihn,H., 39[111], 72[122]
Kuhn, R., 13, 36[75], 37 [83], 39[111],
41 [142], 49 [232, 233], 52 [277], 66 [36, 37],
72[117, 122], 74[131], 75[139, 147],
77 [158], 78 [165, 168, 169, 178, 179],
79 [183], 86 [117, 179, 215, 216, 219],
87 [158, 227, 228], 88 [117], 116[357],
130[400-404],133[404],146[117,158,
165], 149[456, 457], 151 [468], 155[168,
481,482,485,486], 156[487,489],
167[227, 500],168[509, 542], 244[32],
268 [3], 281 [3, 76], 282 [76], 288 [96, 101,
103, 105], 289 [108], 308 [105], 333 [56],
606[217],656[87,88],658[87,88]
Kumamoto, J., 534 [ 443]
Kunisawa, R., 672 [19], 679 [19]
Kupiecki, F.P., 582[50]
Kurmann, B., 749[40]
Kushwaha, S.C., 581 [49], 591 [129],
592 [129a], 596 [147], 605 [129]
Kuzmicky, D. D., 752 [81], 753 [81, 85, 86]
K vakovszky, S., 753 [84]
L
Laber, L.J., 675 [48]
La Face, D., 306 [170]
Lakshmanan, M. R., 718 [7]
Lambertson, G., 724 [59]
Land, D. G., 51 [261], 79 [193], 123 [379],
124[193], 148 [379], 623 [319]
Lang, F., 605 [202], 677 [59]
Lang, K., 14 [23], 757 [120]
Langlet, J., 287 [94]
Lapitskii, G. A., 435 [355], 438 [355]
Larsen, B., 63 [9]
Laurence, J., 36 [67]
Lavery, H., 764 [234]
Laves, F., 276[49]
Law, J., 581 [33]
Lawrence, C. W., 720[24]
Lawrence, D.J., 736[161]
Lawrence, M.A., 725 [70]
Lebreton, P., 762 [217]
Lederer, E., 14[14, 15, 15a], 39 [119],
41 [142], 43 [159, 161], 44 [186, 187, 191],
45 [186, 187, 191], 46 [198], 49 [243],
66 [36, 37, 42], 68 [165], 103 [300], 106 [322,
323], 107 [323], 111 [322, 329, 330],
128 [300], 139 [329], 146 [165], 155 [482],
162[330], 171 [521, 522], 288 [101, 103],
300[141], 656[91]
Lederer, M., 66 [ 42]
Lee, C. P., 696 [196]
Lee, G. F., 68 [83]
Lee, S. W., 344[115], 364[115]
Lee, T.-C., 584 [85], 592 [85]
Lee, W.L., 14[24], 21 [24], 37[89-91],
39 [110, 114], 53 [110], 72 [120], 73 [125],
142[120], 151 [120], 156[120], 657[92],
658 [92, 107, 108], 660[92], 661 [92, 107],
663 [92, 119], 703 [231], 756 [112, 113]
Leermakers, P. A., 728 [78]
Lees, A. D., 703 [229]
Leff, J., 675 [50]
Leftwick, A. P., 40 [123], 79 [182], 85 [182],
101 [287], 134 [412], 141 [182], 143 [182],
171 [287], 251 [ 49], 262 [ 49], 263 [ 49],
456 [373], 463 [373], 464[373], 515 [373],
519 [373], 539 [373], 555 [373], 558 [373],
559[373]
Lehnert, L., 67 [54]
 Author Index
Leibner, G., 482 [ 406], 520 [ 406], 529 [ 406],
531[406], 545[406]
Leinweber,C.L., 675[46], 685[46]
Lemberg, I.K., 600[168]
Lempka, A., 757 [121]
LenhotT,H.M., 71[111], 85[111], 141[432],
142 [111], 663 [120]
LeRosen,A.L., 36[79], 149[461], 269[16],
271 [39], 289 [107], 593 [135], 606 [209],
607 [233], 608 [237]
Leuenberger, F., 257 [58], 259 [58], 642 [32],
645 [37], 649 [80], 650 [84], 655 [85],
658 [105]
Leuenberger, H.J., 49 [239], 168 [507]
Leumann, E., 79 [191], 157 [191]
Levi,A.S., 734[138, 141]
Levin, E., 584 [72]
Lewin, D. R., 730 [93, 96], 732 [93]
Lewis, C. M., 686 [110, 111]
Lewis, M.S., 725 [69], 729 [69]
Liaaen-Jensen,S., 14[17-19], 32[14], 33[14,
34-37], 34[36, 42, 43,46-49, 49a], 37 [49,
49a, 80, 81], 38[93, 96, 97, 99, 104], 39[120,
121], 40[34, 93, 96, 120, 121, 124], 41 [49,
130, 131, 135-137, 139, 139a], 45 [49, 195,
196], 46[49, 199a, 202, 208, 209, 211, 214],
47[49,211,216-219],48[225,226,230],
52 [34, 47, 124], 53 [ 48, 49, 49 a, 96, 226,
283], 63 [5, 6, 16], 65 [6, 17], 66 [6, 41, 46],
68[5, 6, 85, 90], 69[41, 105, 106], 70[85],
71[85, 106,108, 114], 72[124], 74[133],
75 [85, 141, 142, 150, 151, 153], 77 [16, 46,
85, 105, 106, 142, 150, 162-164], 78[85, 106,
142,166, 172, 181], 79[41, 46, 105,142, 172,
184,190, 194,207,217,260],82[85,
194-198, 200], 83 [85, 196-198, 200, 202,
203], 84[204], 85[41, 105,184, 207,212],
86 [190, 217], 87 [17, 41, 142, 184],
88 [41, 105, 106, 212], 91 [46, 150, 200, 212,
253, 255], 92 [46, 150, 253, 262], 93 [271],
98[284], 103[5, 538], 104[46, 108, 197,
206a, 315, 318], 107[124, 326], 111 [340],
114[200], 117[252], 119[46, 252, 253],
120[253], 121 [253, 365], 122[372],
123 [85, 106, 163, 198, 372, 374, 377, 378,
380, 381], 124 [194, 198, 217, 387], 125 [194,
217,260J, 126[133, 184,202,208,217,284,
390, 391], 127 [202, 365], 128 [394],
129 [184, 202, 398], 130 [207, 208], 133 [391],
134[105, 410], 137 [5], 139 [425], 141 [181,
425], 142 [181, 436], 146 [153, 447],
148[196, 380,453,455,510], 149[197,204,
453, 455a], 150[195, 463], 151 [197, 206a,
410,463,464], 152[166], 153[184, 195,196,
202, 453], 155[172, 207], 157[166], 161 [41,
Carotenmds 56
881
493, 549], 163 [549], 164 [212], 166 [17],
167[162, 499], 171 [41, 365, 493], 173[41,
164, 365], 174 [16, 85, 106, 114, 142, 163,
377, 526, 527, 531, 532], 176[262, 531, 532],
177 [16, 262], 202 [12], 233 [24, 25], 246 [37],
248 [40], 250[40, 45, 46], 251 [37, 51],
252 [37, 40], 253 [37], 257 [37, 53, 59],
259[37, 52, 53], 260[37, 59, 60], 261 [37],
262 [37], 263 [52], 270 [30, 31], 271 [37],
272 [31], 273 [31, 42], 278 [58], 285 [31, 81,
84], 289[42], 290[117], 328[25], 414[326],
416[326, 329], 417[330], 452[369a],
456 [330], 465 [383], 500 [326], 512 [369 a],
515[330, 330a], 538[369a], 539[330],
542 [383], 553 [330, 330a], 556 [330a, 369a],
557[326], 559[369a], 618[291, 294],
623[313, 321-323], 624[323], 625[323],
626 [330, 334-336], 627 [341, 343, 344],
647 [ 44-49], 670 [9], 674 [9, 198], 697 [198]
Lichton, I.J., 733 [118]
Lijinsky, W., 74 [130], 582 [51], 583 [51]
Lilly, V. G., 35 [54], 579 [8], 614 [272],
680[75, 76]
Lin, M., 730 [97, 98]
Lincoln,R.E., 32[26], 589[112], 590[112],
594 [112], 608 [112, 241]
Lindberg, M., 586 [103]
Lindberg, 0., 679[71]
Lindemann, 1., 620 [306]
Lindlar, H., 74[134-136], 78 [136], 79[186],
87 [135], 104 [306], 151 [186, 469], 155 [483],
156 [186], 261 [62], 281 [71, 77], 289 [109,
110], 297[71], 328[16], 332[41], 333[60],
340[89], 345[41], 364[41, 187], 372[41,
186, 187, 208], 388 [247], 391 [260],
393 [208, 280], 394 [247], 403 [280],
405 [247], 406 [247], 410[247], 411 [247],
412[60], 420[247], 423 [247], 431 [280],
442 [280], 445 [369], 446 [247], 447 [247],
450 [247], 358 [247], 460 [247], 467 [60,
247], 468 [247], 470[247], 471 [247],
472[247], 480[186], 482[41, 187,208,260,
405], 483[41, 187,208, 405], 489[247],
490 [247], 497 [60], 499 [60], 500 [247],
504 [208], 506 [186, 187, 260], 507 [260],
511 [247], 536 [280], 537 [247, 369],
538 [247], 541 [247], 542 [60, 247], 543 [247],
544 [247], 545 [60, 186, 260], 549 [60],
551 [187], 554 [ 41, 187, 208, 405], 557 [247],
560[208], 563 [247], 564[247], 641 [22]
Lindsay,J.K., 384[228]
Lingen, C., 679[71]
Linner, E., 103 [294], 425 [341], 484[341],
487 [341], 549 [341]
Lipp, M., 412 [324]
882 Author Index
Lippe!, K., 721 [38, 39], 732 [39]
Lippert, M., 44[168], 111 [342]
Liston, J., 174[528], 618 [295]
Little, C. 0., 720 [25, 26]
Little, E. P., 736 [163]
Livingston, A. L., 36 [71], 747 [30, 33, 87],
752 [81, 82], 753 [81, 85, 86], 762 [216],
763 [82, 216, 218, 227]
Livingston, R., 680 [78, 81 ], 685 [78, 81 ],
694[81]
Loeber, D. E., 32 [28], 34 [28], 103 [292],
124[388], 260[61], 261 [61], 303[150],
342 [109], 360 [109], 489 [109], 490[109],
510[109], 551 [109], 552[109], 554[109],
618 [299]
Loeblich,A.R., 53[279], 163[550]
Loewe, L., 87 [223], 146 [223]
Loncin, M., 757 [139]
Loomis, W.D., 581 [38]
Lopatkin, Yu. B., 690 [ 138]
Loran, M. R., 720 [18], 721 [18]
Lord,J.M., 604[189]
Lorenz, R., 352[130]
Losev, A. P., 688 [128]
Lotspeich, F.J., 579[8], 719[16], 720[24],
721 [16]
Low, I., 52 [277], 78 [168], 155 [168]
Lowry, L., 43 [164], 44[164], 111 [332, 336],
123 [336]
Li.ick, H., 761 [202]
Lucy,J.A., 704[237], 735[154, 156]
Luh,B.S., 757[125]
Lui, N. S. T., 735 [155], 736 [155]
Lukton, A., 579 [9], 580 [9, 18], 582 [57, 59],
583 [57, 59]
Lunde, K., 204[13], 275 [46], 278 [46], 280[46]
Lundegardh, H., 691 [160-166], 692 [165, 166]
Lwowski, W., 293 [123]
Lynch, V. H., 691 [157], 692 [172]
Lynen, F., 31 [7], 580[15], 581 [34], 583 [66,
69],584[73, 74, 77]
Lyon, J. L., 527 [ 430, 432, 435]
Lythgoe, B., 87 [221], 119 [221], 139 [221]
Lythgoe, J. N., 724 [55]
M 377 [124], 384 [124], 395 [124], 397 [124],
MacArthur, J. W., 608 [239]
McBeth, J. W., 161 [494], 162[494], 638 [8]
McCall, M.A., 394 [273]
McCance, R.A., 746[15]
McCann, C., 649 [76]
McCarty, R.E., 696[194]
McCollum, E. V., 16
McConnell, D. G., 725 [63]
McCormick, A., 43 [152, 153], 44[153],
75 [146], 77 [162], 95 [146], 107 [146],
110[146], 111 [146], 167[162], 251 [50],
260[50], 289[111], 295[111], 296[111],
297[111], 299[111], 300[111]
McDaniel, E.G., 732[113]
McDermott, J. C. B., 596 [152], 600 [152]
MacDonald, N. S., 337 [72], 344 [115],
364[115]
McDonald, R.N., 428 [342]
McElhaney, R.N., 704 [240]
McGhie, J. F., 91 [250]
MacGillavry,C.H., 14 [27], 285 [85-87, 89, 91],
286 [89, 91], 287 [86, 91]
McKeown, G. G., 244 [34], 758 [143],
759 [147-149]
Mackinney, G., 32[17], 35[57], 506[418],
536[418], 545[418], 579[9], 580[9, 17, 18],
582 [57-59], 583 [57-59], 593 [133],
608[238, 244], 609[242, 244], 612[265],
615[284-286,288],638[8 ]
McLaren, D. S., 722 [ 49, 50], 732 [50],
738 [49]
McLaughlin, C. I., 730 [92], 733 [92],
735 [92]
Maclay, W.D., 747[29]
MacLead, W. D., 63 [13]
McLean, C., 720 [28], 732 [28], 737 [28]
McMeans, J. L., 257 [ 432]
Macmillan, J ., 50 [251]
Macmillan, J.D., 679 [73]
McQuistan, M., 93 [269], 103 [269], 594[140],
608[140], 763[224]
McSweeney, G.P., 581 [35]
McWeeny, D.J., 760[171]
Mader, 1., 244 [36]
Maercker, A., 329 [39]
Maevskaya, A. N., 602 [173, 174], 690[141],
695 [141]
Magid, L., 762 [211]
Magoon, E. F., 269[15]
Mahadevan, S., 720[30], 721 [40, 42, 44],
722[30]
Mainguy, P., 750[42, 44], 751 [42], 753 [88]
Makin, S.M., 351 [124], 370 [124], 376 [124],
398 [124], 400 [124], 403 [124], 405 [124],
412 [124], 435 [355], 438 [355]
Malathi, P., 720[30], 722[30], 734[133]
Malenge, J.P., 46 [200], 612 [270], 614 [270]
Ma!IT, 1., 759 [ 152]
Malhotra, H. C., 32[32], 34[30a, 41],
103[295], 123[544], 148[449], 592[131],
624[324],625[324],626[ 326,327, 332, 333]
 Author Index
Malia by, R., 527 [ 440]
Mallams, A. K., 43 [152, 153, 155, 164],
44[164, 172, 191], 45[191], 75[146],
91 [246, 247], 95 [146, 247], 106 [321, 323],
107 [146, 323], 110 [146], 111 [146, 246, 332],
116[246], 251 [50], 260[50], 270[29],
273 [29], 277 [29], 289 [111, 114], 290 [114],
291[114], 292[114], 295[29, 111], 296[111],
297[111], 299[111], 300[111, 114],
301 [114], 302[29, 114], 305[114], 306[114],
307[114], 308[114], 309[114], 310[114],
311 [114], 312[114], 341[106], 360[106],
508 [106], 546 [106]
Mallory, R. A., 397 [292]
Manchand, P. S., 34 [39], 103 [299], 121 [371],
124[386], 125[389], 148[451], 281 [75],
359 [166], 360 [166], 380 [166], 400 [308,
309], 401 [309], 409[166], 416[309],
422 [166], 423 [309], 450 [166], 452 [166],
454[166], 455 [166, 309, 372], 456 [309],
465 [309], 468 [309], 477 [166], 478 [166],
489 [166, 308], 490 [166, 308], 500 [309],
509 [166, 309], 514[308, 309], 515 [309],
538 [166], 539 [166, 309, 372], 540[309],
542 [309], 543 [309], 547 [166], 548 [166],
549 [166], 553 [309, 372], 556 [308, 309],
560 [308, 309], 561 [309], 563 [308],
Mani, J.-C., 303 [148], 335 [64, 65], 337 [65],
341 [65], 343 [65], 344 [113], 360 [168],
527 [65], 684 [100-102], 710 [101]
Manning, W. M., 32 [19], 44[173], 104[302],
106 [324], 163 [324], 269 [26], 686 [109],
687 [119]
Manske, R. H. F., 51 [276]
Manten, A., 701 [214]
Manz, U., 410[319], 760[172], 761 [203]
Mar bet, R., 261 [62], 339 [82], 340 [84, 85, 89],
388 [247], 394[247], 405 [247], 406 [247],
410[247, 320], 411 [247], 420[247],
423 [247], 446 [247], 447 [247], 450[247],
458 [247], 460 [247], 467 [247], 468 [247],
4 70 [24 7], 4 71 [24 7], 4 72 [24 7], 489 [24 7],
490 [247], 500 [247], 511 [247], 537 [247],
538 [247], 541 [247], 542 [247], 543 [247],
544[247], 557[247], 563[247], 564[247]
Maricq, J., 531 [ 426]
Marion, J ..P., 51 [265, 266], 52 [265]
Mark, E., 758[143], 759[149]
Markham, M.C., 4l [139a], 46 [202], 126[391],
133 [391], 148 [ 453], 149 [ 453], 153 [ 453],
626 [336]
Markovac- Prpic, A.; 438 [360], 494 [360],
546 [360]
Marquering, B., 752[78]
Marquet, A., 295 [129]
883
Marusich, W. L., 641 [26, 28], 745 [10, 11],
749 [73, 89], 750[51], 751 [51, 57-59, 73],
752 [59], 753 [89, 90], 763 [144]
Marx,J. N., 332 [ 42]
Maslova, T.G., 600[168], 695[187]
Matet, J., 355 [149], 386 [149]
Mathews-Roth, M. M., 620 [302], 673 [20,
25-27], 675 [49], 676 [56], 678 [49, 68],
679 [69, 70], 680 [56], 683 [68], 685 [103],
686[104], 704[241], 710[104]
Mathis, P., 680 [82, 83], 685 [82, 83], 694 [82,
181, 182]
Matsuda, K., 675 [ 43]
Matsui, M., 379 [222, 223], 384 [229],
444 [368 a], 457 [377], 458 [368 a],
460[368a], 516[368a], 518[368a],
532 [377], 540 [377], 541 [368 a],
563 [368 a]
Matsuura, T., 344[114]
Matthews, S., 42 [146], 43 [146], 609 [248]
Mauge, R. L. H., 355 [149], 386[149]
Maxwell, W.A., 679[73, 74]
Mayer, H., 146 [ 444], 204 [15], 206 [20],
208 [20], 251 [20], 257 [57], 259 [57],
261 [20], 332 [ 44], 345 [ 44], 347 [ 44],
398 [294], 412 [294], 437 [ 44], 440 [362],
469 [362], 496 [ 44], 534 [ 444], 563 [ 44]
Mayne, B.C., 689[134]
Meaux, R., 757 [129]
Mebane, A. D., 355 [152, 156], 360[152],
376[216], 403 [216], 404[216]
Mechtler, H., 51 [265], 52[265] ·
Mehl, J. W., 658 [102]
Mehner, A., 749[41], 750[41]
Meinwald, J., 50 [254], 303 [145, 146], 332 [52],
535[448], 709[265]
Meinwald, Y.C., 50[254], 303[145], 535[448],
709 [265]
Meissner, G., 596[148], 700[209], 701 [209,
211], 707[211]
Meister, A., 695 [187]
Melin, M., 658[104]
Menger, A., 757 [127]
Menzi, M., 750[53]
Mercer, E. I., 31 [11], 68 [72], 581 [30, 31],
584[90],605[200] ,610[260],612[31]
Merkle, M.G., 675 [ 46], 685 [ 46]
Merrett, M.J., 604[189]
Merrill, A.L., 746[14]
Meth, E.-G., 494[414], 496[414], 520[420],
551 [414]
Meyers, M. B., 706 [251]
Michel, G., 40[125], 52[125], 117[546],
130[452]
Michniewicz, B. M., 706 [249]
884 Author Index
Mieg, W., 13 [10], 155 [ 473]
Mikami, R., 379[222], 444[368a], 457[377],
458[368a], 460[368a], 516[368a],
518 [368a], 532 [377], 540 [377], 541 [368a],
563 [368a]
Mikhailov, B. M., 355 [147]
Miki, K., 116 [362], 168 [362]
Miki, T., 296 [135]
Milas, N.A., 328 [6, 30], 337 [72], 344[115],
364 [115], 369 [198], 484 [6], 486 [6],
545 [6]
Milborrow, B. V., 50[250], 304[156],
305[156, 157], 527[430, 431,433,434,438,
439], 706 [250]
Mildner, P., 281 [73], 433 [352], 434 [352]
Millar, I. T., 87 [220]
Millar, P. G., 725 [68], 728 [83]
Miller, C. K., 66 [ 45]
Millerd, A, 675 [ 44]
Mills, J. A., 305 [ 158]
Mirna, H., 760[161], 761 [182-184, 192,
200]
Miropol'skaya, M.A., 368[195]
Mirsky, A E., 730 [94]
Mitchell, G. E., 720 [25, 26]
Mitchell, G. V., 734 [137]
Mitchell, H. L., 747 [31]
Mitchell, R. L., 674 [38, 39]
Mitchell, R. P., 752 [265]
Mitsui, T., 50 [249]
Miyano, M., 379 [222]
Modi, V. V., 580 [22]
Moffa, D. F., 719 [16], 721 [16]
Moller, E. F., 74 [131], 86 [216]
Moller, F., 364 [183], 379 [183]
Mondelli, R., 615 [278, 279]
Montag, A, 759 [155]
Montavon, M., 19 [35], 74 [135, 136], 78 [136],
79[186], 87[135], 104[306], 129[397],
146[444], 151 [186, 469], 155[483],
156 [186], 169 [397], 206 [20], 208 [20],
251 [20], 261 [20], 281 [71, 77], 289[109,
110], 291 [109, 110], 297 [71], 328 [16, 18,
19], 332 [ 41, 44], 333 [60], 345 [ 41, 44, 119,
120, 122], 347 [ 44], 364 [ 41, 119, 122,
186-188], 365 [119], 372[41, 119, 186, 187,
208], 379 [221], 388 [245], 391 [186, 260],
393 [208, 262, 280], 394 [262], 400 [305],
403 [280], 412 [60], 431 [280], 435 [354],
436 [305], 437 [ 44, 359], 438 [245], 442 [280],
443 [188], 445 [188, 369], 448 [188],
450 [188], 467 [60], 470 [262], 471 [262],
474 [245], 480 [186], 482 [ 41, 187, 208, 245,
260, 405], 483 [ 41, 187, 208, 405], 494 [305,
359], 496 [ 44], 497 [60], 499 [60, 416],
504 [208], 506 [186, 187, 245, 260],
507[245, 260], 517[188], 519[188],
525 [359], 526 [359], 536 [188, 280],
537 [188, 369], 538 [188], 542 [60], 543 [262],
544 [262], 545 [60, 186, 245, 260, 305, 359],
546 [305], 549 [60, 416], 550 [245],
551 [187, 245], 554[41, 187, 208, 359, 405],
557 [188], 560 [208], 563 [44], 564 [359],
641 [22, 23], 755 [101]
Monties, B., 89 [241]
Moor, H., 760[161]
Moore, L.A., 747[22]
Moore, T., 16, 18 [33], 718 [2], 732 [2, 111],
733 [2], 736 [2, 160], 737 [2]
Morf, R., 75 [137], 155 [ 474], 288 [99],
289[112]
Morishima, H., 581 [46]
Morton, R.A., 16 [28], 18 [28], 49 [241],
380 [224], 584 [91], 727 [72], 729 [85],
730[85]
Mose, W. P., 91 [247], 95 [247], 289 [114],
290 [114], 291 [114], 292 [114], 300[114],
301 [114], 302 [114], 305 [114], 306 [114],
307 [114], 308 [114], 309 [114], 310 [114],
311 [114], 312[114], 341 [106], 360[106],
508 [106], 546 [106]
Moshier, S., 46[205], 139[422], 157[422]
Moslein, E. M., 584 [73]
Moss, B. L., 649 [79]
Moss, G.P., 45[191a], 289[115], 311 [176a]
Moudgal, N. R., 733 [116]
Mousseron-Canet, M., 303 [148], 335 [64, 65],
337[65], 341 [65], 343[65], 344[113],
360 [168], 363 [177], 366 [193], 369 [196],
377 [177], 384 [193], 387 [196], 405 [177],
527 [65], 684 [100-102], 710 [101]
Mi.iggler-Chavan, F., 51 [265, 266],
52 [265]
Mi.iller, H., 155 [ 480], 206 [21], 208 [21],
294 [125], 308 [175]
Mi.iller, P., 74 [136], 78 [136], 372 [208],
393 [208], 482 [208], 483 [208], 504 [208],
554 [208], 560 [208], 761 [196]
Mi.iller, R., 262 [64]
Mi.iller, Z., 751 [67]
Mummery, R. S., 48 [231], 589 [119], 605 [208],
610[258]
Mi.inzel, K., 761 [199], 762 [207, 209, 210,
213]
Murata, N., 696[197], 697[197]
Murchison, D. G., 709 [263]
Murphey, M.M., 35[61]
Murray, T. K., 731 [108]
Murthy,S.K., 721[40], 734[129]
Myoga, K., 751 [68]
 Author Index
N Nishimura, M., 68 [78], 693 [179], 704 [236]
Nadai, J., 744 [ 4], 745 [ 4, 13]
Naftali, Y., 606 [224], 607 [224]
Nagao, M., 535[447]
Nagasaki, T., 134[411]
Nagel, W., 606 [215]
Nagy, E., 36[72], 42[144], 44[144], 111 [331],
151[466],6 06[229],60 7[229],690 [140]
Naito, K., 724[57]
Nakagawa, K., 79[188]
Nakata, T., 79 [188]
Nakayama , T.O.M., 31[6], 32[14, 17, 24],
33[14], 34[45], 35[6, 57, 58], 36[6], 38[6],
41 [6], 42 [6], 43 [6], 89 [234], 123 [374],
124[384], 148[450], 579[9, 10], 580[9],
582[53-55, 57, 59], 583 [53-55, 68], 584[68],
593 [134], 594 [137], 600[165, 166],
602[181], 603[181], 606[213], 612[265,
266, 269], 613 [266], 615 [284, 286],
616[290], 623[313], 674[31], 677[62],
678 [62, 97], 682[97], 683 [62], 690[144,
145], 695 [144]
Nandi, D.L., 584[78], 641 [20]
Narozansk i, J. S., 296 [133]
Nath,K., 721[47], 737[47]
Nathanson , N., 657 [93]
Naves, Y.R., 51 [259, 260], 306[168, 169, 171]
Nayler, P., 296[134], 504[417]
Neamtu, G., 42 [150], 95 [272-275], 101 [272],
133 [ 409], 146 [ 448], 149 [275], 150 [ 462],
151 [272], 204[14]
Negelein, E., 686 [107]
Nelan, D. R., 338 [74]
Nelson, G. E. N., 35 [55]
Nelson, J. W., 747 [33, 87], 752 [81], 753 [81,
85], 763 [227]
Nelson, R., 730 [99]
Nelson, T. S., 751 [71]
Nemser, S., 677 [61], 678 [61l
Neujahr, H. Y., 35 [59]
Neumann, J., 696 [193]
Newman, R. H., 51 [269]
Nichols, B. W., 68[81]
Nicoara, E., 37 [84], 38 [108], 66 [ 40],
71[115, 116], 79[187], 81[40], 85[213],
95 [28?, 545], 101 [282], 146 [116, 187, 446,
448], 151[467], 157[40, 490], 204[14],
312 [177]
Nicol, M., 734[130]
Nicolaides, N., 276[49]
Nicolaux, G.J. M., 355 [149], 386[149]
Nielsen, N.C., 725[63], 727 [63]
Nimmo, C. C., 581 [37], 584 [37]
Nishibori, K., 44 [188, 189], 45 [188, 189, 194]
885
Nitsche, H., 43 [158], 44 [158, 169, 185],
46[158, 207], 67[62], 68[71], 76[154, 155],
106[547], 107[547], 111 [154], 112[155],
113[536], 139[421], 269[28], 273[28]
Nockels, C. F., 734[147]
Nomine, G., 397 [293]
Norden, D. A, 657 [101], 658 [101]
Norgard, D. W., 580 [20], 581 [20], 583 [20]
Norgard, S., 47 [217], 53 [283], 63 [16],
68 [85], 70 [85], 71 [85], 75 [85], 77 [16, 85,
164], 78 [85], 82 [85], 83 [85], 92 [262],
123 [85], 151 [464], 161 [549], 163 [549],
173[164], 174[16, 85, 532], 176[262, 532],
177 [16, 262]
Notthafft, A, 156[488]
Nozaki, H., 385[237], 395[289]
Nunez, G., 69[97, 98]
Niirrenbac h, A, 199[7], 341 [100], 354[141],
357 [100], 383 [141], 389 [251], 441 [364,
365], 443 [366], 444 [364, 366], 446 [364],
448 [366], 474[390], 477[141], 536[364-
366],537[3 64,366],54 5[141,390 ]
Nusbaum-C assuto, E., 31 [9], 584[88]
Nybraaten, G., 78 [166], 152 [166], 157 [166]
0
Oediger, H., 328 [35], 345 [117], 351 [35],
352[35, 130], 357[35], 362[35], 364[183],
366 [35], 379 [183, 220], 380 [35], 384 [230],
394[276]
Oeser, A, 604[192, 193]
Ofner, A, 77 [159], 82 [159], 83 [159],
103 [159], 123 [159, 337], 153 [159],
338 [77, 78], 339 [77, 78], 412 [322, 323],
414[323], 416[323], 434[353], 456[374],
489 [353], 499 [323], 500 [323], 513 [374],
514[374], 523[353], 524[353], 531 [426],
539 [374], 545 [353], 549 [323], 553 [374],
555 [323], 556 [374], 557 [323, 374],
564 [353], 755 [103], 761 [103]
Oh, M., 734[136], 736[136]
Ohkuma, K., 527 [ 430, 435-437], 535 [ 445,
446]
Ohloff, G., 306[167], 341 [107], 364[107],
484[107], 486[107], 487[107], 489[107],
546 [107], 547 [107], 548 [107], 597[156]
Ohnoki, S., 581 [43]
Ohta, M., 393 [271], 394[271]
Ohta, T., 41 [134], 174 [530], 177 [530]
Okada, M., 696[197], 697[197]
Okano, S., 355 [145], 379 [222], 444 [368a],
457 [377], 458 [368a], 460 [368a],
886 Author Index

516[368a], 518[368a], 532[377], Patel, D. J., 209 [22], 213 [22], 222 [22],
540[377], 541 [368a], 563[368a] 280[64], 287[64]
Okukado, N., 41 [132], 104 [316] Pathak, M.A., 686 [104], 710 [104]
Okumura, T., 51 [272] Pattenden, G., 167 [503), 276 [54], 281 [54],
Olive, J.-L., 335 [64, 65], 337 [65], 341 [65], 283 [78], 284 [54, 78], 385 [233, 234),
343 [65], 363 [177], 366 [193], 369 [196], 393 [234], 395 [288), 398 [234), 400 [234,
377 [177], 384 [193], 387 [196], 405 [177], 300], 401 [234], 405 [288), 425 [288],
527 [65] 469 [288), 4 71 [288], 544 [288]
Olson, G., 750[52], 751 [52], 752[52] Patwa, D.K., 580[22)
Olson,J.A., 640[18, 123], 719[9, 10, 12], Pauling, L., 271 [39), 272 [ 41 ], 606 [209]
721 [9, 10, 32, 38, 39, 43, 46, 47], 722 [32], Paulus, H., 584 [72)
732[39], 737[47, 172] Pearlman, J. T., 724 [58), 731 [58]
Olson,J.S., 696[194] Penco, S., 41 [140], 75 [144), 78 [144],
Olson, R.E., 734[132] 152 [144], 157 [144), 336 [68], 499 [68],
Oncley, J. L., 658 [103, 104] 552 [68], 555 [68]
Oomen, H. A. P. C., 722 [ 48, 49], 738 [ 49] Pennington, D., 627 [339]
Oppenheimer, J. R., 687 [121] Pennock,J.F., 584[91]
Orchin, M., 273 [45], 274[45] Percy, R. C., 649 [79]
Orihara, K., 581 [46] Perret, M.A., 760[161]
Oroshnik, W., 17, 328 [33], 355 [152, 156], Perumal,A.S., 733[126]
360[152], 377 [216], 379 [218], 397 [292], Peter, A., 757 [136]
403[216,218],404[216] Peters, J., 682 [96], 683 [96], 684 [96], 685 [96]
Orth, A., 747 [23] Petracek, F. J., 71 [109], 78 [170], 79 [170],
Osadca, M., 641 [27], 745 [12], 761 [12], 98 [ 170, 285], 140 [285], 141 [285],
764[237] 155 [170]
Osgan, M., 50 [257, 258] Petter, H. F. M., 174 [525]
O~ianu, D., 95 [545] Petzold, E. N., 36 [70], 93 [269], 103 [269],
Oskay, E., 276 [53] 151 [465], 763 [224]
Osman,H.G., 32[27], 583[61], 589[116], Pfennig, N., 41 [139a], 126[391], 133 [391],
590 [116], 592 [116], 622 [311, 312], 626[330,334,336]
623 [319] Pflugshaupt, R. P., 141 [431]
Oster, R., 394[282] Phaff, H. J ., 612 [265]
Ostroy, S.E., 726[71], 729[71], 730[71] Phagpolngarm, S., 44 [174], 605 [207],
O'Sullivan, D. G., 366 [190], 370 [190] 610[257], 688 [130]
Otting, W., 281 [76], 282 [76] Philips born, W. von, 294 [125]
Oyama, Y., 51 [272] Phillips, A. H., 581 [32, 33, 45]
Phinney, B., 605 [206], 610 [255]
Phipers, R. F., 288 [98]
p Pickett, J. M., 702 [223]
Pieper, B., 288 [99]
Pachler, K., 49 [242] Pinckard, J. H., 36 [64], 140 [426], 269 [17]
Page, H.J., 107[327] Pippert, D. L., 190 [1]
Page, R. M., 700 [210] Pitt, G. A. J., 718 [3, 6], 727 [72], 729 [85],
Painter, T.J., 92[258] 730[3, 85,91-93, 95], 732[93, 110, 114],
Pakshina, E. V., 677 [65] 733 [91, 92, 114, 115, 117], 735 [92, 158],
Palludan, B., 733 [120] 737 [6, 164]
Palmer, L. S., 13 [12], 14 Planta, C. von, 68 [77], 205 [18], 209 [18],
P{mczel, M., 42[144], 44[144], 111 [331], 235 [18], 248 [18], 252 [18], 260[18],
606[229],607[229],690[140) 263 [18], 276 [50], 357 [164, 165], 379 [164,
Papendick, K., 763 [221) 221], 380[164], 385 [165], 398 [165],
Parman, G. K., 760 [173] 403 [312]
Parry, G. R., 306 [168) Platt, J. R., 692 [169], 696 [169]
Pasedach, H., 340 [95, 96), 355 [151), Plempel, M., 614[274]
394 [282, 283) Poincelot, R. P., 725 [65], 728 [83]
Pasternak,C.A., 734[142) Poisson, J ., 50 [252]
 Author Index
Polgar, A., 195 [6], 271 [39], 288 [95],
606[209]
Pollack, 1. D., 704 [240]
Pollard, C.1., 581 [37], 584[37]
Pommer, H., 199[7], 286[93], 328[4, 5, 21,
36], 333 [55, 58, 59, 61], 340[58, 61, 97, 98],
341 [99, 100], 345 [98, 121], 352[98, 129],
354 [138, 140, 141], 355 [148], 357 [100, 158,
159, 161-163], 366[129, 159], 370[36],
371 [159, 206, 207], 373 [36, 209], 374[36,
209], 375 [36, 210], 376 [36, 210, 215],
380[226], 382[36, 140, 227], 383[141],
384[36, 159, 163,231, 232], 387[36],
388 [246], 393 [138, 140, 211, 266],
394 [21, 138, 211, 284-286], 395 [36, 290,
291], 398 [211, 246], 400[58, 211, 227],
401 [58], 402 [206], 403 [36, 148, 316],
406 [211], 412 [36, 58, 211, 316], 428 [21, 58,
61], 430 [347], 435 [21, 61], 436 [357, 358],
438 [21, 61, 97, 98, 129], 441 [364, 365],
443 [366], 444 [364, 366], 446 [364],
448 [366], 467 [21], 470 [21], 471 [21],
474[21, 246], 477[21, 141], 480[206],
483[3, 5, 21], 484[3], 489[21, 159,161,207,
412], 494[414], 496[61, 97, 98, 414],
497 [129], 499 [59], 520 [61, 420, 421],
521 [61], 522[61], 532[98, 129, 159,207,
246, 429], 536 [364-366], 537 [364, 366],
542[21], 543[21], 544[21], 545[3, 5, 21, 61,
97, 98, 129, 141, 159, 161, 206, 207, 246, 412],
549[59], 551 [414]
Pope, C. W., 747 [25]
Popjak, G., 583 [71], 584 [75, 76, 86], 585 [86,
101, 102], 586 [101, 102], 587 [86, 102, 105],
588 [101], 589 [101]
Popova, I. A., 600 [168]
Popova, 0. F., 600 [168]
Poretta, A., 757 [124]
Porte, A. L., 51 [271]
Porter, 1. W., 31 [12], 32 [21, 23, 26], 35 [61],
69 [103, 104], 268 [8, 12], 580 [16, 20],
581 [20, 48, 49], 583 [20, 67, 70], 584[78,
79, 82-84], 585 [84], 589 [110, 112],
590 [112], 591 [122, 124, 128, 129],
592[129a], 594[112], 596[147], 605[129],
608 [112, 241], 641 [20, 21]
Porter, Q. N., 251 [ 49], 262 [ 49], 263 [ 49],
264[65]
Postel, W., 757 [ 117]
Poulsen, H., 649 [77]
Pousset, 1. L., 50 [252]
Povarov, L. S., 355 [147]
Powell, 1. E., 394 [275]
Powell, L. E., 706 [249]
Prabucki, A. L., 752 [77]
887
Pratt, H. K., 606 [226]
Prebble, J., 39 [113], 41 [138], 72 [119],
92 [257], 151 [257], 659 [109]
Prelog, V., 50 [256-258], 312 [182, 183],
313 [182, 183], 316 [182]
Preobrazhenskii, N. A., 368 [195], 388 [245 a]
Prialgauskaite, L. L., 602 [174]
Price, R. 1., 50 [251]
Prieto, A., 614 [275], 615 [278, 279]
Pritchard, H., 49 [241]
Probst, H. P., 750 [50], 764 [50]
Prohaszka, L., 751 [70]
Prominski, W., 757[121]
Pullman, B., 287 [94]
Pummerer, R., 74[128]
Purcell, A. E., 67[50], 580[25], 591 [25],
606 [227], 691 [155]
Q
Quackenbush, F. W., 32 [21, 22], 35 [52],
36[70], 74[126], 75[149], 93[269],
103 [126, 149, 269], 151 [ 465], 268 [8, 9],
275 [9], 580 [21], 591 [123], 594 [136,
140-142], 607[141, 231], 608[140, 142,
243], 609 [243], 751 [76], 753 [76, 84],
763[76,219,224, 226]
Quilico, A., 615 [278, 279]
R
Rabinowitch, E. I., 686 [108], 687 [108],
693 [178]
Rabourn, W. 1., 32 [21, 22], 35 [52], 74 [126],
103 [126, 291], 268 [8, 9], 275 [9], 594 [136],
763 [224]
Radda, G., 696[196]
Radlick, P., 304 [152]
Rafelson, M. E., 615 [281]
Rai, H., 68 [83]
Rajan, T. S., 759 [150]
Ramirez, F., 394[274]
Ramirez,1., 693[173, 180], 694[180],
695 [180]
Randerath, K., 68 [76]
Raper, 1. H., 659 [110]
Rapoport, H., 53 [283], 161 [549], 163 [549]
Rapp, 1., 729 [87]
Raspe, G., 168 [508], 281 [74], 345 [118],
370[118], 391 [118], 433 [350], 470[350,
387], 471 [387], 482 [ 404], 506 [118],
543 [350, 387], 544 [350, 387], 545 [ 404],
546 [118]
888 Author Index
Ratcliff, R. G., 749 [39]
Rau, W., 269 [19], 620[305-310]
Rauch, W., 749[41], 750[41]
Rau-Hund, A., 620[306, 308]
Raymundo, L. C., 594 [143], 607 [235]
Raz, A., 720 [29], 721 [29]
Razin, S., 674 [28], 704 [240], 705 [246]
Rebeiz, C. A., 610[259]
Rebmann, L., 74[128]
Redel, J., 352 [125], 443 [367], 448 [367],
536 [367], 537 [367]
Redfearn, E. R., 638 [1], 640, 641 [24, 25]
Reichel, L., 583 [65]
Reichenbach, H., 38 [94, 95], 40 [95], 52 [95],
117[413]
Reif, W., 341 [99, 100], 357 [100], 380[226],
389[251], 394[284-286], 441 [364, 365],
443 [366], 444 [364, 366], 446 [364],
448 [366], 4 74 [390], 536 [364-366],
537[364, 366], 545[390]
Reiff, H., 355 [144]
Reindel, W., 74[128]
Reppe, W., 394[281]
Reschke, T., 50 [244], 270 [34], 312 [34]
Reyes, P., 35 [58], 616 [290]
Reymond, D., 51 [265, 266], 52[265]
Richardson, R. W., 284 [79]
Rick, C. M., 608 [244], 609 [242, 244]
Ricketts, T. R., 53 [281], 71 [113]
Ridley, M. W., 649 [78, 79]
Riederer, P., 757 [130-132]
Rigassi, N., 205 [19], 209 [19], 210 [19],
213 [19], 226 [19], 239 [19], 245 [19],
248 [19], 249 [19], 250 [19], 251 [19],
257 [55], 259 [19], 261 [62]. 280 [65],
388 [247], 394 [247], 405 [247], 406 [247],
410 [247], 411 [247], 420 [247], 423 [247],
446 [247], 447 [247], 450 [247], 458 [247],
460 [247], 462 [382], 463 [382], 467 [247],
468 [247], 470 [247], 471 [247], 472 [247],
489[247], 490[247], 500[247], 511 [247],
531 [ 425], 535 [ 425], 536 [ 425], 537 [247],
538 [247], 541 [247], 542 [247], 543 [247],
544 [247], 557 [247], 563 [247], 564 [247]
Rilling, H. C., 585 [95], 591 [121], 620 [300,
301,304]
Rinaldini, L. M., 736 [162]
Ringelmann, E., 580[15]
Rispoli, G., 757[135]
Robbins, W.E., 734[151]
Roberts, A. B., 721 [35, 36], 722 [35, 36],
737 [36]
Roberts, D. L., 343 [112], 533 [ 441]
Roberts, J.D., 242 [28]
Robertson, D. S., 605 [199, 201], 674 [34, 37]
Robeson, C. D., 337 [73], 338 [75], 352 [128],
359 [128], 370 [75], 376 [214], 380 [128,
225], 384 [228], 386 [238, 238 a], 425 [339],
454 [371], 457 [376], 462 [75], 464 [75],
476[395, 396],477[339,395],478[339,
396], 480 [339, 403], 520 [395, 403],
522 [395, 403], 529 [73, 75], 531 [75],
538 [371], 540 [376], 545 [395, 396],
549 [339, 395], 554 [339, 403], 560 [339, 403]
Robins<;m, H. V., 720[21]
Robinso~. M. I., 702 [220]
Roels, 0. A., 720 [23], 721 [ 42], 722 [23],
735 [23, 155], 736 [23, 155]
Rogers, D., 295 [130]
Rogers, L.J., 581 [40], 596[146], 604[188],
605 [ 40], 607 [236]
Rogers, W. E., 732 [113], 734 [139, 140, 148],
737 [139, 167]
Rogier, J. C., 753 [84]
Rollins, M.H., 731 [105]
Romagnoli, A., 645 [ 41, 42]
R6nai, A., 36 [73], 43 [155], 78 [173],
91 [246], 111 [246], 116 [246], 155 [173],
270 [29], 273 [29], 277 [29], 295 [29],
302 [29]
Ronco, A., 281 [72), 328[1], 344[1], 379[1,
217],403[1,217,313,314]
Root, D. M., 734 [138]
Rosen, E., 306 [ 165], 341 [ 105]
Rosenthal, A., 86 [218)
Ross, S. T., 393 [265]
Ross, W.A., 91 [250]
Rotfarb, R. M., 600 [ 160]
Roth, H., 75 [139, 147], 76[157], 84[205],
87 [228], 88 [205]
Rother, H., 757 [133], 761 [133],
764 [250-252]
Rottem, S., 674 [28], 705 [246]
Rouques, A., 750[42, 44], 751 [42)
Rousseau, J. E., 747 [22]
Rowland, R.L., 341 [103, 104]
Royals, E. E., 340 [88], 393 [279]
Rubenstein, D., 67 [65]
Rubin, S. H., 755 [103], 761 [103]
Rubinstein, H., 333 [63]
Rudney, H., 580[14]
Riiegg, R., 19 [35], 33 [33], 36 [65], 46 [33,
199], 48 [33], 49 [33], 68 [77, 79], 74 [135,
136], 78 [136], 79 [186], 87 [135], 103 [296,
299], 104 [296, 306], 128 [393], 129 [395,
397], 130[399], 146[444], 151 [186, 469],
155 [ 483], 156 [186], 166 [393], 168 [395],
169 [395, 397], 205 [18], 206 [20], 208 [20],
209 [18], 235 [18], 246 [38], 248 [18],
251 [20], 252[18], 260[18], 261 [20, 38, 62],
 Author Index
263[18], 276[50], 281 [71, 75, 77],
289 [109, 110], 291 [109, 110], 297 [71],
328[13, 16, 22, 23, 37, 41], 332[44],
333[60], 340[94], 345[41, 44, 119, 120,
122], 347 [44], 355 [153], 357 [153, 164,
165], 359[166], 360[153, 166], 364[41,
119, 122, 186-188], 365 [119], 372 [ 41, 119,
186,187, 208], 379[164, 221], 380[164, 166],
385 [165], 388 [245, 247], 391 [186, 260],
393 [208, 262, 280], 394 [247, 262],
398 [165], 400 [298, 303, 305], 403 [280,
312], 405 [247, 298], 406 [247], 409 [166],
410 [247], 411 [247, 298], 412 [60],
420 [247], 421 [303], 422 [153, 166],
423[247],431[280],435[354],436[30 5],
437 [ 44, 359], 438 [245], 442 [280],
443 [188], 445 [188, 369], 446 [247],
447 [247], 448 [188], 450 [166, 188, 247],
452 [166], 454 [166], 455 [166], 457 [375,
378], 458 [247, 298, 375], 459 [13, 378],
460[247, 378], 461 [298, 378], 462[13],
463 [13], 467 [60], 468 [247, 385], 470 [262],
471 [247, 262], 472 [247, 389], 474 [245],
477 [23, 166], 478 [166], 479 [23], 480 [186],
482[41, 187,208,245,260,405],483[41,
187,208, 405], 489[166, 247], 490[166,
247], 494 [305, 359], 496 [ 44], 497 [60],
499 [60, 416], 500[247], 503 [385],
504[153, 208], 506[186, 187, 245, 260],
507 [245, 260], 508 [153], 509 [166],
511 [23, 153], 512 [153], 516 [303],
517 [188, 378], 518 [303], 519 [188, 378],
525 [359], 526 [359], 527 [164], 536 [188,
280], 537 [188, 247, 369], 538 [166, 188,
247], 539[166], 540[375, 378], 541 [13,
247,298,375, 378], 542[60,247,298, 378],
543 [247, 262, 385], 544 [247, 262, 389],
545[60, 153, 186,245,260,305,359],
546 [153, 305], 547 [153, 166, 303],
548[166], 549[60, 166,416], 550[23,245],
551 [187, 245], 554[41, 187, 208, 359, 405],
557[188,247],560[208],563[23,44,2 47,
298, 378, 385], 564 [247, 359], 639 [9],
641 [22, 23], 755 [101]
Riiegger, A., 155 [ 484]
Rummert, G., 286[93], 328 [4], 371 [206],
402 [20~], 480 [206], 531 [ 424], 545 [206]
Riippel, H., 696[195]
Russell, A., 340 [86]
Russell, S. W., 43 [157], 45 [157], 50 [255],
95[280, 281], 109[280], 296[133],
297 [136-138], 299 [138, 139], 302 [136,
138], 303 [139, 150], 304 [136, 139],
305 [139], 307 [139], 309 [139], 310 [139],
311 [139], 342 [109, 110], 360 [109],
889
489 [109], 490 [109], 510 [109], 551 [109],
552[109],554[109],600[163], 709[266]
Rutschmann, J., 46 [197], 87 [224], 93 [266],
103 [266], 116 [351, 358], 119 [364],
121 [364], 128 [224, 392], 288 [104]
Rutter, L., 68 [88]
Ruzicka, L., 13, 339 [83]
Ryback, G., 50[246], 270[36], 304[156],
305[156, 157, 159],527[430,433,434,439,
440], 585 [102], 586 [102], 587 [102],
706[247, 250]
Ryser, G., 36 [65], 46 [199], 74 [136],
78 [136], 103 [296], 104 [296], 128 [393],
129 [397], 166 [393], 169 [397], 281 [77],
333 [60], 355 [153], 357 [153], 360[153],
372 [208], 393 [208, 262], 394 [262],
400[298, 303],405[298],411[298],
412[60], 421 [303], 422[153], 443 [188],
445 [188], 448 [188], 450[188], 457 [375,
378], 458 [298, 375], 461 [298], 467 [60],
470[262], 471 [262], 472 [389], 482 [208],
483 [208], 497 [60], 499 [60], 504 [153, 208],
508 [153], 511 [153, 298], 512 [153],
516[303], 517[188, 378], 518[303],
519[188, 378], 536[188], 537[188],
538 [188], 540 [375, 378], 541 [298, 375,
378], 542 [60, 298, 378], 543 [262],
544[262, 389], 545 [60, 153], 546[153],
547 [153, 303], 549 [60], 554 [208],
557 [188], 560[208], 563 [298, 378],
641 [23]
Ryvarden, L., 34[43], 82[196], 83[196],
148 [196], 153 [196], 627[341]
s
Saakov, B.C., 600[168]
Saakov, V. S., 602 [177, 178], 690 [142, 143]
Sadana, J. C., 606 [225]
Sager, R., 676 [52, 54], 693 [54]
Saijo, S., 355 [145]
Sakal, E., 344 [ 115], 364 [ 115]
Sakan, T., 51 [267, 270], 303 [147]
Sakato, Y., 51 [264]
Salaque, A., 44[191], 45[191], 106[323],
107 [323]
Salomon, H., 36[68], 160[506]
Salomon, K., 657 [96, 97]
Salontai, T., 37 [84], 79 [187], 95 [282],
101 [282], 146 [187, 446]
Salton, M. R. J., 704 [238, 239]
Salvatori, T., 615 [278, 279]
Samokhvalov, G. I., 361 [169], 368 [195],
388 [245a]
890 Author Index
Sander, C., 675 [ 48]
Sandevoir, C. M. T., 355 [149], 386 [149]
Sandoval, A, 32 [20]
Saperstein, S., 38 [103], 47 [215], 63 [14],
583[62], 761[194]
Sapozhnikov, D. I., 600[168], 602[173-176],
690[138, 141, 147], 695 [141]
Sargent, K. H., 430 [346]
Sargent, M. L., 702 [222]
Sarnecki, W., 333 [58, 59], 340[58, 97, 98],
341 [100], 345 [98], 352 [98, 129], 357 [100,
158, 159, 162, 163], 366 [129, 159],
371 [159, 207], 375 [210], 384[159, 163,
231, 232], 388 [246], 389 [251 ], 395 [291 ],
398 [246], 400 [58], 401 [58], 412 [58],
428 [58], 438 [97, 98, 129], 474 [246, 390],
489 [159, 207, 412], 496 [97, 98, 129],
499[59], 532[98, 129, 159,207,246,429],
545[97,98, 129, 159,207,246,390,412],
549[59]
Sa to, T., 379 [222]
Satoh, D., 51 [272, 273]
Sauberli, U., 51 [262]
Saucy, G., 46 [199], 74 [136], 78 [136],
128 [393], 129 [397], 151 [469, 483],
166 [393], 169 [397], 281 [71], 289 [109,
110], 291 [109, 110], 297 [71], 328 [16],
332[41], 339[82], 340[84, 85, 89],
345[41, 122], 364[41, 187, 188], 372[41,
187, 208], 379 [221], 393 [208, 280],
403 [280], 431 [280], 442 [280], 443 [188],
445 [188, 369], 448 [188], 450 [188],
457 [378], 459 [378], 460 [378], 461 [378],
482[41, 187, 208], 483[41, 187, 208],
504[208], 506[187], 517[188, 378],
519 [188, 378], 536 [188, 280], 537 [188,
369], 538 [188], 540[378], 541 [378],
542[378], 551 [187], 554[41, 187, 208],
557[188],560[208],563[378],641[22,23]
Saunders, C., 751 [63]
Savolainen, J. E. T., 754 [99]
Sax, N. W., 338 [77, 78], 339 [77, 78]
Schaeren, S. F., 74 [136], 78 [136], 261 [62],
372 [208], 388 [247], 393 [208, 280],
394 [247], 403 [280], 405 [247], 406 [247],
410 [247], 411 [247], 420 [247], 423 [247],
431 [280], 442 [280], 446 [24 7], 44 7 [24 7],
450 [24 7], 458 [24 7], 460 [24 7], 467 [24 7],
468 [247], 470[247], 471 [247], 472 [247],
482 [208], 483 [208], 489 [247], 490 [247],
500[247], 504[208], 511 [247], 536[280],
537 [247], 538 [247], 541 [247], 542 [247],
543 [247], 544 [247], 554[208], 557 [247],
560 [208], 563 [247], 564 [247], 641 [22]
Schaffner, K., 312 [180]
Schaible, P.J., 74( [25]
Schaltegger, K. H., 68 [82]
Schara, A, 757 [138], 761 [138]
Scharf, S. S., 584 [92]
Scheer, B. T., 44 [192]
Schenck, G.O., 680[84]
Schick, E., 370 [204], 387 [204]
Schiemenz, G. P., 393 [269]
Schillinger, A, 756 [118]
Schimmer, B. P., 43 [164], 44 [164], 91 [245],
111 [332, 333]
Schinz, H., 333 [53]
Schlag, J., 199 [7]
Schlegel, H. G., 41 [139]
Schlegel, W., 760[167]
Schlesinger, H., 435 [356]
Schlesinger, M.J., 582[50]
Schlettwein-Gsell, D., 764 [235]
Schmall, M., 67 [55]
Schmid, G. H., 702 [224]
Schmidt, K., 41 [139], 46 [208], 121 [365],
127 [365], 129 [398], 171 [365], 173 [365],
626[334]
Schmidt, P.J., 754[98]
Schneider, D. F., 123 [376], 385 [233, 235],
414[325], 456[325], 499[325], 500[325],
513 [325], 514 [325], 539 [325], 552 [325],
555[325],556[325],557[325]
Schneider, H. A W., 68 [84]
Schneider, K., 340[95, 96]
Schneider, R., 50[256]
Schnepf, E., 603 [183]
Schoch, T.J., 760[180]
Schodder, H., 66[47]
Schollkopf, U., 333 [55]
Scholtyssek, S., 749[40]
Schon, K., 36[76], 149 [458], 273 [43]
Schopfer, W.H., 578[1-4]
Schor, N.A, 734[144]
Schroder, W., 691 [149, 150]
Schroeder, W.A, 36[77], 75[140], 149[459],
269[20],271[39],606[209]
Schroepfer, G.J., 585 [102], 586 [102],
587 [102]
Schuchardt, W., 760[168]
Schudel, P., 36 [65], 68 [79], 103 [296],
104[296], 191 [4], 281 [68], 328 [22-24],
329 [38], 355 [153], 357 [153], 360 [153],
400[303], 421 [303], 422 [153], 477 [23],
479 [23], 504 [153], 508 [153], 511 [23, 153],
512 [153], 516 [303], 518 [303], 545 [153],
546 [153], 547 [153, 303], 550[23], 563 [23]
Schwang, H., 305 [163]
Schwanzara, S.A, 724[56]
Schwarting, A E., 605 [203], 674 [35]
Schwarze, P., 702 [224]
Schwarzmann, M., 395 [291]
 Author Index
Schwegler, F., 756 [110]
Schwenker, U., 68 [68], 756 [108]
Schwieter, U., 18 [31], 19 [35], 36 [65],
46 [199, 211], 47 [211], 67 [58], 68 [77],
74 [136], 75 [142], 77 [142], 78 [136, 142],
79 [142], 87 [142], 103 [296, 299], 104 [296],
128 [393], 129 [397], 130 [399, 405],
155[405], 166[393], 169[397], 174[142,
527], 205[18, 19], 209[18, 19], 210[19],
213[19],226[19],233[24,251235[18],
239 [19], 245 [38], 248 [18, 19], 249 [19],
250[19], 251 [19], 252[18], 257[56],
259 [19, 52, 56], 260 [18], 261 [38, 62],
263 [18, 52], 276 [50], 278 [58], 280 [65],
281 [75], 328 [8, 10, 13, 37], 345 [118],
355[153], 357[15~ 164, 1651 359[166],
360[153, 166, 167], 364[188], 370[118,
205], 372 [167, 208], 379 [164, 221],
380 [164, 166], 385 [165], 388 [247],
390[167], 391 [118], 393 [208, 280],
394 [247], 398 [165], 400 [303], 403 [280,
312], 405 [247], 406 [247], 408 [167],
409[166], 410[247, 312,320, 321],
411 [247], 420 [247], 421 [303], 422 [153,
166], 423 [247], 431 [280], 442 [280],
443 [188], 445 [188, 369], 446 [247],
447 [247], 448 [188], 450 [166, 188, 247],
452 [166], 454 [166, 370], 455 [166, 370],
456 [370], 457 [370, 378], 458 [247],
459 [13, 378], 460 [247, 378], 461 [378],
462 [13, 382], 463 [13, 382], 467 [247],
468 [247, 385], 471 [247], 472 [247],
477[166, 167], 478[166], 479[167],
480 [167], 482 [208], 483 [208], 489 [166,
167, 247], 490[166, 247], 492[167],
500[247], 503 [385], 504[153, 167, 208],
506 [118, 418], 507 [167], 508 [153],
509[166], 511 [153, 247], 512[153],
515 [370], 516 [303, 321], 517 [188, 378],
518 [303, 321], 519 [188, 378], 523 [167],
524 [167], 525 [3211 526 [321], 527 [164],
531 [ 425], 534 [ 444], 535 [ 425], 536 [188,
280, 418, 425, 451], 537 [188, 247, 369],
538 [166, 188, 247, 370], 539 [166],
540 [370, 378], 541 [13, 247, 378], 542 [247,
378], 543[247, 385],544[247],545[153,
418], 546 [118, 153], 547 [153, 166, 303],
548[166], 549[166], 550[167], 551 [167],
554[208], 555[3~1], 557[167, 188, 247],
558[167], 559[321], 560[167, 208, 370],
561 [167], 562[385], 563 [167, 247, 378,
385], 564 [167, 247' 321], 639 [9], 641 [22,
23], 718 [5], 760 [167]
Schwyzer, R., 477 [399], 545 [399]
Scopes, P.M., 91 [247], 95 [247], 289 [114],
290[114], 291 [114], 292[114], 300[114],
891
301 [114], 302 [114], 305 [114], 306 [114],
307 [114], 308 [114], 309 [114], 310 [114],
311 [114], 312[114], 341 [106], 360[106],
508 [106], 546 [106]
Scott, M.L., 750[52], 751 [52], 752[52]
Sebek, 0., 614[276], 615 [276]
Sebrell, W. H., 18 [30]
Secor, H. V., 51 [269]
Seefelder, M., 355 [151], 394 [283]
Seehafer, M. E., 757 [125]
Seeliger, A., 78 [169]
Seibold, H., 764 [256]
Seidel, C. F., 333 [53]
Sekoguti, Y., 727 [73], 729 [88]
Selva, A., 615 [278, 279]
Semenovskii, A. V., 333 [57]
~erban, M., 38 [108]
Seshadri Sastry, P., 721 [ 44]
Sestak, Z., 68 [86]
Seward,C.R., 734[137]
Sewell, H. B., 720 [25]
Shah, D. H., 583 [67]
Shah, D.O., 735 [155], 736 [155]
Shah, D. V., 69 [103], 584 [84], 585 [84]
Shah, S. P.J., 581 [40], 604[188], 605 [40],
607 [236]
Sharman, I. M., 736 [160]
Sharpless, N. E., 727 [76]
Shaw,C.R., 720[31], 721[31], 723[31]
Shaw, G., 708 [259-262], 709 [260, 264]
Sherma, J., 66 [ 49], 68 [80], 69 [93-95]
Shichi, H., 725 [69], 729 [69]
Shields, J. E., 725 [68]
Shimamoto, H., 111 [339], 277 [56]
Shimomura, 0., 50[253]
Shira tori, T., 719 [15], 720 [19, 22], 721 [15]
Shiryaeva, G. A., 600[168]
Shishido, K., 385 [237], 395 [289]
Shneour, E. A., 34 [ 44], 124 [383], 580 [24],
690[146]
Shropshire, W., 700 [208], 701 [209]
Siddiqui,J.R., 757[137] .
Siddons, P. T., 31 [13], 101 [286], 103 [290,
299], 124 [385], 148 [385], 268 [10],
269 [10], 275 [10], 276 [10], 281 [75],
359[166], 360[166], 380[166], 400[296],
401 [296], 409 [166], 421 [296, 335],
422 [ 166], 423 [296], 426 [296], 428 [296,
335], 450[166], 452[166], 454[166, 296,
335], 455 [166], 477 [166, 296, 335],
478 [166, 296, 335], 489 [166, 296, 335],
490[166, 296, 335], 509[166, 296, 335],
517 [296], 521 [296], 522 [296], 529 [296],
538[166, 296, 335], 539[166], 547[166],
548[166], 549[166,296,335], 550[296,
335], 589 [109], 623 [320]
892 Author Index
Siegel, A., 675 [ 43]
Siekmann, W., 91 [244], 114[244]
Siemer, H., 328 [15]
Simmonet, H., 744 [6]
Simoni, R., 604[197]
Simpson, K.L., 584[92], 594[137, 143],
607 [235], 612 [266, 269], 613 [266],
614[271]
Singer, C. E., 673 [21]
Sissins, M. E., 79 [193], 124[193], 623 [314]
Sistrom, W. R., 671 [12], 672 [12, 16],
673 [20, 26, 27], 677 [61], 678 [61], 684[16]
Skeggs, H. R., 579 [11]
Slates, H. L., 352 [127], 369 [127], 387 [127]
Sly, W. G., 285 [90]
Smallidge, R. L., 751 [76], 753 [76], 763 [76]
Smit,A., 352[126, 133,134, 136], 354[134],
369 [126], 386 [136, 241], 387 [126]
Smit, V.A., 333 [57]
Smith, D., 763 [227]
Smith, E.G., 760[169]
Smith, F.R., 719[11]
Smith, J. H. C., 121 [370], 192 [5], 674 [32]
Smith, L., 692 [171], 693 [173, 180], 694 [180],
695 [180]
Smith, L. W., 67 [65]
Smith, O.E., 527 [430, 432, 435-437],
534[443]
Smith, P. F., 92 [256], 705 [245]
Smith, P. G., 609 [247]
Smith, T. A., 50 [247], 305 [160], 705 [243],
706[243]
Smith, V. E., 38 [101], 39 [101], 53 [279],
63 [13], 163 [550], 638 [5], 649 [81],
652[81], 654[81]
Smith, W.O., 51 [263]
Smith, W. T., 51 [263]
Snatzke, G., 270[35], 305 [163], 312[35, 180],
598 [157], 707 [258]
Sobotka, H., 306[165], 332[47], 337[47],
341 [105]
Sole, P., 757 [119]
Solms, U., 328 [7, 9]
Solmssen, U., 45 [193], 87 [225], 88 [229],
121 [229, 366-368], 122 [229], 123 [229],
133[407, 408], 137[225], 146[445],
148[366], 155[479, 484], 169[484,
512-514], 171 [512], 278 [59]
Sondheimer, E., 706 [249]
Sondheimer, F., 284[79], 332[42], 364[184],
408[318], 429[344], 430[344]
S~rensen, N.A., 45[196], 72[117, 124],
75 [153], 86 [117], 88 [117], 107 [124],
146[117, 153], 251 [51], 656[87, 88],
658 [87, 88]
Sorensen, T. S., 1~1 [3]
Soucek, M., 306[173]
Spalla, C., 41 [140], 75 [144], 78 [144],
152 [144], 157 [144], 336 [68], 499 [68],
552[68], 555[68], 614[275]
Spark, A. A., 43 [151, 153], 44[153], 75 [146],
95 [146], 104[317], 107 [146], 110[146],
111[146], 148[317], 151[317], 168[317],
251 [50], 260[50], 285 [82], 289 [111],
295[111], 296[111], 297[111], 299[111],
300[111], 415 [327], 452 [327], 456 [327],
511 [327], 512 [327], 513 [327], 514 [327],
538 [327], 539 [327], 548 [327], 549 [327],
552 [327], 553 [327]
Spencer, B., 733 [122], 734[122, 149]
Spencer,C.C., 760[180]
Spencer, D., 675 [ 44]
Spicer, F. W., 720[18], 721 [18]
Spiess, W. E. L., 757 [119]
Spinks, A., 344[116], 345 [116], 364[185]
Spiteri, J., 69 [98]
Springall, H. D., 87 [220]
Staab, H. A., 281 [76], 282[76]
Stabursvik, A., 48 [224]
Stahl, E., 67 [52-54], 68 [53, 75]
Stainer, D. W., 731 [108]
Starn, C. H., 285 [87]
Stanier, R. Y., 32 [14], 33 [14], 123 [374],
585 [100], 623 [313, 321], 671 [12], 672 [12,
16, 18, 19], 673[22], 674[29], 676[58],
678 [58], 679 [19], 684 [16]
Starr, M.P., 38 [103], 47 [215], 63 [14],
583 [62]
Stary, Z., 583 [63]
Steele, K.A., 760[170]
Steele, W. J., 579 [7], 580 [7], 581 [7], 583 [7]
Steensland, H., 626 [337]
Steeves, H.R., 734[151]
Steffen, K., 606 [220]
Steffenson, D., 66 [ 45]
Steinegger, P., 750[53-55]
Steinlin, K., 114[349]
Stepp, W., 15
Sterling, C., 285 [88]
Stern, K. G., 657 [96, 97]
Stern, M.H., 352[128], 359[128], 380[128],
389[253], 474[253], 476[396, 397],
478 [396], 480 [397], 538 [397], 545 [253,
396, 397]
Sternhell, S., 213 [23]
Stevens, C. L., 332 [ 49]
Stilz, W., 354[138, 140], 376[211], 382[140],
393 [138, 140, 211], 394[138, 211],
398 [211], 400 [211], 406 [211], 412 [211]
Stobart, A. K., 581 [42], 605 [42]
Stoll, A., 13 [ 11 ], 63 [8], 66 [8]
Stoll, M., 51 [262]
 Author Index
Stolp, F., 394[284]
Stolz, L. P., 51 [263]
Stone, A. L., 725 [69], 729 [69]
Straatsma, B. R., 733 [124]
Strain, H. H., 32 [19], 35 [62], 37 [82],
43[165, 167],44[173, 177-179,181,182,
184], 46[204], 53 [283], 63 [1, 12], 66 [1, 38,
44, 45, 48, 49], 69 [93-95], 83 [201],
89 [238], 91 [248], 104 [302], 106 [248,
324, 325], 107 [325], 111 [1], 113 [537],
139[44,417-419], 156[491], 157[201],
161 [495, 549], 163 [324, 549], 251 [48],
269 [24, 26], 288 [106], 602 [169]
Strang, R.H.C., 46[214], 174[531], 176[531]
Stransky, H., 107 [548]
Straus, W., 756 [109]
Strehler, B., 676 [53], 692 [168, 172],
693 [53, 168]
Streiff, K., 748 [61], 750 [56], 751 [61, 74],
752[83]
Strel'tsov, R. V., 435 [355], 438 [355]
Stricker, K., 74 [136], 78 [136], 372 [208],
393 [208], 482 [208], 483 [208], 504 [208],
554[208],560[208]
Studer, A., 33 [33], 46 [33], 48 [33], 49 [33],
129 [395], 168 [395], 169 [395]
Stumpf, P.K., 604[197]
Subba Rao, K., 733 [125], 734 [125, 128, 132]
Subbarayan, C., 581 [49], 591 [129],
592[129a], 596[147], 605[129]
Suga, K., 369 [199]
Sundaresan, P.R., 733 [123], 734 [123],
737[123, 168, 169]
Sundt, E., 306 [172]
Sunthankar, S. V., 21 [41]
Surma tis, J.D., 77 [159], 82 [159], 83 [159],
103 [159], 123 [159, 337], 153 [159], 328,
333 [62], 338 [77], 339 [77], 341 [102],
342 [62, 111], 343 [102], 347 [102],
360[167], 364[102], 372[102, 111, 167],
373 [111], 379 [219], 380 [219], 389 [252,
254, 255], 390[111, 167, 256], 408 [167],
412 [322, 323], 414 [323], 416 [323],
434 [353], 456 [374], 474 [219, 252, 254, 255],
477[167, 256, 401], 479[167, 256, 401],
480[167], 482[102], 483[102], 489[167,
353], 4941[167], 499[323], 500[323],
504[111, 167], 507[111,167], 513[374],
514 [374], 523 [167, 353, 422], 524 [167,
353, 422], 531 [426], 539[374], 545[252,
255], 549 [323], 550 [167], 551 [167, 252,
401], 553 [374], 554 [102, 256], 555 [323],
556 [374], 557 [167, 323, 374], 558 [167,
252, 401], 560[167, 256], 561 [102, 167],
563[111, 167], 564[167, 353, 422],
755 [10.3], 761 [103]
893
~uteu, M., 66[40], 71 [115, 116], 81 [40],
85 [213], 146 [116], 157 [40, 490], 312 [177]
Sutter, R. P., 615 [281]
Suyama, T., 761 [189-191]
Suzue, G., 580[26, 27], 581 [43, 44, 46, 48, 49],
591 [26, 126, 129], 592[129a], 605[129]
Suzuki, N., 269 [21]
Suzuki, T., 727 [73]
Suzuki, Y., 44[190]
Svec, W.A., 44[178, 179, 181, 182, 184],
46 [204], 53 [283], 66[44], 83 [201],
91 [248], 106 [248, 325], 107 [325],
113[537],139[44],157[201], 161[549],
163[549],251[48]
Swain, T., 51 [261], 163 [497]
Swift, G., 91 [250]
Szabo, D., 137 [415]
Szabolcs, J., 36 [72, 73], 42 [149], 43 [155,
163, 164], 44 [164], 46 [203], 75 [138],
78 [173, 175], 79 [261], 89 [242, 243],
91 [242, 243, 246, 247], 95 [247], 111 [243,
246, 332], 114[243], 116[243, 246],
125 [261], 137 [261, 415], 139 [175],
148[454],151[466], 153[454],155[173],
251 [49], 262[49], 263[49], 264[65],
269 [25], 270 [29], 273 [29], 277 [29],
289 [114], 290[114], 291 [114], 292 [114],
293 [122], 295 [29], 296 [132], 300 [114,
132], 301 [114], 302 [29, 114], 305 [114],
306 [114], 307 [114], 308 [114], 309 [114],
310[114], 311 [114], 312[114], 341 [106],
360[106], 508[106], 516[419], 546[106],
559[419]
T
Tagwerker, F.J., 748 [61], 751 [61]
Taha, M.M., 139[424],140[424]
Tait, G. H., 604[191]
Takagi, M., 729 [88]
Takahashi, H., 393 [271], 394[271]
Takahashi, T., 78 [167], 156 [167]
Takamatsu, K., 704 [236]
Takamiya, A., 696[197], 697[197]
Takeda, T., 761 [191]
Takeda, Y., 41 [133, 134], 104[314], 174[530],
177 [530]
Tlima~. V., 95 [272-279], 101 [272, 278, 279],
114[279], 146[448], 149[275], 150[462],
151 [272], 204[14]
Tamm, R., 74 [136], 78 [136], 372 [208],
393 [208], 482 [208], 483 [208], 504 [208],
554 [208], 560 [208], 760 [167], 761 [196]
Tamura, S., 535 [447]
Tanaka, A., 134[411]
894 Author Index
Tanaka, S., 581 [43, 46, 126]
Tarr, E., 657 [93]
Tasaki, I., 751 [68]
Tavla, M., 680[80]
Tavormina, P.A., 579[12]
Tawney, P. 0., 331 [ 40]
Taylor, H. F., 50[247, 248, 248a], 305[142a,
160, 161], 705 [243], 706 [243, 248]
Taylor, W.C., 681 [88]
Tchen, S.- Y., 352 [125]
Tchen, T. T., 581 [33]
Teale, F. W.J., 687 [122], 688 [122]
Tee,J.L., 43(151-153], 44[153], 75[146],
95 [146], 107 [146], 110 [146], 111 (146],
251[50], 260(50], 289[111], 295[111],
296[111], 297[111], 299[111], 300[111]
Tefft, R.E., 614[271]
Terasaki, M., 760[161], 761 [182-184, 192, 200]
Thaler, H., 757[115]
Theimer, R.R., 620[310]
Thiessen, W.E., 527[430, 437]
Thimann, K. V., 699 [203, 206]
Thirkell, D., 46[214], 174(531], 176[531],
663 [121]
Thomas, A. F., 262 [64]
Thomas, C., 36 [ 67]
Thomas, D. B., 734[142]
Thomas, D.M., 44[172], 106[321], 614[277],
615[277, 282], 707[254]
Thomas, D.R., 581 [42], 605 [42]
Thomas, J. B., 687 [113]
Thomas, M. R., 602 [169]
Thommen, H., 38 [105], 48 [227], 87 [226],
638 [6], 639 [10, 14], 642 [32], 645 [37],
649 [62, 80, 82], 650 [84], 655 [85],
658[105, 106], 746[17, 18], 754[96]
Thommen, R., 123 [337], 333 [62], 341 (102],
342 [62], 343 [102], 347 [102], 360[167],
364[102], 372[102, 167], 389[252, 255],
390[167], 408[167], 456[374], 474[255],
477 [167], 479 [167], 480[167], 482 [102],
483 [102], 489 [167], 492 [167], 504 [167],
507 [167], 513 [374], 514[374], 523 [167],
524 [167], 539 [374], 545 [252, 255],
550 [167], 551 [167, 252], 553 [374],
554 [102], 556 [374], 557 [167, 374],
558 [167, 252], 560[167], 561 [102, 167],
563 [167], 564 [167]
Thompson, A. F., 355 [155]
Thompson, C. R., 66 [20], 747 [29, 30],
762 [216], 763 [218]
Thompson, G. A., 580[25], 591 [25]
Thompson, J. N., 730 [91-93, 96], 732 [93,
110, 114], 733 [91, 92, 114, 115, 117],
735 [92], 737 [164, 173]
Thompson, S. Y., (47 [36]
Thompson, W. W., 606 [223]
Threlfall, D. R., 581 [28]
Tiemann, F., 340[87]
Tiews, J., 744 (7], 747 [21]
Tilak, B.D., 21 [41]
Tilney-Bassett, R., 609 [246]
Tischer, J., 51 [260a], 87 [222], 114[222],
139[429], 141[429]
Tishler, M., 352 [127], 369 [127], 387 [127]
Tobler, E., 424[337], 484[337], 486[337],
487 [337], 547 [337], 549 [337]
Todd, P. H., 758 [145]
Tolbert, N.E., 604[192-194]
Tolibekov, D., 602[175]
Tollin, G., 702[220]
Tomes, M.L., 594[140-142], 607[141, 231,
234], 608 [140, 142, 234, 243], 609 [234,
243], 610[252]
Toogood, J.B., 284[79]
Torto, F.G., 75 [145]
Tortuero, F., 751 [69]
T6th, Gy., 43 [155], 89 [243], 91 [243, 246,
247], 95[247], 111 [243, 246], 114[243],
116 [243, 246], 269 [25], 270 [29], 273 [29],
277[29], 289[114], 290[114], 291 [114],
292[114], 295[29], 296[132], 300(114,
132], 301 [114], 302 [29, 114], 305 [114],
306 [114], 307 [114], 308 [114], 309 [114],
310 [114], 311 [114], 312 [114], 341 [106],
360 [106], 508 [106], 546 [106]
Toube, T.P., 32[28], 34[28], 43[159, 160],
49 [160], 75 [152], 103 [292], 111 [330],
124[388], 162[152, 330], 260[61], 261 [61],
300[142], 303 (150, 151], 305 [142],
342[109], 360[109], 489[109], 490[109],
510[109], 551 [109], 552(109], 554[109],
618 [299]
Tourtellotte, M. E., 704 [240]
Trabesinger, P., 751 [66]
Traettebarg, J., 626 [337]
Trautmann, K. H., 658 [106]
Treharne, K.J., 581 [31], 585 [93], 605 [200],
612 [31]
Trenner, N. R., 352 [127], 369 [127], 387 [127]
Tripett, S., 393 [267, 268], 438 [267]
Trischmann, H., 78[168], 155[168]
Trivedi, A. H., 103 [294], 425 [341], 484[341],
487 [341], 531 [427], 549 [341]
Trout, M.E., 735[155], 736[155]
Truscheit, E., 328 [35], 351 [35], 352 [35],
357[35], 362[35, 175], 363[175, 178],
366 [35], 380 [35], 384 [230], 386 [175],
424 [338], 425 [338, 340], 430 [338],
483 [338, 408], 484 [338], 487 [338],
545[338,408],549[338]
Truscott, T.G., 680[81], 685[81], 694[81]
 Author Index
Tscharner, C., 104[305], 306[166, 167],
341 [107], 364[107, 179a], 484[107, 179a],
486[107, 179a], 487[107], 489[107],
546[107], 547[107, 179a], 548[107, 179a],
597 [156]
Tsoumanis, A., 757[138], 761 [138]
Tsuchiya, Y., 44 [190]
Tsuda, M., 385 [237], 395 [289]
Tsukida, K., 89 [232, 233, 239], 91 [251 ],
111 [338, 339], 113 [239], 269 [21], 277 [56,
57], 304 [154]
Tswett, M., 13 [9], 66 [35]
Tugwell, R. L., 752 [265]
Turchina, V. S., 602 [174]
Turner, J. F., 687 [125]
Tuttobello, L., 645 [41, 42]
Tuzson, P., 48 [228], 74 [127], 82 [127],
121[127], 169[515], 288[100], 289[100],
639[11]
u Villoutreix, J., 31 [9], 35 [53], 129 [396],
Udo, Y., 395 [289]
Uehleke, H., 596[145]
Uhde, G., 306[167], 341 [107], 364[107],
484 [107], 486 [107], 487 [107], 489 [107],
546 [107], 547 [107], 548 [107], 597 [156]
Upadhyay, R. R., 93 [271], 107 [326]
Urion, E., 757 [140]
Usher, C. D., 764[234]
Usher, G., 91 [250]
v Vorob'eva, L. M., 677 [59]
Vacano, E., 731 [105]
Vacheron, M.J., 40[125], 52[125], 117[546],
130[452]
Vail, W. J., 680[76]
Vakulova, L.A., 368 [195], 388 [245a]
Valadon, L. R. G., 48 [231], 589 [119],
605 [208], 610 [258]
Valenzuela, P ., 581 [39], 584 [39]
Vandeberg,J.S., 734[151]
Van den Ende, H., 615 [280]
VandercoQk:, C. E., 49[236], 169 [516]
van der Haak, P.J., 276[51]
van der Merwe, J. P·., 385 [233, 235]
van der Tuin, A. K., 689 [136]
van Dorp, D. A., 16, 340 [93], 353 [93],
366[189],369[189,202],387[189,242,243]
van Geelen, H. E., 369[197]
van ltallie, T. B., 268 [2]
van Leeuwen, P. H., 352 [126], 369 [126],
387 [126], 440[363], 536 [363]
895
van Niel, C. B., 121 [370], 124[382], 623[314,
318], 627 [338], 701 [217]
van Rij, J. H., 352 [126], 369 [126], 387 [126]
Varandani, P. T., 733 [121], 734 [121]
Varma, T.N.R., 581[47], 584[80]
Varsel, C., 51 [269]
Vaughn, J. R., 701 [212]
Verne, J., 656[89, 90]
Vernon, A. F., 662[114]
Vernon, L.P., 662[113, 114]
Vetter, W., 68 [77], 205 [18, 19], 209 [18, 19],
210[19], 213 [19], 226 [19], 235 [18],
239 [19], 245 [19], 248 [18, 19], 249 [19],
250 [19], 251 [19], 252 [18], 259 [19],
260 [18], 263 [18], 280[65], 531 [425],
535[425], 536[425]
Vevers, G., 21 [38]
Viani, R., 51 [265, 266], 52 [265]
Villanueva, V. R., 43 [159], 111 [330],
162[330], 300[141]
Villavicencio, R. U., 751 [60]
584 [88], 594 [138], 612 [267, 268]
Virgin, H. I., 702 [230]
Vishniac, W., 67 [64]
Vlad, P., 306 [173]
Voigt, H., 67[61], 68[61]
Yolk, W.A., 627[339]
Volker, 0., 36 [69], 37 [69], 38 [69], 39 [69],
638 [3], 641, 649 [65-68, 71, 72], 746 [16],
754[16]
Volker, w., 691 [152]
Vollrath, W., 751 [72]
Von Abrams, G.J., 606 [226]
Vose,J.R., 593[131a], 599[158]
Vredenberg, W. J., 695 [185], 696 [185]
Vreeland, J., 641 [26]
Vromen, S., 352 [133]
Vuilleumier,J.P., 747[20], 750[43, 50],
763[223], 764[50,223,235]
w
Wackenroder, H., 13 [6]
Wackernagel, H., 38 [105], 638 [6], 645 [37],
649[62, 82], 650[84], 655[85], 746[17,18]
Wada, T., 51 [272-274], 305 [162]
Wadsworth, W. S., 393 [270]
Wahlbaum, H., 86 [218]
Waight, E. S., 43 [155, 163, 164], 44 [164, 172,
191], 45[191], 46[203], 91 [246], 106[321,
323], 107 [323], 111 [246, 332], 116 [246],
148[454], 153[454], 270[29], 273[29],
277 [29], 295 [29], 302 [29]
 896 Author Index
Wald, G., 16, 17, 270 [32], 657 [93], 704 [234],
705 [244], 720 [27], 721 [34], 723 [34, 52-54],
724 [52, 53, 60], 728 [79], 729 [53, 54],
730 [34, 60], 732 [34], 737 [34], 738 [60]
Waldron, N. M., 91 [250]
Walker, D. M., 393 [267, 268], 438 [267]
Walker, 0., 75 [137], 133 [407], 288 [102],
606[214]
Walker, T., 79[185], 332[48], 337[48],
368[48], 374[48], 375[48], 408[48]
Wallace, R. H., 605 [203, 204],
674[35, 36]
Wallcave, L., 79 [259], 93 [267], 141 [259]
Walles, B., 675 [40, 41], 704[40]
Wallis, M., 583 [65]
Walser, A., 333[62], 342[62, 111], 360[167],
372[111, 167], 373[111], 390[111, 167],
408 [167], 477 [167], 479 [167], 480[167],
489[167], 492[167], 504[111, 167],
507 [111, 167], 523 [167], 524[167],
550[167], 551 [167], 557 [167], 558 [167],
560 [167], 561 [167], 563 [111, 167],
564[167]
Walter, F., 606[220]
Walter, M., 46 [199], 128 [393], 166[393],
457 [378], 459 [378], 460 [378], 461 [378],
517[378], 519[378], 540[378], 541 [378],
542 [378], 563 [378]
Walton, T.J., 46[205], 139[422], 157[422],
311 [176], 592[130], 600[161], 603[161]
Warburg, 0., 686 [107], 691 [149-152]
Wareing, P. F., 50 [246], 270 [36], 305 [159],
527[430,434], 706[247]
Warren, C.K., 42[147], 74[132], 78[174, 180],
137[132], 140[430], 141 [430], 292[118],
293 [118], 337 [71], 415 [328], 419 [328,
331], 466 [328], 497 [331], 501 [70, 71, 328],
505 [331 ], 526 [331 ], 529 [331 ],
558[70, 331], 560[331], 561 [70],
562 [71, 328]
Washiittl, J., 757 [130-132]
Wasmer,A.J.A., 355[149], 386[149]
Wassermann, A., 190[1], 191 [2]
Wassink, E. C., 687[120]
Watt,B.K., 746[14]
Way, J.E., 167 [503], 283 [78], 284[78],
395 [288], 405 [288], 425 [288], 469 [288],
471[288],544[288]
Weaver, L., 682 [96], 683 [96], 684 [96],
685 [96]
Webb, K.E., 720[26]
Weber, J., 338 [77], 339 [77]
Weedon, B.C.L., 14[20, 21], 31 [13], 32[15,
28], 33 [38], 34 [28, 39], 36 [78], 37 [86],
40[123], 41 [38, 126, 130], 42 [38, 147],
43 [151-153, 15(), 156, 157, 159, 160, 163,
164], 44[153, 156, 164,172, 191], 45[156,
157, 191a], 46[212], 49[160, 242], 50[255],
69 [102], 74[132], 75 [102, 145, 146],
78 [174, 175, 177, 180], 79 [182], 85 [182],
89 [242], 91 [242, 246, 247], 95 [146, 247,
280, 281], 101 [177, 286, 287], 103 [290,
292,297, 299], 104[313, 315, 317],
106 [319, 321, 323], 107 [146, 323],
109[280], 110[146], 111 [146, 246,319, 330,
332], 116 [246, 371], 122 [373], 123 [373,
376], 124 [385, 386, 388], 125 [389],
134[412], 137[132, 416], 139[175],
140[430], 141 [182, 430], 142[177, 438],
148 [317, 385, 451], 149 [460], 151 [317],
162 [152, 330], 167 [502, 503], 168 [317],
171[287], 174[524], 202[11], 204[16],
205 [17], 206 [17], 208 [17], 209 [16, 17],
251 [49, 50], 257 [54], 259 [54], 260 [17, 56.
61], 261 [61], 262 [49], 263 [49], 264[65],
268 [10, 11], 269 [10, 22], 270[29], 271 [40],
273 [22, 29, 44], 274 [22], 275 [10, 44, 47],
276[10, 53-55], 277 [22, 29], 278 [62],
279 [62], 280 [62], 281 [ 40, 69, 70, 73, 75],
282 [70], 283 [78], 284 [ 44, 54, 78],
285 [82, 83], 289 [111, 114, 115], 290[114],
291 [114], 292 [114, 118], 293 [118, 124],
295[29, 111,124,126, 127], 296[111],
297[136-138], 299[40, 111, 124, 138, 139],
300[111, 114, 141], 301 [114], 302[29, 114,
136, 138], 303 [139, 150, 151], 304 [ 40,
136, 139], 305[114, 139, 142], 306[114,
164], 307 [114, 139], 308 [114, 174],
309[114, 139], 310[114, 139], 311 [40, 114,
139, 176a], 312 [114], 328 [12, 26-28, 34],
336 [66], 337 [69, 70], 341 [106], 342 [109,
110], 347[123], 350[27], 354[146],
359[166], 360[106, 109, 166], 364[184],
380 [166], 385 [233, 234], 393 [27, 234],
395 [288], 398 [234], 400 [234, 296, 297,
300, 308, 309], 401 [234, 296, 309], 402 [310],
403 [315], 405 [27, 288], 408 [318],
409[166], 414[325], 415[327, 328],
416 [309], 419 [328, 331], 420[333, 334],
421 [296, 335], 422 [166], 423 [296, 309],
425 [288], 426 [296], 428 [296, 335],
429 [344], 430 [344], 431 [333], 433 [352],
434 [352], 439 [334], 441 [123], 444[123],
450[166], 452[166, 297, 327], 453[297],
454 [166, 296, 335], 455 [166, 309, 372],
456 [309, 325, 327, 373], 463 [310, 373],
464[310, 373], 465 [309], 466[315, 328,
384], 467 [315], 468 [309], 469 [27, 288, 310,
315, 386], 470[12], 471 [27, 288, 315],
477 [166, 296, 335], 478 [166, 296, 335],
489[12,27, 109,123, 166,296,308,335],
490 [12, 27, 109, 123, 166, 296, 308, 335],
 Author Index
497[69, 331], 499[66, 325], 500[325],
501 [69-71, 309, 328], 504[310], 505 [331],
508 [28, 106], 509 [28, 166, 296, 309, 335],
510[12, 109, 123], 511 [327], 512[327],
513 [325, 327], 514[308, 309, 325, 327],
515[309, 373], 516[28, 70, 419], 517[296,
310], 518 [12], 519 [310, 373], 521 [296],
522[66, 296], 523 [66], 526[331], 529[296,
331], 534 [ 444], 537 [123], 538 [166, 296,
327,335], 539[166, 309,325,327, 372,373],
540 [309], 542 [309, 315], 543 [12, 309],
544 [27, 288, 315], 546 [66, 106],
547 [28, 66, 166], 548 [28, 66, 166, 327],
549 [66, 166, 296, 327, 335], 550 [296, 335],
551 [109, 123], 552 [12, 109, 325, 327],
553 [12, 27, 123, 309, 327, 372], 554 [109,
123], 555 [123, 325, 373], 556 [12, 308, 309,
325], 557 [325], 558 [70, 310, 331, 373],
559 [12, 28, 373, 419], 560[308-310, 331],
561 [70, 309], 562[69, 71, 328], 563 [308],
589 [109], 600 [163], 618 [293, 299],
623[320], 709[266]
Weeks, 0. B., 32 [15, 31], 33 [15], 46 [210-214],
47 [211], 53 [213], 75 [142], 77 [142],
78 [142], 79 [142], 87 [142], 174 [142, 524,
529, 531], 176[531], 233[24], 257[54],
259[52, 54], 263[52], 268[11], 589[111],
616 [297], 618 [291- 293, 296], 619 [292,
297],620[292,297]
Wegfahrt, P., 53 [283], 161 [549], 163 [549]
Wehrli, H., 36[68], 155[477, 478], 288[99]
Weier, T.E., 675[43]
Weinheimer, A.J., 332 [49]
Weisler, L., 342 [108], 352 [128, 132],
359 [128], 380 [128]
Welzel, P., 305 [163]
Wendler, N. L., 352 [127], 369 [127], 387 [127]
Went, F. W., 593 [135], 607 [233], 608 [237,
240]
Werner, B., 720[19]
Westphal, F., 328 [5], 483 [5], 545 [5]
Wettstein, A., 155 [ 478]
Wettstein, D. von, 675 [42]
Wexler, S., 681 [87, 89]
White, E. P., 51 [275]
White, M. J., 48 [221, 229], 49 [221, 234-237],
85 [2111130 [211], 133 [211], 169 [511, 516,
518], 170[518-520], 171[520],462[379-
381], 464[379-381], 606[228], 610[249,
251]
Whiting, M.C., 296[134], 504[417]
Whittingham, C. P., 6.70[11], 687 [11]
Widdowson,E.M., 746[15]
Widmer, R., 268 [2]
Wie~tkowski, S., 612 [262]
Wiegand,.W., 79[183], l56[489]
Carotenmds 57 a
897
Wiemann, H., 752 [78]
Wiesenfeld, J. R., 729 [87]
Wildfeuer, I., 749[41], 750[41], 751 [65]
Wiley, R. H., 278 [63]
Wilkie, D. W., 37 [87], 155 [471]
Wilkinson, H., 49 [241]
Wilkinson, P. A., 340 [92]
Willhalm, B., 262 [64]
Williams, D. H., 243 [31 ], 252 [31 ], 260 [31]
Williams, R.J. H., 35 [53], 280 [66], 586 [104],
588[107], 589[107, 117], 590[117],
591 [117], 592 [107], 593 [107], 594 [138],
596[153, 154], 597[155], 601 [170],
602 [170], 603 [153-155]
Williams, T. P., 729 [89]
Williams, W. P., 747 [27, 28], 749 [27]
Williamson, I. P., 581 [36]
Willmer, J. S., 591 [120]
Willstatter, R., 13 [10, 11], 63 [8], 66 [8],
107 [327], 155 [ 473]
Willuhn, G., 607 [232]
Wilson, T., 681 [92], 685 [92]
Wilt, F., 724 [57]
Wingerd, W.H., 761 [194]
Winkelmann, K., 430 [347], 436 [357]
Winstanley, D. J., 707 [256]
Winter, M., 306 [172]
Winterstein, A., 33 [33], 35 [60], 46 [33, 199],
48 [33], 49 [33], 66 [37], 68 [74], 79 [183],
87 [228], 103 [298], 128 [393], 129 [395],
155[481,482,485-487], 166[393],
168 [395, 542], 169 [395], 244 [32], 268 [3],
281 [3], 288 [101], 289 [108], 457 [378],
459 [378], 460 [378], 461 [378], 517 [378],
519 [378], 540 [378], 541 [378], 542 [378],
563 [378], 639 [12, 13]
Wiss, 0., 639 [10]
Witt, H. T., 694 [183], 696 [192, 195]
Witt, K., 694 [184], 696 [195]
Wittig, G., 14, 333 [55, 61], 340[61], 352 [131],
355 [144], 357 [161, 162], 373 [209],
374 [209], 382 [227], 393 [263, 266],
400 [227], 428 [61], 435 [61], 438 [61],
489 [161], 496 [61], 520 [61], 521 [61],
522[61], 545[61, 161]
Wohlers, H. C., 344[115], 364[115]
Wolbach, S.B., 732 [112], 736 [112]
Wolf, D. E., 579 [11]
Wolf, G., 536[450], 719[13], 733[121],
734[121, 131, 138, 141, 146], 735 [153],
736 [163], 737 [168, 169]
Wolff, C., 694[183, 184], 696[195]
Wolff, I., 764 [254]
Wolfson, A.A., 38[101], 39[101], 638[5],
649[81], 652[81], 654[81]
Wolken,J., 662[117]
898 Author Index
Wollisch, E.G., 67[55]
Wong, P. S., 615 [285, 288]
Wood,F.W., 759[150]
Woods, R.J., 429[344], 430[344]
Woodward, R.B., 249[42]
Wright,H.F., 344[115], 364[115]
Wright, L. D., 579 [11]
Wright, L. J., 676 [55]
Wuhrmann,J.J., 582[52], 583[52]
Wiirsch, J., 328 [9, 37], 535, 536 [ 449, 451],
615 [287]
Wyckoff, R.W.G., 659[111]
y 141 [259, 414], 149 [459, 461], 153 [414],
Yagami, T., 52 [278], 332 [50]
Yagishita, K., 737 [169]
Yamaguchi, M., 41 [127-129, 132], 74[129],
75[129, 143], 104[129, 143,308-311, 316],
285[80], 364[181, 182], 484[181, 182],
486 [181, 182], 487 [182], 489 [182],
548 [181, 182], 549 [182]
Yamamoto, H. Y., 46 [206], 89 [234, 235],
579 [10], 582 [53-56], 583 [53-56],
600[165-167], 610[250], 690[144, 145],
695 [144]
Yamashita, K., 379 [222]
Yamazaki, R. K., 604[192, 193]
Yanovskaya, L.A., 355 [154]
Yeadon, A., 708 [259]
Yokota, M., 89 [232, 233], Ill [338, 339],
277 [56, 57], 304 [154]
Yokoyama, H., 43 [164], 44 [164], 46 [206],
48 [221-223, 229], 49 [221, 234-237],
85 [211], 111 [332, 336], 123 [336], 130 [211],
133[211, 406], 143[441], 169[511, 516, 518],
170[518-520], 171 [520], 308[167a],
462[379-381], 464[379-381], 579[9],
580 [9, 17], 582 [52, 57-59], 583 [57-59, 68],
584[68],596[150],606[228],610[249,251]
Yonemoto, T., 734 [135, 136], 736 [136]
Yonge, C. M., 647 [51-54]
Yoshikami, S., 724[58], 731[58], 732[109]
Yoshizawa, T., 723[54], 729[54, 86]
Young, R. W., 312[179]
Yuan, C., 586 [103]
Yudelevich, A., 581 [39], 584 [39]
z Zucker, H., 747 [21]
Zabin, 1., 580 [24]
Zachman, R. D., 721 [ 43, 46]
Zagalsky, P.F., 14[24], 21 [24], 39[110,
115-117], 53 [\10], 73 [125], 657 [92, 95, 98,
99], 658 [92, 98], 659 [95, 99, 112], 660 [92,
112], 661 [92], 663 [92, 112, 119], 703 [231,
233], 756 [112, 113]
Zakharkin, L. I., 361 [169], 388 [245 a]
Zalokar, M., 165 [540], 620 [303], 676 [52],
700[209], 701[209]
Zanetti, G., 750[54, 55]
Zechmeister, L., 12 [3], 14 [13], 31 [2], 32 [20],
36 [77, 79], 46 [201], 48 [228], 70[107],
71 [109], 74[127, 130], 75[140], 78[170],
79 [170, 259], 82 [127], 89 [239],
93 [263-265, 267, 268], 95 [263, 283],
98 [170, 263, 285], 101 [263, 283], 104 [268],
113[239], 121 [127], 137[413a], 140[285],
155[170], 169[515], 195[6], 204[13],
249 [ 44], 268 [6], 269 [6, 13, 15-17, 20],
270 [6], 271 [6, 38, 39], 272 [6], 273 [6],
274 [6], 275 [6, 46], 278 [ 46, 60], 279 [60],
280[ 46], 281 [6], 284 [6], 285 [6], 288 [95,
97, 100], 289 [97, 100, 107], 328 [11],
593 [135], 606 [209], 607 [233], 608 [237,
240], 639 [11]
Zehender, C., 269 [19], 620 [305]
Zeller, P., 74 [135, 136], 78 [136], 79 [186],
87[135], 104[306], 151[186,469], 155[483],
156 [186], 168 [508], 281 [71, 74, 77],
289 [109, 110], 291 [109, 110], 297 [71],
328[16, 17], 332[41], 333[60], 345[41, 119,
120, 122], 364[41, 119, 122, 186, 187],
365[119], 372[41, 119,187, 208], 388[245],
391 [186, 260], 393 [208, 262], 394 [262],
400 [305], 412 [60], 433 [350], 435 [354],
436 [305], 437 [359], 438 [245], 467 [60],
470[262, 350], 471 [262], 474[245],
480[186], 482[41, 187,208,245,260, 405],
483 [ 41, 187, 208, 405], 494 [305, 359],
497 [60], 499 [60, 416], 504 [208],
506 [186, 187, 245, 260], 507 [245, 260],
525 [359], 526 [359], 542 [60], 543 [262, 350],
544[262,350],545[60, 186,245,260,305,
359], 546 [305], 549 [60, 416], 550 [245],
551 [187, 245], 554[41, 187,208,359, 405],
560[208], 564[359]
Zhikhareva, L. T., 388 [245 a]
Zile, M., 720 [33], 722 [33], 734 [143],
737[33, 170, 171]
Zimmermann, G., 757 [118]
Zimmermann, M., 615 [287]
Zubrys, A., 155 [474], 289 [112]
Zurcher, P., 74 [136], 78 [136], 372 [208],
393 [208], 482 [208], 483 [208], 504 [208],
554[208], 560[208]
Zurzycki, J., 606 [219]
Zweig, G., 68 [80]
 899

Subject Index

A P-C 15 -Aldehyde ({J-ionylideneacetaldehyde)

cis isomer, 353, 380
Abscisic acid syntheses of, 350-355, 361
absolute configuration, 304, 305, 318 syntheses from, 369, 379, 380, 382, 384, 489,
biosynthesis, 50, 706 520,522
derivatives, 50 I)(-C 19 -Aldehyde, 370
glucoside, 50 P-C 19 -Aidehyde, 20
methyl ester, 304 syntheses of, 370-372
function, 706 syntheses from, 388, 442, 448, 480, 506, 507,
geometrical isomers, 270, 527, 533, 535 532
labelled compounds, 527, 536 Aldehydes, see also Apocarotenals, Carotenals
occurrence, 50,270 p.m.r., 233, 234
syntheses, 335, 343, 52~ 533-535 reactions, 74, 85-87, 128
Abscisin II, see Abscisic acid structures, 128
Abscisyl P-o-glucopyranoside, 50 Aleuriaxanthin (49), 36, 40, 150, 781
Absolute configuration, 288-319 Allenic carotenoids (69, 70, 86, 122, 124, 131,
Absorption maxima, 191, 192, 200 178,179,189, 190,246,273)
3-Acetoxyretinaldehyde, 480 biosynthesis, 43, 303
4-Acetoxyretinaldehyde, 390, 477, 479, 480 occurrence, 43
4-Acetoxyretinol, 390 reactions, 75, 106
3-Acetoxytorulene, 153, 155 structure, 106
Acetylenic carotenoids (41, 65, 66, 72, 94, 118, Alloxanthin (65), 784
166, 200, 202, 247, 252) absolute configuration, 291, 305
geometrical isomers, 271, 273 diacetate, 291
occurrence, 44 9,9'-di-cis isomer, 271, 281
reactions, 74, 106, 161 conformation, 284
structure, 106, 161 synthesis, 106, 281, 328, 350, 489, 490, 553
Actidione, 616, 620 i.r., 202, 204
Actinidiol, 51 occurrence, 44
Actinidiolide, 51 structure, 106
Actinioerythrin (272), 830 Amino acids, 582, 583
conformation, 285 Anchovyxanthin, 830
derivatives, 171, 172 Anhydroazafrinone amide, 167
occurrence, 39 Anhydroeschscholtzxanthin (35). 778
reactions, 85, 88 occurrence, 36, 37
structure, 161 syntheses, partial, 93, 95, 157, 160
Actinioerythrol, 39, 40, 171, 172 Anhydrofucoxanthols, 110
Adonirubin (196), 38, 143, 157, 814 Anhydrolutein (38), 779
Adonixanthin (167), 38, 143, 644, 807 Anhydrookenol, 126
I)(-C 14-Aldehyde, 344, 345, 370 Anhydrorhodovibrin (97), 791
P-C 14-Aldehyde, 20 biosynthesis, 621
cis isomer, 233, 234 m.s., 252, 256
derivatives, 345, 347 occurrence, 34
3-acetoxy compound, 345, 494, 496 reactions, 123
3-acetoxy retroeehydro compound, 345,347 structure, 123
diethyl acetal, 364, 370, 492, 494 synthesis, 513, 515, 556
enol ether, 345 Anhydrovitamin A
p.m.r., 234 Carr- Price reaction, 190
syntheses of, 344 p.m.r., 212, 218
syntheses from, 361, 364, 368, 370, 371, 379, syntheses of, 374, 477
494,496,520,528,530 synthesis from, 388, 474

Carotenmds 57 b
900 Subject Index

Anhydrowarmingol (63), 784 syntheses from, 21, 169-171, 450,460, 464,

Anhydrowarmingone, see Lycopen-20-al 505, 511, 516, 522
Annatto, 49, 755, 758 vitamin A activity, 744, 745
Antheraxanthin (119), 796 P-Apo-10'-carotenal (256), 827
biosynthesis, 600 biosynthesis, 639
diacetate, 252, 258 genetic studies, 609
geometrical isomers, 269 light absorption, visible, 193
metabolism, 602,603,607,663 metabolism, 639,640,719
m.s., 252, 258 occurrence, 48
occurrence, 41, 42,44 p.m.r., 226,228
reactions, 91 structure, 169
structure, 114 syntheses,444,445,447,450,537
Antioxidants, 63, 747, 757 P-Apo-12'-carotenal (262), 828
Aphanicin, see Canthaxanthin biosynthesis, 639, 641
Aphanin, see Echinenone derivatives, 87, 445
Aphanizophyll (94a), 117,791 geometrical isomers, 226, 228, 232, 239
Apo-P-carotenal, see P-Apocarotenal light absorption, visible, 193
Apo-ljl-carotenal, see Apolycopenal p.m.r., 226-228, 232
cx-Apo-8'-carotenal, 87 syntheses of, 440-444, 536
cx-Apo-12'-carotenal, 281,450,452, 509, 510, syntheses from, 447,448,450,458,460,461,
538 508,510
P-Apo-2-carotenal, see P-Apo-8'-carotenal vitamin A activity, 744, 745
P-Apo-3-carotenal, see P-Apo-10'-carotenal P-Apo-14-carotenal, 440, 536,639
P-Apo-4-carotenal, see P-Apo-12' -carotenal Apocarotenals (233, 242, 248, 249, 254, 256-
P-Apo-2'-carotenal (233), 823 259, 262-264, 267, 268), see also the
light absorption, visible, 193 individual Apolycopenals, Hydroxy-
m.s., 252, 256 didehydroapocarotenals, Hydroxy-
occurrence, 48 didehydrodihydroapolycopenals,
reactions, 87 Hydroxydihydroapolycopenals
syntheses,450,451, 538 analogous compounds, 451
P-Apo-4' -carotenal biosynthesis, 639, 640
light absorption, visible, 193 light absorption, visible, 193
metabolism, 640,719 metabolism, 639
syntheses of, 166,448-450, 537 occurrence, 639
syntheses from, 516, 518 p.m.r., 226, 228, 280
P-Apo-6'-carotenal, 193,448,449, 537 reactions, 168
P-Apo-8'-carotenal (248), 826 structure, 168
biosynthesis, 639 syntheses, 440-455, 536-538
colouring agent, 19, 749-751, 753, 755, 759, Apo-P-carotenoic acid, see P-Apocarotenoic
761, 762, 764 acid
derivatives, 87, 169 P-Apo-2'-carotenoic acid, 128
determination, 753, 764 methyl ester, 455, 461, 542
genetic studies, 609 P-Apo-4'-carotenoic acid, see Neurospora-
geometrical isomers xanthin
9-cis, 228, 230, 233 ethyl ester, 455, 460, 541
15,15'-cis, 203,204 methyl ester, see Neurosporaxanthin
labelled compound, 536 methyl ester
light absorption P-Apo-6'-carotenoic acid
i.r., 203, 204 methyl ester, 455, 460, 541
u.v., 196 P-Apo-8'-carotenoic acid (250), 826
visible, 193, 196 ethyl ester, see Ethyl P-apo-8' -carotenoate
metabolism, 639-641, 719 metabolism, 639-641
occurrence, 48 methyl ester, see Methyl P-apo-8'-
p.m.r., 208, 228-230, 233 carotenoate
syntheses of, 21, 139, 169, 445-449, 537' synthesis, 458, 541
 Subject Index 901

P-Apo-10'-carotenoic acid (260), 457, 541, 639, Apolycopenoic acid esters (243, 255), 464, 465,
827 824,827
methyl ester, 455, 457, 540 Apo-6'-lycopenol, 167
P-Apo-12'-carotenoic acid (263a), 828 Apo-4'-okenal, 452, 453, 518, 538
isoprenologous compounds, 527, 532 Apo-10'-violaxanthal (259), 163, 827
metabolism, 639, 641 Apo-12'-violaxanthal (263), 163, 828
methyl ester, 444, 455, 457, 540 Apo-2-zeaxanthinal, see P-Citraurin
synthesis, 457, 540 Apo-3-zeaxanthinal, see 3-Hydroxy-P-apo-
P-Apo-14'-carotenoic acid, 639 10'-carotenal
methyl ester, 228,440,455,457, 540 Aromatic carotenoids (6, 13-16, 52-54,79,
P-Apocarotenoic acid esters (235, 251), see 164, 180, 181), 40, 104, 285
also the individual P-Apocarotenoic Ascorbic acid, 603, 757
acids and 15,15'-Didehydro-P-apo- Asperxanthin, 830
carotenoic acid esters, Ethyl Astacein (199), 815
15,15'-didehydro-P-apo-carotenoate, Astacene (198), 815
Methyl P-apo-10' -carotenoate biosynthesis, 646, 650, 651
5,6-epoxide, Methyl15,15'-dide- derivatives, 77, 87, 146
hydro-P-apo-8' -carotenoate diacetate, 146
p.m.r., 228 dipalmitate (199), 815
syntheses, 455, 457-461 15,15'-didehydro-, 101
1/t-Apocarotenoic acid esters (243, 255) occurrence,39,40, 146
Apocarotenoic acids (234, 250, 260, 261, partial syntheses of, 88, 101, 141, 146
263a), see also the individual partial syntheses from, 160, 171
Apocarotenoic acids and Hydroxy- reactions, 101
P-apo-8' -carotenoic acid Astaxanthin (203), 816
biosynthesis, 639, 640 absolute configuration, 290, 312
metabolism, 639, 640 biosynthesis 612, 628, 638, 642-646,
Apocarotenoids (232-264), see also Diapo- 648-651,653,684
carotenoids in carotenoproteins, 39, 53, 657-659, 663.
occurrence, 48, 49 755-757
p.m.r., 226-235 derivatives
reactions, 160-169 dimethyl ether, 482, 483, 561
structures, 160-169 esters, 39, 160, 642, 649, 650, 756
syntheses,440-464,536-542 isolation, 72
P-Apo-2'-carotenol (232), 169, 443, 823 light absorption, visible, 290, 291
P-Apo-4'-carotenol, 166, 443 metabolism, 40, 653
P-Apo-6'-carotenol, 443 m.s., 252, 258
P-Apo-8'-carotenol, 169, 195, 197, 228, 443 occurrence,38-40,45,642,649,650,652,655.
P-Apo-10'-carotenol, 169,443 656, 754
P-Apo-12'-carotenol, 228,443,444 o.r.d., 290, 291
P-Apo-14'-carotenol, 440 reactions, 81, 85, 88, 143, 146
P-Apocarotenones, 455, 462-464, 519 Asterinic acid (200, 202), 815, 816
Apo-2-lycopenal, see Apo-6'-lycopenal isolation, 72
Apo-3-lycopenal, see Apo-8'-lycopenal occurrence, 45
Apo-15-lycopenals, 453, 454, 538-540 structure, 107, 162
Apo-4'-lycopenals, 455,457, 540 Asteroidenone (154), 38, 142, 804
Apo-6'-ly<;.openal (242), 48, 167, 168, 824 Asym. (-carotene, see 7,8,11,12-Tetrahydro-
Apo-8'-lycopenal (254), 826 lycopene
occurrence, 48 Aurochrome (139), 800
structure, 168 m.s., 252, 258
synthesis of, 455, 456, 539 structure, 14, 113, 114
syntheses from, 465, 513 Auroxanthin (141), 116, 602, 800
Apo-8'-lycopenals, 455, 456, 513-515, 539, 540 Azafrin (261), 828
Apo-12'-lycopenal, 454, 455, 509, 539 absolute configuration, 288
Apo-6'-lycopenoic acid, 167 derivatives, 167
902 Subject Index

Azafrin {261) (continued) caroten-1,1'-diol, see Bisanhydro-

derivatives bacterioruberin
methyl ester, see Azafnn methyl ester 2,2'-Bis{3-methyl-2-butenyl)-1,2,1 ',2' -tetra-
methyl ester methyl ether, 308 hydro-1/1,1/J-carotene-1 ,1 '-diol, see
i.r., 308 3,4,3',4'-Tetrahydrobisanhydro-
m.s., 252, 256 bacterioruberin
occurrence, 49 Bixin {265), 828
p.m.r., 206 colouring agent, 755, 758
reactions, 77, 78, 167, 168 geometrical isomerism, 268, 278-280,469
structure, 167, 168 isolation, 13
Azafrinaldehyde {258), 49, 169, 827 light absorption, visible, 278, 279
Azafrin methyl ester methyl ester, see Methylbixin
i.r., 308 occurrence, 31,49
m.s., 252, 256 p.m.r., 278-280
p.m.r., 206, 226, 230 reactions, 167
synthesis, partial, 167 structure, 167
Azafrinone, 167 synthesis, 469
Azafrinone amide, 167 Bixindial {diapo-6,6'-carotenedial)
derivatives, 87
m.s., 252, 256
B reactions, 86
syntheses
Bacterial phytoene, 830 partial, 121, 122
Bacteriochlorophyll, 33, 671-673, 680, 689, total, 466, 468, 543
690, 701 Blue-light effects, 698
Bacterioerythrin a, 793 Boron trifluoride-carotenoid complexes, 95,
Bacterioerythrin p, see Rhodopin 98
Bacteriopurpurin IX, 793
Bacteriopurpurin fJ, 830
Bacterioruberin {231), 822 c
biosynthesis, 618, 619
derivatives, 78, 123 Caloxanthin {69), 785
geometrical isomerism, 271 Camphor, 293
occurrence, 47 Camphoronic acid, 293
reactions, 174 Canaryxanthophyll, 642, 831
structure, 174 Canthaxanthin {193), 813
IX-Bacterioruberin, see Bacterioruberin analogue, 526
{3-Bacterioruberin, 830 biosynthesis, 612, 628, 644-647, 649, 651.
"IX-Bacterioruberin monomethyl ether", 663,665,684
see OH-Spirilloxanthin in carotenoproteins, 39, 658, 661, 663, 756
Bisanhydrobacterioruberin {228), 47, 174, colouring agent, 19,749-751,753-755,758,
822 759, 761, 762
Bisdehydro-, see also Dehydro-, Didehydro-, conformation, 285, 287
Tetradehydro- derivatives, 87
3,4,3',4'-Bisdehydro-{3-carotene, see 3,4,3',4'- determination, 764
Tetradehydro-fJ,fJ-carotene 15,15'-didehydro-, 93, 503, 504
Bisdehydrolycopene {17), see 3,4,3',4'-Tetra- geometrical isomers, 281, 328
dehydrolycopene labelled compound, 536
Bisdihydrocanthaxanthin, 655, 830 light absorption
2,2'-Bis{4-hydroxy-3-methy1-2-butenyl )-E.E- u.v., visible, 194, 196
carotene, see Decaprenoxanthin metabolism, 651, 653, 663, 665
2,2'-Bis{3-methyl-2-butenyl)-E,E-carotene, m.s., 252, 258
see Sarcinene occurrence,38,39,644,645.647,650,652,
2,2'-Bis{3-methyl-2-butenyl)-3,4,3' ,4' -tetra- 655,656,754
dehydro-1,2, 1',2' -tetrahydro-1/J ,1/J- p.m.r., 235-237
 Subject Index 903

reactions, 93, 95, 99 Capsorubol, 81, 137

structure, 98, 141 Capsorubone (210), 137, 818
syntheses Carangoxanthin, 831
partial syntheses of, 78, 98, 141 Carbon-13 nuclear magnetic resonance, 239,
partial syntheses from, 101, 141 242,243
total syntheses, 20, 21, 390, 402, 469, 477, Carbon-carbon double bond. 74. 75
479,480,482,483,497,500,503,560 Carbon-carbon triple bond. 74
X-ray crystallography, 285-287 16'-Carboxyl-3',4' -dehydro-J>-carotene,
Capsanthin (170), 808 see Torularhodin
absolute configuration, 288, 292-295, 308 Carboxylic acids. 85, 87, 128
biosynthesis, 42, 602, 609 see also Apocarotenoic acids,
chirality, 292 Torularhodin
colouring agent, 755, 759 Carcinoxanthin, see Flavoxanthin
derivatives, 87 Caricaxanthin, see Cryptoxanthin (39)
acetate, 137 Carophyll red, 749
3'-epi-, 292 Carophyll yellow, 749
genetic studies, 609 "P-Carotenal", see P-Apo-8'-carotenal
i.r., 202, 293 1/J,I/J-Caroten-20-al, see Lycopen-20-al
m.s., 252, 258 Carotenals (142-147), 68, 69
occurrence,31,42,52,606.607 see also Aldehydes, Apocarotenals
o.r.d., 295, 308 Carotene, 13
reactions, 137 K-Carotene, 831
structure, 137, 292-295 Carotene X, see P-Zeacarotene
synthesis, 337, 516, 559 a-Carotene (5), 773
Capsanthin 5,6-epoxide (188), 42, 114, 116, 812 absolute configuration, 288, 306, 315
Capsanthinone, see Capsanthone biosynthesis, 32, 594, 595, 597, 608
Capsanthol, 137 chirality, 292
Capsanthone (197), 814 enantiomer, synthesis of, 341,487, 489, 546
absolute configuration, 292 genetic studies, 608
m.s., 252, 258 geometrical isomerism, 271
occurrence, 42 i.r., 275
structure, 292 metabolism, 51, 308, 654, 684, 719
synthesis, partial, 137, 292 m.s., 252, 258
Capsochrome (191), 812 occurrence, 32,35
Capsolutein, 831 p.m.r., 235-237,239, 241
Capsolutein epoxides, 831 racemate, synthesis of, 487,489, 506, 508.
Capsorubin (205), 816 546
absolute configuration. 289.291-295,308, reactions, 75, 93, 101, 116, 133
309, 316 structure, 104,
biosynthesis, 42, 609 syntheses
chirality, 292 partial syntheses of, 114, 116
colouring agent, 755, 759 partial syntheses from, 93, 116, 133
dipalmitate, 289 total syntheses, 341, 360,428, 487,489.
epimer, 501, 562 506,508,564
genetic studies, 609 vitamin A activity, 744, 745
m.s., 252, 258 vitamin A, conversion to, 719
occurrepce,42,607 P-Carotene (3), 773
o.r.d., 291, 308, 309 biosynthesis, 312, 578, 580, 583, 584, 589-
racemates, 501, 562 592,594-597,604,608,610,612-615,
reactions, 87, 137 627
structure, 137 carbon-13 n.m.r., 242, 243
synthesis, 337, 41&, 466, 497, 501, 562 in carotenoproteins, 53, 640, 658, 704
Capsorubin isomer, see 2,2'-Dihydroxy- colouring agent, 19, 747, 748, 750, 755,
K,K-carotene-6,6' -dione 758-762
Capsorubindione, see Capsorubone conformation, 284, 285, 287
904 Subject Index

P-Carotene (3) (continued) y-Carotene (8), 774

determination, 763, 764 biosynthesis, 32, 35, 589, 594, 595, 615
functions, 676, 677, 681-683, 686, 688, conformation, 284
691-693,701 genetic studies, 608
genetic studies, 608, 610 geometrical isomers, 269, 271
geometrical isomers, 271, 682 light absorption
9-cis, 274 u.v., 193, 194
11-cis, synthesis of, 474, 545 visible, 193, 194, 284
13-cis, synthesis of, 474, 545 metabolism, 612, 613
15-cis occurrence,32,35
light absorption, 195, 198, 274 p.m.r., 235-237, 239, 241
m.s., 251 reactions, 75, 76, 93, 95, 103, 151
p.m.r., 236,237, 239 syntheses
stereomutation, 271, 682 partial syntheses, 75, 93, 103, 151
synthesis of, 281, 328, 481, 482, 489, 545 total syntheses, 428, 487, 489, 508, 509,
11,11 '-di-cis, synthesis of, 281, 483, 494, 495, 511, 512, 547
546 vitamin A activity, 744, 745
neo-B,269, 745 o-Carotene (11), 774
neo-U, 745 biosynthesis, 35, 591, 595, 608
poly-cis, 269 genetic studies, 608
history, 13 light absorption
isoprenologous compounds, 522 u.v., visible, 194, 195
labelled compound, 15,15'-02 occurrence, 35
m.s., 250, 251 p.m.r., 235-237, 239, 241
p.m.r., 235-237, 239, 242, 243 reactions, 75, 93, 95, 103
synthesis, 536 structure, 103
light absorption syntheses
u.v., 194-198, 200,274 partial syntheses of, 93, 103
visible, 194-198,200,274,284 partial syntheses from, 75, 95, 103
metabolism, 48, 51, 270, 628, 638-642, total syntheses, 450, 477, 478, 509, 548
645-649,651,707-709,71 8,719 8-Carotene (10), 774
m.s., 244, 246-252, 258,260 absolute configuration, 306
13 C n.m.r., 242, 243 biosynthesis, 595
occurrence,30-32,35,607 ,642,644,645, enantiomer, synthesis of, 341, 484,486, 548
647,650,652,655,748,75 4,755 light absorption
p.m.r., 205, 235-237, 239, 240 u.v., visible, 194, 195
reactions, 75, 93, 95, 98, 101, 113, 130, 133, m.s., 252, 258
140,141,156,169,304 occurrence, 35, 655
stability, 757 p.m.r., 206, 236, 237,239, 240
syntheses racemate, syntheses of, 477,478, 489, 490,
partial syntheses of, 91, 114 547
partial syntheses from, 93, 95, 98, 101, structure, 104
113, 130, 133, 140, 141, 156, 169 syntheses,341,428,484,4 86, 547
total syntheses, 19-21, 364, 428, 436-438, 81 -Carotene, see 8-Carotene
474-477,480-486,488,48 9,492-498, 8,8-Carotene, see 8-Carotene
504,506,508,545 8,1/f-Carotene, see o-Carotene
vitamin A activity, 744-746 (-Carotene (26), 777
as vitamin A precursor, 16, 17,718,719,744 biosynthesis, 93, 589-591, 605, 615, 622, 626
X-ray crystallography, 285, 287 function, 673, 677
p,p-Carotene, see P-Carotene genetic studies, 605, 608
p,y-Carotene (3a), 104, 151, 773 geometrical isomer, 269, 489,490, 550
p,8-Carotene, see <X-Carotene metabolism, 32, 589-591, 616, 618, 622,
p,cp-Carotene, see P- Isorenieratene 626
P.x-Carotene, see P-Renierapurpurin occurrence, 31,32
p,l/f-Carotene = p.m.r., 276, 277
 Subject Index 905

structure, 102 reactions, 102-105

synthesis, 489, 490, 522, 550
(-Carotene, asym., see 7,8, 11,12-Tetrahydro-
lycopene
( 1 -Carotene, 831
17-Carotene (4), 35, 104, 773
9-Carotene, 831
K-Carotene, 831
~-Carotene, see Aurochrome, (-Carotene
q>,q>-Carotene, see Isorenieratene
q>,x-Carotene, see Renieratene
q>,l/J-Carotene, see Chlorobactene
x,x-Carotene, see Renierapurpurin
x,l/J-Carotene, see y- Renierapurpurin
1/J,l/1-Carotene, see Lycopene
P-Carotene 5,6,5',6'-diepoxide (133), 91, 113,
114, 719, 799
p,p-Carotene-3,4-diol, see 3,4-Dihydroxy-
P-carotene
p,p-Carotene-3,3' -diol, see Zeaxanthin
p,p-Carotene-3,4'-diol, 101, 142
p,p-Carotene-4,4' -diol, see Isozeaxanthin
/J,E-Carotene-3,3' -diol, see Lutein
E,E-Carotene-3,3' -diol, see Tunaxanthin
K,K-Carotene-3,3'-diol, 831
q>,q>-Carotene-3,3' -diol, see 3,3' -Dihydroxy-
isorenieratene
1/1,1/J-Carotene-16,16'-diol, see Lycophyll
p,p-Carotene-3,4-dione, see 3-Ketoechinenone
p,p-Carotene-3,3' -dione
m.s., 252, 258
partial synthesis of, 86, 146
partial syntheses from, 86, 146
total synthesis, 489, 491, 560
p,p-Carotene-4,4' -dione, see Canthaxanthin
/J,E-Carotene-3,4-dione (156), 804
/J,E-Carotene-3,3' -dione, see 4,6' -Dihydro-
rhodoxanthin
p,l/J-Carotene-4,4'-dione (194b), 813
E,E-Carotene-3,3' -dione, see 6,6' -Dihydro-
rhodoxanthin
K,K-Carotene-6,6'-dione, see 3,3'-Dideoxy-
capsorubin
1/J,l/J-Carotene-4,4'-dione (194c), 814
IX-Carotene 5,6-epoxide (115), 42, 116, 795
IX-Carotene 5,8-epoxide, 42
P-Carotene 5,6-epoxide (114), 42, 113, 114, 719,
795
P-Carotene 5,8-epoxjde, 42, 95
Carotene-lipoprotein complex, 640
P-Carotene 15,15'-peroxide, 719
Carotenes (1-37), 7727779
see also Carotenoids
biosyntheses, 584-599
occurrence, 31-36
related compounds, 526
syntheses,474-490,492-499,506-509,
511-513, 545-551
p,p-Carotene-3,4,3',4' -tetrol, see Crustaxanthin
p,p-Carotene-3,4,3',4'-tetrone, see Astacene
K,K-Carotene-3,6,3',6'-tetrone (210), 137, 818
p,p-Carotene-3,4,3' -trio!, 143, 144, 642, 643,
831
p,p-Carotene-3,4,4' -trio!, see
3,4,4'-Trihydroxy-P-carotene
/J,E-Carotene-2,3,2' -trio!, 831
/J,E-Carotene-3,4,3' -trio!, 831
/J,E-Carotene-3,19,3' -trio!, see Loroxanthin
/J,E-Carotene-3,20,3' -trio!, see Pyrenoxanthin
P-Carotene-3,8,3' -trio! 5,6-epoxide, see
5,6-Epoxy-3,3' -dihydroxy-5,6, 7,8-
tetrahydro-p,p-caroten-8-one
p,p-Carotene-3,4,4' -trione, see Phoeniconone
Carotenoic acids, see Carboxylic acids
Carotenoid F1 , 161
Carotenoids
biosynthesis, 577-636
characterization, 69-72
colouring agents, 19-21, 746-764
definition, 48, 852
functions, 669-705, 710
geometrical isomerism, 268-284
isolation, 63-73
light absorption, 192-204
metabolism, 637-665
m.s., 243-265
n.m.r., 204-243
nomenclature, 22-24,851-864
reactions
group reactions, 73-92
related compounds, 526-531
vitamin A activities, 744, 745
as vitamin A precursors, 718, 719, 744-746
C 20 -Carotenoids, 517
C 30 -Carotenoids, 520-522
C 34 -Carotenoids, 522, 523
C 37 -Carotenoids, 522, 523
C 40 -Carotenoids (1-216), see also Carotenes,
Xanthophylls
aryl analogues, 522
occurrence, 30-46
reactions, 102-160
related compounds, 526
structures, 102-160
syntheses
partial, 93-101
total, 474-517, 545-564
C 43 -Carotenoids, 173
C 44 -Carotenoids, 526, 530, 531
906 Subject Index

C 45 -Carotenoids (216a-219), 819, 820 acetate, see 3-Acetoxytorulene

biosynthesis, 47, 616-619 occurrence, 36
occurrence, 46 structure, 149, 150
reactions, 173 syntheses, partial, 119, 120, 150, 153
structures, 173 Chirality, 292, 307-311
C 50 -Carotenoids (220-231), 820-822, 831 Chlorellaxanthin (193), 813
biosynthesis, 47, 616-619 Chlorobactene (15), 775
isolation, 71 biosynthesis, 622, 627
occurrence, 46 conformation, 285
reactions, 174 light absorption, 285
structures, 174 m.s., 252, 256
syntheses,522-526, 564 occurrence, 41
C 60 -Carotenoids, 47, 525, 526, 564 reactions, 104
C 70 -Carotenoids, 47 structure, 104
p,p-Caroten-3-ol, see Cryptoxanthin syntheses
p,p-Caroten-4-ol, see Isocryptoxanthin partial synthesis, 151
p,e-Caroten-3-ol, see Zeinoxanthin total syntheses, 451, 512, 513, 548
p,e-Caroten-4-ol, 101, 141, 151 Chlorophyll, 33, 65, 304, 612, 645, 671, 674, '
p,e-Caroten-3'-ol (43), 151, 780 675,677,680,682,683,685-692,694~
p,t/J-Caroten-3-ol, see Gazaniaxanthin, 697, 706
Rubixanthin Chloroxanthin (58), 783
p,t/J-Caroten-4-ol, 133, 134 biosynthesis, 623
p,t/J-Caroten-3'-ol, 150 occurrence, 34
e,t/J-Caroten-3-ol (51), 782 structure, 148
cp,cp-Caroten-3-ol, see Hydroxyisorenieratene synthesis, 514, 553
t/J,t/J-Caroten-16-ol, see Lycoxanthin Chromatography, 66-69, 273
t/J,t/J-Caroten-20-ol (63), 784 Chrysanthemaxanthin (130), 114, 116, 288, 798
Carotenol esters, see Esters Chrysophlein, 831
Carotenols Circular dichroism (c.d.), 289
reactions, 76-84 Cis peak, 195, 200, 273
Carotenonaldehyde (264), 828 Citranaxanthin (237), 823
P-Carotenone (216), 819 biosynthesis, 49, 610
derivatives, 87 colouring agent, 19, 749, 751
m.s., 252, 256 genetic studies, 610
occurrence, 48 occurrence, 49
p.m.r., 235-237 reactions, 464
reactions, 130 structure, 169
structure, 130, 131 syntheses,20,21, 169,170,462,464
P-Caroten-3-one, 492, 557 vitamin A activity, 744, 745
p,p-Caroten-4-one, see Echinenone Citranaxanthol, 463, 464
p,e-Caroten-4-one, see Phoenicopterone a-Citraurin, 155
p,t/J-Caroten-4-one = P-Citraurin (249), 826
y-Caroten-4-one, see 4-Keto-y-carotene absolute configuration, 307
Carotenoproteins, 656-665 derivatives, 87
chromoproteins, 53 genetic studies, 610
function, 662, 663, 703 history, 14
isolation, 72, 659 occurrence, 48
light absorption, 660, 661 o.r.d., 307
lipoproteins, 640, 657, 658, 756 structure, 169
molecular weight, 659 syntheses of, 137, 155, 169
occurrence,39,45,53, 755-757 synthesis from, 516
Carr-Price reaction, 16, 18, 190, 191 Citroxanthin, see Mutatochrome
C.d., see Circular dichroism Colourants, Colouring agents, see Pigmenters
Celaxanthin (44), 780 Colour measurement, 750, 752
absolute configuration, 289 Compound X, 831
 Subject Index 907

Conformation, 284-288 biosynthesis, 42

Corynexanthin (227), 46, 53, 174, 618, 821 occurrence, 42
C.p. 435, 831 o.r.d., 291
C.p. 450 (220), 47, 177, 618, 619, 820 reactions, 139
C.p. 450-mono-ol, 831 structure, 139
C.p. 473 (221), 47, 174, 618, 820 synthesis (racemate), 516, 559
C.p. 473-mono-ol, 831 Cryptocapsone, 139
Crocetin (269), 829 Cryptochrome (140), 114, 800
colouring agent, 21, 755 Cryptoflavin (127), 114, 797
derivatives 1/J-Cryptoflavin, 831
diethyl ester, 195, 197, 469, 470, 543 Cryptomonaxanthin (65), 784
dimethyl ester, see Dimethylcrocetin Cryptoxanthin (39), 779
geometrical isomer, 268, 269, 281 absolute configuration, 288, 289, 291, 307
history, 14 biosynthesis, 311, 600,607,628,647, 648
occurrence, 49 colouring agent, 752
reactions, 168 derivatives
structure, 160, 168 acetate, 114
synthesis, 469 palmitate, 95, 101, 151
a-Crocetin, see Crocetin geometrical isomer
y-Crocetin, see Dimethylcrocetin 15-cis, 281, 328
Crocetindial = metabolism, 42, 48, 649, 651
Crocetindialdehyde (267), 829 m.s., 252, 258
bis(diethyl acetal), 467, 542 occurrence, 36,607,647,752
light absorption o.r.d., 291, 307
u.v., visible, 195, 196 reactions, 95, 101, 114, 151
m.s., 252, 256 structure, 151
occurrence, 49 syntheses
structure, 168 partial syntheses from, 95, 101, 114,
syntheses of, 466, 467, 542 151
syntheses from, 446, 451, 452, 455, 456, 469, total syntheses
471,472,496-501,521-523,525,526 15-cis, 281
Crocetindiol, 195, 197 racemate, 504, 506, 510, 551
Crocetinsemialdehyde (268), 49, 168, 829 vitamin A activity, 744, 745
Crocin (271), 49, 52, 116, 168, 759, 829 as vitamin A precursor, 744
Crocoxanthin (41), 780 a-Cryptoxanthin (42, 43), 151, 780
absolute configuration, 310, 311 /3-Cryptoxanthin, see Cryptoxanthin
geometrical isomer Cryptoxanthin 5,6,5',6'-diepoxide (134), 114,
9-cis, 106, 271, 281, 328, 510, 552 799
occurrence, 44 Cryptoxanthin 5,6,5',8'-diepoxide (137), 799
o.r.d., 310, 311 Cryptoxanthin 5,8,5',8'-diepoxide, see
structure, 106 Cryptochrome
synthesis (9-cis), 106, 281, 328, 510, 552 Cryptoxanthin 5,6-epoxide (117), 42, 114, 795
Crustacyanin, 39, 657, 659-661 Cryptoxanthin 5,8-epoxide, see Cryptoflavin
Crustaxanthin (93), 790 Cucurbitaxanthin (73), 786
absolute configuration, 160, 312 Cyanopsin, 725
biosynthesis, 646 14-Cyanovitamin A acid, 386
conformation, 157 Cyclic (-carotene, 594
derivatives, 157 Cynthiaxanthin (65), 44, 45, 106, 306, 784
metabolism, 64~, 644, 646, 649, 651
occurrence, 38,645
reactions, 79, 81, 157, 160 D
structure, 157
synthesis, partial, 160 Damascenone, 51
Cryptocapsin (157), 805 1,2,7,8,11,12,7',8',11 ',12' -Decahydro-1/J,i/1-
absolute configuration, 291, 295, 308 carotene, see 1,2-Dihydrophytoene
908 Subject Index

7,8,11,12,15,7',8',11',12',15' -Decahydro- 7',8' ,11 ',12'-Dehydrononaprenoxanthin

1/1,1/J-carotene, see Lycopersene ( = P452) (216a), 617-619, 819
1,2,7,8,11,12,7' ,8',11 ',12'-Decahydro- 11 ', 12'-Dehydrononaprenoxanthin (217), 819
1/1,1/J-caroten-1-ol, see 1-Hydroxy- biosynthesis, 617, 618
1,2-dihydrophytoene m.s., 252, 256
Decahydrolycopene, 31 occurrence, 46
see also 1,2-Dihydrophytoene, structure, 174
Lycopersene 3',4'-Dehydro-17'-oxo-y-carotene,
Decapreno-P-carotene see Torularhodinaldehyde
light absorption, 2'-Dehydrophleixanthophyll, 117
u.v., visible, 200,201 2'-Dehydroplectaniaxanthin (162), 806
syntheses,392,522-526,564 m.s., 252, 256
Decapreno-e-carotene, 522, 524, 564 occurrence, 40
Decaprenoxanthin (226), 821 reactions, 130
absolute configuration, 312 structure, 130
biosynthesis, 617-619 synthesis, 516, 518, 559
geometrical isomerism, 277, 278 3-Dehydroretinaldehyde
m.s., 252, 258 formation, 719
occurrence, 46 function, 724, 725
p.m.r., 278 geometrical isomers
reactions, 78, 79, 174 9-cis, 380, 382
structure, 174 11-cis, 18, 270, 379, 380
Decaprenoxanthindialdehyde, 87, 174,233, 13-cis, 380, 382
234 9,13-di-cis, 380, 382
Decaprenoxanthin monoglucoside, 11,13-di-cis, 379, 380
see Corynexanthin history, 18
Deepoxyneoxanthin (86), 44, 76, 107, 112, 789 physical data, 18
Degraded carotenoids, 48, p.m.r., 216
see also Apocarotenoids, Seco- syntheses,379,380,382
carotenoids, Terpenoids 3-Dehydroretinoic acid, see Vitamin A 2 acid
Dehydro-, see also Bisdehydro-, Didehydro-, 3-Dehydroretinol, 718, 731
Monodehydro- geometrical isomers, 276
Dehydroadonirubin (195), 143, 814 9-cis, 220, 224, 380, 382
Dehydroadonixanthin (16Sa), 143, 144, 806 11-cis, 379, 380
Dehydrocarotene III (1), 772 13-cis, 222, 274, 380, 382, 385
Dehydro-P-carotene, see Retrodehydro- 9,13-di-cis, 380, 382
carotene 11,13-di-cis, 222, 225, 379, 380
3',4'-Dehydro-y-carotene, see Torulene history, 18
Dehydrocarotenes,93,95 physical data, 18
Dehydroflexixanthin, 88, 134 p.m.r., 216
Dehydrogenans-C-carotene-di-OH, 831 syntheses,379, 380,382,385
Dehydrogenans-C-carotene-mono-OR, 831 Dehydroretrocarotene, see Retrodehydro-
Dehydrogenans-P 373, see Nonaprenoxanthin carotene
Dehydrogenans-P 422, see 11',12'-Dehydro- 3,4-Dehydrorhodopin (55), 782
nonaprenoxanthin biosynthesis, 621
Dehydrogenans-P 439, see Decaprenoxanthin occurrence, 34
Dehydrohydroxyechinenone, see 3-Keto- reactions, 82
echinenone structure, 148
3'-Dehydrolutein (156a), 155, 252, 258, 804 synthesis, 513, 552
"Dehydrolycopene", see 3,4,3',4'-Tetrade- Dehydrorhodovibrin, see OH-Spirilloxanthin
hydrolycopene 3' ,4'-Dehydrorubixanthin, see Celaxanthin
3,4-Dehydrolycopene (18), 775 3-Dehydrovitamin A acid, see Vitamin A 2
occurrence, 33 acid
partial syntheses, 93, 103, 124, 149 Demethylated okenone (164), 41, 806
15,15'-Dehydrolycopersene, see Phytoene Demethylated spheroidene (57 a), 623, 783
 Subject Index
Demethyl carotenoids, see Norcarotenoids
9-Demethylretinaldehyde, 730
13-Demethylretinaldehyde, 730
9-Demethylvitamin A acid, 214, 225
13-Demethylvitamin A acid, 214,225
4'-Deoxookenone, 505, 512, 556
3'-Deoxycryptocapsin, 516, 558
Deoxyflexixanthin (158), 38, 134, 805
see also Myxobactone
Deoxylutein I (38), 779
Deshydroxydecaprenoxanthin (223), 821
biosynthesis, 617
m.s., 252, 258
occurrence, 46
structure, 174
4,4'-Diacetoxy-p,p-carotene, see Isozea-
xanthin, derivatives
Diadinochrome, 76
Diadinoxanthin (118), 795
m.s., 251, 258,265
occurrence, 44
reactions, 91
structure, 106, 113
Dianhydrobacterioruberin, see Bisanhydro-
bacterioruberin
Dianhydro-P-carotenone, 130
Dianhydroeschscholtzxanthin,
see Anhydroeschscholtzxanthin
Dianhydro-3,4,3' ,4' -tetrahydrobacterioruberin,
see Tetrahydrobisanhydrobacterio-
ruberin
Diapo-2,2'-carotenedial, 466,468, 543
Diapo-4,4'-carotenedial, 121, 122,466, 468,
472,503,543
Diapo-4,8'-carotenedial(C 25 ), 121, 122,452,
453
Diapo-6,6' -carotenedial, see Bixindial
8,8'-Diapocarotene-8,8'-dial, see Crocetin-
dialdehyde
Diapo-6,6'-carotenedioic acid, see Norbixin
Diapo-8,8' -carotenedioic acid, see Crocetin
Diapocarotenedioic acid esters (265-266a, 270),
469-472,828,829
see also Bixin, Crocetin, Diethyl
diapo-2,2' -carotenedioate, Diethyl
diapo-4,4' -carotenedioate, Diethyl-
norbixin
Diapo-8,8' -carotenediol, see Crocetindiol
Diapocarotenedion.es, 466, 469, 473, 503, 504
Diapocarotenoids (265-271), 828, 829
see also Retro diapocarotenoids
occurrence, 49
syntheses,464,466-473,542-544
Diatoxanthin (66), 785
absolute configuration 309, 310
909
occurrence, 44
o.r.d., 310
structure, 106
syntheses
partial synthesis, 76, 112
total synthesis, 504, 510, 554
Didehydro-, see also Dehydro-, Monodehydro-
3,4-Didehydro-P-C 14-aldehyde
p.m.r., 234
syntheses of, 345, 346
syntheses from, 379,494
3,4-Didehydro-P-C 15 -aldehyde (3,4-didehydro-
P-ionylideneacetaldehyde), 357, 369,
380,384
3',4'-Didehydro-/1-apo-2' -carotenal,
see P-Apo-2'-carotenal
15,15'-Didehydro-p-apo-2' -carotenal, 193, 450,
451, 517, 519
15,15'-Didehydro-P-apo-4'-carotenal, 193, 448,
450,461,516,518
15,15'-Didehydro-P-apo-6'-carotenal, 193,448,
460
15, 15'-Didehydro-P-apo-8' -carotenal
geometrical isomer, 233, 234
light absorption, visible, 193
p.m.r., 228, 233, 234
synthesis of, 448
syntheses from, 448,460,461,464, 512
15,15'-Didehydro-P-apo-10' -carotenal
light absorption, 193
p.m.r., 228
syntheses of, 444, 445, 447, 448, 537
syntheses from, 445, 447, 448, 459
11 ',12'-Didehydro-p-apo-10'-carotenal diethyl
acetal, 405, 444, 445
15,15'-Didehydro-p-apo-12' -carotenal
derivatives, 442, 445
light absorption, visible, 193
p.m.r., 228
syntheses of, 441, 442
syntheses from, 442,457,458, 460, 504, 508
15,15'-Didehydro-P-apocarotenals, 193
15,15'-Didehydro-/1-apocarotenoic acid esters,
228,231,235,458,459
3',4'-Didehydro-2' -apo-P-caroten-2' -ol (232),
823
15,15'-Didehydroapocarotenones, 463,464,
517,519
15,15'-Didehydro-apo-8'-lycopenal, 456,464,
539
15,15'-Didehydroastacene, 101
7,8-Didehydroastaxanthin (202), 816
15,15'-Didehydrocanthaxanthin
conformation, 285, 286
syntheses, 93, 503, 504, 560
910 Subject Index
3' ,4'-Didehydro-p,ift-caroten-16' -a!,
see Torularhodinaldehyde
3,4-Didehydro-a-carotene, 93
3,4-Didehydro-P,P-carotene (2), 772
occurrence, 36, 104
syntheses, 93, 504, 506, 507, 545
4,4' -Didehydro-p-carotene, see Retrodehydro-
carotene
15, 15'-Didehydro-P-carotene
carbon-13 n.m.r., 242
conformation, 285
syntheses, 481, 482, 488, 489, 493, 494,
496-498,504
15, 15'-Didehydrocarotenes, 209
3',4' -Didehydro-P,I/t-carotene =
(3',4')-Didehydro-y-carotene,
see Torulene
3',4'-Didehydro-.5-carotene, 95
3,4-Didehydro-1/t,l/t-carotene,
see 3,4-Dehydrolycopene
7,8-Didehydro-p,p-carotene-3,3'-diol,
see Diatoxanthin
7,8-Didehydro-p,e-carotene-3,3' -diol,
see Monadoxanthin
3',4'-Didehydro-P,ift-caroten-16'-oic acid,
see Torularhodin
3',4'-Didehydro-p,p-caroten-3-ol (38), 779
7,8-Didehydro-p,e-caroten-3-ol, see Croco-
xanthin
3' ,4'-Didehydro-P,I/t-caroten-3-ol,
see Celaxanthin
3',4'-Didehydro-P,ift-caroten-16'-ol (50), 46,
128,517,781
4,4' -Didehydro-P-caroten-3-one, 492, 557
3',4' -Didehydro-p,ift-caroten-4-one, see
4-Ketotorulene
3',4'-Didehydrochlorobactene, 92, 104, 120
15,15' -Didehydrocrocetindialdehyde, 466-469,
471,472,496-500,542
7,8-Didehydrocryptoxanthin, 504, 510, 551
3',4'-Didehydrocryptoxanthin, 95, 151
19, 19'-Didehydrodecapreno-P-carotene,
523-526,564
15, 15'-Didehydrodianhydro-P-carotenone,
235-237
15,15' -Didehydrodiapo-4,4' -carotenedial, 466,
468,503,543
15,15'-Didehydrodiethylcrocetin, 469, 470,
544
3,4-Didehydro-7,7'-dihydro-P-carotene, 504,
507,550
4,4' -Didehydro-4,7-dihydro-p-carotene, 474,
504, 506,.551
5,18-Didehydro-5,6-dihydro-p,p-carotene,
see p, y-Carotene
3,4-Didehydro-1 ,2-dihydro-1/t ,ift-carotene,
see 3,4-Didehydro-1 ,2-dihydro-
lycopene
6, 7-Didehydro-5,6-dihydro-p,p-carotene-
3,3' -diol (69), 785
3',4'-Didehydro-1 ',2' -dihydro-P,ift-carotene-
3,1 '-diol, see Saproxanthin
3',4'-Didehydro-1 ',2' -dihydro-P,ift-carotene-
1',2'-diol, see Plectaniaxanthin
3',4' -Didehydro-1 ',2' -dihydro-p,ift-carotene-
4,2'-dione (194a), 813
7' ,8'-Didehydro-5,6-dihydro-p,p-carotene-
3,5,6,3'-tetrol, see Heteroxanthin (94)
6,7-Didehydro-5,6-dihydro-p,p-carotene-
3,5,3'-triol, see Deepoxyneoxanthin
7',8' -Didehydro-5,6-dihydro-p,p-carotene-
3,5,3'-trio!, 790
1',16'-Didehydro-1',2' -dihydro-p,ift-caroten-
2'-ol, see Aleuriaxanthin
3',4'-Didehydro-1 ',2' -dihydro-P,ift-caroten-
1'-ol (47a), 781
3',4' -Didehydro-1',2' -dihydro-P,ift-caroten-
2'-ol, 831
3,4-Didehydro-1 ,2-dihydro-1/t ,1/t-caroten-1-ol,
see 3,4-Dehydrorhodopin
3,4-Didehydro-1,2-dihydrolycopene (20), 32,
103, 776
15,15'-Didehydrodimethylcrocetin 469,477,
544
15,15'-Didehydroechinenone, 517, 519, 558
3',4'-Didehydroechinenone, 141
"3',4' -Didehydro-18' -hydroxy-y-carotene"
(50),46, 128,517,781
3,4-Didehydroionene, 51
3,4-Didehydro-P-ionylideneacetaldehyde,
see 3,4-Didehydro-P-C 15 -aldehyde
3,4-Didehydro-P-C 18 -ketone, 369
11,12-Didehydro-P-C18 -ketone, 368
7,8-Didehydrolutein, see Monadoxanthin
3,4-Didehydrolycopene, see 3,4-Dehydro-
lycopene
15,15'-Didehydrophillipsiaxanthin, 503
3,4-Didehydroretinaldehyde, see 3-Dehydro-
retinaldehyde
11,12-Didehydroretinaldehyde, 377
3,4-Didehydroretinol, see 3-Dehydroretinol
7,8-Didehydroretinol, 375
11,12-Didehydroretinol, 379
11,12-Didehydroretinol methyl ether, 376, 404
4' ,5'-Didehydro-4,5' -retro-p,p-carotene,
see Retrodehydrocarotene
4' ,5'-Didehydro-4,5' -retro-p,p-carotene-
3,3' -diol, see Eschscholtzxanthin
4' ,5'-Didehydro-4,5' -retro-P.P-carotene-
3,3' -dione, see Rhodoxanthin
 Subject Index
4' ,5' Didehydro-4,5' -retro-{1,{1-caroten-3-one,
492,557
3,4-Didehydrorhodopin, see 3,4-Dehydro-
rhodopin
3',4'-Didehydrorubixanthin palmitate, 95
3',4'-Didehydro-7,8,1',2' -tetrahydro-{1,1/1-
carotene-3,4,1',2'-tetrol, 831
3,4-Didehydro-1,2,7' ,8' -tetrahydro-r/1 ,r/J-
caroten-1-ol, see Demethylated
spheroidene
15,15'-Didehydrotorularhodinaldehyde, 193,
517
15,15'-Didehydrotorularhodin methyl ester,
517
3,4-Didehydrotorulene, 92, 117, 119, 130,
134
7,8-Didehydrozeaxanthin, see Diatoxanthin
9,13-Didemethyl-9-ethylvitamin A acid, 216,
225
9,13-Didemethylretinaldehyde, 730
Didemethylspirilloxanthin, see 3,4,3',4'-Tetra-
dehydro-1,2,1 ',2' -tetrahydro-
r/1 ,r/1-carotene-1,1 '-diol
9,13-Didemethylvitamin A acid, 214, 225
3,3'-Dideoxycapsorubin, 501, 561
Diepoxy-{1-carotene =
5,6,5',6'-Diepoxy-5,6,5' ,6' -tetrahydro-{1,{1-
carotene, see {1-Carotene
5,6,5',6' -diepoxide
5,6,5',8'-Diepoxy-5,6,5' ,8' -tetrahydro-
p,p-carotene, see Luteochrome
5,8,5',8'-Diepoxy-5,8,5',8'-tetrahydro-
p,p-carotene, see Aurochrome
5,6,5',6'-Diepoxy-5,6,5' ,6' -tetrahydro-
{1,{1-carotene-3,3'-diol,
see Violaxanthin
5,6,5',8'-Diepoxy-5,6,5' ,8' -tetrahydro-
{1,{1-carotene-3,3'-diol,
see Luteoxanthin
5,8,5',8'-Diepoxy-5,8,5' ,8'-tetrahydro-
{1,{1-carotene-3,3'-diol,
see Auroxanthin
5,6,5' ,6'-Diepoxy-5,6,5' ,6' -tetrahydro-
p,p-caroten-3-ol, see Cryptoxanthin
5,6,5',6' -diepoxide
5,6,5',8'-Diepoxy-5,6,5',8'-tetrahydro-
PJ-caroten-3-ol (137), 799
5,8,5',8'-Diepoxy-5,8,5' ,8' -tetrahydro-
p,p-carote.n-3-ol, see Cryptochrome
5,8,5',8'-Diepoxyzeaxanthin, see Auroxanthin
3,3'-Diethoxy-3,4,3',4' -tetradehydro-
p,p-carotene, 489,491
Diethylcrocetin, see Crocetin, derivatives
Diethyl diapo-2,2'-carotenedioate, 469, 472,
544
911
Diethyl diapo-4,4'-carotenedioate, 469, 472,
544
Diethyl diapo-6,6' -carotenedioate =
Diethylnorbixin, 469, 471, 544
Digiprolactone, see Loliolide
Dihydroactinidiolide, 51
3,4-Dihydroanhydrorhodovibrin, see
1-Methoxy-1 ,2-dihydrolycopene
7,8-Dihydroapo-8'-lycopenal, 455, 514
7,8-Dihydroapo-12'-lycopenal, 454, 455, 465,
509,539
7,8-Dihydroastaxanthin, see Guaraxanthin
Dihydrobixin, see Eugorgiaenoic acid, 831
P-Dihydrocarotene, see 7,7'-Dihydro-
P-carotene
7,8-Dihydro-P-carotene, 650
7,7'-Dihydro-{1-carotene (37), 779
syntheses,482,494,496,551
15,15'-Dihydro-{1-carotene, 438, 494,496, 546
7',8' -Dihydro-{1,1/J-carotene =
7',8'-Dihydro-y-carotene, see P-Zeacarotene
7',8'-Dihydro-.5-carotene =
7',8' -Dihydro-e,r/J-carotene, see cx-Zeacarotene
1,2-Dihydro-r/J,r/J-carotene, see 1,2-Dihydro-
lycopene
7,8-Dihydro-r/J,r/J-carotene, see Neurosporene
5,6-Dihydro-{1,{1-carotene-5,6-diol, 78
1,2-Dih ydro-1/J ,r/J-carotene-1 ,20-dio I,
see Rhodopinol
5,6-Dihydro-1/J ,r/J-carotene-5,6-diol, 101
5,6-Dihydro-{1,{1-carotene-3,5,3' -trio!, see
5-Hydroxy-5,6-dihydrozeaxanthin
5,6-Dihydro-{1,{1-caroten-5-ol, 82, 114
1',2'-Dihydro-{1,1/J-caroten-1' -ol, see
1'-Hydroxy-l' ,2' -dihydro-y-carotene
1',2'-Dihydro-cp,r/J-caroten-1 '-ol, see
OH-Chlorobactene
1,2-Dihydro-r/J,r/J-caroten-1-ol, see Rhodopin
Dihydro-{1-carotenone, 130
Dihydrocrocetin, 831
1,2-Dihydro-3,4-dehydrolycopene, see
3,4-Didehydro-1,2-dihydrolycopene
1,2-Dihydro-3' ,4' -dehydrolycopene, 831
1,2-Dihydro-3,4-dehydro-1-0H -lycopene,
see 3,4-Dehydrorhodopin
Dihydrodianhydro-f:l-carotenone, 130
1',2'-Dihydro-1 ',2' -dihydroxy-4-ketotorulene
(171a), 808
1',2'-Dihydro-4,2'-diketotorulene (194a), 813
3'-Dihydro-cx-doradecin, see 4-Ketolutein
1',2'-Dihydro-1'-hydroxy-y-carotene (48), 781
1',2'-Dihydro-1' -hydroxychlorobactene,
see OH-Chlorobactene
1',2'-Dihydro-2'-hydroxy-3',4'-dehydro-
4-keto-y-carotene (161), 805
912 Subject Index
1',2' -Dihydro-1' -hydroxy-3,4-dehydrotorulene
glucoside, see Myxobactin
1',2' -Dihydro-1' -hydroxy-4-keto-y-carotene,
see 1'-Hydroxy-1',2'-dihydro-
y-caroten-4-one
1',2' -Dihydro-1 '-hydroxy-4-ketotorulene,
see Deoxyflexixanthin
1',2' -Dihydro-1' -hydroxy-4-ketotorulene
glucoside, see Myxobactone
1,2-Dihydro-1-hydroxylycopene, see Rhodopin
1',2'-Dihydro-1'-hydroxytorulene (47a), 781
1,2-Dihydrolycopene (21), 776
biosynthesis, 622, 626
occurrence, 32
structure, 103
7,8-Dihydrolycopene, see Neurosporene
1,2-Dihydroneurosporene (23), 32, 103, 626,
776
13', 14'-Dihydro-20' -nor-13' -apo-P-caroten-
14'-one, 392,462, 464
2'-Dihydrophillipsiaxanthin (177), 126, 130,
809
Dihydrophytoene, see Lycopersene
1,2-Dihydrophytoene (33), 32, 626, 778
1,2-Dihydrophytofluene =
"1',2' -Dihydrophytofluene" (31), 32, 626, 777
5,6-Dihydroretinaldehyde, 730
7,8-Dihydro-8, 7' -retro-P,P-carotene, see
7, 7' -Dihydro-P-carotene
7,8-Dihydro-8,1 0-retro-p,p-caroten-1 0-one,
see 10-Keto-7,10-dihydro-P-carotene
7',8'-Dihydrorhodovibrin, see OH-Spheroidene
4,4'-Dihydrorhodoxanthin, see p,p-Carotene-
3,3'-dione
4,6'-Dihydrorhodoxanthin, 489, 491, 561
6,6'-Dihydrorhodoxanthin, 489,491, 561
3,4-Dihydrospheroidene (100), 123, 792
11',12'-Dihydrospheroidene (101), 34, 123,
124,624,792
3,4-Dihydrospirilloxanthin (109), 124, 794
1',2'-Dihydrotrihydroxy-4-ketotorulene, 831
3,3'-Dihydroxycanthaxanthin, see Asta-
xanthin
3,3'-Dihydroxy-IX-carotene, see Lutein
3,4-Dihydroxy-P-carotene (64), 38, 142, 784
3,3'-Dihydroxy-P-carotene, see Zeaxanthin
4,4'-Dihydroxy-P-carotene, see Isozeaxanthin
3,3'-Dihydroxy-E-carotene, see Tunaxanthin
Dihydroxy-(-carotene, 831
3,3'-Dihydroxy-p,p-carotene-4,4' -dione,
see Astaxanthin
2,2'-Dihydroxy-K,K-carotene-6,6' -dione
(capsorubin isomer), 501, 562
3,3'-Dihydroxy-K,K-carotene-6,6' -dione,
see Capsorubin
Dihydroxycarotenoid C.wH 54 0 2 , 831
3,3'-Dihydroxy-p,p-caroten-4-one (167), .642,
643,647-649,651,807
3,3'-Dihydroxy-p,E-caroten-4-one,
see 4-Ketolutein
3,3'-Dihydroxy-p,K-caroten-6' -one,
see Capsanthin
Dihydroxy-3,4-dehydro-IX-carotene, 831
3,3'-Dihydroxydehydro-P-carotene,
see Eschscholtzxanthin
3,3 '-Dih ydroxy-2,3-didehydro- P,p-carotene-
4,4' -dione (201 ), 815
3,3'-Dihydroxy-7,8-didehydro-p,p-carotene-
4,4'-dione (202), 816
3,3 '-Dih ydrox y-2,3-didehydro-p,p-caroten-
4-one (165a), 143, 144, 806
3,3' -Dihydroxy-7',8' -didehydro-p,p-caroten-
4-one (166), 807
3,4'-Dihydroxy-2,3-didehydro-p,p-caroten-
4-one (168), 807
3,3 '-Dih ydrox y-2,3-dideh ydro- P,E-caroten-
4-one, see IX-Doradecin
4,1 '-Dihydroxy-15, 15'-didehydro-1 ',2' -dihydro-
y-carotene, 504, 505, 515, 555
3,1' -Dihydroxy-3',4' -didehydro-1',2' -dihydro-
p,ljf-caroten-4-one, see Flexixanthin
1',2'-Dihydroxy-3' ,4' -didehydro-1 ',2' -dihydro-
{J,Ijl-caroten-4-one (17la), 808
3,5-Dihydroxy-6, 7-didehydro-5,6, 7' ,8'-
tetrahydro-7' -apo-P-caroten-8' -one,
see Paracentrone
3,3'-Dihydroxy-5,8,5',8' -diepoxy-P-carotene,
see Auroxanthin
5,6-Dihydroxy-5,6-dihydro-10' -apo-
P-caroten-10'-al, see Azafrinal-
dehyde
5,6-Dihydroxy-5,6-dihydro-10' -apo-
P-caroten-10'-oic acid, see
Azafrin
3,3'-Dihydroxy-7,8-dihydro-p,p-carotene-
4,4'-dione, see Guaraxanthin
1',2'-Dihydroxy-1 ',2'-dihydrotorulene,
see Plectaniaxanthin
3,3'-Dihydroxy-4,4' -diketo-P-carotene,
see Astaxanthin
1,1 '-Dihydroxy-6,6' -diketo-1,2,5,6,1',2',5',6'-
octahydrolycopene,497,501,562
1,1 '-Dihydroxy-2,2' -diketo-1, 1',2,2' -tetrahydro-
3,3' ,4,4' -tetradehydrolycopene,
see Phillipsiaxanthin
3,3'-Dihydroxy-2,2' -dinor-p,p-carotene-
4,4' -dione 3,3'-diacylate,
see Actinioerythrin
3,3'-Dihydroxyechinenone, see Adonixanthin,
Hydroxyasteroidenone
 Subject Index
3,3'-Dihydroxy-5,6-epoxy-fi-carotene,
see Antheraxanthin
2-(Dihydroxyisopentenyl)-2' -isopentenyl-
fi-carotene (220), 47, 177, 618, 619,
820
3,3'-Dihydroxy isorenieratene (79), 787
occurrence, 41
reactions, 78, 157
structure, 157
synthesis, 336, 497, 499, 555
"3,3'-Dihydroxy -4' -keto-1)(-carotene ",
see 4-Ketolutein
3,3'-Dihydroxy-4-keto-fi-carotene (167), 807
3,3'-Dihydroxy luteochrome, see Luteo-
xanthin
Dihydroxylyco pene (81), 788
biosynthesis, 626
m.s., 252, 256
occurrence, 34
reactions, 82, 153
structure, 153
synthesis, 497, 499, 555
Dihydroxyneur osporene, 831
Dihydroxypirar dixanthin, 831
Dihydroxyretro -fi-carotene =
Dihydroxyretro dehydrocaroten e, see
Eschscholtzxan thin
2,2'-Dihydroxys pirilloxanthin, 79, 125
3,3'-Dihydroxy-2,3,2',3 '-tetradehydro-/i,/i-
carotene-4,4' -dione, see Astacene
3,3' -Dihydroxy-7,8,7',8' -tetradehydro-/i ,/i-
carotene-4,4' -dione (200), 815
3,3'-Dihydroxy-2,3,2' ,3' -tetradehydro-/i ,/i-
carotene-4,4' -dione dipalmitate (199),
815
1,1 '-Dihydroxy-3,4,3',4' -tetradehydro-1,2,1 ',2'-
tetrahydro-1{1 ,!{!-carotene-2,2' -dione,
see Phillipsiaxanthi n
1,1 '-Dihydroxy-3,4,3',4' -tetradehydro-1 ,2, 1',2'-
tetrahydrolycop ene, see 3,4,3',4'-
Tetradehydro-1 ,2,1',2' -tetrahydro-
1{!,1{1-carotene-1,1 '-diol
3,8'-Dihydroxy-5',6', 7',8' -tetrahydro-5' -apo-
18'-nor-fi-caroten- 6' -one, see
8' -Hydroxy-7',8' -dihydroreticul ata-
xanthin
1,1 '-DihyQroxy-1 ,2,1 ',2' -tetrahydro-
(-carotene (82), 153, 788
3,3'-Dihydroxy-7,8,7',8' -tetrahydro-K,K-
carotene-6,6' -dione (206), 817
1,1 '-Dihydroxy-1,2, 1',2' -tetrahydrolyco pene,
see Dihydroxylycop ene
16,16' -Diisopropyl-/i,fi-carotene, 526, 530,
531
Diketo-, see also Dioxo- or -dione
913
"Diketobacteri oruberin ", see Phillipsia-
xanthin
3,4-Diketo-IJ(-carotene (156), 804
3,4-Diketo-fi-carotene, see 3-Ketoechineno ne
4,4'-Diketo-fi-carotene, see Canthaxanthin
4,4'-Diketo-y-c arotene (194b), 813
2,2'-Diketo-15,15' -didehydrospiri lloxanthin,
502,503
4,10'- Diketo-7',10' -dihydro-fi-caro tene, 477,
479,561
4,4' -Diketolycopen e (194c), 814
Diketopirardix anthin (194), 141, 663, 665, 813
3,3' -Diketoretro-fi- carotene =
3,3'-Diketoretro dehydro-fi-caro tene,
see Rhodoxanthin
2,2'-Diketospiri lloxanthin (208), 817
biosynthesis, 628
m.s., 246, 247, 252, 256
occurrence, 34
reactions, 125, 126
structure, 125, 126
syntheses,489,4 90, 501-503,563
10,1 0'-Diketo-7 ,1 0, 7' ,1 0' -tetrahydrodeca preno-
fi-carotene, 523, 524, 564
6,6'-Diketo-5,6, 5',6' -tetrahydrolyco pene, 496,
501,561
1,1' -Dimethoxy-3,4-didehydro-1,2,1 ',2', 7',8'-
hexahydro-1{!,1{1-carotene, see 3,4,7,8-
Tetrahydrospir illoxanthin
1,1 '-Dimethoxy-3,4-didehydro-1,2,1 ',2'-
tetrahydro-1{1,1{1-carotene, see
3,4-Dihydrospi rilloxanthin
1,1 '-Dimethoxy-1 ,2, 7,8,1 ',2'-hexahydro-1{1 ,!{!-
carotene (112), 124, 794
1,1 '-Dimethoxy-1 ,2, 7,8,11,12,1 ',2' -octahydro-
1{1,1{1-carotene, see 3,4,3',4',7',8',11',12'-
0ctahydrospiril loxanthin
1,1 '-Dimethoxy-3,4,3',4'-tetradehydro-1,2,1 ',2'-
tetrahydro-1{1,1{1-carotene, see
Spirilloxanthin
1,1 '-Dimethoxy-3,4,3' ,4' -tetradehydro-1 ,2, 1',2'-
tetrahydro-1{1 ,!{!-carotene -2,2' -dione,
see 2,2'-Diketospiri lloxanthin
1,1 '-Dimethoxy-1 ,2,1 ',2' -tetrahydro-1{1 ,1/1-
caroten-20-al, see 3,4,3',4'-
Tetrahydrospiri lloxanthin-20-a l
1,1 '-Dimethoxy-1,2,1 ',2'-tetrahydro-1{1 ,!{!-
carotene=
1,1 '-Dimethoxy-1,2,1 ',2' -tetrahydrolyco pene,
see 3,4,3',4'-Tetrahy drospirilloxanth in
1,1 '-Dimethoxy-1,1 ',2,2'- tetrahydroneur o-
sporene, see 3,4,3',4',7',8'-
Hexahydrospiri lloxanthin
2,2'-Dimethyl-/i,/i-carotene, 526, 530, 531
16,16'-Dimethyl-fi,fi-carotene, 526, 530, 531
914 Subject Index

2,2'-Dimethyl-a-carotene= occurrence,38,644,645,647,649,651,
2,2'-Dimethyl-e,e-carotene, 235-237, 526, 530, 652,655
531 p.m.r., 235-237
Dimethylcrocetin (270), 829 reactions, 87, 139, 140
geometrical isomers structure, 98, 139
9-cis, 281, 328, 469, 470, 543 syntheses
15-cis, 281, 469, 470, 544 partial, 140, 151
m.s., 244 total, 390,402,464,477,479,505,517,
structure, 168 519,558
syntheses,281,469,470,543 vitamin A activity, 745
Dimethylcyclodecapentaene, 249 Eicosahydrolycopene, see Tetrahydro-
Dimethyl 6,6' -diapocarotene-6,6'-dioate, see phytoene
trans-Methylbixin (266) Electronic spectra, see Light absorption
Dimethyl 9-cis-6,6' -diapocarotene-6,6'-dioate, Eloxanthin (120), 796
see 9-cis-Methylbixin (266a) Enzymes, 17,583
Dimethyl 8,8' -diapocarotene-8,8'-dioate, see Carotene cleavage enzyme, 640
Dimethylcrocetin P-Carotene 15,15'-oxygenase, 640,719
Dimethyl retro-diapo-3,3' -carotenedioate, 252, Glucuronyl transferase, 732
256,469,473 Retinaldehyde isomerase, 17, 724, 731
Dimethyl retro-diapo-7,7' -carotenedioate, 469, Retinaldehyde oxidase, 732
473 Retinaldehyde reductase, 719
Dinorcrocetin, 439 Retinol isomerases, 731
Dinoxanthin, 831 Squalene synthetase, 31
Diosphenols, 39, 65, 101, 107, 134, 142, 263 Violaxanthin deepoxidase, 662
Dioxo-, see also Diketo- or -dione Enzymic hydrolysis, 92, 117
5,6-Dioxo-10' -apo-5,6-seco-P-caroten-1 0' -al, Epoxy-, see also epoxide
see Carotenonaldehyde Epoxy carotenoids (114-141, 187-191, 259,
3,4-Dioxo-4' -hydroxy-P-carotene, see 263, 273)
4'-Hydroxy-p,p-carotene-3,4-dione biosynthesis, 602, 606
5,6,5',6' -Diseco-p,p-carotene-5,6,5',6' -tetrone, de-epoxidation, 91, 662
see P-Carotenone epoxidation, 95, 332
Dodecahydro-P-carotene, 831 function, 672, 690
Dodecahydrolycopene (30), 777 metabolism, 602, 662
Dodecapreno-P-carotene m.s., 264
light absorption, 200, 201 occurrence,41,42,610
m.s., 248 p.m.r., 206,228,230
reactions, 95 reactions, 42, 71, 88-91, 113-116, 163, 296
syntheses, 525, 526, 564 structure, 113 -116, 163
IX-Doradecin (168a), 143, 807 syntheses, partial, 95, 332
P-Doradecin (16Sa), 143, 144, 806 9,10-Epoxycanthaxanthin, 95, 99
IX-Doradexanthin, see 4-Ketolutein 13,14-Epoxycanthaxanthin, 95, 99
P-Doradexanthin (167), 807 5,8-Epoxy-IX-carotene, see Flavochrome
Dormin, see Abscisic acid 5,8-Epoxy-p-carotene, see Mutatochrome
1,2-Epoxy-1,2, 7,8,11,12,7' ,8' ,11 ',12' -decahydro-
1/1,1/J-carotene, see Phytoene
E 1,2-epoxide
5,6-Epoxy-7',8' -didehydro-5,6-dihydro-P,P-
Echinenone (148), 803 carotene-3,3'-diol, see Diadino-
biosynthesis, 612, 628, 629, 647-649, 651 xanthin
derivatives, 87 5',6'-Epoxy-6, 7-didehydro-5,6,5',6' -tetrahydro-
geometrical isomers, 281 p,p-carotene-3,5,19,3'-tetrol, see
light absorption Vaucheriaxanthin
u. v., visible, 194-196 5',6'-Epoxy-6, 7-didehydro-5,6,5',6' -tetrahydro-
metabolism, 645, 646, 663, 665 p,p-carotene-3,5,3' -trio!, see
m.s., 252, 258 Neoxanthin
 Subject Index
5' ,8'-Epoxy-6, 7-didehydro-5,6,5' ,8' -tetrahydro-
fJ,fJ-carotene-3,5,3' -trio!, see
Neochrome
5,6-Epoxy-5,6-dihydro-fJ,fJ-carotene, see
fJ-Carotene 5,6-epoxide
5,8-Epoxy-5,8-dihydro-fJ,fJ-carotene, see
Mutatochrome
5,6-Epoxy-5,6-dihydro-fJ,e-carotene, see
IX-Carotene 5,6-epoxide
5,8-Epoxy-5,8-dihydro-fJ,e-carotene, see
Flavochrome
5,6-Epoxy-5,6-dihydro-fJ,fJ-carotene-3,3 '-diol,
see Antheraxanthin
5,8-E poxy-5,8-dihydro-fJ,fJ-carotene-3,3'-diol,
see Mutatoxanthin
5,6-Epoxy-5,6-dih ydro-fJ,e-carotene-3,3 '-diol,
see Lutein epoxide, Taraxanthin
5,8-Epoxy-5,8-dihydro-fJ,e-carotene-3,3 '-dio I,
see Flavoxanthin
9,10-Epoxy-9,10-dihydro-fJ,fJ-carotene-
4,4'-dione, 95, 99
13,14-Epoxy-13, 14-dihydro-fJ,fJ-carotene-
4,4'-dione, 95,99
5,6-Epoxy-5,6-dihydro-fJ,e-carotene-3,3',6'-trio!
(123), 797
5,8-Epoxy-5,8-dihydro-fJ,e-carotene-3,3 ',6'-trio!
(132), 798
5,6-Epoxy-5,6-dihydro-fJ,fJ-caroten-3-ol, see
Cryptoxanthin epoxide
5,8-Epoxy-5,8-dihydro-fJ,fJ-caroten-3-ol, see
Cryptoflavin
5',8' -Epoxy-5',8' -dihydro-fJ,fJ-caroten-4-ol, 101
5,8-Epoxy-5,8-dihydro-fJ,t/J-caroten-3-ol, see
Rubichrome
5',6'-Epoxy-5' ,6' -dihydro-fJ,t/J-caroten-4-one,
831
5,8-Epoxy-3,3' -dihydroxy-IX-carotene, see
Flavoxanthin
5,6-Epoxy-3,3' -dihydroxy-5,6-dihydro-fJ,K-
caroten-6'-one, see Capsanthin
5,6-epoxide
5,8-Epoxy-3,3'-dihydroxy-5,8-dihydro-fJ,K-
caroten-6'-one (191), 812
5,6-Epoxy-3,3'-dihydroxy-5,6,7,8-tetrahydro-
fJ,fJ-caroten-8-one (187), 106, 107, 811
5,8-Epoxy-3-hydroxy-y-carotene, see
Rubichrome
5,6-Epoxy-3-hydroxy-5,6-dihydro-1 0' -apo-
fJ-caroten-10'-al, see Apo-10'-
violaxanthal
5,6-Epoxy-3-hydroxy-5,6-dihydro-12' -apo-
fJ-caroten-12'-al, see Apo-12'-
violaxanthal
5,8-Epoxylutein, see Flavoxanthin
5,6-Epoxyzeaxanthin, see Antheraxanthin
915
Eschscholtzxanthin (84), 788
absolute configuration, 288
biosynthesis, 602
dipalmitate, 95
geometrical isomerism, 275
i.r., 275
occurrence,37,656
reactions, 79, 146, 156
structure, 156, 469
syntheses, partial, 146, 156
Eschscholtzxanthone (192), 37, 146, 812
Esters(ofcarotenols)(68, 74,121,175,179,190,
199, 272, 273), 65, 68, 77, 84, 85, 146
Ethoxyquin, 747
Ethyl fJ-apo-8' -carotenoate
determination, 764
colouring agent, 19, 749-751, 753, 758-762
labelled compound, 536
light absorption
u.v., visible, 195, 197-199
syntheses, 455, 458, 459, 541
vitamin A activity, 744, 745
Euglenanone, 141, 803, 813, 831
Euglenarhodone (198), 815
Eugorgiaenoic acid, 831
F
Farnesal, 421
Farnesol, 276
Farnesyl pyrophosphate, 31, 47, SO, 583, 584,
586, 588, 611, 641
Fenicotterina, 831
Flavacin (125), 114, 797
Flavochrome (126), 114, 797
Flavorhodin (22), 776
Flavoxanthin (130), 114, 116,288, 798
Flexixanthin (171), 38, 88, 134, 808
Foliachrome (131), 252, 258, 798
Foliaxanthin, see Neoxanthin
Fourier transform technique, 239
Fucochromes, 108,109,295,296
Fucoxanthin (190), 812
absolute configuration, 288,289, 295-300,
314, 317
biosynthesis, 43, 602, 684
derivatives, 87, 107, 108
function, 686, 687
i.r., 202, 204
isolation, 13, 65
metabolism, 49
m.s., 251, 258, 265
occurrence, 31, 43, 44
p.m.r., 235-237
916 Subject Index

Fucoxanthin (190) (continued) 7,8,11,12,7',8'-Hexahydro-{J,{J-carotene, see

reactions, 75, 106-110, 162, 163,295,297 Phytofluene, cyclic analogue
structure, 46, 106-110, 113,295 1,2,7,8,11,12-Hexahydro-Y,,Y,-carotene, see
Fucoxanthinol (189), 812 1,2,7,8,11,12-Hexahydrolycopene
absolute configuration, 300 1,2,7,8,1 ',2'-Hexahydro-Y, ,Y,-carotene, see
function, 688 1,2,1 ').'-Tetrahydroneurosporene
occurrence, 43 1,2,7,8,7',8'-Hexahydro-Y,,Y,-carotene, see
p.m.r., 277 7,8,1';1.',7',8'-Hexahydrolycopene
reactions, 107, 109, 295 7,8,11,12,7',8'-Hexahydro-Y,,Y,-carotene, see
structure, 107, 109, 113, 295 Phytofluene
Fucoxanthols, 108-110,295 1,2,7',8' ,11 ',12' -Hexahydro-Y, ,Y,-caroten-1-ol,
see 1-Hydroxy-1,2,7',8',11',12'-
hexahydrolycopene
G 1,2,7,8,11,12-Hexahydrolycopene (27), 32, 777
Hexahydrolycopene =
Galloxanthin, 831 7,8,11,12,7',8'-Hexahydrolycopene, see
Gazaniaxanthin (46), 781 Phytofluene
absolute configuration, 307 7,8,1',2',7',8'-Hexahydrolycopene (29), 32, 777,
conformation, 284 3,4,3',4',7',8'-Hexahydrospirilloxanthin (112),
geometrical isomerism, 269, 274, 276, 277 124, 794
isolation, 273 1-Hexosyl-1,2-dihydro-3,4-didehydroapo-
light absorption, 274, 284 8'-lycopenol (253), 117, 165,826
occurrence, 36 Hill reaction, 687, 691, 702
p.m.r., 276, 277 Hopkinsianone, 161
structure, 149 Hopkinsiaxanthin (247), 161, 825
Geraniol, 276, 584 Hydroxy-, see also OH- or -ol
Geranylgeranyl pyrophosphate, 31, 584, 585, Hydroxyactinioerythrin, 831
588,589,592,593,611 3-Hydroxy-IX-apo-12' -carotenal, 510
Geranyl pyrophosphate, 47, 583, 584, 586, 611, 3-Hydroxy-{J-apo-8' -carotenal, see {3-Citraurin
641 3-Hydroxy-{J-apo-10'-carotenal (257), 48, 169,
Glover-Redfearn degradation, 638, 639 827
IX-Glycols, 70, 78, 79, 81 3-Hydroxy-{J-apo-12'-carotenal, 510
Glycosides (47, 54, 57, 77, 77a, 90-92, 94a-96, Hydroxy-{J-apo-8'-carotenoic acid, 831
145, 159, 172, 176,225, 227, 253, 16-Hydroxyapo-8'-lycopenal, 455,456, 515,
255, 271), 53, 79, 91, 116, 164, 260 539
Glycosidic apocarotenoids (253, 255, 271), 164 Hydroxyasteroidenone (166, 167), 45, 143, 807
Glycymerin, 45, 807, 831 Hydroxycamphonanic acid, 293, 295
Grasshopper ketone 3-Hydroxycanthaxanthin, see Adonirubin,
absolute configuration, 303 Phoenicoxanthin
biosynthesis, 304, 709 Hydroxycapsanthin-like compound, 831
function, 709 Hydroxycapsolutein, 831
occurrence, 50, 709 Hydroxycapsolutein 5,6-epoxide, 831
syntheses, 332, 535 Hydroxy-IX-carotene, 831
Guaraxanthin (204), 146, 650, 653, 655, 816 3-Hydroxy-IX-carotene, see Zeinoxanthin
4-Hydroxy-IX-carotene, 101, 141, 151
3'-Hydroxy-IX-carotene (43), 151, 780
H 3-Hydroxy-{J-carotene, see Cryptoxanthin (39)
4-Hydroxy-{J-carotene, see lsocryptoxanthin
Haematoxanthin, 831 3-Hydroxy-y-carotene, see Rubixanthin
Helenien (74), 155, 786 4-Hydroxy-y-carotene, 133, 134
Heteroxanthin (94), 44, 49, 106, 790; (187), 811 3'-Hydroxy-y-carotene, 150
Hexadecahydrolycopene, see Phytoene 3-Hydroxy-15-carotene (51), 782
2,3,2' ,3',4',5'-Hexadehydro-4,5' -retro-{3,{3- Hydroxy-(-carotene, 831
carotene, see Anhydroeschscholtz- 3-Hydroxy-{J,{J-carotene-4,4' -dione, see
xanthin Adonirubin, Phoenicoxanthin
 Subject Index 917
3'-Hydroxy-,B,,B-carotene-3,4-dione (165a), 143, 1'-Hydroxy-3',4'-didehydro-1',2'-dihydro-
144,806 ,B,Y,-caroten-2'-one, see
4'-Hydroxy-,8,,8-carotene-3,4-dione (168), 645, 2'-Dehydroplectaniaxanthin
807 2'-Hydroxy-3' ,4' -didehydro-1 ',2' -dihydro-
3'-Hydroxy-,B,e-carotene-3,4-dione (168a), 143, ,B,Y,-caroten-4-one (161), 805
807 3-Hydroxy-3',4'-didehydrolycopene, 95, 149
3-Hydroxy-,B,K-carotene-3' ,6' -dione, see 3'-Hydroxy-4' ,5' -didehydro-4,5' -retro-
Capsanthone ,B,,B-caroten-3-one, see
Hydroxy-IX-carotene epoxides, 831 Eschscholtzxanthone
3'-Hydroxy-,8,,8-carotene-3,4,4'-trione (201), 815 3-Hydroxy-5,8,5',8' -diepoxy-,8-carotene, see
3-Hydroxy-,8,,8-caroten-4-one, see 3-Hydroxy- Cryptochrome
echinenone 3-Hydroxy-7',8'-dihydro-7' -apo-,B-caroten-
3'-Hydroxy-,B,,B-caroten-4-one, see 8'-one, see 3-Hydroxysintaxanthin
3'-Hydroxyechinenone 1-Hydroxy-1,2-dihydroapo-8'-lycopenal, 455,
4'-Hydroxy-,8,,8-caroten-4-one (155), 804 456,514,539
biosynthesis, 629, 645 3-Hydroxy-5',6' -dihydro-5' -apo-18' -nor-
metabolism, 663, 665 ,B-caroten-6'-one, see
occurrence, 142,645 Reticulataxanthin
reactions, 141 1-Hydroxy-l ,2-dihydro-Y,,Y,-caroten-20-al, see
x- Hydroxy-,8,,8-caroten-y-one, 831 Rhodopin-20-al
3-Hydroxy-,B,e-caroten-3'-one, see 5-Hydroxy-5,6-dihydro-,B-carotene, 82, 114
3'-Dehydrolutein 1'-Hydroxy-l ',2' -dihydro-y-carotene (48), 781
3'-Hydroxy-,B,K-caroten-6' -one, see occurrence, 40
Cryptocapsin reactions, 151
Hydroxy-,B,Y,-caroten-4-one, 831 structure, 151
3-Hydroxy-,B,Y,-caroten-4'-one, see synthesis, 505, 511, 552
Rubixanthone 1'-Hydroxy-l' ,2' -dihydro-,B,Y,-caroten-4-one =
3'-Hydroxy-1/t,Y,-caroten-4-one (165), 806 1'-Hydroxy-1',2' -dihydro-y-caroten-4-one
3-Hydroxycitranaxanthin, see Reticulata- (160), 805
xanthin occurrence, 38
3'-Hydroxy-3,4-dehydro-,8-carotene (38), 779 reactions, 133
3-Hydroxy-7 ,8-didehydro-,8-apo-12' -carotenal, structure, 13 3
441, 444, 510, 537 synthesis, 516, 519, 559
4-Hydroxy-15,15' -didehydro-,B-apo- 1'-Hydroxy-l ',2' -dihydro-x,Y,-caroten-4' -one,
8' -carotenal, 515 see OH-Okenone
3-Hydroxy-2,3-didehydro-,B,,B-carotene- 8'-Hydroxy-7',8'-dihydrocitranaxanthin (239),
4,4'-dione, see Dehydroadonirubin, 824
Phoeniconone occurrence, 49
3-Hydroxy-2,3-didehydro-,B,,B-caroten-4-one, reactions, 170
see 3-Ketoechinenone structure, 170
3-Hydroxy-2,3-didehydro-,B,e-caroten-4-one synthesis, 170, 462, 464
(156), 804 1'-Hydroxy-l' ,2' -dihydro-2-isopentenyl-
3-Hydroxy-7 ,8-didehydro-7',8' -dihydro- 2' -(hydroxyisopentenyl)torulene
7' -apo-,B-carotene-4,8' -dione, see (221), 47, 174, 618, 820
Hopkinsiaxanthin 1'-Hydroxy-l ',2' -dihydro-4-keto-y-carotene,
3-Hydroxy-7 ,8-didehydro-7',8' -dihydro- see 1'-Hydroxy-1',2'-dihydro-
7' -apo-,B-caroten-8' -one, see y-caroten-4-one
Triophaxanthin 1-Hydroxy-1,2-dihydrolycopene, see
1-Hydroxy-3,4-didehydro-1,2-dihydroapo- Rhodopin
8'-lycopenal, 455,456, 514, 539 1-Hydroxy-1,2-dihydroneurosporene, see
1-Hydroxy-15,15' -didehydro-1,2-dihydroapo- Chloroxanthin
8'-lycopenal,.455, 456, 464, 539 1-Hydroxy-1,2-dihydrophytoene (61), 34, 146,
1'-Hydroxy-3',4'-didehydro-1',2'-dihydro- 626, 783
,8,1/t-caroten-4-one, see 1-Hydroxy-1,2-dihydrophytofluene (60), 34,
Deoxyflexixanthin 146,626,783

Carotenmds 58
918 Subject Index
8'-Hydroxy-?' ,8'-dihydroreticulataxanthin
(240), 49, 824
1'-Hydroxy-l' ,2' -dihydrospheroidene, see
OH-Spheroidene
2' -Hydroxy-l' ,2' -dihydrotorulene, 832
5-Hydroxy-5,6-dihydrozeaxanthin (87), 157,
789
"3-Hydroxy-3',4'-diketo-a-carotene ", see
a-Doradecin
3-Hydroxy-4,4'-diketo-P-carotene (196), 814
3'-Hydroxy-3,4-diketo-P-carotene (16Sa), 143,
144,806
2'-Hydroxy-3,1 '-dimethoxy-3',4' -didehydro-
1',2'-dihydro-P,Y,-caroten-4-one (186),
811
4-Hydroxy-3',4'-dioxo-P-carotene (168), 807
3-Hydroxyechinenone (153), 38, 141,647-649,
651,804
3'-Hydroxyechinenone (154), 101, 142, 804
4'-Hydroxyechinenone, see 4'-Hydroxy-
p,p-caroten-4-one
Hydroxyflavoxanthin, 832
1-Hydroxy-1,2,7',8',11',12'-hexahydrolycopene
(59), 34, 624, 626, 783
2-(4-Hydroxy-3-hydroxymethyl-2-butenyl)-
2' -(3-methyl-2-buteny!)-p,p-carotene,
see 2-(Dihydroxyisopentenyl)-
2'-isopentenyl-P-carotene
3-Hydroxyisorenieratene (52), 782
occurrence, 41
reactions, 78, 152
syntheses,336,496,499,552
3-Hydroxy-3'-keto-a-carotene, see
3'-Dehydrolutein
3-Hydroxy-4-keto-P-carotene, see
3-Hydroxyechinenone
4-Hydroxy-4'-keto-P-carotene, see 4'-Hydroxy-
p,p-caroten-4-one
1'-Hydroxy-4-keto-1 ',2'-dihydro-y-carotene,
see 1'-Hydroxy-1',2'-dihydro-
y-caroten-4-one
1'-Hydroxy-2-keto-1' ,2' -dihydrotorulene, see
2'-Dehydroplectaniaxanthin
19-Hydroxylutein, see Loroxanthin
3-Hydroxylycopene, 148
16-Hydroxylycopene, see Lycoxanthin
1'-Hydroxy-1-methoxy-3,4-didehydro-
1,2,1',2',7',8'-hexahydro-Y,,!/f-caroten-
2-one, see OH-Spheroidenone
1'-Hydroxy-1-methoxy-3,4,3',4' -tetradehydro-
1,2,1 ',2' -tetrahydro-Y,,Y,-caroten-
2-one (184), 811
2-(4-Hydroxy-3-methyl-2-butenyl)-ll,Y,-
carotene, see 7',8',11',12'-
Dehydrononaprenoxanthin
2-(4-Hydroxy-3-methyl-2-butenyl)-
7',8'-dihydro-ll,Y,-carotene, see
11',12'-Dehydrononaprenoxanthin
2-(4-Hydroxy-3-methyl-2-butenyl)-
2' -(3-methyl-2-butenyl)-ll,ll-carotene,
see Deshydroxydecaprenoxanthin
2'-(4-Hydroxy-3-methyl-2-butenyl)-2-
(3-methyl-2-butenyl)-3',4' -didehydro-
1',2' -dihydro-p,Y,-caroten-1 '-ol, see
1'-Hydroxy-1',2'-dihydro-
2-isopentenyl-2'-(hydroxy-
isopentenyl)torulene
2-(4-Hydroxy-3-methyl-2-butenyl)-7' ,8',11 ', 12'-
tetrahydro-ll,Y,-carotene, see
~onaprenoxanthin
2-(3-Hydroxy-3-methylbutyl)-2'-(3-methyl-
2-butenyl)-3,4,3',4' -tetradehydro-
1,2,1 ',2'-tetrahydro-Y,,Y,-carotene-
1,1'-diol, see Monoanhydro-
bacterioruberin
4-Hydroxymyxoxanthophyll (94a), 117,791
15-Hydroxy-7',8',9',10',11',12',13',14'-
octahydro-6'-apo-P-caroten-7'-one
(241), 824
3'-Hydroxy-4-oxo-P-carotene, see
Asteroidenone, 3'-Hydroxy-
echinenone
3-Hydroxy-4-oxo-2,3-dehydro-P-carotene,
see 3-Ketoechinenone
4'-Hydroxy-3-oxoechinenone (168), 807
4-Hydroxyphleixanthophyll, 119
Hydroxyphytoene (61), 783
Hydroxyphytofluene (60), 783
Hydroxypirardixanthin, 832
Hydroxy-Red, see OH-Spheroidenone
4-Hydroxyretinal, 390
3-Hydroxy-4,5'-retro-5'-apo-P-caroten-5' -one,
see Tangeraxanthin
3'-Hydroxy-5,6-seco-p,p-carotene-5,6-dione =
3-Hydroxysemi-P-carotenone, see
Triphasiaxanthin
3-Hydroxysintaxanthin (245), 171, 825
Hydroxyspheroidene, see OH-Spheroidene
Hydroxyspheroidenone, see
OH-Spheroidenone
8'-Hydroxy-5' ,6', 7',8'-tetrahydro-5' -apo-
18'-nor-P-caroten-6' -one, see
8' -Hydroxy-7',8' -dihydrocitrana-
xanthin
1-Hydroxy-1,2,7' ,8'-tetrahydrolycopene, see
Chi oro xanthin
3-Hydroxytorulene, see Celaxanthin
3-Hydroxy-3' ,4,4' -triketo-P-carotene (201),
815
Hypervitaminosis A, 735
 Subject Index 919
3,3'-Isorenieratenediol, see 3,3'-Dihydroxy-
isorenieratene
Idoxanthin (173), 38, 642,643,649, 651, 809 3-Isorenieratenol, see 3-Hydroxyisorenieratene
Illumination, see Light Isozeaxanthin (71), 786
Infrared light absorption, 202 biosynthesis, 629, 645, 646
Intermediates, syntheses of, 331-440 colouring agent, 750
Iodopsin, 725, 731 derivatives
IX-Ionone, 34, 51, 306, 337, 341, 371, 533, 534, diacetate, 98, 100, 477, 480, 554, 751
707 diethyl ether, 98, 100
P-Ionone dimethyl ether, 78, 141, 480, 554, 750
biosynthesis, 707 metabolism, 645
as carotenoid precursor, 35, 591, 615, 616 m.s., 252, 258
occurrence, 51 occurrence, 37
syntheses of, 19, 338-341 reactions, 78, 98, 141, 156
syntheses from, 340, 341, 344, 352, 354, 355, structure, 156
357,362,366,376,520 syntheses
P-Ionone 5,6-epoxide, 51 partial, 98, 141
P-Ionylideneacetaldehyde, see P-C 15 -Aldehyde total,390,477,479,482,483,554
P-Ionylidenecrotonic acid, 285,287, 366, 369
l.r., see Infrared light absorption
lrone, 51 K
Isoanhydrovitamin A, 17
Isobixin (265), 828 Kerogen, 708
Isocarotene, see Retrodehydrocarotene Keto-, see also Oxo- or -one
Isocryptoxanthin (40), 779 Ketocapsanthin, see Capsanthone
biosynthesis, 629, 645, 646, 684 Ketocapsorubin, see Capsorubone
derivatives 4-Keto-IX-carotene, see Phoenicopterone
acetate, 477 4-Keto-P-carotene, see Echinenone
metabolism, 629, 645, 646 4-Keto-y-carotene (151), 803
occurrence, 37, 629 occurrence, 38
reactions, 140, 141, 151 reactions, 133, 134
structure, 151 syntheses, 505, 515, 516, 519, 558
syntheses Keto carotenoids (148-210, 213-216, 236-241,
partial, 140 244-247,252,264,272)
total, 477, 479, 551 formation, 628, 629
Isocryptoxanthin 5,8-epoxide, see 5',8'-Epoxy- metabolism, 641-656, 658
5',8'-dihydro-P,P-caroten-4-ol reactions, 71, 85-88, 129-147
Isofucoxanthin (179), 43, 107, 108, 110, 810 structures, 129-147
Isofucoxanthinol (178), 43, 107, 108, 111, 810 10-Keto-7,10-dihydro-P-carotene, 474, 558
Isolutein (120), 796 4-Keto-1',2'-dihydro-1'-hydroxytorulene, see
2-Isopentenyl-3,4-dehydrorhodopin (219), Deoxyflexixanthin
47,173,820 3-Ketoechinenone (152), 141, 142, 803
Isorenieral, 104, 451, 452, 512, 538 4'-Keto-3-hydroxy-y-carotene, see Rubi-
Isorenieratene (13), 775 xanthone
see also Leprotene 4-Keto-3'-hydroxylycopene (165), 806
conformation, 285 Ketohydroxypirardixanthin, 832
light absorption, 285 4-Ketolutein (169), 143, 808
occurrence, 41 2-Keto-1-methoxy-3,4-didehydro-1,2-
structure, 104 dihydroapo-15-lycopenal, 453-457,
syntheses,336,428,484,486,496,499,548 538
P-Isorenieratene (6), 773 2-Keto-1-methoxy-3,4-didehydro-1,2-
occurrence, 41 dihydroapo-4'-lycopenal, 455, 457, 540
phenyl analogue, 522 2-Keto-1-methoxy-3,4-didehydro-
structure, 104 1,2-dihydroapo-8'-lycopenal, 455,
synthesis, 504, 511, 546 456,515,540
920 Subject Index

4-Ketomyxol2'-(5-C-methylpentoside) (176), colouring agent, 749, 752, 753

117, 120, 647, 809 derivatives, 155
<X-C 18 -Ketone, 369 acetate, 311
P-C18 -Ketone, 366, 368, 369, 383, 386-388, 482 bis(trichloroacetate), 78, 155
9-cis, 369, 387 diacetate, 252, 258
11-cis, 387 dimethyl ether, 78, 155
Ketones, see Keto carotenoids dipalmitate, see Helenien
2-Keto-OH-spirilloxanthin (184), 126, 811 3-(methyl ether), 155
4-Ketophleixanthophyll (172), 38, 53, 117, 808 3'-(methyl ether), 78, 155, 252, 258
3-Ketoretrobisdehydrocarotene, 146, 160 function, 677, 688
2-Ketorhodovibrin isolation, 13
methyl ether, 515, 560 metabolism, 642, 644, 650, 654, 655, 658
m.s., 252, 256 m.s., 252, 258
structure, 126 occurrence, 31, 36, 41, 607, 642, 650, 654,
synthesis, 454, 515, 516, 560 655
"2-Ketorhodovibrin" (184), 811 o.r.d., 290, 311
4'-Ketorubixanthin, see Rubixanthone protein complex, 53, 658
"2-Ketospirilloxanthin" (208), 817 reactions, 75, 78, 79, 155, 311
4-Ketotorulene (150), 119, 803 structure, 155
Kitol, 49 Lutein epoxide (120), 796
Kwashiorkor, 722 see also Eloxanthin, Taraxanthin,
Tareoxanthin
absolute configuration, 288, 290, 311
L geometrical isomerism, 269
occurrence, 41
Labelled carotenoids, 535, 536 reactions, 116, 311
Laetiporxanthin, 48, 832 synthesis, partial, 116, 155, 311
Leprotene (13), 41, 104, 775 Luteochrome (136), 113, 114, 799
see also Isorenieratene Luteoxanthin (138), 114, 800
Light absorption spectroscopy Lycopen-16-al, 148, 149, 252, 256
infrared, 202-204 Lycopen-20-al (Lycopenal) (143), 126, 129, 270,
ultraviolet, 192-202, 273 801
visible, 192-202, 273 Lycopene (19), 776
Lipids, 65 biosynthesis, 581, 589-592, 594-596, 605,
Lipoprotein, 39, 657 606,608,610,615,622,626,641
see also Carotenoproteins colouring agent, 21, 759
Loliolide, 51, 305 conformation, 284
Loroxanthal, 157 function, 677, 701
Loroxanthin (88), 789 genetic studies, 608, 610
derivatives geometrical isomerism, 269, 271, 272,
dimethyl ether, 157 275-277,281,593,608
methyl ether, 157 history, 13
triacetate, 139, 157 labelled compounds
m.s., 251 7,7'-D 2 , 250
occurrence, 46 15,15'-D 2 , 210,211, 235-237, 536
reactions, 157 light absorption
structure, 157 u.v., visible, 192-195
synthesis, partial, 139 metabolism, 35, 48, 616, 618, 621, 623, 626
Luciferin, 50 m.s., 245, 246, 252, 256, 260
Lumirhodopsin, 726 occurrence,30-34,606
Lusomycin (211), 818 p.m.r., 210-212, 236, 237, 239, 240, 277
Lutein (73), 786 reactions, 74, 83, 93, 95, 98, 102, 103, 148,
absolute configuration, 288-290, 292, 311 153, 168
biosynthesis, 311, 600, 602, 642, 654 syntheses
chirality, 292 partial, 83, 93, 148, 153
 Subject Index 921

total, 424, 428, 477, 478, 484, 487, 489, 1'-Methoxy-3',4' -didehydro-1 ',2' -dihydro-
490,497,499,505,513,516,549 y-carotene (96a), 627, 791
Lycopene-16,16' -diol, see Lycophyll 1-Methoxy-3,4-didehydro-1 ',2' -dihydro-
Lycopene epoxide, 95, 832 l/l,l/1-carotene, see Anhydro-
Lycopen-16-ol, see Lycoxanthin rhodovibrin
Lycopen-20-ol (63), 784 x- Methoxy-3,4-didehydro-7',8' -dihydro-
Lycopersene (34), 778 p,l/f-caroten-2-one, 832
biosynthesis, 584, 585 1-Methoxy-3,4-didehydro-1,2, 7',8',11',12'-
m.s., 256, 261 hexahydro-l/J,l/1-carotene, see
occurrence, 31, 102 11',12' -Dihydrospheroidene
synthesis, 477, 479, 550 1'-Methoxy-3',4'-didehydro-1,2, 7,8, 1',2'-
Lycophyll (83), 788 hexahydro-l/J,l/f-caroten-1-ol, see
derivatives OH-Spheroidene
diacetate, 153 1-Methoxy-3,4-didehydro-1,2, 7' ,8' -tetrahydro-
dipalmitate, 153 l/J,l/f-carotene, see Spheroidene
m.s., 252, 256 1'- Methoxy-3',4'-didehydro-1,2, 1',2'-tetrahydro-
occurrence, 46 l/1,1/J-caroten-1-ol, see Rhodovibrin
structure, 153 1-Methoxy-3,4-didehydro-1,2, 7' ,8'-tetrahydro-
synthesis, 556 l/l,l/f-caroten-2-one, see Spheroidenone
Lycoxanthin (62), 784 1-Methoxy-1,2-dihydro-l/J,l/f-caroten-20-al,
biosynthesis, 602 see Methoxylycopenal
derivatives 1-Methoxy-1,2-dihydro-l/f,l/f-carotene, see
acetate, 48, 148 1-Methoxy-1 ,2-dihydrolycopene
palmitate, 95, 149 1'-Methoxy-1',2' -dihydro-qJ,I/J-caroten-4' -one,
tetrahydropyranyl ether, 148, 417, 515, see Thiothece-4 78
553 1'-Methoxy-1',2' -dihydro-x,l/f-caroten-4'-one,
m.s., 252, 256 see Okenone
occurrence, 46 1' -Methoxy-1',2' -dihydro-3',4' -dehydro-
reactions, 79, 148, 149 y-carotene (96a), 627, 791
structure, 47, 148 1-Methoxy-1,2-dihydro-3,4-didehydro-
synthesis, 149, 505, 515, 516, 553 lycopen-20-al (145a), 127, 802
Lycoxanthin epoxide, 832 1-Methoxy-1,2-dihydrolycopene (98), 123, 792
1-Methoxy-1,2-dihydroneurosporene,
see 3,4-Dihydrospheroidene
M 1-Methoxy-1,2-dihydrophytoene (104), 626,
793
Mannosides (253, 255), 53, 165, 826, 827 see also Methoxyphytoene
Manool, 306 1-Methoxy-1,2-dihydrophytolluene (103), 626,
Mass spectrometry, 68, 243-265, 289 792
Membrane stabilization, 703, 704 see also Methoxyphytolluene
Metabolism, 605, 638-665 1'-Methoxy-1 ',2' -dihydrospheroidene, see
Metarhodopsins, 726, 728, 729 3,4,7,8-Tetrahydrospirilloxanthin
Methoxy-, see also Monomethoxy- 1-Methoxy-1,2, 7',8',11',12' -hexahydro-
3-Methoxy-/1-Cwaldehyde, 347 l/J,l/f-carotene, see 3,4,11',12'-
Methoxy-p,l/f-caroten-4-one, 832 Tetrahydrospheroidene
1-Methoxy-1,2, 7,8,11,12, 7',8',11',12' -decahydro- 1-Methoxy-2-keto-7',8' -dihydro-3,4-
1/J.,I/J-carotene, see 1-Methoxy- dehydrolycopene, see Spheroidenone
1,2-dihydrophytoene Methoxylycopenal (146), 126, 270, 802
1-Methoxy-3,4-didehydro-1 ,2-dihydroapo- 1-Methoxy-1,2, 7,8,7',8',11 ', 12'-octahydro-
8'-lycopenal, 455, 456, 515, 540 l/J,l/f-carotene, see 1-Methoxy-
1-Methoxy-3,4-didehydro-1,2-dihydro- 1,2-dihydrophytolluene
l/J,l/f-caroten-20-al (145a), 127, 1-Methoxy-2-oxo-3,4-didehydro-1,2-dihydro-
802 15-apo-1/J-caroten-15-al, see 2-Keto-
1'-Methoxy-3' ,4' -didehydro-1',2' -dihydro- 1-methoxy-3,4-didehydro-1 ,2-
/1,1/1-carotene = dihydroapo-15-lycopenal
922 Subject Index
1-Methoxy-2-oxo-3,4-didehydro-1,2-dihydro-
4' -apo-tjt-caroten-4' -a!, see 2-Keto-
1-methoxy-3,4-didehydro-
1,2-dihydroapo-4' -lycopenal
1-Methoxy-2-oxo-3,4-didehydro-1,2-dihydro-
8'-apo-tjt-caroten-8'-al, see 2-Keto-
1-methoxy-3,4-didehydro-
1,2-dihydroapo-8' -lycopenal
Methoxyphytoene (104), 123, 793
see also 1-Methoxy-1,2-dihydro-
phytoene
Methoxyphytofluene (103), 123, 792
see also 1-Methoxy-1,2-dihydro-
phytofluene
1'-Methoxy-3,4,3' ,4' -tetradehydro-1,2,1 ',2'-
tetrahydro-tjt,tjt-caroten-1-ol,
see OH-Spirilloxanthin
1-Methoxy-1,2, 7' ,8' -tetrahydro-1/t,tjt-carotene,
see 3,4-Dihydrospheroidene
Methyl apo-P-carotenoate, see Methyl
P-apocarotenoate
Methyl P-apo-2'-carotenoate, 455,461, 542
Methyl P-apo-4' -carotenoate (neurospora-
xanthin methyl ester) (235), 165, 455,
460,541,823
Methyl P-apo-6'-carotenoate, 455, 460, 541
Methyl P-apo-8'-carotenoate (251), 455, 459,
541,826
Methyl P-apo-10'-carotenoate, 455, 457, 540
Methyl P-apo-12'-carotenoate, 444, 455, 457,
540
Methyl P-apo-14'-carotenoate, 440,455,457,
540
Methyl6'-apo-tjt-caroten-6'-oate, see Methyl
apo-6' -lycopenoate
Methyl P-apo-10'-carotenoate 5,6-epoxide,
209,228,252,256
Methyl apo-6'-lycopenoate (243), 824
occurrence, 48
reactions, 166, 167
structure, 166, 167
synthesis, 464, 465, 542
Methylbixin (266), 829
geometrical isomerism, 271, 278, 279
9-cis, see 9-cis-Methylbixin
15-cis, 281, 469,471, 544
p.m.r., 278, 279
stereomutation, 271, 279
synthesis, 403, 469, 471, 544
9-cis-Methylbixin (266a), 829
geometrical isomerism, 271, 278, 279
p.m.r., 278, 279
stereomutation, 271, 279
synthesis, 167, 283, 284, 405, 425, 469, 471,
544
2-(3-Methyl-2-butenyl)-3,4-didehydro-
1,2-dihydro-tjt,tjt-caroten-1-ol, see
2-Isopentenyl-3,4-dehydrorhodopin
2'-Methyl-oc-carotene, 252, 258
16-Methyl-p,p-carotene, 526, 530, 531
Methyl carotenoates, see Carboxylic acids,
Methyl16-lycopenoate
2-Methyl carotenoids, 51
Methyll5,15' -didehydro-P-apo-8' -carotenoate,
455,458,459,541
Methyl 3',4' -didehydro-p,tjt-caroten-16'-oate,
see Torularhodin methyl ester
Methyl 7,8-dihydroapo-8'-lycopenoate, 455,
464,465,542
Methyll-hexosyloxy-3,4-didehydro-1,2-
dihydroapo-8' -lycopenoate (255), 48,
53, 117, 164, 827
Methyl hydrogen 9' -cis-6,6' -diapocarotene-
6,6' -dioate, see Bixin
3'-0-Methyllutein, 258
Methyl16-lycopenoate, 505, 515
Methyl-cis(natural)-bixin, see 9-cis-
Methylbixin
Methyl y-retinoate, 453, 454, 464
Mevaldic acid, 580
Mevalonic acid, 311, 578-588, 591-594,
596-600,602,605,610,611,614,641
Mevalonic acid 5-pyrophosphate, 581, 582,
585
Micronone, 832
Microxanthin, 832
Mill's rule, 305
Monadoxanthin (7,8-didehydrolutein) (72), 786
absolute configuration, 312
occurrence, 44
structure, 106
synthesis, 504, 510, 555
Monoanhydrobacterioruberin (230), 47, 174,
822
Monodehydrolycopene, see 3,4-Dehydro-
lycopene
Monodemethylated spirilloxanthin =
Monodemethylspirilloxanthin, see
0 H -Spirilloxanthin
Monohydroxy-4-keto-y-carotene, 832
Monomethoxy-4-keto-y-carotene, 832
M.s., see Mass spectrometry
Muconic acid, 278
Mutatochrome (125), 14, 113, 114,252, 258,
797
Mutatoxanthin (129), 114, 116, 798
Mycoplasma laidlawii B carotenol, 832
Mycoxanthin, 832
Mytiloxanthin, 45, 832
Myxobactin (47), 40, 52, 117, 781
 Subject Index 923

Myxobactone (159), 38, 52, 117, 805 biosynthesis from, 612, 616, 618, 622, 623,
Myxol, 119 625,626
Myxol2'-glucoside (92), 117, 119, 790 function, 673, 677
Myxol 2' -(0-methyi-5-C-methylpentoside) genetic studies, 605, 608
(91), 37, 117, 120, 647, 790 geometrical isomers, 269, 276, 277, 593, 608
Myxol2'-rhamnoside, see Myxoxanthophyll occurrence, 31, 32, 34, 610
Myxoxanthin, see Echinenone p.m.r., 276, 277
Myxoxanthophyll (90) reactions, 93
m.s., 252, 256 structure, 102
occurrence, 37, 53 syntheses,426,477,478,504,509,549
reactions, 92, 117, 119, 120 Nomenclature rules, 22-24, 851-864
structure, 119 Nonaprenoxanthin (218), 820
biosynthesis, 617, 618
m.s., 252, 256
N occurrence, 46
structure, 174
Natural methylbixin, see 9-cis-Methylbixin Norbixin (diapo-6,6' -carotenedioic acid), 86,
Nea-P-cryptoxanthin A, see Physoxanthin (39) 167, 758, 759
Neochrome (131), 798 see also Diethylnorbixin
absolute configuration, 302 Norcarotenoids (272, 273), 160, 171, 527, 531,
occurrence, 44 830
reactions, 107, 111, 113, 302 Nostoxanthin (70), 785
structure, 112 Nuclear magnetic resonance, see Proton
Neoxanthin (122), 796 magnetic resonance
absolute configuration, 302, 314, 317
biosynthesis, 600, 684
colouring agent, 752, 753 0
function, 706
geometrical isomers, 269, 273, 277, 291 1,2, 7,8,11 ,12, 7' ,8' -Octahydro-1/t,l/t-carotene,
metabolism, 50, 304, 706 see" 1',2'-Dihydrophytofluene"
m.s., 252, 258 7,8,11, 12,7',8', 11 ', 12'-Octahydro-1/t,l/t-carotene,
occurrence, 31, 41, 43, 44, 52, 607 see Phytoene
o.r.d., 273, 291 1,2, 7,8, 1',2', 7',8' -Octahydro-1/t,1/t-carotene-
p.m.r., 277, 302 1,1'-diol, see 1,1'-Dihydroxy-
reactions, 75, 76, 91, 111, 302, 305 1,2, 1',2' -tetrahydro-( -carotene
structure, 44, 106, 107, 111, 113 1,2,7,8,11,12, 7',8' -Octahydro-1/t,l/t-caroten-1-ol,
Neurosporaxanthin (234), 823 see 1-Hydroxy-1,2-dihydrophyto-
derivatives fluene
ethyl ester, 455, 460, 541 Octahydrolycopene, see Phytoene
methyl ester, see Neurosporaxanthin "Octahydrolycopene", see 17-Carotene
methyl ester "5,6,7,8,5',6', 7',8' -Octahydrolycopene ",
m.s., 252, 256 see (-Carotene
occurrence, 48 7,8,11,12,7',8',11',12' -Octahydrolycopene,
reactions, 165 see Phytoene
structure, 165 3,4,3',4', 7',8', 11 ', 12'-Octahydrospirilloxanthin
syntheses, 455, 460, 541 (113), 123, 124, 794
Neurosporaxanthin methyl ester (235), 823 OH, see also Hydroxy- or -ol
reactions, 165 OH-Chlorobactene (53), 782
structure, 165 occurrence, 41, 117
syntheses reactions, 82, 151
partial, 165 structure, 150, 151
total, 455, 460, S41 synthesis, 415, 514, 516, 552
Neurosporene (22), 776 OH-Chlorobactene glucoside (54), 41, 151, 782
biosynthesis of, 32, 34, 590, 592-595, 605, OH-Dihydrocitranaxanthin, see 8'-Hydroxy-
608, 613, 623 7',8' -dihydrocitranaxanthin
 924 Subject Index

OH-Lycopene, see Rhodopin 3-0xocanthaxanthin (195), 814

OH-Neurosporene, see Chloroxanthin 8' -Oxo-8,8' -diapocaroten-8-oic acid, see
OH-Okenol, 133 Crocetinsemialdehyde
OH-Okenone (164), 133, 806 4-0xo-15,15' -didehydro-P-apo-8' -carotenal,
OH-P 481, see Rhodovibrin 515
OR-Pigment Y, see OH-Spheroidene 4-0xo-4'-hydroxy-p-carotene, see 4'-Hydroxy-
OH-R, see OH-Spheroidenone p,p-caroten-4-one
OH-Rhodopin, see Dihydroxylycopene 3-0xo-iso-C14-aldehyde, 345
OH-Spheroidene (107), 793 4-0xoretinaldehyde, 480, 730
biosynthesis, 628
occurrence, 34
reactions, 82, 124 p
structure, 124
OH-Spheroidenone (185), 811 p 412, 792
biosynthesis, 628 P 426 (p-Zeacarotene?), 677
occurrence, 34 P 444, see p,y-Carotene
reactions, 82, 124 P 450, see Spheroidene
structure, 124, 125 P 452, see 7',8',11',12'-Dehydrononapreno-
OH-Spirilloxanthin (105), 793 xanthin
biosynthesis, 621, 625 P 476, see Myxol2'-(0-methyl-5-C-m ethyl-
m.s., 252, 256 pentoside)
occurrence, 34 P 481, see Anhydrorhodovibrin
reactions, 123 P 500, see 3,4,3',4'-Tetrahydrospirill oxanthin-
structure, 122, 123 20-al
synthesis, 514, 516, 556 p 512, 126
OH-Y, see OH-Spheroidene P 518, see 2,2'-Diketospirilloxanthin
Okenol, 126, 516, 518, 556 1'-[(x-0- Palmitoyl-P-o-glucopyranosyl)oxy]-
Okenone (181), 810 3',4' -didehydro-1 ',2' -dihydro-
biosynthesis, 626, 628 p,tjJ-caroten-2' -ol (77 a), 787
m.s., 252, 256 Papilioerythrins, 832
occurrence, 41 Paracentrone (246), 825
reactions, 126 absolute configuration, 300
structure, 121, 126, 127 biosynthesis, 49
syntheses, 451, 453, 504, 512, 516, 518, 559 derivatives
Oleoresin, 759 acetate, 162
Opsins, 17, 723-730 occurrence, 43, 49
Optical activity, 288 reactions, 75, 162
Optical rotatory dispersion, 273, 289 structure, 162
O.r.d., see Optical rotatory dispersion Paracentrotin A, B, 832
Oscillaxanthin (95), 791 Pararhodopsin, 726
occurrence, 34 Pectenolone (166), 45, 107, 807
reactions, 92, 117, 121 Pectenoxanthin (65), 44, 45, 106, 306, 784
structure, 53, 120, 121 Pentaxanthin (178), 43, 111, 810
Oscillol, 121, 130 Perhydroazafrin, 167
Oscillol 2,2' -bis(O-methyl-5-C-methyl- Perhydroazafrinone, 167
pentoside) (96), 791 Perhydro-P-carotene,69
metabolism, 647 Perhydro-y-carotene, 69
occurrence, 34, 647 Perhydro carotenoids, 74, 289
reactions, 117, 121 Perhydrocryptoxanthin, 289
structure, 121 Perhydrofucoxanthin, 289
Oscillol 2,2' -dirhamnoside, see Oscillaxanthin Perhydrolutein, 289
Overhauser effect, 242 Perhydrolycopene,69
Ovorubin, 39, 657, 659-661 Perhydronorbixin, 167
Ovoverdin, 39, 656, 657, 659, 661 Perhydrophytoene,69
Oxo-, see also Keto- or -one Perhydrosqualene, 69
 Subject Index 925

Perhydrotrollichrome acetate, 111 occurrence, 31, 32,268,671,673,676

Perhydrozeaxanthin, 155,156,289,291 p.m.r., 276
Peridinin (273), 44, 53, 161, 163, 171, 830 reactions, 93
Persicachrome, 832 structure, 102
Persicaxanthin, 832 synthesis (trans), 426, 477, 478, 517, 550
Petaloxanthin, 832 Phytoene 1,2-epoxide =
Phaseic acid, 50 Phytoene 1,2-oxide (116), 34, 113, 116, 610, 795
Phillipsiaxanthin (207), 817 Phytoenol, see 1-Hydroxy-1,2-dihydrophytoene
occurrence, 34, 52 Phytofluene (30), 777
reactions, 130 biosynthesis, 589-592,605,608,616,617,622
structure, 130 cyclic analogue, 35
synthesis, 502, 503, 562 function, 673, 676, 677
Philosomaxanthin, see 3' -Dehydrolutein genetic studies, 605, 608
Phleixanthophyll (77) geometrical isomers
occurrence, 40, 53 all-trans, 268, 477, 478, 550
reactions, 92, 117-119 occurrence, 31, 32, 673, 676
structure, 92, 117-119 p.m.r., 276
Phoeniconone (195), 814 reactions, 93
metabolism, 649, 651, 655 structure, 102, I 03
occurrence, 143,655 synthesis (trans), 426, 477, 478, 550
reactions, 101, 142 Phytofluenol, 783, 832
structure, 142 Phytol, 276
Phoenicopterin, 832 Picrocrocin, 51, 52
Phoenicopterone (149), 803 Pigment R, see Spheroidenone
metabolism, 645, 650, 652-655 Pigment X (/J-Zeacarotene?), 612, 774
occurrence, 38 Pigment Y, see Spheroidene
reactions, 141 Pigmenters
structure, 141 definition,746
Phoenicoxanthin (196), 814 determination, 762
metabolism, 645, 646, 649-652 in feed, 746-754
occurrence, 38 in food, 754-762
reactions, 142, 143 Pigments, natural, 21
structure, 143 Plectaniaxanthin (76), 787
Photofunctions, 670-703 methyl ether, 516, 518, 555
Photoinhibitions, 702 m.s., 252, 256
Photoperiodism, 703 occurrence, 40, 52
Photoprotection, 671-686 reactions, 79, 130, !55
Photoreception, 702, 703 structure, !53, 155
Photostimulation, 702 synthesis, 516, 518, 555
Photosynthesis, 686-698 Poly-cis-ljf-carotene, see Neurosporene
Phototaxis, 701, 702 Polytomaxanthin, 832
Phototropism, 699-701 Porphyrin, 620, 672
Physalien (68), 785 Porphyropsin, 725, 731
colouring agent, 750 Porter-Lincoln pathway, 589
reactions, 95, 156, !57 Prelumirhodopsin, 726, 729
synthesis, 482, 483, 554 Pro-y-carotene, 269
Physoxant)lin (39, 43), 151, 779, 780 Prolycopene, see also Lycopene
Phytoene (32), 778 biosynthesis, 593, 608
biosynthesis, 31, 32, 34, 280, 584-586, 588- geometrical isomerism, 271, 275, 276
592,605,608,609,615,618,622,623 occurrence, 269
function, 671, 673, 676, 677 Proneurosporene,269,593,608
genetic studies, 608, 609 see also Neurosporene
geometrical isomers Protein deficiency and vitamin A, 720
all-trans, 268, 275, 477, 478, 550 Protein complexes, see Carotenoproteins
light absorption, i.r., u. v., 275 Protein stabilization, 703, 704
926 Subject Index

Proton magnetic resonance 9-cis

apocarotenoids, 226-233 p.m.r., 218, 224
C 40 -carotenoids, 235, 236-242 syntheses, 380, 381, 387
chemical shifts 11-cis, 270
aldehydic protons, 233, 234 biogenesis, 738
end-group protons, 205-208 conformation, 287
'in-chain' methyl groups, 209 function, 17, 723, 724, 726, 729
olefinic protons, 209-213 physical data, 18
in conformational analysis, 213, 287 p.m.r., 220, 224
of geometrical isomers, 224, 225, 275, 280, synthesis of, 378, 379
287 synthesis from, 474
vitamin A compounds, 213-225 13-cis
Protoplasmic viscosity, 703 p.m.r., 220, 224
Protoporphyria, 686 synthesis of, 380, 381
Provitamin A, 13, 15-19, 744 synthesis from, 474, 476
Pyrenoxanthin (89), 46, 789 9, 13-di-cis
Pyrroxanthin, 832 p.m.r., 222, 225
synthesis, 380, 381
11, 13-di-cis
R conformation, 287, 288
p.m.r., 222, 225
Renieral history, 16, 17
m.s., 252, 256 metabolism, 638-640, 718, 719
reactions, 104 occurrence, 49
synthesis of, 451, 452, 538 physical data, 18
synthesis from, 512 p.m.r., 214
Renierapurpurin (16), 775 syntheses of, 351, 376, 377, 380, 381, 384,
conformation, 285 386,387,390,405
occurrence, 41 syntheses from, 440-447, 457, 458, 464,
structure, 104, 105 474-477,479,523,524,526,528
syntheses, 336,484,486,497,499,549 cx-Retinaldehyde, 382, 383,477, 730
{3-Renierapurpurin, 504, 511, 516, 522, 547 y-Retinaldehyde, 216, 454, 477, 478, 538
y-Renierapurpurin, 504, 513, 516, 549 Retinaldehyde 5,6-epoxide, 216, 730
Renieratene (14), 775 Retinaldehyde 5,8-epoxide, 730
conformation, 285 Retinene, Retinene 1 , see Retinaldehyde
occurrence, 41 Retinene 2 , see 3-Dehydroretinaldehyde
structure, 104, 105 Retinoic acid, see Vitamin A acid
syntheses, 487, 489, 497, 499, 548 Retinol, see also 4-Acetoxyretinol,
Renieraxanthin, 832 Anhydrovitamin A, Vitamin A
Reticulataxanthin (238), 824 analogous compounds, 209, 213-225, 718
absolute configuration, 307 colour reaction, 190
biosynthesis, 610 derivatives, 718, 721
genetic studies, 610 acetate, 17
occurrence, 49 carbon-13 n.m.r., 242
o.r.d., 307 colour reaction, 190
reactions, 169 geometrical isomers
structure, 170 9-cis, 379, 384
Retinal= 11-cis, 220, 224, 384
Retinaldehyde 13-cis, 220, 224
conformation, 288 11, 13-di-cis, 222, 225
derivatives labelled compound, 536
enol acetate, 390 p.m.r., 213,214
retinylidene-opsin, 726-730 syntheses of, 374, 375, 378, 379, 382,
function, 703, 723-730, 732 384,387
geometrical isomers, 270, 280 syntheses from, 389, 474
 Subject Index 927
esters, 658, 720, 721 occurrence, 34
ethyl ether, 376, 405 reactions, 82, 148
glucuronide, 721 structure, 148
methyl ether, 214, 228, 376, 377, 388, 403, syntheses
474 partial, 129
palmitate, 17, 708 total, 415, 505, 513, 514, 553
retinyl-opsin, 728 Rhodopinal (144), 801
p-toluenesulphonate, 477 biosynthesis, 626-628
formation, 718, 719 derivatives, 87, 129
function, 731, 732 geometrical isomerism, 270, 272, 273
geometrical isomers, 209, 276, 280 light absorption
7-cis, 536 u.v., visible, 273
9-cis, 17, 281, 380, 381 occurrence, 46
11-cis, 281, 378, 379,724 reactions, 126, 129
13-cis, 17, 274, 381 structure, 129
labelled compound, 536 Rhodopinal n-glucoside (145), 117, 121, 801
p.m.r., 220, 224 Rhodopin glucoside (57), 117, 121,783
syntheses,281, 380,388 Rhodopinol (80), 788
9,13-di-cis, 281, 380, 381 derivatives, 129
11,13-di-cis, 222, 225, 281, 378 geometrical isomerism, 270, 272
labelled compound, 536 occurrence, 46
metabolism, 720-722 reactions, 79, 129
occurrence, 49 structure, 129
physical data, 18 Rhodopsin, 17,270,725-727,731
p.m.r., 213, 214, 276 Rhodopurpurin, 776
syntheses of, 19, 373,378-381, 385-387, Rhodovibrin (106), 793
403,412 biosynthesis, 621, 625
syntheses from, 380, 388-390, 444, 474, 477 m.s., 252, 256
oc-Retinol, 382, 383, 477, 735 occurrence, 34
y-Retinol, 216, 423, 454 reactions, 123
Retroanhydrovitamin A, 17 structure, 123
Retro apocarotenoids, 203, 204 synthesis, 505, 514, 557
see also Tangeraxanthin Rhodoviolascin, see Spirilloxanthin
Retrobisdehydro(-{J-)carotene, 93, 151 Rhodoxanthin (209), 817
4,7' -Retro-P,P-carotene, see 4,4' -Didehydro- biosynthesis, 602
4,7-dihydro-P-carotene derivatives, 87
Retro carotenoids (35-37, 84, 192, 209, 236), metabolism, 642, 656
37,79,204,206,563 m.s., 251,258
Retrodehydrocarotene (36), 778 occurrence, 37
syntheses p.m.r., 206
partial, 93, 95, 141, 477 reactions, 86, 146, 157
total,477, 480,482,550 syntheses
Retrodehydrocryptoxanthin, 95 partial, 86, 95, 146
Retrodehydrododecaprenocarotene, 95 total, 408, 437, 469, 496, 497, 504, 507, 563
Retro-diapo-3,3' -carotenedial, 252, 256, 469, 473 Riboflavin, 591
Retro-diapo-7, 7' -carotenedial, 469, 473 Rosaxanthin, 832
Retro-diapo-5,5'-carotenedione, 252, 256, 469, Rubichrome (128), 114, 798
473 Rubixanthin (45), 780
Retro diapocarotenoids, 440, 469, 473 absolute configuration, 292, 307
Retroretinol, acetate, 218 derivatives
Retrovitamin A acid ethyl ester, 388 acetate, 155, 252, 256
Rhamnosides (90, 94a, 95), 53 palmitate, 95
Rhodopin (56), 782 geometrical isomerism, 274, 276, 277
biosynthesis, 621, 626 5' -cis, see Gazaniaxanthin
m.s., 252, 256 isolation, 273
928 Subject Index

Rubixanthin (45) (continued) mass spectrometry, 243-265

light absorption nuclear magnetic resonance spectroscopy,
u.v., visible, 274 204-243
m.s., 252, 256 ultraviolet absorption spectroscopy,
occurrence, 36 192-202
o.r.d., 307 visible light absorption spectroscopy,
p.m.r., 276, 277 192-202
reactions, 76, 149 Sphaerobolin, see y-Carotene
structure, 149, 150 Spheroidene (99), 792
Rubixanthin 5,6-epoxide, 114 biosynthesis, 623-625, 627
Rubixanthone (163), 133, 806 occurrence, 32, 34
reactions, 124
s structure, 124
synthesis, 505, 514, 556
Salmon acid (198), 815 Spheroidenol, 79, 124, 505, 514, 556
Saproxanthin (75), 786 Spheroidenone (182), 810
acetate, 82, 153, 155 biosynthesis, 620, 622,627, 628
occurrence, 37 m.s., 252, 256
reactions, 153 occurrence, 34
structure, 153 reactions, 79, 124
synthesis, partial, 120 structure, 125
Sarcinaxanthin (224), 47, 174, 176, 821 syntheses,416,423,504,505,509,514,560
Sarcinaxanthindialdehyde, 176 Spirilloxanthin (108), 793
Sarcinaxanthin mono-o-glucoside (225), 177, biosynthesis, 620, 621, 624-628
821 geometrical isomerism, 271
Sarcinene (222), 47, 177, 820 function, 689, 693, 701
5,6-Seco-p,p-carotene-5,6-dione, see Semi- m.s., 248, 252, 256
P-carotenone occurrence,31,32,34
5,6-Seco-P,e-carotene-5,6-dione, see Semi- reactions, 121-123
IX-carotenone structure, 122
Secocarotenoids (213-216, 264), 48 synthesis, 497, 500, 557
Secondary carotenoids, 645 Sporopollenin, 708
Semi-IX-carotenone (214), 48, 133, 308, 819 Squalene
Semi-P-carotenone (213), 48, 78, 133, 610, 818 biosynthesis, 21, 584-588
Semifucoxanthols, 108 geometrical isomerism, 275, 280
S.g. 434, 832 occurrence, 31
S.g. 460, 832 perhydro compound, 69
s g 500. 832 S.t. 483, 832
IX-Sinensal, 233, 234 Stanier hypothesis, 671
P-Sinensal, 233, 234 Stereochemistry, 268-319
Sinensiachrome, 832 absolute configuration, 288-319
Sinensiaxanthin, 832 conformation, 284-288
Sintaxanthin (244), 825 geometrical isomerism, 268-284
biosynthesis, 49, 610 Steric distortion, 202
genetic studies, 610 Steric hindrance, 77, 272, 273, 275, 284, 287
occurrence, 49 Steroids, 65
reactions, 170, 171 Sulcatoxanthin (273), 53, 830
structure, 171
synthesis, 462, 464
Sintaxanthol, 171, 462, 464 T
Siphonaxanthin (174), 46, 139, 602, 809
Siphonaxanthol, 138, 139 Tangeraxanthin (236), 49, 169, 823
Siphonein (175), 46, 139, 809 Taraxanthin (120), 41, 114, 116, 288, 796
Spectroscopic methods, 190-265 see also Lutein epoxide
infrared spectroscopy, 202-204 Taraxanthin dipalmitate=
 Subject Index
Taraxien (121), 116, 796
Tareoxanthin (120), 269, 796
Terpenoids,21,22,610,611
7,8,7',8'-Tetradehydroastaxanthin, see
Asterinic acid
Tetradehydro-P-carotene =
3,4,3',4'-Tetradehydro-p,p-carotene (1 ),
772
occurrence,36, 104
syntheses
partial, 93
total,334,437,482,492,494,496,504,
507,545
as vitamin A precursor, 719
11,12,11',12'-Tetradehydro-P-carotene, 484,
494,495
3,4,3',4'-Tetradehydro-P,i/1-carotene =
3,4,3',4'-Tetradehydro-y-carotene, see
3',4'-Didehydrotorulene
3,4,3',4'-Tetradehydro-1/1,1/1-carotene, see
3,4,3',4'-Tetradehydrolycopene
7,8, 7' ,8'-Tetradehydro-P,P-carotene-3,3' -diol,
see Alloxanthin, Cynthiaxanthin,
Pectenoxanthin
11,12,11',12'-Tetradehydrocrocetindialdehyde,
466,467,471,542
3,4,3',4'-Tetradehydro-7,7' -dihydro-
P-carotene,482,550
3,4,3',4'-Tetradehydrolycopene (17), 775
biosynthesis, 619
m.s., 256, 261
occurrence, 33
reactions, 93, 103
syntheses, 103, 412, 497, 499, 549
3,4,3',4'-Tetradehydro-16,16' -lycopenedial, 489,
490,497,500,557
3,4,3',4'-Tetradehydro-16,16' -lycopenedioic
acid diethyl ester, 497, 500, 564
10,11,10',11'-Tetradehydrorhodoxanthin, 504,
507,563
6, 7,6', 7'-Tetradehydro-5,6,5',6' -tetrahydro-
p,p-carotene-3,3'-diol (70), 785
3,4,3',4'-Tetradehydro-1,2,1 ',2' -tetrahydro-
1/1,1/1-carotene-1,1'-diol
(didemethylspirilloxanthin)
m.s., 252, 256
reactions, 82, 120, 121
synthesis
partial, 120, 121
total, 497, 499, 555
7,8, 7',8'-Tetradehydrozeaxanthin, 444, 489,
490,504,51.0,553
see also Alloxanthin
(3,4,3',4'-)Tetrahydrobisanhydrobacterio-
ruberin (229), 174, 822
929
5,6,5',6'-Tetrahydrocanthaxanthin, see
Diketopirardixanthin
7,8,7',8'-Tetrahydrocapsorubin (206), 817
Tetrahydro-P-carotene =
7,8, 7' ,8'-Tetrahydro-p,p-carotene,
see 11-Carotene
1,2,7,8-Tetrahydro-1/1 ,i{!-carotene,
see 1,2-Dihydroneurosporene
1,2,1 ',2'-Tetrahydro-1/1 ,i{!-carotene,
see 1,2,1',2'-Tetrahydrolycopene
5,6,5',6'-Tetrahydro-1/1 ,1/1-carotene,
see 5,6,5',6'-Tetrahydrolycopene
7,8, 11,12-Tetrahydro-1/1 ,1/1-carotene,
see 7,8,11,12-Tetrahydrolycopene
7,8,7',8'-Tetrahydro-1/1,1/1-carotene,
see (-Carotene
1,2,1 ',2'-Tetrahydro-1/1 ,i{!-carotene-1,1 '-diol,
see Dihydroxylycopene
5,6,5',6'-Tetrahydro-p,p-carotene-4,4' -dione,
see Diketopirardixanthin
1,2, 7' ,8'-Tetrahydro-i{!,i{!-caroten-1-ol,
see Chloroxanthin
1,2,1 ',2'-Tetrahydro-3,4,3',4' -dehydrolycopene,
832
(7,8, 7' ,8'-)Tetrahydrolycopene, see (-Carotene
"Tetrahydrolycopene", see Neurosporene
1,2,1',2'-Tetrahydrolycopene (24), 32, 626, 776
5,6,5',6'-Tetrahydrolycopene, 103,425, 484,
487,549
7,8,11,12-Tetrahydrolycopene (25), 776
biosynthesis, 34,617,618,622-624,626
occurrence, 32, 103
structure, 103
synthesis, 477,478, 549
1,2,1',2'-Tetrahydroneurosporene (28), 32, 626,
777
Tetrahydrophytoene, 589, 832
16-(Tetrahydropyranyloxy)apo-8'-lycopenal,
455,456,515,539
3,4,11',12'-Tetrahydrospheroidene (102), 34,
123,124,624,626,792
3,4,7,8-Tetrahydrospirilloxanthin (111), 124,
794
3,4,3',4'-Tetrahydrospirilloxanthin (110), 794
m.s., 252, 256
occurrence, 34
structure, 122, 123
synthesis, 497, 500, 557
3',4',7' ,8'-Tetrahydrospirilloxanthin (111 ), 124,
794
3,4,3',4'-Tetrahydrospirilloxanthin-20-al (147),
126,270,802
3,4,3',4'-Tetrahydroxy-P-carotene, see
Crustaxanthin
3,4,3',4'-Tetraketo-P-carotene, see Astacene
930
Theaspirone, 51
Thiothece-478 (180), 41, 810
Thiothece-484, 832
Thiothece-polar-484, 832
Titanium tetrachloride-carotenoid complexes,
95
Torularhodin (211), 818
biosynthesis, 612-614
derivatives
ethyl ester
m.s., 252, 256
labelled compound, 535, 536
synthesis, 505, 511, 563
methyl ester, see Torularhodin methyl
ester
occurrence, 46
reactions, 128
structure, 128
syntheses, 516-519, 563
vitamin A activity, 744, 745
Torularhodinalcohol (SO), 46, 128,517,781
Torularhodinaldehyde (142), 800
biosynthesis, 613
light absorption, visible, 193
metabolism, 640, 719
occurrence,46, 129
structure, 128
syntheses
partial, 128
total, 505, 511, 517, 519, 557
as vitamin A precursor, 719
Torularhodin methyl ester (212), 818
occurrence, 46
reactions, 128
syntheses,505,511,517,519,563
Torulene (7), 774
biosynthesis, 612-614
occurrence, 36
structure, 103, 104
syntheses
partial, 95
total, 516, 518, 547
"Torulenecarboxylic(l6') acid", see Torula-
rhodin
Total syntheses, 325-575
apocarotenoids,440-464,536-542
carotenoids, 474-526, 545-564
carotenoid analogues, 526-535
diapocarotenoids, 464-473, 542-544
intermediates, 331-440
acyclic components, 392-428
central components, 428-440
cyclic components, 331-392
Tricyclocrocetin dimethyl ester, 244
2,3,2'-Trihydroxy-cx-carotene, 832
Subject Index
3,4,3'-Trihydroxy-cx-carotene, 832
3,4,4'-Trihydroxy-fi-carotene (85), 38, 143, 157,
645,646,789,831
3,3',4'-Trihydroxy-fi,fi-caroten-4-one,
see Idoxanthin
3,19,3'-Trihydroxy-7 ,8-dihydro-fi,e-caroten-
8-one, see Siphonaxanthin
3,19,3'-Trihydroxy-7,8-dihydro-fi,e-caroten-
8-one 19-laurate, see Siphonein
3,3',19-Trihydroxy-7 ,8-dihydro-8-oxo-
cx-carotene, see Siphonaxanthin
"3,3',5'-Trihydroxy-6'-hydro-7,8-dehydro-
fi-carotene" (94), 790
3,3',4'-Trihydroxy-4-keto-fi-carotene,
see Idoxanthin
1,1 ',2'-Trihydroxy-3,4,3',4'-tetradehydro-
1,2,1 ',2' -tetrahydro-t/l,t/1-caroten-
2-one, see 2'-Dihydrophillipsiaxanthirl.
3,4,4'-Triketo-fi-carotene, see Dehydroadoni-
rubin, Phoeniconone
3,1',2'-Trimethoxy-3',4'-didehydro-1',2'-
dihydro-fi,t/1-caroten-4-one (183), 811
2,2,6-Trimethylcyclohexanone, 52
Triophaxanthin (252, 244a), 162, 825
Triphasiaxanthin (215), 48, 133, 819
Trisporic acids, 35, 50,270, 312, 578, 598,
614-616, 705, 706
Triticoxanthin, 832
Trivial names, 24, 772, 830-833, 864
Trollein (86), 44, 112, 789
see also Deepoxyneoxanthin
Trollichrome (131, 132), 107, 111, 113, 798
Trolliflavin-like compound, 832
Trolliflor, 832
Trollixanthin (122, 123), 44, 107, 111, 113,
796, 797
Tugali-carotenoid X, 832
Tunaxanthin (78), 37, 155, 787
u
Unsym. (-carotene, see 7,8,11,12-Tetrahydro-
lycopene
v
Valenciachrome, 832
Valenciaxanthin, 832
Vaucheriaxanthin (124), 43, 44, 46, 107, 113,
114, 797
Violaxanthin (135), 799
absolute configuration, 288, 299-301,
317
 Subject Index 931

biosynthesis, 42, 50, 600, 602, 607, 706 history, 17

colouring agent, 752, 753 isoprenologues, 527
function, 677, 695, 706 labelled compound, 536
geometrical isomerism, 269, 277 metabolism, 638, 639, 721, 722
9-cis, see Violeoxanthin nor compounds, 214, 216, 224, 225
metabolism, 662 physical data, 18
m.s., 252, 258 p.m.r., 213, 214, 225
occurrence, 31,41,607 synthesis, 373, 375, 380, 381, 384
o.r.d., 301 Vitamin A aldehyde, see Retinaldehyde
reactions, 91, 116, 300, 301, 305 Vitamin A compounds
synthesis, partial, 300 p.m.r., 205, 209, 213-225
Violeoxanthin (135), 269, 799 tx-Vitamin A, see IX- Retinol
Violerythrin tx-Vitamin A acid, 214, 220,222, 224, 380, 382,
derivatives, 87 383
formation, 40, 88, 172 tx-Vitamin A aldehyde, see tx-Retinaldehyde
m.s., 252, 256 y-Vitamin A, see }'-Retinol
occurrence, 40 }'-Vitamin A acid, 216,453,454,464
reactions, 171-17 3 Vitamin A 2 , see 3-Dehydroretinol
structure, 40, 172 Vitamin A 2 acid
Violerythrol, 79, 171, 252, 258 derivatives
Visual pigments, 723-731 methyl ester, 276, 384
Visual purple, see Rhodopsin nitrile, 384
Vitamin A, see also 3-Dehydroretinol, geometrical isomers
Retinol 9-cis, 380, 382, 384
biological activity, 19, 74, 718, 744, 745 13-cis, 380, 382, 385
deficiency, 722,732-736 9,13-di-cis, 380, 382
determination, 17-19, 762-764 physical data, 18
formation, 638, 639, 718, 719 p.m.r., 216
functions, 704, 722-738, 752 syntheses, 380, 382, 384, 385
history, 15-17 Vitamin A 2 aldehyde, see 3-Dehydroretinal-
metabolism, 16, 17, 658, 719-723, 732 dehyde
nomenclature, 17, 718, 723 Vomifoliol, 50
Vitamin A acid (retinoic acid)
conformation, 285
derivatives w
amide, 387
chloride, 388 Warmingol, see Rhodopinol
14-cyano-, 386 Warmingone, see Rhodopinal
ethyl ester, 373-376, 384, 385, 387, 388
9-cis, 376
p-glucuronide, 721, 722 X
methyl ester, 214, 228, 276, 380, 387
9-cis, 380 Xanthomonas-carotenoid, 832
11-cis, 387 Xanthophyll, see Lutein
13-cis, 276, 380 Xanthophyll dipalmitate, see Helenien
9,13-di-cis, 380 Xanthophyll epoxide, see Lutein epoxide
nitrile, 382, 383, 386-388 Xanthophylls (38-216), see also Carotenoids
formation, 639, 719, 720 absolute configuration, 307-312
function, 723, 733, 735, 737 biosynthesis, 600-603, 627, 628
geometrical isomerism, 276, 280, 385 nomenclature, 852, 857-860
9-cis, 218, 224, 375, 376, 380, 381, 384 occurrence, 33, 34, 36-40
11-cis, 220, 224 . reactions, 105-160
13-cis, 220, 222, 224, 380, 381, 385 structures, 105-160
9,13-di-cis, 222, 225, 380, 381 X-ray crystallography, 77, 109, 285, 292, 297,
11,13-di-cis, 276, 385 299,303
932 Subject Index

z geometrical isomer
15-cis, 281, 328
0(-Zeacarotene (12), 774 history, 13
biosynthesis, 35, 594, 595 light absorption, u.v., 289, 290
occurrence, 35 metabolism, 629, 642-644, 649, 651, 662-664
reactions, 93, 103 m.s., 252, 258
structure, 96 occurrence, 31, 36
synthesis, 103, 504, 509, 548 o.r.d., 289-292, 307
P-Zeacarotene (9), 774 p.m.r., 235-237
biosynthesis, 35, 594, 595, 613 reactions, 75, 78, 91, 95, 111, 116, 146,
function, 677 155-157,169,300
occurrence, 35 structure, 156
reactions, 76, 93, 103 syntheses
structure, 96 partial, 91, 109, 111, 116, 146, 155, 157,
synthesis, 103, 504, 508, 547 296,300
vitamin A activity, 744, 745 total, 437, 482, 483,489, 490,494,496,
P1 -Zeacarotene, see P-Zeacarotene 504,510,554
Zeaxanthin (67), 785 Zeaxanthin diepoxide, see Viola-
absolute configuration, 288, 289-292, xanthin
295-300,307,314 Zeaxanthin furanoxide, see Mutato-
biosynthesis, 42, 45, 48, 600, 602, 603, 606, xanthin
607,629,662-664,695 Zeaxanthin-like carotenoid, 647
c.d., 289, 290 Zeinoxanthin (42), 780
colouring agent, 750, 752, 753 absolute configuration, 309
derivatives, 156 metabolism, 654
acetate, 300 occurrence, 36
diacetate, 116, 480, 554 o.r.d., 309
dimethyl ether, 78, 156, 482, 483, 554 reactions, 151
dipalmitate, see Physalien structure, 151
methyl ether, 78, 156 synthesis (racemate), 504, 510, 552