Biochemical Systematics and Ecology, Vol. 18, No. 6, pp. 435-438, 1990. 0305-1978/90 $3.00+ 0.

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Printed in Great Britain. © 1990 Pergamon Press plc.

Cyanohydrin Glucosides of Turneraceae*

ELIN S. OLAFSDOTTIR,t JERZY W. JAROSZEWSKIt and MARIA MERCEDES ARBO~

tDepartment of Organic Chemistry, Royal Danish School of Pharmacy, Universitetsparken 2, DK-2100 Copenhagen, Denmark;
:~lnstituto de Botanica del Nordeste, Sargento Cabral 2131, Casilla de Correo 209, 3400 Corrientes, Argentina

Key Word Index--Piriqueta; Turnera; Wormskioldia; Turneraceae; cyanogenesis; cyclopentanoid cyanohydrin glycosides;
linamarin.
Abstract--Cyanohydrin glucosides from Piriqueta taubatensis, R viscosa ssp. australis, Tumera angustifolia, T. caerulea, T.
chamaedryfolia, T. hassleriana and Wormskioldia Iongipedunculata were isolated and identified. Members of the Turneraceae
produce cyclopentanoid cyanohydrin glucosides as well as linamarin. This is the first report of linamarin from the Turneraceae.
Both types of 1,4-dioxygenated cyclopentanes (cis and trans) were represented. The relationship of the Turneraceae to the
Passifloraceae and Flacourtiaceae is discussed briefly.

Introduction listed in Table 1. The compounds isolated and

The plant family Turneraceae [1-6] consists of 10 identified are: linamarin (1), the nonhydroxylated
genera and approximately 170 species. These cyclopentenes deidaclin (2) and tetraphyllin A
are perennial or annual herbs, shrubs or rarely (3), and the cis- as well as the trans-l,4-dioxy-
trees; six genera with only a few species are genated cyclopentenes, epivolkenin (4), tarakto-
African, one monotypic genus is American, and phyllin (5), volkenin (6) and tetraphyllin B (7). This
three genera have representatives in the is the first time that linamarin (1) has been
Americas as well as Africa or Madagascar. reported from the Turneraceae.
Turnera is the largest genus, with over 100 The plants were extracted in hot 80%
species distributed mainly in tropical and sub- methanol, and the cyanohydrin glucosides
tropical America. Morphological studies suggest isolated by chromatography as described in the
a close relationship between the Turneraceae Experimental. The compounds were analysed by
and Passifloraceae, and to some extent to
Flacourtiaceae [7], especially the cyanogenic
TABLE 1. SUMMARY OF STRUCTURES OF CYANOHYDRIN GLUCO-
part of this heterogeneous family [8]. SIDES AND THEIR DISTRIBUTIONIN THE TURNERACEAE*
The first report of cyanogenesis from the
Turneraceae was made by Gibbs in 1954 [9], and Species Compounds References
later confirmed by others [10, 11], but without Piriqueta taubatensis Epivolkenin (4),
identifying the cyanogenic compounds present. taraktophylin (8) This work
About 10 years ago, some species of Turner- R viscose ssp. australis Deidaclin (2)t, 4, 5 This work
Turnera angustifolia 2, tetraphyllin A (3),
aceae were shown to produce cyclopentanoid
vofkenin (6), tetraphylin B
cyanohydrin glycosides [12-14], as do the related (7} This work
families Passifloraceae [15], Flacourtiaceae [16] 7: caerulea 4, 5 This work
and Malesherbiaceae [17]. T chamaedryfolia Linamarin (1) This work
T. diffusa 75 [13]
7; hassleriana 1, 4t This work
Results and Discussion T. ulmifolia 2, 3, 75 [12, 22]
The species of Turneraceae containing cyano- Wormskioldia Iongipedunculata 4-7 This work
hydrin glycosides with known structures are
*Data from ref. [14] are not included because the structures reported
*Part Xl in the series "Cyclopentanoid Cyanohydrin Glyco- are uncertain (see text).
sides". For Part X see Jaroszewski, J. W. and Fog, E. (1989) tBecause only a small amount (ca 2 rag) of this compound was iso-
Phytochemistry 28, 1527. lated, the presence of a trace amount of its stereoisomeric partner could
not be confirmed.
(Received 29 May 1990) 5Co-occurrence of 6 was not excluded.

435
436
NC OGlu
GluO CN NO OGlu
2 3
OH OH
NC OGlu GluO CN
4 5
OH OH
GluO CN NC OGlu
6 7
FIG. 1. STRUCTURES OF CYANOHYDRIN GLUCOSIDES FOUND IN
TURNERACEAE AND RELATED FAMILIES: LINAMARIN (1), DEIDACLIN
(2), TETRAPHYLIN A (3), EPIVOLKENIN (4), TARAKTOPHYLLIN (5),
VOLKENIN (6) AND TETRAPHYLLIN B (7). Glu~f]-D-glucopyranosyl.
high field 1H NMR spectra of the pure glucosides
as well as their acetates. Comparison with the
spectra of reference compounds [18-20] assured
unambiguous identification. The cyanohydrin
glucosides of six cyanogenic species investi-
gated have not previously been reported. In
agreement with our previous results, plants
producing 4-hydroxylated cyclopentanoid gluco-
sides almost always produce a pair of either
cis-l,4-dioxygenated (4 and 5) or trans-l,4-dioxy-
genated isomers (6 and 7) [15, 18, 19, 21]. The
results are given in Table 1.
A study of cyanogenic constituents of Turner-
aceae, claiming identification of cyanohydrin
glucosides from 17 species (these reports are not
listed in Table 1), was published previously [14].
However, the authors report all compounds
E.S. OLAFSDOTTIR ETAL
isolated to be either nonhydroxylated (2 and 3)
or trans-l,4-dioxygenated cyclopentenes (4 and
5), and the presence of cis-l,4-dioxygenated
isomers (6 and 7) was not considered. This is in
contrast with our identification of the cis isomers
(6 and 7) in five out of seven cyanogenic species
investigated. Since the previous authors did not
recognize the presence of the cis isomers in spite
of striking differences between 1H NMR spectra
of the latter and of the trans isomers (4 and 5)
[18, 19], all the species reported in that work [14]
need to be reinvestigated. Indeed, our examin-
ation of Wormskioldia Iongipedunculata resulted
in identification of the cis as well as the trans
isomers, with the cis isomers predominating
(Table 1), whereas only the trans isomers were
assumed in the previous work [14]. The presence
of both structural types (cis and trans) in the
same plant is highly unusual, and may imply the
presence of two different hydroxylating enzymes
in one plant. This situation has been
encountered only once before, in the Passi-
floraceae [15], where, however, the evidence for
the presence of only three isomers exists [15, 23].
Wormskioldia Iongipedunculata is thus the first
example of a plant clearly documented to
produce all four stereoisomers in significant
amounts.
The types and pattern of cyanohydrin gluco-
sides in the Turneraceae, established in the
present work, confirm the relationship of this
family to the Passifloraceae and Flacourtiaceae.
The Passifloraceae is a highly diversified family
in its ability to produce cyanohydrin glucosides
but, considering the stereochemistry of
4-hydroxylated cyclopentenes (4-7), it may be
dominated by the trans-l,4-dioxygenated
isomers [15]. A characteristic feature of Passi-
flora, the largest genus of the Passifloraceae,
i s a coexistence of cyclopentanoids with the
valine and isoleucine-derived cyanohydrins, 1
and Iotaustralin [15]. On the other hand, the
cyanogenic tribes of Flacourtiaceae predomi-
nantly produce the cis-l,4-dioxygenated cyclo-
pentenes [16]. Species of Turneraceae syn-
thesize 1 as well as both types of the stereo-
isomeric pairs of 4-hydroxylated cyclopentenes,
apparently with a preference for the cis isomers.
In this respect, the Turneraceae could possibly
be considered intermediate between the Passi-
floraceae and Flacourtiaceae, although more
CYANOHYDRINGLUCOSIDESOF TURNERACEAE
species should be investigated before reaching
definitive conclusions. Turnera hassleriana, in a
similar manner to Passiflora lutea [15, 24], com-
prises a rare example of the co-occurrence of 1
and cyclopentanoid cyanohydrins in the same
species.
Finally, it should be emphasized that about
one-third of the Turneraceae species investi-
gated to date, including those tested by Spencer
et al. [14], were noncyanogenic. The absence of
cyanogenesis in plant material may in general be
related to sample preparation, and is usually con-
sidered less significant than its presence. How-
ever, the lack of cyanogenesis in the Turneraceae
seems to be more frequent than in other exten-
sively studied families, e.g. Passifloraceae [15, our
unpublished observations] and Malesherbiaceae
[17], and the Turneraceae may indeed be more
variable in its ability to produce cyanide. The
apparently noncyanogenic species of Turner-
aceae belong mainly to Tumera and Wormskiol-
dia, whereas material of Piriqueta species ob-
tained from similar sources and processed using
similar procedures was consistently cyanogenic
(13 species) and subspecies were investigated,
which is about one-third of the total). Whether
this apparent absence of cyanogenesis in a
significant number of Turneraceae species is
genotypic or ecotypic in nature, is yet to be
determined.
Experimental
Plant material. Aerial parts of Piriqueta taubatensis (Urban)
Arbo (Arbo 2544), P. viscosa Grisebach ssp. australis Urban
(Ferraro 2969), Tumera caerulea DC. (Krapovickas 38740); T.
chamaedryfolia Cambess. (Noblick 3175), T. hassleriana Urban
(Vanni 347), cultivated at Corrientes, Argentina, T. angustifolia
Miller, obtained from the Botanical Garden, University of
Copenhagen, Copenhagen, and Wormskioldia Iongipeduncu-
lata Mast. [ Tricliceras Iongipedunculatum (Mast.) R. Fernandes
var. Iongipedunculatum], obtained from South Africa (Van
Wyk 8004), were used.
Aerial parts of T. concinna Arbo (Vanni 213), T. aft. coriacea
Urban (Krapovickas 38793), T. grandidentata (Urban) Arbo
(Arbo 1674), T. krapovickasiiArbo (Ahumada 4549, Arbo 2706,
Krapovickas 40303), T. orientalis (Urban) Arbo (Cabral, E. 358),
all cultivated in Corrientes, Argentina, and T. angustifolia Miller
and T. subulata Smith, obtained from The Royal Botanic
Gardens, Kew, were not cyanogenic.
General isolation procedure. The dried plant material was
pulverized and boiled for 5 rain with 80% aq. MeOH, filtered,
the filter cake boiled in a fresh portion of aq. MeOH, filtered
again and the liquid evaporated. The residue was adsorbed on
silica gel in MeOH, and the material fractionated by column
chromatography on silica gel (Merck silica gel 60, 0.066-0.2
437
ram), using EtOAc-Me2CO-CH2CI2-MeOH-H20 (20:15:6:5:4)
as the eluent. The fractions (25 ml) were monitored by TLC
(Merck silica gel 60 plates) with the same solvent, using
naphthoresorcinol [25] as well as the cyanide-specific picrate
sandwich method [26] of visualizing the spots. The cyanogenic
fractions were combined, evaporated and purified further by
preparative HPLC on Lichrosorb RP-18, 5 Bm (1.6×25 cm
column), using 4 ml rain 1 H20_MeOH (ratio 9:1 or 4:1 for
fractions containing 4-hydrox'ylated and nonhydroxylated
cyclopentanoids, respectively). The compounds were detected
with a differential refractometer and tested by TLC [25, 26]. The
individual glucosides with enantiomeric aglycones were
usually not separated from each other, but were identified and
their ratio measured by means of 500 MHz 1H NMR spectra in
CD3OD. The spectra of the pure glucosides were used as ref-
erences [18-20]. The identification was in each case confirmed
by 500 MHz 1H NMR spectra in CDCI3 of acetates obtained by
overnight treatment (room temperature) with pyridine-Ac20.
Piriqueta taubatensis. Dry material (1.85 g) yielded 16 mg of
a mixture of epivolkenin (4) and taraktophyllin (5; 1:2).
R viscosa ssp. australis. Dry material (4.8 g) yielded 1.6 mg
of deidaclin (2) and 83 mg of a mixture of 4 and 5 (1:9).
T. angustifolia. Fresh material (6.9 g) yielded ca 10 mg of 2
and tetraphyllin A (3; 7:1) and a trace amount of cyanogenic
compound with TLC properties identical to those of volkenin
(6) and tetraphyllin B (7).
T. caerulea. Dry material (4.82 g) yielded 20 mg of a mixture
of 4 and 5 (12:1).
T. chamaedryfolia. Dry material (5.2 g) yielded ca 100 mg of
linamarin (1).
T. hassleriana. Dry material (6.5 g) yielded 3 mg of 1 and 1
mg of 4.
Wormskioldia Iongipedunculata. Dry material (100.8 g)
yielded 30.6 mg of a mixture of 6 and 7 (2:1), and 270 mg of a
mixture of 4 and 5 (1:4).
Acknowledgement--We thank Dr A. E. Van Wyk, University of
Pretoria and Dr J. L. S. Keesing, The Royal Botanic Gardens,
Kew, for plant material used in this work. Financial support to
E. S. O. from the Alfred Benzon Fund (Reykjavik), and the
Nordic Council of Ministers, is gratefully acknowledged.
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