Four new triterpenoids isolated from the resin of Garcinia hanburyi

Article abstract of DOI:10.1080/10286020.2013.875012

Four new triterpenoids, 2-O-acetyl-3-O-(4′-O-acetyl)-α-l- arabinopyranosylmaslinic acid (1), 2-O-acetyl-3-O-(3′-O-acetyl)-α-l- arabinopyranosylmaslinic acid (2), 2-O-acetyl-3-O-(3′,4′-O-diacetyl) -α-l-arabinopyranosylmaslinic acid (3), and 3-O-(3′-O-acety

Full text of DOI:10.1080/10286020.2013.875012

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Four new triterpenoids isolated from
the resin of Garcinia hanburyi
Hong-Min Wanga, Qun-Fang Liua, Yi-Wu Zhaobc, Shuang-Zhu Liua,
Zhen-Hua Chena, Ru-Jun Zhanga, Zhen-Zhong Wangbc, Wei Xiaobc
Wei-Min Zhaoa
&
a Department of Natural Products Chemistry, Shanghai Institute of
Materia Medica, Chinese Academy of Sciences, Shanghai, 201203,
China
b Jiangsu Kanion Pharmaceutical Co., Ltd., Lianyungang, 222001,
China
c State Key Laboratory of Pharmaceutical Process New-Tech for
Chinese Medicine, Lianyungang, 222001, China
Published online: 07 Jan 2014.
To cite this article: Hong-Min Wang, Qun-Fang Liu, Yi-Wu Zhao, Shuang-Zhu Liu, Zhen-Hua Chen,
Ru-Jun Zhang, Zhen-Zhong Wang, Wei Xiao & Wei-Min Zhao (2014) Four new triterpenoids isolated
from the resin of Garcinia hanburyi, Journal of Asian Natural Products Research, 16:1, 20-28, DOI:
10.1080/10286020.2013.875012
To link to this article: http://dx.doi.org/10.1080/10286020.2013.875012
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Journal of Asian Natural Products Research, 2014
Vol. 16, No. 1, 20–28, http://dx.doi.org/10.1080/10286020.2013.875012
Four new triterpenoids isolated from the resin of Garcinia hanburyi
Hong-Min Wanga, Qun-Fang Liua, Yi-Wu Zhaob,c, Shuang-Zhu Liua, Zhen-Hua Chena,
Ru-Jun Zhanga, Zhen-Zhong Wangb,c, Wei Xiaob,c* and Wei-Min Zhaoa*
aDepartment of Natural Products Chemistry, Shanghai Institute of Materia Medica, Chinese
Academy of Sciences, Shanghai 201203, China; Jiangsu Kanion Pharmaceutical Co., Ltd.,
b
c
Lianyungang 222001, China; State Key Laboratory of Pharmaceutical Process New-Tech for
Chinese Medicine, Lianyungang 222001, China
(Received 7 October 2013; final version received 10 December 2013)
Four new triterpenoids, 2-O-acetyl-3-O-(40-O-acetyl)-a-L-arabinopyranosylmaslinic
acid (1), 2-O-acetyl-3-O-(30-O-acetyl)-a-L-arabinopyranosylmaslinic acid (2), 2-O-
acetyl-3-O-(30,40-O-diacetyl)-a-L-arabinopyranosylmaslinic acid (3), and 3-O-(30-O-
acetyl)-a-L-arabinopyranosyloleanolic acid (4), together with six known triterpenoids,
3-O-(40-O-acetyl)-a-L-arabinopyranosyloleanolic acid (5), maslinic acid (6), 2-O-
acetylmaslinic acid (7), 3-O-acetylmaslinic acid (8), betulinic acid (9), and 2a-
hydroxy-3b-O-acetylbetulinic acid (10), were isolated from the EtOAc extract of
Garcinia hanburyi resin. Their structures were elucidated by analysis of the
spectroscopic data and chemical methods.
Keywords: Garcinia hanburyi; Guttiferae; triterpenoid; saponin
1. Introduction
C39H60O10 was established by HR-TOF-
MS (m/z 711.4089 [M þ Na]þ), implying
10 degrees of unsaturation. Its 1H (Table 1)
spectrum indicated the presence of one
triterpene aglycone [dH 1.10 (s, 3H), 1.08
(s, 3H); 1.03 (s, 3H); 0.92 (s, 3H); 0.91 (s,
3H); 0.90 (s, 3H); 0.73 (s, 3H)], one sugar
moiety (dH 3.51–4.33), and two acetyl
methyl groups [dH 2.11 (s, 3H), 1.99 (s,
3H)]. The 13C NMR (Table 1) spectrum of
1 exhibited 39 carbon signals, of which
nine carbon signals were consistent with
the presence of an arabinopyranosyl
moiety [dC 104.5 (d), 72.6 (d), 71.5 (d),
70.4 (d), 63.5 (t)] [9–12] and two acetyl
methyl groups [dC 171.1 (s), 171.1 (s),
21.6 (q), 21.2 (q)]. The remaining 30
carbon signals strongly resembled those of
maslinic acid, except for the downfield
shift of C-3 (from dC 83.5 to 89.2), due to
the glycosylation of 3-hydroxy group in 1.
Moreover, the HMBC correlations from
H-10 (dH 4.33) to C-3 confirmed the
Gamboge, the pulverized golden color resin
collected primarily from Garcinia hanburyi
(Guttiferae), and to a lesser extent from
G. Morella, has a particularly long and rich
history in the arts and sciences [1–4]. It has
been used as dye and folk medicine for the
treatment of infected wound and tumors
[1–4]. Previous phytochemical investi-
gations of gamboge have mainly resulted
in the identification of caged polypreny-
lated xanthones [1–8]. In this study, 10
triterpenoids including four new triter-
penoids (1–4) and six known ones (5–10)
were isolated from the resin of G. hanburyi
(Figure 1). The structures of these com-
pounds were elucidated by spectroscopic
analysis and chemical methods.
2. Results and discussion
Compound 1 was obtained as white
amorphous powder. Its molecular formula
*Corresponding authors. Email: wmzhao@mail.shcnc.ac.cn; fang-yu520@126.com
q 2014 Taylor & Francis

Journal of Asian Natural Products Research
21
29
20
12
H
COOH
1
H
COOH
R1
O
R1
R2
10
R2
H
H
27
3
4'
R3
O
1'
H
23
H
OH
6 R1 = R2 = OH
7
R1 = OAc, R2 = OH
8 R1 = OH, R2 = OAc
1
1a
2
R1 = R2 = OAc, R3 = OH
R = R = R = OH
R1 = R3 = OAc, R2 = OH
R1 = R2 = R3 = OAc
1
2
3
3
R1 = H, R2 = OH, R3 = OAc
R = H, R = R = OH
R1 = H, R2 = OAc, R3 = OH
4
4a
5
H
1
2
3
COOH
H
R1
R2
H
H
9
R1 = H, R2 = OH
10
R1 = OH, R2 = OAc
Figure 1. Structures of compounds 1–10, 1a, and 4a.
connection between the maslinic acid
moiety and the arabinopyranosyl moiety
(Figure 2). Furthermore, the locations of
two acetyl groups at 2- and 40-hydroxy
groups were established based on the
HMBC correlations from H-2 (dH 5.14) to
an acetyl carbonyl carbon (dC 171.1), and
from H-40 (dH 5.08) to the other acetyl
carbonyl carbon (dC 171.1) (Figure 2). The
arabinopyranosyl unit in 1 was determined
to connect to the aglycone in a-configur-
comparison of their spectral data with
those in the literature and co-TLC [15,16].
Derivation of the arabinose hydrolysate
from 1 with L-cysteine methyl ester gave a
GC-MS peak at 16.09 min, same as that of
a standard L-arabinose derivative. Accord-
ingly, the structure of 1 was established as
2-O-acetyl-3-O-(40-O-acetyl)-a-L-arabi-
nopyranosylmaslinic acid.
Compound 2, a white amorphous
powder, possessed the same molecular
formula as 1, based on HR-TOF-MS (m/z
711.4090 [M þ Na]þ). Its 1H and 13C
NMR data (Table 1) were very similar to
those of 1 and displayed signals belonging
to one maslinic acid moiety, one arabino-
pyranosyl unit, and two acetyl groups. The
HMBC correlations from H-10 (dH 4.89) to
C-3 (dC 89.9) established the connection
between the maslinic acid moiety and the
arabinopyranosyl moiety (Figure 2).
Assignment of two O-acetyl groups at C-
2 and C-30 was evidenced from the HMBC
ation in 4C1 form on the basis of the
3
0
0
JH-1,H-2 value (6.4 Hz) and the NOE
correlations between H-10 and H-30 and
between H-10 and H-50 in the NOESY
experiment (Figure 2) [9–12]. The absol-
ute configuration of the arabinopyranosyl
unit in 1 was found to belong to L series by
GC-MS analysis of its peracetylated
thiazolidine derivative [13,14]. Hydrolysis
of 1 with 10% KOH yielded 1a, which was
further hydrolyzed with acid to give
maslinic acid and arabinose identified by

22
H.-M. Wang et al.
d
d
d
d
d
d
d
d

Journal of Asian Natural Products Research
23
d
d
d
d
d
d
d
d

24
H.-M. Wang et al.
29
29
20
20
18
12
12
18
O
O
H
COOH
O
COOH
H
1
1
O
O
O
10
10
O
H
H
3
3
O
1'
OH
27
27
OH
4'
HO
O
O
4'
H
23
H
23
1'
O
OH
O
1
2
29
20
18
12
O
COSY
H
COOH
1
O
O
10
HMBC
NOE
H
3
27
O
4'
O
1'
O
H
O
23
OH
O
3
1
Figure 2. Selected H–1H COSY, HMBC, and NOE correlations of compounds 1–3.
correlations between H-2 (dH 5.58) and
one acetyl carbonyl carbon at dC 171.2 and
between H-30 (dH 5.49) and the other
acetyl carbonyl carbon at dC 171.3
(Figure 2). In addition, hydrolysis of 2
with 10% KOH yielded 1a, which
confirmed that the absolute configuration
of arabinopyranosyl moiety in 2 was
L. Thus, the structure of 2 was character-
ized as 2-O-acetyl-3-O-(30-O-acetyl)-a-L-
arabinopyranosylmaslinic acid.
diacetyl)-a-L-arabinopyranosylmaslinic
acid.
Compound 4 was obtained as white
amorphous powder and exhibited a quasi-
molecular ion peak at m/z 653.4065
[M þ Na]þ, corresponding to the molecu-
lar formula C37H58O8. The 1H and 13C
NMR spectra of 4 were very close to those
of 5 [17] and showed one oleanolic acid
moiety, one arabinopyranosyl moiety, and
one acetyl group. Hydrolysis of 4 with
10% KOH yielded 4a [17], suggesting that
the structural difference between 4 and 5
laid in the location of the acetyl group. The
connectivity of the acetyl group at 30-
hydroxy of the sugar residue was deter-
mined by the downfield chemical shift of
C-30 and upfield chemical shift of C-20 and
C-40 compared with those of 4a. Accord-
ingly, the structure of 4 was determined as
3-O-(30-O-acetyl)-a-L-arabinopyranosylo-
leanolic acid.
Compound 3, a white amorphous
powder, gave the molecular formula as
C41H62O11 according to its HR-TOF-MS
(m/z 753.4197 [M þ Na]þ), 42 Dalton
higher than that of 2. Comparison of its 1H
and 13C NMR spectra with those of 2
revealed that the only difference was the
presence of an additional acetyl group. The
downfield shift of H-40 (dH 5.66) assigned
by 1H–1H COSY spectrum of 3 (Figure 2)
indicated that the 40-hydroxy group was
acetylated in 3. In addition, hydrolysis of 3
with 10% KOH yielded 1a, suggesting that
the arabinopyranosyl moiety in 3 belongs
to L series. Therefore, compound 3 was
characterized as 2-O-acetyl-3-O-(30,40-O-
Six known compounds, namely 3-O-
(40-O-acetyl)-a-L-arabinopyranosylolea-
nolic acid (5) [17], maslinic acid (6) [15],
2-O-acetylmaslinic acid (7) [18], 3-O-
acetylmaslinic acid (8) [19], betulinic acid

Journal of Asian Natural Products Research
25
(9) [20], and 2a-hydroxy-3b-O-acetylbe-
tulinic acid (10) [17], were also isolated
from the EtOAc extract of G. hanburyi
Shimadu GC-MS-QP2010E (Shimadzu,
Kyoto, Japan) using a RXIw-5 ms capillary
column (30 m £ 0.25 mm, 0.25 mm)
(RESTEK, Bellefonte, PA, USA).
1
resin. Their H and 13C NMR data were
identical to those reported in the literature.
To evaluate the biological activities of
the four new compounds, compounds 1–4
were tested in an array of bioassay (see
Bioassay section in the Supporting infor-
mation). Unfortunately, compounds 1–4
showed no cytotoxicity against NCI-H460
cell lines at 1 mM and no inhibitory effect
against KDR protein tyrosine kinase at 10
mM. They were also inactive in the T-cell
immunomodulatory bioassay at 1, 10, and
100 mM, respectively.
3.2 Plant material
The resin of G. hanburyi (Guttiferae)
collected in Vietnam was purchased from
the Bozhou Herbal Medicine Market,
Anhui Province, in December 2010, and
was authenticated by Professor Qinan Wu
of Nanjing University of Traditional
Chinese Medicine. A specimen (No.
SIMM20101228) is deposited at Herbar-
ium of Shanghai Institute of Materia
Medica, Chinese Academy of Sciences.
3. Experimental
3.1 General experimental procedures
3.3 Extraction and isolation
Optical rotations were measured with a
Perkin-Elmer 341 polarimeter (Perki-
nElmer, Waltham, MA, USA). IR spectra
were recorded using a Perkin-Elmer 577
spectrometer (PerkinElmer, Waltham,
MA, USA). NMR spectra were recorded
Dried and ground resin (1.5 kg) was
successively extracted with EtOAc (9 l)
three times at room temperature. After
combination and removal of the solvent
under vacuum, the residue (392 g) was
subjected to column chromatography over
silica gel (200–300 mesh) and eluted with
a petroleum ether–EtOAc gradient (from
50:1 to 0:1, v/v) to yield 11 fractions (Fr.
A–Fr.K). Test with vanillin–sulfuric acid
solution on TLC plates suggested the
presence of triterpenoids in Fr.G, Fr.I, and
Fr.K. The three fractions were analyzed by
LC-PDA-MS, revealing the existence of
four compounds with molecular weights
688, 688, 730, and 630 not reported in the
genus Garcinia in the Reaxys database.
Fr.G (7 g) was subjected to RP-18
column chromatography eluted with a 70–
100% aqueous MeOH gradient to give six
subfractions (Fr.G1–Fr.G6). Fr.G2 (0.9 g)
was further subjected to repeated chroma-
tography over silica gel (300–400 mesh)
to afford compounds 7 (123 mg), 8
(243 mg), 9 (58 mg), and 10 (31 mg). Fr.I
(12 g) was separated by column chroma-
tography over silica gel (300–400 mesh)
eluted with a CHCl3–MeOH gradient
(from 50:1 to 10:1, v/v), affording four
on
a
Varian-MERCURY Plus-300
(Varian, Palo Alto, CA, USA), Bruker
AM 400 (Bruker, Ettlingen, Germany),
Varian-MERCURY Plus-400 (Varian,
Palo Alto, CA, USA), Bruker Advance
III 500 (Bruker, Ettlingen, Germany), or
Bruker Advance III 600 (Bruker, Ettlin-
gen, Germany) spectrometer with TMS as
internal standard. ESI-MS and HR-ESI-
MS data were measured using a Shimadzu
LC-MS-2020 (Shimadzu, Kyoto, Japan)
and a Bruker Daltonics microTOF QII
mass spectrometer (Bruker, Ettlingen,
Germany), respectively. Column chroma-
tographic separations were conducted by
using silica gel H60 (3002400 mesh;
Qingdao Haiyang Chemical Co., Ltd.,
Qingdao, China) and Sephadex LH-20
(Pharmacia Biotech AB, Uppsala, Swe-
den) as packing materials. HSGF254 silica
gel TLC plates (Yantai Chemical Indus-
trial Institute, Yantai, China) were used for
analysis. GC analysis was carried out on a

26
H.-M. Wang et al.
1250, 1084, 1049 cm21
subfractions (Fr.I1–Fr.I4). Fr.I2 (1.5 g)
was chromatographed over silica gel
(300–400 mesh) eluted with CHCl3–
MeOH (40:1, v/v), followed by RP-18
column chromatography eluted with a 50–
75% aqueous CH3CN gradient to give
compounds 3 (198 mg), 4 (43 mg), and 5
(135 mg). Fr.I4 (1.5 g) was subjected to
column chromatography over silica gel
(300–400 mesh) and Sephadex LH-20 gel
(95% EtOH), affording compound 6
(35 mg). Fr.K (2.5 g) was subjected
repeatedly to chromatography over silica
gel (300–400 mesh) eluted with CHCl3–
MeOH (30:1, v/v) to give compounds 1
(98 mg) and 2 (18 mg).
;
1H NMR
(300 MHz, CDCl3): dH 5.25 (br s, 1H),
5.19 (br s, 1H), 5.13 (m, 1H, H-2), 4.93
(dd, J ¼ 9.9, 3.4 Hz, 1H, H-30), 4.36 (d,
J ¼ 7.2 Hz, 1H, H-10), 3.93 (d,
J ¼ 12.5 Hz, 1H, H-50a), 3.80 (dd,
J ¼ 9.9, 7.2 Hz, 1H, H-20), 3.57 (d,
J ¼ 12.5 Hz, 1H, H-50b), 3.27 (d,
J ¼ 9.8 Hz, 1H, H-3), 2.80 (dd, J ¼ 13.0,
3.5 Hz, 1H, H-18), 2.09 (s, 3H, OAc), 2.06
(s, 3H, OAc), 2.00 (s, 3H, OAc), 1.10 (s,
3H), 1.07 (s, 3H), 1.02 (s, 3H), 0.91 (s,
1
3H), 0.89 (s, 6H), 0.72 (s, 3H). H NMR
(500 MHz, C5D5N): dH 5.66 (br s, 1H, H-
40), 5.58 (m, 1H, H-2), 5.55 (br s, 1H, H-
30), 5.51 (s, 1H, H-12), 4.85 (d, J ¼ 6.8 Hz,
1H, H-10), 4.36 (dd, J ¼ 8.0, 6.8 Hz, 1H,
H-20), 4.17 (d, J ¼ 12.6 Hz, 1H, H-50a),
3.86 (d, J ¼ 12.6 Hz, 1H, H-50b), 3.52 (d,
J ¼ 9.8 Hz, 1H, H-3), 3.32 (d, J ¼ 13.0,
2.9 Hz, 1H, H-18), 2.27 (s, 3H, OAc), 2.01
(s, 6H, OAc), 1.31 (m, 6H), 1.02 (s, 3H),
0.99 (s, 3H), 0.97 (s, 9H). For 13C NMR
(125 MHz, C5D5N) spectroscopic data, see
Table 1; ESI-MS: m/z 753 [M þ Na]þ;
HR-TOF-MS: m/z 753.4197 [M þ Na]þ
(calcd for C41H62O11Na, 753.4190).
3.3.1 2-O-Acetyl-3-O-(30-O-acetyl)-a-L-
arabinopyranosylmaslinic acid (1)
22
White amorphous powder; ½aꢀD þ 35.0
(c ¼ 0.20, MeOH); IR(KBr): nmax 3373,
2945, 2908, 1738, 1720, 1695, 1641, 1454,
1367, 1252, 1095, 1034 cm21; for 1H
NMR (300 MHz, CDCl3) and 13C NMR
(100 MHz, CDCl3) spectroscopic data, see
Table 1. ESI-MS: m/z 711 [M þ Na]þ;
HR-TOF-MS m/z 711.4089 [M þ Na]þ
(calcd for C39H60O10Na, 711.4084).
3.3.4 3-O-(30-O-Acetyl)-a-L-
arabinopyranosyloleanolic acid (4)
22
3.3.2 2-O-Acetyl-3-O-(40-O-acetyl)-a-L-
arabinopyranosylmaslinic acid (2)
22
White amorphous powder; ½aꢀD þ 43.1
(c ¼ 0.20, MeOH); IR(KBr): nmax 3340,
White amorphous powder; ½aꢀD þ 31.0
2941, 2877, 2858, 1724, 1705, 1649, 1464,
1
(c ¼ 0.21, MeOH); IR(KBr): nmax 3429,
2947, 1720, 1695, 1642, 1365, 1252, 1142,
1080, 1047 cm21; for 1H NMR (300 MHz,
C5D5N) and 13C NMR (100 MHz, C5D5N)
spectroscopic data, see Table 1. ESI-MS:
m/z 711 [M þ Na]þ; HR-TOF-MS m/z
711.4090 [M þ Na]þ (calcd for
C39H60O10Na, 711.4084).
1377, 1267, 1086, 1022 cm21; H NMR
(300 MHz, CDCl3): dH 5.27 (br s, 1H, H-
12), 4.87 (d, J ¼ 9.7 Hz, 1H, H-30), 4.32
(d, J ¼ 7.2 Hz, 1H, H-10), 4.02 (br s, 1H,
H-40), 3.97 (d, J ¼ 13.2 Hz, 1H, H-50a),
3.85 (dd, J ¼ 9.7, 7.2 Hz, 1H, H-20), 3.58
(d, J ¼ 13.2 Hz, 1H, H-50b), 3.17 (dd,
J ¼ 10.9, 4.2 Hz, 1H, H-3), 2.81 (d,
J ¼ 12.3 Hz, 1H, H-18), 2.16 (s, 3H,
OAc), 1.12 (s, 3H), 0.99 (s, 3H), 0.92 (s,
6H), 0.90 (s, 3H), 0.81 (s, 3H), 0.74 (s,
3H). 1H NMR (300 MHz, C5D5N): dH 5.51
(m, 2H, H-12 and H-30), 4.88 (d,
J ¼ 6.6 Hz, 1H, H-10), 4.64 (t,
J ¼ 7.8 Hz, 1H, H-20), 4.60 (br s, 1H, H-
3.3.3 2-O-Acetyl-3-O-(30,40-O-diacetyl)-
a-L-arabinopyranosylmaslinic acid (3)
26
White amorphous powder; ½aꢀD þ 16.4
(c ¼ 0.20, MeOH); IR(KBr): nmax 3435,
2945, 2864, 1732, 1693, 1641, 1464, 1369,

Journal of Asian Natural Products Research
27
40), 4.34 (dd, J ¼ 12.2, 3.4 Hz, 1H, H-50a),
3.91 (d, J ¼ 11.4 Hz, 1H, H-50b), 3.35 (m,
2H, H-3 and H-18), 1.97 (s, 3H, OAc),
1.32 (s, 6H), 1.03 (s, 3H), 1.02 (s, 3H),
0.98 (s, 3H), 0.95 (s, 3H), 0.87 (s, 3H). For
13C NMR (100 MHz, C5D5N) spectro-
scopic data, see Table 1; ESI-MS: m/z 653
[M þ Na]þ; HR-TOF-MS m/z 653.4065
[M þ Na]þ (calcd for C37H58O8Na,
653.4029).
NMR (300 MHz, C5D5N): dH 5.41 (br s,
1H, H-12), 4.71 (d, J ¼ 7.6 Hz, 1H, H-10),
4.48 (dd, J ¼ 8.5, 7.6 Hz, 1H, H-20), 4.26
(m, 2H, H-30 and H-40), 4.12 (dd, J ¼ 11.4,
3.3 Hz, 1H, H-50a), 3.99 (dt, J ¼ 11.4,
3.3 Hz, 1H, H-2), 3.84 (d, J ¼ 11.4 Hz, 1H,
H-50b), 3.24 (m, 2H, H-3 and H-18), 2.21
(dd, J ¼ 12.5, 4.4 Hz, 1H), 1.36 (s, 3H),
1.24 (s, 3H), 0.97 (s, 6H), 0.94 (s, 3H),
0.91 (s, 3H), 0.86 (s, 3H); for 13C NMR
(100 MHz, C5D5N) spectroscopic data, see
Table 1.
3.4 Hydrolysis and derivatization
Compounds 1–4 (10 mg) were hydrolyzed
with 10% KOH (2 ml) at room temperature
for 2 h. The reaction products of 1–3 were
diluted with water and extracted with n-
BuOH (3 £ 5 ml) to yield compound 1a
(6 mg). The reaction product of 4 was
treated in the same way to give 4a (5 mg).
The structure of 1a was elucidated by
detailed spectroscopic analysis and chemi-
cal methods, while 4a was identified by
comparison of NMR data with those in the
literature [17].
3.5 GC analysis
The peracetylated derivative was analyzed
by GC under the following conditions:
Column, RXIw-5 ms, 30 m £ 25 mm, 0.25
mm; carrier gas: He (17.8 ml/min), injec-
tion temperature at 2808C, detection
temperature at 2008C, column temperature
at 1008C, initial temperature was main-
tained at 1008C for 5 min, and then raised
to 2808C at the rate of 208C/min and
maintained at 2808C for 10 min; injection
volume: 1.0 ml, split ratio: 1/10. The
absolute configuration of sugar moiety was
determined by comparing their retention
times with the derivatives of authentic D-
and L-arabinose prepared in the same
manner [13,14]. The derivative of arabi-
nose hydrolyzed from compound 1a gave a
peak with retention time of 16.09 min,
while the derivatives of standard D- and
L-arabinose had GC retention time of
16.36 and 16.10 min, respectively.
Compound 1a (5 mg) was refluxed
with 1 M HCl (2 ml) for 2 h under constant
stirring. The reaction product was diluted
with water and extracted with EtOAc (3
£ 5 ml). The aqueous layer and EtOAc
layer were lyophilized to dryness, respect-
ively, to give arabinose and maslinic acid
which was identified by comparison with
NMR data in the literature and co-TLC
with authentic sample [15,16]. The arabi-
nose hydrolyzed from 1a was dissolved in
dry pyridine (1 ml), and L-cysteine methyl
ester hydrochloride (3 mg) was added. The
mixture was stirred at 608C for 2 h, 150 ml
of Ac2O was added, and stirred at 908C for
another 1 h. The reaction product was
evaporated to dryness to give methyl 3-
acetyl-2R-(L-arabino-10,20,30,40-tetraace-
toxybutyl)-thiazolidine-4R-carbonylate
[13].
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