New 19α-hydroxyursane-type triterpenes from the leaves of Meyna spinosa (= Vangueria spinosa), Rubiaceae

Article abstract of DOI:10.1016/j.phytol.2014.03.009

Two new 19α-hydroxyursane-type triterpenes, 2α,3α, 19α,24,28-pentahydroxyurs-12-ene (1) and meyanthic acid, 3β-acetoxy-2β,19α,23-trihydroxyurs-12-en-28-oic acid (2) along with one new aliphatic ester, myricyl pentadecanoate (3) and five known compounds, 1

Full text of DOI:10.1016/j.phytol.2014.03.009

Phytochemistry Letters 9 (2014) 7–10
Contents lists available at ScienceDirect
Phytochemistry Letters
journal homepage: www.elsevier.com/locate/phytol
New 19
a
-hydroxyursane-type triterpenes from the leaves of Meyna
spinosa (= Vangueria spinosa), Rubiaceae
Prasenjit Rudrapaul a, Niranjan Das b, Utpal Chandra De a, Biswanath Dinda a,c,
*
a Department of Chemistry, Tripura University, Suryamaninagar 799022, Tripura, India
b Department of Chemistry, Netaji Subhash Mahavidyalaya, Udaipur 799120, South Tripura, India
c Department of Chemistry, National Institute of Technology, Agartala, Jirania 799055, Tripura, India
A R T I C L E I N F O
A B S T R A C T
Article history:
Two new 19
meyanthic acid, 3
aliphatic ester, myricyl pentadecanoate (3) and five known compounds, 19
oleanolic acid (5), myricyl alcohol (6), -sitosterol (7) and its glycoside (8) were isolated from the
a
-hydroxyursane-type triterpenes, 2
a
,3
a
,19
a
,24,28-pentahydroxyurs-12-ene (1) and
Received 7 October 2013
Received in revised form 7 March 2014
Accepted 25 March 2014
Available online 13 April 2014
b
-acetoxy-2 ,19
b
a
,23-trihydroxyurs-12-en-28-oic acid (2) along with one new
-hydroxyasiatic acid (4),
a
b
methanolic leaf extract of Meyna spinosa Roxb. ex Link (= Vangueria spinosa Roxb., Rubiaceae). The
structures of the new compounds were elucidated on the basis of extensive spectroscopic (including 2D
NMR) analysis and comparison with literature. Except oleanolic acid, isolation of known compounds was
reported for the first time from this plant.
Keywords:
Meyna spinosa
Vangueria spinosa
Rubiaceae
ß 2014 Phytochemical Society of Europe. Published by Elsevier B.V. All rights reserved.
19a-Hydroxyursane-type triterpenes
1. Introduction
2. Results and discussion
Meyna spinosa Roxb. ex Link (= Vangueria spinosa Roxb.,
Rubiaceae), a medium sized thorny bushy shrub, is distributed
in East and South India, and in neighboring countries Bangladesh,
Nepal, Java, Thailand, Vietnam and Myanmar (Deb, 1983). In Assam
(India), the plant is known as ‘‘Kutkura’’ and in Bangladesh as
‘‘Moyna’’. Ripe fruits have slightly sweet in taste and are eaten raw.
In traditional medicine, the mature fruits are used for treatment of
gastritis, cracked heels, piles, biliary and hepatic congestions
(Buragohain, 2008; Yusuf et al., 2009). Leaves are used for the
treatment of bone-fracture, skin irritation, abortion, diphtheria
and renal diseases (Pullaiah, 2002). Previous studies on the plant
The powdered leaves of M. spinosa were successively extracted
with methanol at room temperature and the methanolic extract
was fractionated into petroleum ether, chloroform, ethyl acetate
and n-butanol fractions. Compounds 3, 6, 7 were isolated from the
petroleum ether extract and compounds 1, 2, 4, 5 and 8 (Fig. 1)
from chloroform extract after extensive silica gel column
chromatography. Compounds 4 and 5 were also obtained from
ethyl acetate extract.
Known compounds 4, 5, 6, 7 and 8 were identified as 19
hydroxyasiatic acid (2 ,3 ,19 ,23-tetrahydroxyurs-12-en-28-oic
acid) (Gao et al., 1985), oleanolic acid (Mahato and Kundu, 1994;
a-
a
b
a
reported the isolation of (-) epicatechin-3-O-
from leaves (Chatterjee et al., 2011), oleanolic acid from fruits
(Buragohain, 2008) and saponin, 3 -O-[ -rhamnopyranosyl-
(1!2) -xylopyranosyl]-oleanolic acid from leaves (Gogoi and
Sarma, 1997). We chemically investigated the leaves of the plant
and isolated two new 19 -hydroxyursane-type triterpenes 1 and 2
as well as a new aliphatic ester 3 and five known compounds, 4–8
from the methanolic extract. We report in this paper, the isolation
and structure elucidation of these compounds.
b
-
D
-glucopyranoside
Maillard et al., 1992), myricyl alcohol (Dinda et al., 1999),
b
-
-
sitosterol (Zhang et al., 2006) and 3-O- -glucopyranosyl-b
b
-D
b
a
-L
sitosterol (Paulo et al., 2000), respectively, by comparison of their
1H and 13C-NMR spectra and mass spectral fragmentation patterns
with the reported data.
b
-D
a
Compound 1 (Fig. 1) was obtained as colorless amorphous
powder. Its molecular formula was determined as C30H50O5 from
the HR-FAB-MS at m/z 491.3733 [M+H]+ (Calcd. for C30H51O5,
491.3737) and 13C-NMR spectral data. The IR spectrum showed
characteristic absorptions for hydroxyl (3436 (br) cmꢀ1) and
olefinic (1636 cmꢀ1) functions. The 1H-NMR spectrum (Table 1)
showed one proton singlet at dH 2.68 along with five tertiary
methyl singlets at dH 1.05, 1.18, 1.34, 1.45 and 1.69, a secondary
*
Corresponding author at: Department of Chemistry, Tripura University,
Suryamaninagar 799022, Tripura, India. Tel.: +91 9436502137.
methyl at dH 0.93 (d, J = 6.6 Hz, H3-30) and an olefinic proton signal
E-mail address: dindabtu@gmail.com (B. Dinda).
http://dx.doi.org/10.1016/j.phytol.2014.03.009
1874-3900/ß 2014 Phytochemical Society of Europe. Published by Elsevier B.V. All rights reserved.

8
P. Rudrapaul et al. / Phytochemistry Letters 9 (2014) 7–10
spectrum suggested the presence of a hydroxymethyl group at 23 or
24 position. The 13C-NMR spectrum (Table 1) showed 30 carbons
including six methyl carbons ( 16.3, 17.1, 17.8, 24.4, 25.0, 27.4) and
two sp2 carbons of a trisubstituted olefin at
C 127.2 and 138.4, one
tertiary carbon at 73.0, two hydroxymethyl carbons at
68.1andtwomethinecarbonsbearinga hydroxylgroupat
73.8 suggesting its 19
d
d
d
d
d
65.3 and
68.0and
a-hydroxyurs-12-ene-type triterpenoid
structure with two secondary hydroxyl groups and two hydro-
xymethyl groups for the compound (Cheng and Cao, 1992). The
presence of carbon signals in 1 at dC 24.4 (CH3) and 65.3 (CH2)
suggested the presence of hydroxyl group at C-24 position (Kojima
and Ogura, 1989; Zhang and Yang, 1994). The additional two proton
signalsatdH 3.30(1H, d,J = 10.2 Hz)and 3.62(1H, d, J = 10.2 Hz)inits
1H-NMR spectrum, and a methylene carbon signal at dc 68.1
suggested the presence of a hydroxymethyl group. Its location at C-
28 position was assigned by HMBC correlation of H-18 with dC at
Fig. 1. Structures of compounds 1–4 and 6.
68.1. The HMBC correlation of H-3 with
location of another hydroxymethyl group at 24-position. The 13C-
NMR data were very similar to that of 2 ,3 ,19 ,24-tetrahydrox-
yurs-12-en-28-oic-28- -glucopyranosylester except at C-28
dC at 65.3 also supported the
at
d
5.50 (br s), characteristic of 19
a
-hydroxyursane type-
a
a
a
triterpene (Jung et al., 2004). The 1H-NMR spectrum also exhibited
b-D
signals for two carbinol methines at
d
H 4.21 (1H, ddd, J = 10.0, 5.4,
and H-3
(Jung et al., 2004). The NOESY correlations between H-2 and H-
25; H-3 and H-24 and between H-18 and H-28 of 1 also supported
3.0 Hz) and 3.83 (1H, d, J = 3.0 Hz), assignable to H-2
b
b,
respectively (Kojima and Ogura, 1989). The coupling constants 3JH-
1/H-2 (trans) = 10.0 Hz, 3JH-1/H-2 (cis) = 5.4 Hz and 3JH-2/H-3 (cis) = 3.0 Hz
the assignments of 2
Furthermore, NOESY relations between H-18andH-19indicated the
-orientation of C-19 hydroxyl group. Selected NOESY correlations
in 1 are shown in Fig. 2. Thus, compound 1 was determined to be
a,3a,24,28-hydroxyl groups in the compound.
were consistent for 2
a
,3a
-dihydroxyl substituents in ring A
a
(Kojima and Ogura, 1989). Furthermore, two proton signals at dH
3.72 (1H, d, J = 10.8 Hz) and 4.17 (1H, d, J = 10.8 Hz) in the 1H-NMR
2a,3a,19a,24,28-pentahydroxyurs-12-ene (1).
Table 1
NMR spectroscopic dataa of compounds 1 and 2 (in C5D5N) (
d, ppm; J, Hz).
Position
1/CH2
1
2
dC
dH
dC
dH
42.4
3.11 (m), 1.25 (m)
42.9
2.31 (dd, 12.6, 4.2)
1.25 (dd, 12.6, 2.4)
4.27 (ddd, 4.2, 4.0, 2.4)
5.06 (d, 2.4)
2/CH
68.0
73.8
42.8
48.9
19.0
33.5
40.2
48.3
38.8
24.3
127.2
138.4
42.6
27.9
24.2
48.6
54.8
73.0
43.2
30.3
37.5
4.21 (ddd, 10.0, 5.4, 3.0)
69.2
78.6
42.5
48.3
19.0
33.5
40.8
48.6
38.8
24.5
128.3
140.3
42.5
29.6
26.7
48.2
54.9
73.0
42.7
27.3
38.7
3/CH
3.83 (d, 3.0)
–
4/C
–
5/CH
1.60 (m)
2.05 (m)
6/CH2
7/CH2
8/C
–
–
9/CH
1.74 (m)
1.77 (m)
10/C
11/CH2
12/CH
13/C
5.50 (br s)
5.60 (t-like)
–
–
–
–
14/C
15/CH2
16/CH2
17/C
–
–
18/CH
19/C
2.68 (s)
–
3.06 (s)
–
20/CH
21/CH2
22/CH2
1.87 (br d, 11.4)
1.74(dd, 11.4, 4.2)
3.74 (d, 10.2)
4.22 (d, 10.2)
1.43 (s)
23/CH3/CH2
24/CH2/CH3
24.4
65.3
1.69 (s)
66.6
14.7
4.17 (d, 10.8)
3.72 (d, 10.8)
1.05 (s)
25/CH3
26/CH3
27/CH3
28/CH2/C
17.8
17.1
25.0
68.1
17.7
17.6
1.11 (s)
1.09 (s)
1.15 (s)
–
1.18 (s)
1.45 (s)
25.0
3.62 (d, 10.2)
3.30 (d, 10.2)
1.34 (s)
181.0
29/CH3
30/CH3
19/OH
3/OAc
27.4
16.3
27.4
17.1
1.67 (s)
0.93 (d, 6.6)
5.82 (br s)
1.12 (d, 6.0)
6.29 (s)
21.7, 170.2
2.05 (s)
a
Assignment based on HSQC, HMBC and DEPT experiments; 600 MHz for 1H- and 150 MHz for 13C-NMR data.

P. Rudrapaul et al. / Phytochemistry Letters 9 (2014) 7–10
9
Fig. 2. Selected NOESY correlations of 1.
Fig. 3. Selected NOESY correlations of 2.
Compound (2) named meyanthic acid, was isolated as colorless
powder. Its molecular formula was determined as C32H50O7 from
its quasi-molecular ion peaks at m/z 547.3632 [M+H]+ (Calcd for
C32H51O7, 547.3635) and 569.3450 [M+Na]+ (Calcd for C32H50O7Na,
569.3454) and 13C-DEPT NMR spectral data. Its IR spectrum
showed the presence of hydroxyl (3467 cmꢀ1), ester (1732 cmꢀ1),
carboxyl (1692 cmꢀ1) and olefinic (1639 cmꢀ1) functionalities. The
1H-NMR spectral data (Table 1) showed five tertiary methyl
br s), 1.58 (2H, quint), 1.62 (2H, quint), 2.32 (2H, t, J = 6.8 Hz), 4.05
(2H, t, J = 6.8 Hz), attributable for a long linear aliphatic ester chain.
The 13C-NMR spectrum with DEPT experiments showed carbon
signals at dC 14.1 (CH3), 22.7 (CH2), 25.7 (CH2), 29.2 ꢀ 29.7 (CH2),
32.8 (CH2), 63.1 (CH2) and 174.6 (C) also corroborating its long
chain saturated fatty acid ester structure. The acid moiety was
assigned as pentadecanoic acid from the presence of a strong
fragment ion at m/z 243 in its FAB-MS due to McLafferty cleavage.
Its saponification with 1 N methanolic KOH under N2 atmosphere
gave myricyl alcohol (1-triacontanol), CH3(CH2)29OH and penta-
decanoic acid, CH3-(CH2)13-COOH (Dinda et al., 1999). Based on
this evidence, compound 3 was identified as myricyl-1-pentade-
canoate (Fig. 1). It was isolated for the first time from nature.
It may be noted that although we have isolated oleanolic acid
from this plant but failed to isolate its saponin. Possibly saponins
may be present in the polar butanol fraction, work on this fraction
is in process. The plants of family Rubiaceae contain both
oleananes and ursanes (Zhao et al., 1995, 1996).
singlets (
1.12, d, J = 6.0 Hz), one trisubstituted olefinic proton (
and one methine proton ( 3.06, s), characteristic of 19
hydroxyurs-12-ene type triterpenoid (Cheng and Cao, 1992).
The signals of two methine protons [ 5.06 (1H, br s) and 4.27 (1H,
ddd, J = 4.2, 4.0, 2.4 Hz)], a hydroxymethyl signal [ 3.74 (1H, d,
d
1.09, 1.11, 1.15, 1.43 and 1.67), one secondary methyl (
d
d
5.60, t-like)
d
a-
d
d
J = 10.2 Hz) and 4.22 (1H, d, J = 10.2 Hz)] and one acetoxymethyl
(3H, s, 2.05) in the 1H-NMR spectrum indicated the presence of two
hydroxyl groups and one acetoxy group in ring A (Kojima and
Ogura, 1989; Cheng and Cao, 1992). The location of the acetoxy
group at C-3 position was suggested by the downfield shift of H-3
(
d
5.06) and the location of the hydroxyl group at C-23 position was
Studies on the evaluation of bioefficacy of the isolated
chemicals in bone fracture, renal disorders and others will be
carried out after isolation of sufficient amount of the compounds
from these fractions of medium polarity as well as from butanol
fraction.
deduced by the chemical shifts of C-23 and C-24 (Table 1) (Mahato
and Kundu, 1994). The 13C-NMR spectral data (Table 1) recorded
32 carbon signals consisting of seven methyl, nine methylene,
seven methine and nine quaternary carbons. The presence of two
olefinic carbons (
73.0), one methylene carbon (
carbons ( 69.2, 78.6), a methyl carbon (
181.0) and carbons of an acetoxy group (
d
128.3 and 140.3), one quaternary carbon (
66.6), two hydroxymethine
14.7), a carboxyl carbon
21.7 and 170.2) also
d
d
3. Experimental
d
d
(
d
d
3.1. General experiment
corroborated its 3-acetoxy,2,19,23-trihydroxyurs-12-en-28-oic
acid-like structure for it (Zhou et al., 1992). The HMBC correlation
of H-18 with dC at 73.0 confirmed the location of one hydroxyl
group at C-19 position. Similarly, HMBC correlation of H-1 with dC
at 69.2 and of H-2 with dC at 78.6 and of H-3 with dC at 66.6
supported the position of hydroxyl group at C-2, acetoxy group at
C-3 and hydroxyl group at C-23. HMBC correlation of H-18 with dC
181.0 confirmed the location of carboxyl group at 28 position. The
signal of H-2 was observed as a doublet of double doublet with the
coupling constants of 4.2, 4.0 and 2.4 Hz, for one diequatorial and
Melting points were determined on a Ko¨fler block type
apparatus. The IR spectra were measured on a Perkin Elmer
FTIR-100 spectrometer. NMR spectra were measured on a Bruker
Avance DRX-600 and DRX-400 spectrometers operating at 600 and
400 MHz for 1H-NMR, and 150 and 100 MHz for 13C-NMR. EI, HR-
FAB and FAB-MS were recorded on
a JEOL JMS-303 mass
spectrometer. Optical rotation was measured on a Perkin Elmer
digital polarimeter. Column chromatography (CC) was performed
with silica gel (60–120 mesh, Merck, India) and TLC with silica gel
G (Merck, India).
two axial-equatorial couplings, characteristic for 2b, 3b-dihy-
droxyurs-12-en-28-oates (Kojima and Ogura, 1989). The NOESY
correlations between H-3 and H-23, H-2, H-1 supported the
3.2. Plant material
stereochemical assignment of 3
groups. Similarly, the NOESY relations between H-18 and H-29 and
H-20 confirmed the -configuration of hydroxyl group at C-19
position(Fig. 3). Thus, the structure of meyanthic acid 2 was
b-acetoxy, 2b and 23-dihydroxyl
The leaves of M. spinosa Roxb. were collected from Dharma-
nagar, North Tripura in June 2011. The plant was identified by Prof.
B.K. Datta, Taxonomist, Department of Botany, Tripura University.
A voucher specimen (TU/H/1535) was deposited in the laboratory
of B.K.D.
a
elucidated as 3b-acetoxy,2b,19a,23-trihydroxyurs-12-en-28-oic
acid (Fig.1).
Compound 3 was isolated as colorless amorphous powder. Its
molecular formula was assigned as C45H90O2 from the quasi-
molecular ion peak at m/z 663.7022 [M+H]+ (Calcd for C45H91O2,
663.7019) in HR-FAB-MS. Its IR spectrum revealed the presence of
an ester (1740 cmꢀ1) function. Its 1H-NMR spectrum showed
3.3. Extraction and isolation
Air-dried leaves (2 kg) of M. spinosa were extracted with
methanol by maceration at room temperature (6 L ꢁ 2, 6 d each
time) and the methanolic extract was concentrated in vacuo using a
proton signals at dH 0.84 (3H, t), 0.88 (3H, t, J = 6.6 Hz), 1.26 (76H,

10
P. Rudrapaul et al. / Phytochemistry Letters 9 (2014) 7–10
rotavapour to a residue (550 g). This crude residue was suspended
in water (80 mL) and partitioned against petroleum ether,
chloroform, ethyl acetate and n-butanol successively. The residue
from petroleum ether extract (20 g) on silica gel CC afforded
compounds 3 (30 mg), 6 (42 mg) and 7 (60 mg) on elution with
different ratios of petroleum ether–ethyl acetate mixture. Elution
of the column with petroleum ether–ethyl acetate (9:1) afforded a
solid residue purified on repeated silica gel CC using mixture of
petroleum ether–ethyl acetate (15:1, 12:1 and 10:1) to give 3, 5
and 6. The chloroform extract (80 g) on silica gel CC gave
compounds 1 (45 mg), 2 (20 mg), 4 (50 mg), 5 (100 mg) and 8
(60 mg). Elution of the column with CHCl3–EtOAc (9:1) mixture
afforded 5 and CHCl3–EtOAc (6:1) afforded 1, while CHCl3–EtOAc
(4:1) afforded a mixture of two compounds, purified on silica gel
CC eluted by CHCl3–EtOAc (5:1) and (4:1) mixtures to give 2 and 4,
respectively. The purification of the fractions eluted by CHCl3–
EtOAc (1:1) on silica gel CC using a gradient of CHCl3–EtOAc (3:1,
2:1 and 1:1) mixtures afforded 8. The fractionation of the ethyl
acetate extract on silica gel CC gave compounds 5 and 4 in fractions
eluted by CHCl3–EtOAc (4:1) and (3:1), respectively.
CH3(CH2)29-OH, mp 89 8C (lit. 88 8C, Weast, 1976) and pentade-
canoic acid (1.3 mg), mp 538C (lit. 53–54 8C, Weast, 1976).
Acknowledgments
The authors are grateful to Prof. S. Ray, Director, IICB, Kolkata for
spectral facilities and Prof. B.K. Datta, Department of Botany,
Tripura University for identification of plant material. This work
was financially supported by a grant (No. SR/S1/OC-75/2009) from
DST, New Delhi, India. One of the authors, P.R. is thankful to CSIR,
New Delhi, India for the award of Junior Research Fellowship
(Project no. 09/714(0013)/2011-EMR-1).
References
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Publishers, New Delhi, , pp. 64.
2
a
,3
amorphous powder; [
m/z: 491.3733 [M+H]+; IR (KBr)
a
,19
a
,24,28-Pentahydroxyurs-12-ene (1): Colorless
a
]
D
24 + 21.2 (c 0.03, MeOH); HR-FAB-MS
max cmꢀ1: 3436, 1636, 1565, 1468,
Dinda, B., Das, S.K., Hajra, A.K., Bhattacharya, A., De, K., Chel, G., Achari, B., 1999.
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n
1418, 1380, 1178, 1102, 800, 779; 1H and 13C-NMR data (C5D5N),
Table 1.
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xyursane-type triterpene glucosyl esters from the roots of Rubus suavissimus S.
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Gogoi, J., Sarma, P.K., 1997. Chemical investigation of Meyna laxiflora Robyns syn.
Vangueria spinosa Hook. II. Indian Drugs 34, 610–611.
3
b
-Acetoxy-2
b
,19
a
,23-trihydroxyurs-12-en-28-oic
acid
(= Meyanthic acid) (2): Colorless powder; mp 286 ꢀ 288 8C
(dec); [a]
25 + 10.6 (c 0.2, MeOH); FAB-MS m/z: 547 [M+H]+,
D
569 [M+Na]+; 1H and 13C-NMR data (C5D5N), Table 1.
Jung, H.A., Chung, H.Y., Jung, J.H., Choi, J.S., 2004. A new pentacyclic triterpenoid
glucoside from Prunus serrulata var. spontanea. Chem. Pharm. Bull. 52, 157–159.
Kojima, H., Ogura, H., 1989. Configurational studies on hydroxyl groups at C-2, 3 and
23 or 24 of oleanene and ursene-type triterpenes by NMR spectroscopy.
Phytochemistry 28, 1703–1710.
Mahato, S.B., Kundu, A.P., 1994. 13C-NMR Spectra of pentacyclic triterpenoids – a
compilation and some salient features. Phytochemistry 37, 1517–1575.
Maillard, M., Adewunmi, C.O., Hostettmann, K., 1992. A new triterpene glycoside
from the fruits of Tetrapleura tetraptera. Phytochemistry 31, 1321–1323.
Paulo, A., Jimeno, M.L., Gomes, E.T., Houghten, P.J., 2000. Steroidal alkaloids from
Cryptolepis obtuse. Phytochemistry 53, 417–422.
Pullaiah, T., 2002. Medicinal Plants in India. Regency Publications, New Delhi, pp. 1–4.
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C-531.
Yusuf, M., Begum, J., Yahab, M.A., Chowdhury, J.U., 2009. Medicinal Plants of
Bangladesh. BCSIR Laboratories, Bangladesh, pp. 435.
Myricyl pentadecanoate (3): Amorphous powder; mp 76–
78 8C; FAB-MS m/z (rel. int.): 663 [M+H]+ (33), 635 (2), 437 (3), 243
(4), 225 (5), 111 (12), 97 (21), 83 (41), 69 (6), 57 (100); IR (KBr)nmax
cmꢀ1: 1752, 1416, 1075; 1H-NMR (600 MHz, CDCl3):
d 0.84 (3H,
distorted t, Me), 0.88 (3H, t, J = 6.6 Hz, Me), 1.26 (ꢂ76H, br s,
CH2 ꢁ 38), 1.58 (2H, quin, CH2 adjacent to OCH2), 1.62 (2H, quin,
CH2), 2.32 (2H, t, J = 6.8 Hz, –CH2CO–), 4.05 (2H, t, J = 6.8 Hz, OCH2);
13C-NMR (150 MHz, CDCl3): 174.6 (C-1), 63.1 (C-10), 32.8 (C-2, 20),
d
29.2–29.7 (C-4–13, 40–280), 25.7 (C-3, 30), 22.7 (C-14, 290), 14.1 (C-
15, 300).
19
78 8C; FAB-MS m/z (rel. int.): 505 [M+H]+; 1H-NMR (600 MHz,
C5D5N): 4.27 (ddd, J = 10.0, 9.0, 4.2 Hz, H-2), 4.25 (d, J = 10.0, Hz,
a-Hydroxyasiatic acid (4): Colorless powder; mp 276–
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H-3), 5.60 (t-like, H-12), 3.06 (s, H-18), 3.74 (d, J = 10.2 Hz, H-23),
4.21 (d, J = 10.2 Hz, H-23), 1.0 (s, H3-24), 1.15 (s, H3-25), 1.11 (s, H3-
26), 1.41 (s, H3-27), 1.67 (s, H3-29), 1.12 (d, J = 6.6 Hz, H3-30). 13C-
NMR (150 MHz, C5D5N): similar to reported data (Gao et al., 1985).
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