全 文 ：第 47卷 第 6期
2011年 12月
兰 州 大 学 学 报（自然科学版）
Journal of Lanzhou University (Natural Sciences)
Vol. 47 No. 6
Dec. 2011
Articlcal ID: 0455-2059(2011)06-0120-07
Lupane-triterpenoids from the methanol extracts of
leaves of Acanthopanax gracilistylus W.W. Smith
ZOU Qin-peng 1, LIU Xiang-qian 1,2, Lee Hyeong-kyu 3, OH O-jin 4
1. School of Chemistry and Chemical Engineering, Central South University, Changsha 410083, China
2. School of Pharmacy, Hunan University of Chinese Medicine, Changsha 410208, China;
3. Korea Research Institute of Bioscience and Biotechnology, Chungbuk 363-883, Republic of Korea;
4. Department of Herbal Medicines Management, Catholic Sangji College, Gyeongsangbuk-do 760-711,
Republic of Korea
Abstract: Nine lupane-triterpeniods were isolated from hot methanol extract of the leaves of Acanthopanax
gracilistylus W.W. Smith. Based on the chemical-physical properties and spectroscopic data including HR-MS,
1D and 2D-NMR, their chemical structures were determined as impressic acid(1), 3α, 11α-dihydroxy-23-
oxo-lup-20(29)-en-28-oic acid(2), 3α, 11α, 23-trihydroxy-lup-20(29)-en-28-oic acid(3), acankoreagenin(4),
3-O-β-D-glucopyranosyl 3α, 11α-dihydroxylup-20(29)-en-28-oic acid(5), acantrifoside A(6), acankoreoside
D(7), acankoreoside B(8), acankoreoside A(9), respectively. To our best knowledge, compounds 1−3 and
compound 5 were obtained for the first time from this plant.
Key words: Acanthopanax gracilistylus; lupane-triterpenoids; araliaceae; acankoreagenin; acankoreoside A
CLC number: R284.1 Document code: A
细柱五加叶甲醇提取物中的羽扇豆烷型三萜成分
邹亲朋 1, 刘向前 1,2, 李炯奎 3, 吴旿真 4
1. 中南大学 化学化工学院, 长沙 410083
2. 湖南中医药大学 药学院, 长沙 410208
3. 韩国生命工学院,韩国 首尔 363-883
4. Catholic上智大学传统药材管理科,韩国 庆尚北道 760-711
摘 要: 从细柱五加叶中分离得到了四个羽扇豆烷型三萜苷元及五个对应的苷类化合物.根据其理化性质和
波谱数据,鉴定出其化学结构分别为 impressic acid (1), 3α, 11α-dihydroxy-23-oxo-lup-20(29)-en-28-oic acid(2),
3α, 11α, 23-trihydroxy-lup-20(29)-en-28-oic acid(3),五加苷元 (acankoreagenin, 4), 3-O-β-D-glucopyranosyl 3α,
11α-dihydroxylup-20(29)-en-28-oic acid(5), acantrifoside A(6), acankoreoside D(7), acankoreoside B(8)和五加
苷 (acankoreoside A, 9), 其中化合物1−3和化合物5是首次从细柱五加中分离得到.
关键词: 细柱五加; 羽扇豆烷型三萜;五加属; 五加苷元; 五加苷
中图分类号: R284.1 文献标识码: A
Acanthopanax gracilistylus W.W. Smith (AGS) is
a deciduous shrub of the Araliaceae family which
is widely distributed in China. Its dried roots
and stem barks, known as Acanthopanacis Cortex,
are used as medicine for the treatment of rheuma-
tism, paralysis, arthritis, sinew, bone pains and
Received date: 2011-06-04
Found term: Supported by Hunan Provincial Natural Science Foundation of China(11JJ2042); Scientific Research Foundation of
Hunan University of Chinese Medicine (48010101-37); Graduate Degree Thesis Innovation Foundation of Central
South University(2011ssxt080)
Biography:LIU Xiang-qian(1967-), male, born in Changsha, Hunan Province, professor, doctor, e-mail: lxq0001cn@163.com,
majoring in active components from natural plants, correspond author.
DOI:10.13885/j.issn.0455-2059.2011.06.012
No. 6
ZOU Qin-peng, et al: Lupane-triterpenoids from the methanol extracts of
leaves of Acanthopanax gracilistylus W.W. Smith 121
as a tonic in traditional Chinese medicine [1−2].
However, little is known about their mechanism
and chemical nature. It is further known in re-
cent years that the leaves of this plant contain
diterpenoids, lignans, triterpenoids, polyacetylenes,
phenylpropanoids, and flavo-noids [2−8]. Previous
pharmacological study on Acanthopanax gracilistylus
reported anti-inflammatory [9] and suppressive effect
on human lymphocytes [10]. Recently, the research of
Zhang, et al [11] indicated that acankoreagenin from
AGS could significantly attenuate the release of high
mobility group box chromosomal protein 1 (HMG-
B 1) and suggests this component as a candidate
therapy for fulminant hepatitis. In our previous pa-
pers, several studies on the triterpenoids constituents
of AGS have been reported [3−7, 12].
In this manuscript, as a result of an on-going
search for the phytochemical constituents in Acan-
thopanax genus, we report on the isolation and
structural determination of four lupane-triterpenes
and their five glycosides from the leaves of AGS,
which were identified as impressic acid(1), 3α, 11α-
dihydroxy-23-oxo-lup-20(29)-en-28-oic acid(2), 3α,
11α, 23-trihydroxy-lup-20(29)-en-28-oic acid(3), a-
cankoreagenin(4), 3-O-β-D-glucopyranosyl 3α, 11α-
dihydroxylup-20(29)-en-28-oic acid(5), acantrifoside
A(6), acankoreoside D(7), acankoreoside B(8), and
acankoreoside A(9) by comparing their spectrosco-
pic data with those previously reported. A hot
MeOH extract of the leaves of AGS was suspended
in water and then partitioned with petroleum ether
(60∼90 ◦C), ethyl acetate, and n-butyl alcohol suc-
cessively. Further separation of each fraction with a
combination of silica gel, MDS, and Sephadex LH-20
columns gives compounds 1−9.
1 Experimental
1.1 General experimental procedures
Melting points (uncorrected) were measured us-
ing a Boetius micromelting point apparatus. Opti-
cal rotations were determined on a Jasco DIP-1000
KUY polarimeter (l=0.5). NMR spectra were mea-
sured in pyridine-d5 on a JEOLα-500 spectrometer
and chemical shifts were relative to tetramethylsi-
lane. Column chromatography was carried out on
silica gel 60(0.040∼0.063 mm, Merck), Sephadex LH-
20(Pharmacia Biotech Co) and MDS. TLC was per-
formed on a precoated silica gel G(Merck) and RP-
18F254S(Merck) plates.
1.2 Plant material
The leaves of AGS were collected in Changsha,
Hunan province of China, in September 2009 and
were botanically identified by Prof. Yook Chang-soo,
Kyung Hee University, Korea.
1.3 Extraction and isolation
The dried leaves of the plant (1 100 g) were ex-
tracted three times with hot MeOH. The combined
solution was evaporated under reduced pressure to
give an MeOH extract (157 g), which was suspend-
ed in water and then partitioned with petroleum
ether, ethyl acetate, and n-butyl alcohol, succes-
sively giving petroleum ether (7 g), ethyl acetate
(65 g), and n-butyl alcohol (77.1 g) extracts after
removal of the solvent in vacuo. Part of the residue
of ethyl acetate fraction (30 g) was decolorized on
a silica gel column by using gradient elution with
V(CH3Cl)
:V(MeOH) = 25 : 1 → 10 : 1, and then sub-
sequently chromatographed on a silica gel column
(∅ 56 mm×150 mm) by using gradient elution with
V(CH3Cl)
:V(MeOH) = 25 : 1→ 10 : 1 to give four frac-
tions. Fraction 1 was chromatographed on a silica
gel column using V(CH3Cl) :V(MeOH) = 25 : 1 to ob-
tain compound 1. Fraction 2 was chromatographed
on silica gel and MDS column again to give com-
pounds 2−4. Fraction 3 was chromatographed on
Sephadex LH-20 and purified by silica gel column
using V(CH3Cl) :V(MeOH)=25 :1 to give compound 3
and 4. Part of the residue of ethyl acetate fraction
(4 g) was chromatographed on a silica gel column
(∅ 25 mm×100 mm) using V(CH3Cl) :V(MeOH)= 10 : 1
to give compound 5. The residue of n-butyl alco-
hol fraction (2 g) was chromatographed on a MDS
column (∅ 30 mm×150 mm) using V(CH3Cl) :V(MeOH)
=5 : 5 to give compounds 6−9.
1.3.1 Impressic acid (1)
White crystal, m.p. 232∼234 ◦C(from dil. MeO-
H); negative ESI-MS m/z: 471.35 [M-H]−; 1H-
NMR(400 MHz, pyrindine-d5) δ: 3.64(1H, brs, H-
3), 2.21(1H, m, H-5), 1.88(1H, m, H-9), 4.25(1H,
122 Journal of Lanzhou University (Natural Sciences) Vol. 47
dt, J =2.5 Hz, 10.5 Hz, H-11), 2.26(1H, m, H-13),
1.76(1H, m, H-18), 3.53(1H, m, H-19), 1.26(6H, s,
H-23 and H-25), 0.97(3H, s, H-24), 1.15(3H, s, H-
26), 1.02(3H, s, H-27), 4.66(1H, brs, H-29a), 4.88(1H,
brs, H-29b), and 1.71(3H, brs, H-30); 13C NMR
(125 MHz, pyrindine-d5): see Table 1.
triterpenes R1 R2 R3 R4
1 OH CH3 OH H
2 OH CHO OH H
3 OH CH2OH OH H
4 OH COOH H H
glycosides R1 R2 R3 R4
5 -O − β-D-glc CH3 OH H
6 OH CH3 OH S
7 OH CHO OH S
8 OH CH2OH OH S
9 OH COOH H S
Figure 1 Structures of compounds 1−9
1.3.2 3α, 11α-dihydroxy-23-oxo-lup-20(29)-en-28-
oic acid (2)
White crystal, m.p. 189∼191◦C(from dil. MeOH);
negative ESI-MS m/z: 485.33[M-H]−; 1H NMR
(400 MHz, pyrindine-d5) δ: 4.02(1H, brs, H-3), 2.58
(1H, d, J =12.4 Hz, H-5), 4.26(1H, dt, J =5.2, 16 Hz,
H-11), 1.75(1H, m, H-18), 3.53(1H, m, H-19), 10.09
(1H, s, H-23), 1.18(3H, s, H-24), 1.25(3H, s, H-25),
1.13(3H, s, H-26), 1.05(3H, s, H-27), 4.66(1H, brs,
H-29a), 4.88(1H, brs, H-29b), and 1.71(3H, s, H-30);
13C NMR(125 MHz, pyrindine-d5): see Table 1.
1.3.3 3α, 11α, 23-trihydroxy-lup-20(29)-en-28-oic
acid (3)
White crystal, m.p. 183∼184 ◦C(from dil. MeOH);
negative ESI-MS m/z: 487.34 [M-H]−; 1H NMR
(400 MHz, pyrindine-d5) δ: 3.92(1H, brs, H-3), 2.21
(1H, m, H-5), 1.99(1H, d, J =10.4 Hz H-9), 4.27
(1H, dt, J =2.5, 10.5 Hz, H-11), 2.95(1H, m, H-
13), 1.76(1H, m, H-18), 3.53(1H, m, H-19), 3.70(1H,
d, H-23a), 3.89(1H, d, H-23b), 0.84(1H, s, H-24),
1.29 (3H, s, H-25), 1.17(3H, s, H-26), 1.05(3H, s, H-
27), 4.66(1H, brs, H-29a), 4.88(1H, brs, H-29b), and
1.71(3H, brs, H-30); 13C NMR(125 MHz, pyrindine-
d5): see Table 1.
1.3.4 Acankoreagenin (4)
White crystal, m.p. 232∼234 ◦C(from dil. MeOH);
[α]23
D
= −3.1(EtOH, c 0.5); HR-MS m/z: 486.33[M]+;
1H-NMR(400 MHz, pyrindine-d5) δ: 4.27(1H, brs, H-
3), 2.56(1H, m, H-5), 1.67(1H, m, H-9), 1.84(1H, m,
H-11), 2.75(1H, m, H-13), 1.71(1H, m, H-18), 3.53
(1H, m, H-19), 1.43(3H, s, H-24), 0.93 (6H, s, H-25
and H-27), 1.11(3H, s, H-26), 4.78(1H, brs, H-29a),
4.93(1H, brs, H-29b), and 1.76(3H, brs, H-30); 13C
NMR(125 MHz, pyrindine-d5): see Table 1.
1.3.5 3-O-β-D-glucopyranosyl 3α, 11α-dihy-droxy-
lup-20(29)-en-28-oic acid (5)
White amorphous solid, m.p. 273∼275 ◦C(from
dil. MeOH); negative ESI-MS m/z: 633.40 [M-H]−;
1H-NMR (400 MHz, pyrindine-d5) δ: 3.73(1H, brs,
H-3), 1.70(1H, m, H-5), 1.77(1H, d, H-9), 4.21(1H,
m, H-11), 2.90(1H, dt, H-13), 1.73(1H, m, H-18),
3.54(1H, dt, H-19), 1.28(1H, s, H-23), 0.90(1H, s, H-
24), 1.21(3H, s, H-25), 1.11(3H, s, H-26), 0.87(3H, s,
H-27), 4.72(1H, brs, H-29a), 4.90(1H, brs, H-29b),
and 1.74(3H, brs, H-30); C-3 O-glc: 4.90(1H, d,
J = 10.4 Hz, H-1), 4.04(1H, t, J = 8.0 Hz, H-2),
4.25(1H, m, H-3), 4.23(1H, m, H-4), 3.96(1H, m, H-
5), 4.58(1H, dd, J = 2.4 Hz, 8.8 Hz, H-6). 13C NMR
(125 MHz, pyrindine-d5): see Table 1.
1.3.6 Acantrifoside A (6)
White powder, m.p. 265∼268 ◦C(from dil. MeO-
H); [α]23
D
=−42.6(EtOH, c 0.5); 1H NMR(400 MHz,
pyrindine-d5): 3.64(1H, m, H-3), 1.83(1H, d, J =
11.0 Hz, H-9), 4.29 (1H, m, H-11), 2.84(1H, m, H-
13), 3.38(1H, m, H-19), 1.23(3H, s, H-23), 0.97(3H, s,
H-24), 1.26(3H, s, H-25), 1.21(3H, s, H-26), 0.98(3H,
s, H-27), 4.62(1H, brs, H-29a), 4.81(1H, brs, H-29b),
1.64(3H, s, H-30); C-28-O-inner glc: 6.31(1H, d, J=
7.9 Hz, H-l), 4.07(1H, m, H-2), 4.18(1H, m, H-3),
4.29(1H, m, H-4), 4.09(1H, m, H-5), 4.29(H-6), and
4.68(1H, d, J = 11.0 Hz, H-6); glc: 4.93(1H, d, J =
8.0 Hz, H-l), 3.93(1H, brs, H-2), 4.11(1H, m, H-3),
4.36(1H, t, J=9.6 Hz, H-4), 3.64(1H, d, J=9.0 Hz,
No. 6
ZOU Qin-peng, et al: Lupane-triterpenoids from the methanol extracts of
leaves of Acanthopanax gracilistylus W.W. Smith 123
H-5), 4.08(H-6) and 4.18(H-6); rha: 5.82(1H, brs,
H-l), 4.64(1H, brs, H-2), 4.52(1H, d, J = 9.6, H-3),
4.33(H-4), 4.95(H-5) and 1.68(3H, d, J =6.3 Hz, H-
6). 13C NMR data (125 MHz, pyrindine-d5): see
Table 1.
1.3.7 Acankoreoside D (7)
White powder, m. p. 221∼223 ◦C(from dil. MeO-
H); [α]23
D
=−40.8◦(EtOH, c 0.5); ESI-MS m/z: 941,
779, 617, 471; 1H NMR(400 MHz, pyrindine-d5):
4.12(1H, brs, H-3), 2.54(1H, m, H-5), 1.95(1H, m,
H-9), 4.36(m, H-11), 2.89(1H, m, H-13), 3.38(1H,
m, H-19), 10.06(1H, brs, H-23), 1.19(3H, s, H-24),
1.26(3H, s, H-25), 1.23(3H, s, H-26), 1.01(3H, s,
H-27), 4.61(1H, brs, H-29a), 4.80(1H, brs, H-29b),
1.64(3H, s, H-30); C-28-O-inner glc: 6.34(1H, d,
J = 7.9, H-1), 4.11(1H, m, H-2), 4.22(1H, m, H-3),
4.31(1H, m, H-4), 4.11(1H, m, H-5), 4.30(H-6) and
4.70(1H, m, H-6); glc: 4.96(1H, d, J = 8.0 Hz, H-
1), 3.92(1H, t, J = 9.7 Hz, H-2), 4.15(1H, m, H-3),
4.25(1H, t, J=9.6 Hz, H-4), 3.67(1H, d, J=10.0 Hz,
H-5), 4.09(H-6) and 4.22(H-6); rha: 5.86(1H, brs,
H-1), 4.68(1H, brs, H-2), 4.55(1H, d, J = 9.6, H-
3), 4.34(H-4), 4.99(H-5) and 1.72(3H, d, J =6.0 Hz,
H-6). 13C NMR data(125 MHz, pyrindine-d5): see
Table 1.
1.3.8 Acankoreoside B (8)
White powder, m. p. 219∼220 ◦C(from dil. MeO-
H); [α]23
D
=−37.2◦(EtOH, c 0.5); 1H NMR(400 MHz,
pyrindine-d5): 3.92(1H, brs, H-3), 2.24(1H, m, H-
5), 1.95(1H, m, H-9), 4.34(m, H-11), 2.89(1H, m, H-
13), 3.38(1H, m, H-19), 3.69(1H, d, H-23a), 3.90(1H,
d, H-23b), 0.84(3H, s, H-24), 1.29(3H, s, H-25),
1.26(3H, s, H-26), 1.00(3H, s, H-27), 4.61(1H, brs, H-
29a), 4.80(1H, brs, H-29b), 1.64(3H, s, H-30); C-28-
O-inner glc: 6.34(1H, d, J=7.9, H-l), 4.11(1H, m, H-
2), 4.22(1H, m, H-3), 4.31(1H, m, H-4), 4.11(1H, m,
H-5), 4.30(H-6) and 4.70(1H, m, H-6); glc: 4.96(1H,
d, J = 8.0 Hz, H-l), 3.92(1H, t, J = 9.7 Hz, H-
2), 4.15(1H, m, H-3), 4.25(1H, t, J = 9.6 Hz, H-
4), 3.67(1H, d, J = 10.0 Hz, H-5), 4.09(H-6) and
4.22 (H-6); rha: 5.86(1H, brs, H-l), 4.68(1H, brs, H-
2), 4.55(1H, d, J = 9.6, H-3), 4.34(H-4), 4.99(H-5)
and 1.72(3H, d, J = 6.0 Hz, H-6). 13C NMR data
(125 MHz, pyrindine-d5): see Table 1.
1.3.9 Acankoreoside A (9)
White powder, m. p. 226∼228 ◦C(from dil. MeO-
H); [α]23
D
=−41.0◦(EtOH, c 0.5); 1H-NMR (400 MHz,
pyrindine-d5): 4.28(1H, brs, H-3), 1.67(1H, d, J =
11.6 Hz, H-9), 2.67(1H, m, H-13), 3.38(1H, m, H-
19), 1.46(3H, s, H-24), 0.96(3H, s, H-25), 1.21(3H,
s, H-26), 0.87(3H, s, H-27), 4.72(1H, brs, H-29a),
4.86(1H, brs, H-29b), 1.71(3H, s, H-30); C-28-O-
inner glc: 6.34(1H, d, J=7.9 Hz, H-l), 4.08(m, H-2),
4.21(m, H-3), 4.30(m, H-4), 4.09(m, H-5), 4.28(H-6)
and 4.67(1H, d, J = 9.8 Hz, H-6); glc: 4.95(1H, d,
J = 7.9 Hz, H-l), 3.93(1H, t, J = 9.2 Hz, H-2), 4.13
(H-3), 4.39(1H, t, J = 9.4 Hz, H-4), 3.65(1H, d,
J =9.2 Hz, H-5), 4.07(1H, d, J =10.2 Hz, H-6) and
4.19(H-6); rha: 5.84(1H, brs, H-l), 4.66(1H, brs, H-
2), 4.54(1H, d, J =9.0 Hz, H-3), 4.34(H-4), 4.96(H-
5) and 1.69(3H, d, J=6.1 Hz, H-6). 13C NMR data
(125 MHz, pyrindine-d5): see Table 1.
2 Results and discussion
A hot MeOH extract of the leaves of AGS was sus-
pended in water and then partitioned with petroleum
ether, ethyl acetate, and n-butyl alcohol successively.
Further separation of each fraction with a combina-
tion of silica gel, MDS, and Sephadex LH-20 columns
gave compounds 1−9.
Compound 1 was given a positive reaction in the
Liebermann-Burchard and Molish tests. The 1H N-
MR spectrum displayed signals due to six tertiary
methyl groups at δ 0.97, 1.02, 1.15, 1.26, 1.26, and
1.71, two couples of olefinic protons at δ 4.66(1H,
brs) and δ4.88(1H, brs), two oxygen-bearing me-
thine protons at δ 3.64(1H, brs) and δ 4.25(1H, dt,
J = 2.5 Hz, 10.5 Hz). On the other hand, carbon
signals observed in the 13C NMR spectrum showed
the presence of one carboxyl group at δ 179.2, 1-
disubstituted double carbon at δ 110.5 and δ 151.2,
two oxygen-bearig methine carbons at δ 72.3 and δ
75.6, five methine carbons, nine methylene carbons
and six methyl carbons at δ 15.1, 17.2, 18.1, 19.9,
23.3, and 30.3. Therefore, it was deduced to be a
lupane-triterpene compound. Moreover, all assign-
ments of its 1H-NMR and 13C-NMR confirmed by
1H-1H COSY, REOSY, HMBC and HMQC were in
good agreement with the literature values[13] of im-
pressic acid. From the above evidences, compound 1
124 Journal of Lanzhou University (Natural Sciences) Vol. 47
was identified to be an impressic acid isolated from
this plant for the first time.
Compound 2 was given a positive reaction in the
Liebermann-Burchard and Molish tests. The 1H N-
MR spectrum displayed signals due to five tertiary
methyl groups at δ 1.05, 1.13, 1.18, 1.25, and 1.71,
two couples of olefinic protons at δ 4.66(1H, brs)
and δ 4.88(1H, brs), two oxygen-bearing methane
protons at δ 4.02(1H, brs) and δ 4.26(1H, dt, J =
5.2, 16 Hz) while the 13C NMR spectrum showed
30 carbon signals including five methyl, ten methy-
lene, eight methine and seven quaternary carbons
(Table 1). Therefore, it was deduced to be a lupane-
triterpene compound. Furthermore, in the HMBC
spectrum, the aldehydic proton signal at δ 10.09
(1H, s, H-23) correlated with carbons C-3 (δ 73.5),
C-4(δ 53.4), and with C-5(δ 44.6), the proton sig-
nal at δ 1.18(3H, s, H-24) correlated with carbons
C-3 (δ 73.5), C-4(δ 53.4), and C-23(δ 210.5). In
the rotating frame Overhauser effect spectroscopy
(ROESY) spectrum, cross-peaks between H-24 and
H-25 as well as H-3, which indicated that the methyl
group (H-24) was axial, which in turn suggested
that the aldehyde group at C-4 was α-positioned.
Moreover, all assignments of its 1H-NMR and 13C-
NMR confirmed by 1H-1H COSY, REOSY, HMBC
and HMQC were in good agreement with the litera-
ture values of 3α, 11α-dihydroxy-23-oxo-lup-20(29)-
en-28-oic acid[14]. From the above evidences, com-
pound 2 was identified to be 3α, 11α-dihydroxy-23-
oxo-lup-20(29)-en-28-oic acid isolated from this plant
for the first time.
Compound 3 formed white crystals and gave a
positive reaction in the Liebermann-Burchard and
Molish tests. Its basic ion peak at m/z 487.34[M-
H]− was observed in the negative-ion electrospray
ionization (ESI)-MS which suggested the molecular
formula to be C30H48O5. The
13C NMR and distor-
tionless enhancement by polarization transfer (DEP-
T) spectra revealed that 30 carbon signals were as-
signed to a triterpenoid sapogenol with the typical
olefinic carbons at δ 155.9(C-20) and 107.7(C-29),
five methyl carbons at δ 15.2, 17.4, 18.1, 18.8, and
19.9, two oxymethine carbons at δ 70.3 and 76.3,
one oxymethylene carbon at δ 72.3, and carboxyl
signals at δ 179.2. The 1H NMR spectrum of 3 (in
pyrindine-d5) showed five tertiary methyl groups at δ
0.84, 1.05, 1.17, 1.29, and 1.71, two couples of olefinic
protons at δ 4.66(1H, brs) and δ 4.88(1H, brs), t-
wo oxygen-bearing methine protons at δ 3.92(1H,
brs) and δ 4.27((1H, dt, J = 2.5, 10.5 Hz), and two
oxygen-bearing methylene protons at δ 3.70(1H, d),
3.89(1H, d). Assignment of the α-hydroxyl group at
C-3 and C-11 was performed by comparing its spec-
tral data with literature values[15]. Consequently,
the structure of 3 was determined as 3α, 11α, 23-
trihydroxy-20(29)-en-28-oic acid isolated from this
plant for the first time.
Compound 4 was obtained as a white crystal and
gave a positive reaction in the Liebermann-Burchard
and Molish tests. It was identified as acankoreagenin
by comparing the NMR and mass spectral data with
the literature values[3].
Compound 5 was obtained as a white amorphous
powder from the methanolic extract of AGS leaves
by the various chromatographic techniques and gave
a positive reaction in the Liebermann-Burchard
and Molish tests. The IR spectrum of 5 indicat-
ed the presence of OH, C=O, and C-O groups at
3 432, 1 705, and 1 176 cm−1, respectively. Its basic
ion peak at m/z 633 [M-H]− was observed in the
negative-ion ESI-MS, which suggested the molec-
ular formula to be C36H58O9(calcd for C36H57O9:
633.400 3). The 1H NMR spectrum of 5 showed sig-
nals due to six tertiary methyl groups at δ 0.87, 0.90,
1.11, 1.21, 1.28 and 1.74(each 3H, s), two olefinic
protons at δ 4.72(1H, brs) and 4.90(1H, brs), two
oxymethine protons at δ 3.73(1H, brs) and 4.21(1H,
m), and one anomeric proton due to a hexosyl residue
at δ 4.90(1H, d, J = 10.4 Hz). The carbon signals
of 5 observed on the 13C NMR spectrum (Table 1)
showed 36 carbon signals including the presence of
one carboxyl group, one disubstituted double bond,
two oxygen-bearing methine carbons, five methine
carbons, nine methylene carbons and six methyl
carbons. Furthermore, heteronuclear multiple bond
correlation (HMBC) from the glucose H-1 at δ 4.90
(1H, d, J = 10.4 Hz) to C-3 at δ 81.8 of the agly-
cone was observed. This evidence indicated the
location of the sugar linkage of 5. Consequently, the
structure of 5 was determined as 3-O-β-D-glucopyr-
anosyl 3α, 11α-dihydroxylup-20(29)-en-28-oic ac-
No. 6
ZOU Qin-peng, et al: Lupane-triterpenoids from the methanol extracts of
leaves of Acanthopanax gracilistylus W.W. Smith 125
Table 1 13C NMR data(125 MHz, pyridine-d5) of compounds 1−9
aglycone 1 2 3 4 5 6 7 8 9
1 36.6 35.9 36.3 33.3 36.6 36.5 35.6 36.3 33.6
2 27.4 27.6 27.6 26.5 22.2 27.2 27.5 27.5 26.4
3 75.6 73.5 76.3 73.1 81.8 75.6 73.4 76.3 73.1
4 38.9 53.4 41.6 52.3 38.2 38.9 53.4 41.5 51.9
5 49.9 44.6 44.3 45.4 51.0 49.9 44.6 44.2 45.7
6 18.9 21.7 18.7 21.4 18.8 18.9 21.7 18.7 21.9
7 36.3 35.8 36.0 35.1 36.1 36.2 35.8 35.8 34.8
8 43.1 43.2 43.2 42.2 43.1 43.0 43.1 43.2 42.0
9 56.5 56.4 56.6 51.3 56.2 56.5 56.3 56.6 51.2
10 40.2 39.4 40.0 37.7 40.1 40.2 39.4 40.0 37.6
11 70.3 70.2 70.3 22.2 70.3 70.2 70.1 70.2 21.2
12 38.8 38.7 38.8 26.4 38.7 38.6 38.7 38.7 26.3
13 38.0 40.0 38.0 38.9 38.0 37.8 37.8 37.8 38.8
14 43.3 43.7 43.2 43.3 43.3 43.3 43.7 43.3 43.2
15 30.5 30.5 30.5 30.4 30.6 30.4 30.4 30.4 31.2
16 33.3 33.2 33.2 33.1 33.2 32.6 32.6 32.6 32.7
17 56.9 56.9 56.9 56.9 56.9 57.3 57.3 57.3 57.3
18 49.8 49.8 49.8 50.0 49.8 49.8 49.8 49.8 50.0
19 47.9 47.9 47.9 48.1 47.9 47.5 47.5 47.5 47.7
20 151.2 151.2 151.3 151.3 151.3 150.8 150.8 150.8 150.9
21 31.6 31.6 31.6 31.5 31.6 31.2 31.3 31.2 31.4
22 37.8 37.8 37.8 37.9 37.8 37.1 37.1 37.1 37.3
23 30.3 210.5 72.3 179.7 30.3 30.2 210.5 72.3 179.7
24 23.3 15.3 18.8 18.3 23.4 23.3 18.7 18.7 18.6
25 17.2 17.2 17.4 17.1 17.2 17.2 17.2 17.5 17.3
26 18.1 18.2 18.1 17.2 18.0 18.0 18.2 18.1 16.9
27 15.1 15.1 15.2 15.1 15.1 15.1 15.3 15.2 15.2
28 179.2 179.2 179.2 179.2 179.2 175.4 175.4 175.4 175.3
29 110.5 110.5 110.5 110.3 110.4 110.5 110.6 110.6 110.7
30 19.9 19.9 19.9 19.8 20.0 19.8 19.9 19.9 19.8
C-28 O-inner glc 1 2 3 4 5 6 7 8 9
1 95.6 95.7 95.6 95.6
2 74.3 74.4 74.3 74.4
3 78.6 78.6 78.6 78.6
4 71.2 71.2 71.2 71.3
5 77.4 77.5 78.4 78.4
6 69.7 69.8 69.8 69.8
glc′(1→6)glc 1 2 3 4 5 6 7 8 9
1′ 105.4 105.5 105.4 105.4
2′ 75.6 75.7 75.7 75.7
3′ 76.8 76.8 76.8 76.8
4′ 79.0 79.1 79.1 79.0
5′ 78.4 78.4 77.5 78.4
6′ 61.7 61.7 61.7 61.7
rha(1→4)glc′ 1 2 3 4 5 6 7 8 9
1′′ 103.1 103.1 103.0 102.9
2′′ 72.8 72.9 72.9 72.8
3′′ 73.0 73.1 73.1 73.0
4′′ 74.4 74.4 74.4 74.4
5′′ 70.6 70.7 70.7 70.6
6′′ 18.8 18.9 18.9 18.8
C-3 O-glc 1 2 3 4 5 6 7 8 9
1 102.3
2 75.5
3 79.3
4 72.5
5 78.7
6 63.6
126 Journal of Lanzhou University (Natural Sciences) Vol. 47
id[14, 17] which was isolated from this plant for the
first time.
Compounds 6−9 were isolated as white amor-
phous solid and given positive reactions in the
Liebermann-Burchard and Molish tests. They were
identified as acantrifoside A(6)[13, 16], acankoreo-
side D(7)[14], acankoreoside B(8)[15], acankoreoside
A(9)[15, 17] by comparing the NMR and mass spec-
tral data with the literature values[13−17](Figure 1).
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