Pergamoa Phytcx~y. Vol. 36, No. 3. pp.

81141f 195’4
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SHORT REPORTS

DIHYDROABSCISIC ALCOHOL FROM A F’ERRHOA CARAMBOLA FRUIT

ANDREA LUTZ and PETER WINTERHALTER

Lehrstuhl fir Lebensmittelchemic, Universitat Wiirzburg, Am Hubland, 97074 Wiirzburg, Germany

(Received in revised form 11 January 1994)

Key Word Index-Averrhoa carambola; Averrhoaceae; starfruic dihydroabscisic alcohol; abscisic

alcohol; MLCCC.

Ahatract+l’S,4E)-2,SDihydroabscisic alcohol has been identified in an ether extract of starfruit (Awrrhw car-
umbola). After purification by multilayer coil countercurrent chromatography (MLCCC) and HPLC, the structure of
the new ABA derivative was elucidated by ‘H and 13CNMR spectroscopy.

INTRODUCl’ION RESULTS AND DKXUSSION

In previous studies [l-3], numerous C, ,-apocarotenoid
metabolites have been identified in Averrhoa carambola
L. fruit (starfruit; syn. carambola). More recently, struc-
tures l-3 relatedto abscisic acid have been isolated from
A. carambola with ABA glucose ester as the major
compound [4]. ABA derivatives l-3 were accompanied
by small amounts of an unknown C, ,-conjugate which,
after enzymatic hydrolysis, liberated an aglycone ex-
hibiting an apparent M, of 252. This latter structure was
found in higher concentrations in non-conjugated form in
starfruit juice, thus enabling its structural elucidation
spectroscopic techniques.
D~C%D&-
The 10000 g supernatant of homogenized starfruit was
extracted with ether. The ethereal isolate was concen-
trated and subjected to flash chromatography. The un-
known constituent was eluted along with the isomeric
abscisic alcohols (ABA-OH) 3. Although analytical
MLCCC provided base-line separations of 2-cis and 2-
tram L, the later eluting c&isomer of 3a could not be
completely separated from the target compound 4. A final
purification of 4 was achieved by normal phase HPLC.
iH NMR spectral data revealed two three-proton singlets
by at 6 1.00 and 1.08 and two three-proton doublets at 6 1.05
and 1.89, the latter showing a long range coupling with an
olefinic proton at 65.90. The proton spectrum also in-
chided signals at 65.52 and 5.62 for an (Q-configured
double bond (J = 15.6 Hz), an AB-type signal at 62.23 and
2.43 (J = 17 Hz) for the CS’-protons, and a hydroxy-
methylene signal at 63.63. The 13C NMR spectrum show-

Table 1. ‘HNMR spectral data

(400 MHG CIBCl,) of compound 4
(b relative to TMS)

Proton 6

Me-6 1.00 and 1.08 2s

Me-3 1.05 d (6.8)
Hz-2 1.6m
2 R=H. R’=Sugrr &R=H, R’=H
Me-2’ 1.89 d (1.5)
3 R-OH, R’-Sugar 3R R-OH, R’-H
H,-5’ 223 d (17.0)
H-3 2.4 m
H,-5 2.43 d (17.0)
H,-1 3.63 m
H-5 5.55 d (15.6)
H-4 5.62 dd (15.6, 7.5)
H-3’ 5.90 br s

J (in Hz) in parentheses.

811
812 Short Reports
ed 15 signals, with the majority of them being similar to
those reported for abscisic alcohol [S]. Instead of two
signals for olefinic carbons in 3n, the DEPT spectrum of
the unknown exhibited signals for a CH- as well as a
CH,-group at 633.6 and 39.6, respectively, which indi-
cated saturation of the C-2/C-3-double bond. The UV-
spectrum showed an absorption maximum at 234 nm,
consistent with an r,b-unsaturated ketone structure.
Comparison of the CD spectrum of 4 with that reported
for (S)-ABA suggested S-configuration at C-l’ [6]. Due to
the tiny amounts of 4, the configuration at the remaining
asymmetric centre, C-3, could not be clarified. Thus, the
structure of the unknown starfruit constituent was estab-
lished to be that of (l’S,4E)-2,3-dihydroabscisic alcohol
(4).
EXPERIMENTAL
General. NMR: 4OOMHz (‘H) and 1OOMHz (13C);
Aash chromatography (FC); Merck silica gel 60
(0.032-0.063 mm); MLCCC [7]: Ito multilayer coil
separator-extractor (P.C. Inc., Potomac) equipped with a
160 m x 1.6 mm PTFE tubing HRGC: J&W fused silica
DB-5 capillary column (30 m x 0.25 mm i.d., film thick-
ness 0.25 pm). The temp. program was from 60” to 300’ at
5” min- I.
Plant material. Fresh ripe starfruit (A. carambola L.)
was obtained by air-freight from Malaysia and was
worked-up immediately after arrival.
Isolation and purification of4. Intact starfruit (24 kg)
was first washed with pentane and then homogenized
with 0.2 M Pi buffer (10 1,pH 7.0). After centrifugation at
10000 g (30 min), the supernatant was continuously
(16 hr) extracted with Et,O. The ethereal extract was
coned and sepal by FC; eluent: Et,O. Frs with 3%and 4
(HRGC control) were further sepd by analytical MLCCC
183 using hexane-EtOAc-MeOH-H,O (0.6: 1.4: 1 :I) as
biphasic solvent system [9] to give trans-3a (250 mg), and
cis-3a (21 mg) and crude 4 (3 mg). The latter was further
purified by normal phase HPLC (LiChrosorb Si60,
250 mm x 4 mm (5 pm), Knauer, Berlin; eluent: Et,O;
UV-detection: 230 nm).
Compound 4 (1.8 mg). UV: 234 nm; [LY]~ +49.7”
(EtOH; ~0.0018); ‘HNMR: Table 1; 13C NMR
(100 MHz, CDCI,): 6 18.9 and 20.8 (Me-2’/Me-3), 22.9
and 24.1 (2 x Me-6’); 33.6(C-3), 39.6 (C-2), 41.1 (C-6’), 49.8
(C-5’), 61.0 (C-l), 79.7 (C-l’), 126.9 (C-3’), 128.5 and 137.4
(C-4/C-5), 162.9 (C-2’). 198.0 (C-4). MS (70 eV) m/z (rel.
int.): 234 [M-H,O]+ (5), 209 (4), 196 (38), 178 (40), 163
(57), 151 (84), I35 (20). 124 (loo), 111 (57), 85 (37), 69 (36),
55 (62). 43 (88); HRGC ret. index: 2033 (DB-5).
Acknowledgements-We wish to thank Dr Scheutzow
and E. Ruckdeschel (Institute fur Organ&he Chemie,
Universitlt Wiit-zburg) for NMR analyses, and Deutsche
Forschungsgemeinschaft, Bonn, for funding the research.
REFERENCFS
I. MacLeod, G. and Ames, J. M. (1990) Phytochemistry
29, 165.
2. Herderich, M., Neubert, C., Winterhalter, P., Schreier,
P. and Skouroumounis, G. K. (1992) F&our Fragr. J.
7, 179.
3. Lutz, A., Winterhalter, P. and Schreier, P. (1993) Nat.
Prod. Letters 3, 95.
4. Winterhalter, P., Lute A., Herderich, M. and Schreier,
P. (1994) in Advances in Fruit Flavors (Leahy, M. and
Rouseli, R., eds), American Chemical Society:
Washington, DC. (in press).
5. Lutz, A. and Winterhalter, P. (1992) J. Agric. Food
Chem. 40, 1116.
6. Ohloff, G., Otto, E., Rautenstrauch, V. and Snatzke, G.
(1973) He/u. Chim. Acta 56, 1874.
7. Ito, Y. (1986) CRC Crit. Rer;. Analyt. Chem. 17, 65.
8. Roscher, R. and Winterhalter, P. (1993) J. Agric. Food
Chem. 41, 1452.
9. Mandava, N. B. and Ito, Y. (1984) J. Liq. Chromatogr.
7, 303.