Separation of Enantiomers in Thin-Layer Chromatography
Margot Mack*/H. E. Hauck
E. Merck, FO REAG CHROM, Postfach 41 19, D-6100 Darmstadt, F RG
Key Words
Thin-layer chromatography
Retention mechanisms for enantiomer separation
Application of ligand exchange chromatography
Summary
Up to now in thin-layer chromatography a series of re-
tention mechanisms for the determination of enantio-
rneric compositions have been applied. Subject of this
paper is a corresponding survey with a focal point on
ligand exchange chromatography. In connection with
this retention mechanism most successfully used in TLC
for the separation of optical antipodes, some additional
findings of the authors are included.
Retention mechanisms in question are:
- Formation of inclusion complexes within the cavities
of cyclodextrins. Substance classes separated by this
system are e.g. dansyl amino acids or metallocenes.
- Interaction with so-called "brush type" phases. In
this case, chiral groups with rr-acceptor or ~ l o n o r
properties are chemically or physically bonded to an
aminomodified silica gel matrix. Chiral compounds,
like aromatic alcohols, were separated on these
stationary phases.
-Differences in retention of diastereomers formed
previously or during the chromatographic process
on an achiral stationary phase. Some examples for
-the manifold applicability of this principle are the
separation of chiral amines from biological matrices,
hydroxy-fatty acids, and amino acids.
- Selective interactions with the helical structures of
cellulose or cellulose triacetate. Referring to this
technique, separations of several amino acids and of
the analgesic agent Oxindanac in the R- and S-species
are described.
- Enantioselective ligand exchange, based on reversed
phase silica gels impregnated with copper salts and
optically active amino acid derivatives. In thin-layer
chromatography this is the most important principle
used mainly for the separation of racemic mixtures
Chromatographia,Vol. 26 (1988)
0009-5893/88 0197-09 ~ 03.00/0 9 1988 Friedr. Vieweg & Sohn VerlagsgesellschaftmbH
of free amino acids and amino acid derivatives. The
recently developed HPTLC pre-coated plate CHIR
with concentrating zone is also based on this reten-
tion mechanism. Aside from the applications mention-
ed above, dipeptides, hydroxy-carboxylic acids, ~z-
halogenated carboxylic acids, amino lactones, and
thiazolidine derivatives have been separated on this
plate up to now. Furthermore, this new pre-coated
layer is particularly suitable for an optical in situ
evaluation and quantitative determinations of optical
antipodes.
Introduction
In recent years there has been an increasing demand for the
separation of enantiomers with chromatographic techniques.
This is expressed by a rapidly growing number of publi-
cations and innovations in this field. Up to now research
work here is concentrated on HPLC and GC [ 1 - 5 ] . Besides
these techniques, which require an expensive apparatus,
there is demand for a simple, economic and fast method for
the determination of the enantiomeric composition. Here
thin-layer chromatography, with an enantioselective station-
ary phase, is the method of choice.
The importance of enantiomeric separation increased when
it became evident that corresponding optical antipodes
often differ in respect to their effects in a biological organ-
ism. As exemplified in Fig. 1, enantiomers can deviate in
flavor, pharmacological effectiveness or even in toxicity.
In spite of this, many chiral substances, like pharmaceuticals
or plant protective agents, are still used as racemic mix-
tures [6, 7].
For the chromatographic separation of enantiomers three
different methods can be used in principle:
a) Direct separation of optical antipodes on chiral station-
ary phases (CSP).
This separation principle has the advantage of being fast;
a derivatization step is not necessary and the danger of
racemization is negligible. A disadvantage is the high
price of most CSPs. Additionally, the prediction of a
suitable CSP is often difficult and empirical.
197
 CHIRALITY-EFFECT

CH3 CH=
Limonene 9 R

H o ~ C H= H=C~CH3

Lemon odor Orange odor

. S~
SS Ethambutol RR
"OH H H HO/
Tuberculostatic Blindness

H3C CH= H3C CH3

S Hs,~COOH Penlclllamine HOOC~s H R
NH2 HzN
Antlarthrltlc Extremly toxic
J J

Thyroxine HOOCH2N~'~O"~ JOH .R

S HO,,j~~ jt NHz
1
J J

Thyroid gland hormone Ant ihypocholesterlnlc

CH3 H3C

s OH Propanolol HO R

p-Blocker Contraceptive

Teratogen
i- o Thalidomid
(Contergan~))

Sedative
R

Fig. 1
Different effects in respect to the absolute configuration of chiral substances.

b) Direct separation of enantiomers on achiral stationary
phases which are impregnated with a chiral substance,
the so-called chiral selector. Impregnation can also be
realized by adding the chiral selector to the mobile
phase.
Here conventional sorbents as carriers for the chiral
stationary phases can be use. The sample molecules do
not have to be derivatized in this case. A certain dis-
advantage is the possible elution of the selector which
is not covalently bonded.
c) Indirect separation of chiral substances on achiral
stationary phases after forming diastereomeric deriva-
tives of the sample molecules.
This widely applied method has an important disad
vantage: The possible racemization as a consequencec!
the derivatization step. The same problem appears whe:
splitting diastereomers for regaining pure enantiomer~
after chromatographic separation.
Mechanisms for Enantiomer Separation
After discussing these basic techniques for optical res01u
tion, the mechanisms for enantiomeric separation usedin
TLC will be treated in detail. The mechanisms of ch[ra'
recognition can be subdivided into five groups:

198 Chromatographia, Vol. 26 (19~!

Formation of Inclusion Complexes Charge-Transfer C o m p l e x a t i o n w i t h PirkleoType CSP

Certain cyclic chiral molecules are capable of including
guest molecules in their cavities. A group of suitable mole-
cules for building these inclusion complexes are cyclo-
dextrins. In Fig. 2 the a-, ~- and *f-cyclodextrins are shown,
which are built up from 6, 7 or 8 (D)-(cz)-glucopyranose
units in an a-l,4-bonding. Because of their increasing
cavitydiameters they are able to form host guest complexes
with molecules of increasing size. The separation of enanti-
0mers is caused by the formation of diastereomeric in-
clusion complexes during the chromatographic process,
which leads to differences in retention behavior of the
respectiveoptical isomers.
Previously, cyclodextrins were used in TLC to improve
the selectivity for certain isomers like substituted phenols,
quinones and benzoic acids [ 8 - 1 0 ] . This technique was
named "pseudophase liquid chromatography" by D.W.
Armstrong. In 1986 the same author prepared a silica
gel chemically modified with /~-cyclodextrin [11]. This
01igocarbohydrate was fixed via a spacer onto the matrix.
He was able to separate the optical antipodes of several
dansyl amino acids, alanine-/3-naphthylamides, and some
metallocenes [12] with handmade plates of this modified
silica gel. I. D. Wilson [13] impregnated silica gel pre-coated
plates with a solution of ~-cyclodextrins solved in ethanol
and DMSO. With these chiral layers he succeeded only in
a partial separation of R- and S-mandelic acid.
In liquid chromatography, a widely used mechanism for the
separation of enantiomers is based on fundamental re-
search on chiral recognition by W. Pirkle [5, 14-16]. He
created a series of chiral selectors with ~-donor or ~-ac-
ceptor properties. The most important selectors of "'PirMe-
type" are 3,5-dinitrobenzoyl amino acids. The carboxylic
groups of these chiral selectors can be bonded ionically or
covalently to an NH2-grou p of aminopropyl modified
silica gel forming so-called brush type phases.
The main interaction in the chiral recognition process is
the formation of charge transfer complexes between the
~-acid, in this case the 3,5-dinitrobenzoyl group of the
selector, with a suitable ~T-base in the chiral molecule to
be separated.
According to the "three point rule" of C. Dalgliesh [17], at
least three interactions are necessary for the discrimina-
tion of two optical isomers. In the "Pirkle selector" there
are binding sites for the formation of hydrogen bondings,
dipol-dipol, and other interactions [15].
I. Wainer et al. used this principle in TLC. The chiral selec-
tor (R)-N-3,5-dinitrobenzoyl phenylglycine was bonded
ionically to 3,-aminopropyl silanized silica gel plates. The
resulting chiral TLC-plate was used for the separation of a
racemic mixture of 2,2,2-trifluoro-l-(9-anthryl) ethanol
[18].

Cyclodextrins
structure and C h a r a c t e r i s t i c s

OH

OH @ O~o ~-~-~
o~ o§ o. o (
" O
o ~ O NO 0 o.o'2' " ~
o -~247 "o o,1 # ~
o ~o

or=
% = o
o_'~,...,
~,o>'~'~~ --o, ~
V
o~oy-
g

WoZj o
~o ~0 HO

,~oCyclodextrin /~-Cyclodextrin 7-Cyclodextrin

1,53 ~ '

Glucose units 6 7 8

Mol. wt. 972 1135 1297

o 150-'O,5 162,5• 177,4-+ 0,5

Cavity diameter/~, 4,7 - 5,3 6 , 0 - 6,5 7,5 - 8 ~

Fig. 2
Chemical and physical properties of cyclodextrins [46, 47].

Chromatographia,Vol. 26 (1988) 199

Formation of Diastereomers
A widespread mechanistic principle used in TLC for the
separation of optical antipodes is based on differences in
retention of diastereomers built up by the reaction with
chiral agents before or during the chromatographic pro-
cess.
The racemates of 22 amino acids were derivatized by Mar-
fey's reagent, a derivative of L-alanine amide. The so form-
ed diastereomers were separated on a TLC RP-18 plate by
K. Ruterbories and D. Nurok [19].
H. Weber et al. estimated the enantiomeric composition of
several pharmaceutically relevant amines extracted from a
biological matrix, like amphetamine or methamphetamine.
By forming the diastereomeric amides with (-) or (+)-ben-
oxaprofen a separation of the enantiomers was performed
on silica gel plates [20].
The separation of racemates of some anti-inflammatory
drugs based on a 2-arylpropionic acid structure like keto-
profen, indoprofen, and suprofen was described by V.
Rosetti et al. [21]. In this case, a resolution of the analytes
was performed on an HPTLC pre-coated plate silica gel 60
after derivatization to the diastereomeric 1-phenylethyl
amides.
The formation of diastereomeric salts during the chroma-
tographic process was used by R. Bhushan and I. All for
the separation of amino acid enantiomers. They prepared
silica gel plates containing the optically active alcaloids (-)-
brucine [22] or (-)-berberine hydrochloric acid [23]. The
resolution of PTH-amino acids was performed on silica gel
plates impregnated with (+)-tartaric acid [24].
I. Wilson impregnated silica gel plates with D-campher
sulfonic acid and with quinine. Only with the latter, a
partial separation of mandelic acid was possible [13].
Selective Interactions with Organic Polymers
As early as during the 1950ies, M. Kotake and other au-
thors observed that certain racemates, first of all amino
acids, were separated into two spots by the chromato-
graphic development on paper or cellulose layers [ 2 5 - 2 9 ] .
In 1952, C. Dalglish postulated his "three point rule" for
the chiral recognition in the chromatographic separation
of optical antipodes [17]. His findings were based on sys-
tematic investigations of the paper-chromatographic separa-
tion of the enantiomers of different dihydroxy phenyl-
alanines.
The mechanism of chiral recognition on cellulose is not
clarified completely. It is assumed that several helical
chains of cellulose, which are built up from /3-D-glucose
units, are forming secondary structures containing chiral
cavities. Sample molecules to be separated interact select-
ively with these cavities. In recent years, the chromato-
graphic properties of cellulose have been improved by
chemical modification, e.g. by acetylation obtaining
cellulosetriacetate.
A new, commercially available pre-coated plate built from
microcrystalline cellulosetriacetate was introduced in 1987.
With this plate M. Faupel separated the optical antipodes
of the benzylester of the analgesic agent "Oxindanac" [30].
200
Up to now, no further results have been published base
on this mechanistic principle in TLC.
Chiral Ligand Exchange
The most important principle for the separation of enanti0.
mers in TLC is the chiral ligand exchange mechanisrn,i
method first introduced by Davankov in column liquic
chromatography [31, 32]. The mechanism is based onth~
copper complex formation of a chiral selector (in m0~
cases an amino acid derivative) and the optically activ,,
selectand. This is shown in Fig. 3 through the examp1:
of amino acid. Differences in retention of the antip0de~
to be separated are caused by dissimilar stabilities of the~
intermediately formed diastereomeric copper complexes.
Applications based on ligand exchange mechanism at!
performed either with handmade plates or with commerc
ially available pre-coated TLC-plates.
R. Marchelli et al. impregnated an HPTLC pre-coated plat~.
RP-18 with a copper salt and a chiral selector consistin!
of two amino acids, which were combined by the reacti0r
with e.g. ethylene diamine and simultaneous formati0r
of amide bonds. Through this chiral stationary phase,i
separation of racemic dansyl amino acids was performr
[33].
Dansyl amino acids were also separated by S. Weinstei"
with a N,N-dialkyl amino acid as chiral selector [34].
Some applications of the commercially available Chiral
plate | were published by K. GLinther and coworkers. It,
this case, enantio-selectivity was achieved by using a ~l-
dodecyl derivative of L-hydroxyproline and a copper salt
The advantages of ligand exchange mechanism have beer
shown in the field of the separation of a large number of
optically active compounds, like amino acids, a number QI
their derivatives, and some other related compounds [35-
42]. The optical resolution of the enantiomers of N-car.
bamyl tryptophan with this plate was performed at 10~
temperature by L. Gont and S. Neuendorf [43]. With thi~
chromatographic system the (~- and ~-anomers of several
nucleobases were separated by R. Feldberg and L. Repucci
[44].
D or L-aminoacid ~ H chiralselector
~, /
s / S /" / '~
S i-O-Si-O-Si-O-Si-O-Si
Fig. 3
Ligand exchange chromatography -- intermediate formed copier
complex with the chiral selector and an amino acid [45].
Chrornatographia, Vol. 26 (1s
HPTLC Pre-Coated Plate CHIR with Concentrating Zone

The recently developed HPTLC pre-coated plate CHIR with

(I-Alk]yl- i halogonat,I ajld- N-AlkylI-
concentrating zone (developed under licence of Degussa) is I
basedon chiral ligand exchange mechanism.
The composition and characteristics of this enantioselec-
Formyl.
Amino acid
Me~hoxy-
and HydtOxy-
I
derivatives
t{ve plate are as follows:
- The matrix is built up from silica gel 60 in high perfor-
mance quality, which requires a very narrow particle
N-Acetyl. I Nitro- I
size distribution and a small mean particle diameter of
about 5#m. This in turn causes a smooth surface struc-
ture. Therefore, the HPTLC pre-coated plate CHIR with
concentrating zone is especially suitable for qualitative Amino acids [~ Di- andTri-

pepUdes
and quantitative in-situ evaluations.
- The silica gel matrix is chemically modified to an RP-18 /
phase with a defined degree of modification optimized
for this special purpose.
- I n order to obtain enantioselectivity, the modified
matrix is impregnated with a copper salt and a chiral
I Butyrolactone HPTLC pre-coated plate

CHIR I Thlazolldlne

proline derivative as optimized chiral selector. derlvatlvea derivatives

with concentrating zone
-Additionally, this chiral separation layer is combined
with a so-called concentrating zone. This small band,
consisting of silicon dioxide with extremely wide pores

t
and a very small specific surface area, is placed in front
of the separation layer. Sample molecules do not inter-
act with this inert silicon dioxide. During the chromato- Mandelacilcd/ I ct-Hydroxy (r-halogenated
graphic development, sample molecules are transported arid derivatives carboxyllc acids carboxyllc acids
with the solvent front. Reaching the borderline of the
two layer sections, the spots are concentrated to small
Fig. 4
bands. Hence an improved starting position and a higher
Application range of the HPTLC pre-coated plate CHIR with con-
efficiency of the separation process following is ob- centrating zone [45].
tained, especially in the case of large sample volumes.

Qualitative Applications
A survey of optically active substance classes separated up
to now with the HPTLC pre-coated plate CHIR with con-
centrating zone is shown in Fig. 4. All substances listed
here are able to form complexes with copper ions, a pre-
condition for the possible applicability of ligand exchange
mechanism.
The most important field of application of this special plate
is the separation of amino acid enantiomers. This is demon-
strated in the case of the racemates of the proteinogenic
amino acids leucine, phenylalanine, and proline (Fig. 5).
The amino acids can be separated directly without derivat-
ization. The visualization is performed by staining the
acids with ninhydrin. The background of the plate is not
affected by this reagent. Therefore, in-situ evaluations with
a TLC-scanner can be performed advantageously.
However, the HPTLC pre-coated prate CH|R with concen-
trating zone is not only very suitable for free amino acids,
but also for a variety of their derivatives. Here sometimes
even better selectivities for the optical isomers are obtained.
This is shown by the optical resolution of the racemates of
the halogenated amino acid D L-4-chlorophenylalanine (Fig.
6a) and the naphthyl-derivatized DL-3-(2-naphthyl)alanine
(Fig. 6b). The substitution of sulfur by selenium does not
affect the enantiomeric separation, as demonstrated in
Fig. 6c by the example of DL-selenomethionine. A methyl-
group at the asymmetric carbon atom instead of the hydro-
gen does not remove enantioselectivity. This is shown by
the separation of e-methyI-DL-methionine into the optical
antipodes (Fig. 7 ).
As demonstrated in Fig. 8, the important class of dipeptides
can be separated on this special pre-coated plate into opti-
cal isomers and diastereomers. The concrete example here
is the resolution of D-Leu-L-Leu, L-Leu-L-Leu, and L-Leu-
D-Leu.
A biochemically relevant substance class, which can be
separated with the HPTLC pre-coated plate CHIR with
concentrating zone into the enantiomers, is e-hydroxy
carboxylic acid. The enantioselectivity for DL-vanillin
mandelic acid is demonstrated in Fig. 9. Despite the stain-
ing method with manganese chloride and sulfuric acid used
here, an in-situ evaluation is possible.
The suitability of this chiral plate for the optical resolution
of e-halogenated carboxylic acids is shown in Fig. 10 for
D L-2-chloro-6-benzoylamino caproic acid.
Apart from the substance classes mentioned, which can be
separated directly, there are resolution problems, which can
only be solved after a suitable derivatization of the sample
molecules with achiral agents. A concrete application of

Chromatographia,Vol. 26 (1988) 201

 L

L
D

U~

ee

t ! t
0 7cm o 10cm 0 7cnn

a. b. c.

Fig. 5
Separation of racemic amino acids
plate: HPTLC pre-coated plate CHIR with concentrating zone eluent: methanol/water/acetonitrile 50/50/30
(v/v/v)
migration distance: 7cm (a, c}; 10cm (b}
compounds:
a} DL-leucine b} DL-phenylalanine c) DL-proline
detection:
dipped in 0.5% ninhydrin in ethanol/acetic acid glacial 98/2, heating up to 120~ for 5min, in-situ evaluation
with TLC/HPTLC scanner (Camag)
UV: 520nm (a, b } ; 4 1 0 n m (c).

Fig. 6
Separation of amino acid derivatives
6a}
plate: see Fig. 5
eluent: methanol/water/acetonitrile 50/50120 (v/v/v}
compounds:
a) D-4-chlorophenylalani ne
b) L-4-chlorophenylalanine (all 0.1%}
application volume: 5/~1
migration distance; 7cm
chamber: normal chamber without chamber saturati0~
detection: see Fig. 5
UV: 410nm.
=.
6b)
03
compounds:
M
~tJ a) D-3-(2-Napht hyl)-alani ne
b) L-3-(2-Naphthyl)-alanine
conditions of chromatography and detection: seeDL.
4-chlorophenylalanine.

6c)
f t ~ 1 ! compound: Selenium-DL-methionine
7cm 0 7crn 0 7crn
conditions of chromatography and detection: seeDL.
a. b. c. 4-chlorophenylalanine

202 Chromatographia, Vol. 26 (1988:

 Fig. 7
Separation of m-methyl amino acids
plate: see Fig. 5
eluent: methanol/weter/acetonitrile 50/50/30 (v/v/v)
compound: c~-methyl-DL-methionine (0.1%)
application volume: 5#1
migration distance: 7cm, normal chamber without chamber satura-
tion
detection: ninhydrin (see Fig. 5)
UV 520nm.

! t
Fig. 9 0 7r
Separation of e-hydroxy carboxylic acids
plate: see Fig. 5
eluent: chloroform/methanol 90/10 (v/v), 80% saturated with water
compound: DL-vanillin mandelic acid (4-Hydroxy-3-methoxyman-
delic acid) (0.05%)
migration distance: 7cm, normal chamber without chamber satura-
tion
application volume: 5/~1
detection: dipped in MnCI2-sulfuric acid heating up to 110~ for
10min, in-situ evaluation with T L C / H P T L C scanner (Camag)
UV 460 nm.

!
7cm

3 Fig. 8
Separation of dipeptides
plate: see Fig. 5
eluent: methanol/1-propanol/water 50/10/40 (v/v/v)
compou nds:
1 ) D-Leu-L-Leu
2) L-Leu-L-Leu
3) L-Leu-D-Leu (all 0.05%)
application volume: 5#1
detection: ninhydrin (see Fig. 5)
UV 410nm.

t,,
0 7crn
Fig. 10
Separation of ~-halogenated carboxylic acid
plate: see Fig. 5
eluent: methanol/water/acetonitrile 50/50/20 (v/v/v)
compou nd: D h-2-chloro-6-benzoylam inocaproic acid
application volume: 5/zl
migration distance: 7cm, normal chamber with chamber saturation
t detection: ninhydrin (see Fig. 5)
0 lOcrn UV 410nm.

Chromatographia, Vol. 26 (1988) 203

 this method is shown in the example of the amino alcohol
noradrenaline reacted w i t h salicylaldehyde forming a Schiff
base (Fig. 1 1 ).
Quantitative Analysis
The calibration curve of D-tryptophan determined in the
racemic mixture shows that there is a linear relationship
between sample concentration and peak area in the range
of 50ng to 1,5/1g per spot (Fig. 12).
The determination of the enantiomeric composition is
possible even in trace level. The determination of L-trypto-
phan as a minor component in presence of an excess of D-
tryptophan can be realized in such extreme relative pro-
portions as 1 : 100 to 1 : 1000. This means a relative pro-
portion of 1% to 0.1%, as shown in Fig. 13.
A d d i t i o n a l l y , it does not matter if the minor component
follows or precedes the main enantiomer. This is basically
attributed to the benefits of the concentrating zone, which
can also fulfill a certain clean-up function.
Conclusion
In TLC some different retention mechanisms are applied
for the separation of optical antipodes. Here chiral ligand
exchange mechanism is the best established method fore
defined field of application.
In order to extend the applicability of TLC in the field of
enantiomeric separations, further retention mechanisms
have to be developed and existing methods should be ira.
proved. Here a considerable backlog demand exists compar.
ed to HPLC. This, too, can be said of the possible use of
T L C as a pilot technique for column liquid chrornat0-
graphy.
References
[1] W. Lindner, Chimia 35,294 (1981).
[2J R. Db'ppen, H. Arm, V. Meyer, J. Chromatogr. 373, 1 (1986).
[3] G. GiJbitz, GIT Suppl. Chromatogr. 4, 6 (1985}.
[4] G. Blaschke, Angew. Chem. 92, 14 (1980).

j
u~
IU

60.
1

t t
0 7cm

Fig. 11 Fig. 12
Separation of c~-aminoalcohols Calibration curve of D-tryptophan [45]
derivatization: Substances were derivatized with an equimolar a- (received under the conditions of separation of racemic tryptophanl
mount of salicylic aldehyde [48] plate: see Fig. 5
plate: see Fig. 5. eluent: methanol/water/acetonitrile 50/50/30 (v/v/v)
eluent: chloroform/methanol 90/10 (v/v), 80% saturated with migration distance: 7cm, normal chamber with chamber saturation
water application volume: 0.75/~1
compounds: concentration of D-tryptophan samples:
1) S(+)-Noradrenaline 1O0ng/spot
2) R (-)-Noradrenaline 250 ng/spot
3) Salicylic aldehyde (all 0.1%) 500 ng/spot
application volume: 5/41 750 ng/spot
migration distance: 7cm, normal chamber with chamber saturation 1000 ng/spot
detection: in-situ evaluation with TLC/HPTLC scanner (Camag) 1500 ng/spot
UV 410nm detection: in-situ evaluation with TLC/HPTLC scanner (Camag)
UV 510 nm

204 Chromatographia, Vol. 26 (1986~

 1 1 [12] A. Alak, D. Armstrong, Anal. Chem. 58,582 (1986).
[13] I. Wilson, Method. Surv. Biochem. Anal. 16,277 (1986).
[14] W. Pirkle, J. Finn, L. Schreiner, B. Hamper, J. Am. Chem.
5oc. 103, 3964 (1981).
[ 15] W. Pirkle, C. Welch, J. Org. Chem. 49, 138 (1984).
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[17] C. Dalgliesh, J. Chem. Soc. 137,3940 (1952).
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(1983).
[19} K. Ruterbories, D. Nurok, Anal. Chem. 59, 2735 (1987).
[20] H. Weber, H. Spahn, E. Mutschler, J. Chromatogr. 307, 145
(1984}.
[21 ] V. Rosetti, A. Lombard, M. Buffa, J. Pharm. Biomed. Anal.
4,673 (1986).
[22] R. Bhushan, I. Aft, Chromatographia 23,141 (1987).
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(1988).
[24] R. Bhushan, I. A l i , J. Chromatogr. 392,460 (1987).
[25] M. Kotake, J. Am. Chem. Soc. 73, 2973 (1951).
[26] R. We[chert, Acta Chem. Scand. 9,547 (1955).
[27] C. Dalgliesh, Biochem. J. 52, 3 (1952).
J [28] J. Lambooy, J. Am. Chem. Soc. 76,133 (1953).
[29] M. Mason, C. Berg, J. Biol. Chem. 188,783 (1951).
130] M. Faupel in: Proceedings of the Fourth International
Symposium on instrumental HPTLC, Selvino, 1987, 147.
~J [31 ] V. Davankov, S. Rogozhin, A. Semechkin, T. Sachkova, J.
Chromatogr. 82,359 (1973).
a. [32J [I. Davankov, Adv. Chromatogr. 18, 139 (1960).
bo C~ [33] R. Marchelli, R. Virgil[, E. Armani, A. Dossena, J. Chroma-
Fig. 13 togr. 355, 354 (1986).
Determination of the enantiomeric composition of DL-tryptophan [34] S. Weinstein, Tetrahedron Lett. 25,985 (1984).
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