全 文 ： 药学学报 Acta Pharmaceutica Sinica 2012, 47 (1): 77−83 · 77 ·




Triterpenes from Callicarpa integerrima Champ
ZHU Chen-chen1, 2, GAO Li1, 2, ZHAO Zhong-xiang1, LIN Chao-zhan1, 2*
(1. School of Chinese Materia Medica, Guangzhou University of Chinese Medicine, Guangzhou 510006, China;
2. Guangzhong Jianfeng Research Lab for Natural Products, Guangzhou 510006, China)
Abstract: A new triterpenoid saponin and fourteen known triterpenoids were isolated from the methanol
extract of the stems and leaves of Callicarpa integerrima Champ, which is used in Chinese folk medicine
for stopping bleeding, expelling the wind, dissipating stagnation, and treating scrofula, by using various
chromatographies, such as silica gel, Sephadex LH-20 and RP-C18 column chromatography. Their structures were
identified as a new compound 2α, 3β, 19α, 23-tetrahydroxy-olean-12-en-28-oic acid-28-O-β-D-glucopyranosyl-
(1→4)-β-D-glucopyranoside (1), together with fourteen known compounds: oleanolic acid (2), 3-acetyl oleanolic
acid (3), 3β-O-acetyl ursolic acid (4), 2α-hydroxy-ursolic acid (5), 2α, 3β, 19α, 23-tetrahydroxy-urs-12-en-28-
oic acid (6), α-amyrin-3-O-β-D-glucopyranoside (7), pomolic acid (8), betulinic acid (9), ursolic acid (10), 2α,
3β, 19α, 23-tetrahydroxy-olean-12-en-28-oic acid (arjungenin) (11), 2α-hydroxy-oleanolic acid (12), hederagenin
(13), 2α, 19α-dihydroxy-ursolic acid (14) and pruvuloside A (15), by the spectroscopic techniques of NMR,
HMBC, IR and MS, separately. All these compounds were obtained from this plant for the first time, and
compounds 3, 4 and 15 were isolated from genus Callicarpa L. for the first time.
Key words: Callicarpa L.; Callicarpa integerrima Champ; triterpenes
CLC number: R284.1 Document code: A Article ID: 0513-4870 (2012) 01-0077-07
全缘叶紫珠三萜类成分研究
祝晨蔯 1, 2, 高 丽 1, 2, 赵钟祥 1, 林朝展 1, 2*
(1. 广州中医药大学中药学院, 广东 广州 510006; 2. 广中尖峰天然产物研发共建实验室, 广东 广州 510006)

摘要 : 为了对全缘叶紫珠的茎和叶进行化学成分研究 , 采用硅胶、凝胶等柱色谱方法分离和纯化化合
物, 根据理化性质和波谱方法鉴定化合物结构。从全缘叶紫珠的甲醇提取物中分离得到了 15 个三萜类化合物,
包括 1 个新化合物: 2α, 3β, 19α, 23-tetrahydroxy-olean-12-en-28-oic acid-28-O-β-D-glucopyranosyl-(1→4)-β-D-
glucopyranoside (1) 和 14 个已知化合物: oleanolic acid (2)、3-acetyl oleanolic acid (3)、3β-O-acetyl ursolic acid
(4)、2α-hydroxy-ursolic acid (5)、2α, 3β, 19α, 23-tetrahydroxy-urs-12-en-28-oic acid (6)、α-amyrin-3-O-β-D-
glucopyranoside (7)、pomolic acid (8)、betulinic acid (9)、ursolic acid (10)、2α, 3β, 19α, 23-tetrahydroxy-olean-12-
en-28-oic acid (11)、2α-hydroxy-oleanolic acid (12)、hederagenin (13)、2α, 19α-dihydroxy-ursolic acid (14) 和
pruvuloside A (15)。所有化合物均为首次从全缘叶紫珠中分离得到, 其中化合物 3、4 和 15 为首次从紫珠属中
分离得到。
关键词: 紫珠属; 全缘叶紫珠; 三萜类

Received 2011-08-08, Accepted 2011-11-15.
Project supported by the National Science Foundation Committee of China (81001613); International Cooperation Fund in Guangdong Province
(2008A050200005).
*Corresponding author Tel / Fax: 86-20-39358325, E-mail: linchaozhan@sina.com
· 78 · 药学学报 Acta Pharmaceutica Sinica 2012, 47 (1): 77−83


Callicarpa integerrima Champ (family Verbenaceae),
distributes in southwest China[1]. Stems and leaves of
this plant have been used as a hemostatic drug for various
internal and external hemorrhages in traditional
Chinese medicine. During our systematic research on
the bioactive chemical constituents of C. integerrima,
a new triterpenoid saponin, named 2α, 3β, 19α,
23-tetrahydroxy-olean-12-en-28-oic acid-28-O-β-D-
glucopyranosyl-(1→ 4)-β-D-glucopyranoside (1), together
with fourteen known compounds: oleanolic acid (2),
3-acetyl oleanolic acid (3), 3β-O-acetyl ursolic acid
(4), 2α-hydroxy-ursolic acid (5), 2α, 3β, 19α, 23-
tetrahydroxy-urs-12-en-28-oic acid (6), α-amyrin-3-
O-β-D-glucopyranoside (7), pomolic acid (8), betulinic
acid (9), ursolic acid (10), 2α, 3β, 19α, 23-tetrahydroxy-
olean-12-en-28-oic acid (11), 2α-hydroxy-oleanolic
acid (12), hederagenin (13), 2α, 19α-dihydroxy-ursolic
acid (14) and pruvuloside A (15) were isolated. In this
paper, the isolation and structural elucidation of these
compounds are described.

Results and discussion
Compound 1 (Figure 1) was obtained as a white
amorphous powder, [α] 25D −5.2 (c 0.06, MeOH), which
was considered to be a triterpenoid saponin due to
the positive results for both Libermann-Burchard and
Molish reactions. Its molecular formula was established
to be C42H68O16 by HR-ESI-MS, which showed a
pseudomolecular ion peak at m/z 851.443 0 [M+Na]+,
calcd. 851.440 5. The IR spectrum of 1 indicated the
presence of hydroxyl (3 409 cm−1) and carbonyl (1 736
cm−1) groups.
Compound 1 displayed 42 carbon signals in its
13C NMR spectrum, of which 30 could be assigned to


Figure 1 Key HMBC and 1H-1H COSY correlations of
compound 1
the signals of the aglycone. It was evident that 1 was
a triterpenoid saponin related to an oleanane skeleton
based on the 1H NMR spectral signals (Table 1) assigned
to six tertiary methyl groups at δH 0.67, 0.69, 0.94, 0.96,
1.01, and 1.23, together with six corresponding sp3 carbon
signals in the 13C NMR spectrum (Table 1). The
chemical shifts of C-12, C-13 and C-18 also provided
some evidence for the aglycone related to an oleanane
skeleton[2−6]. In addition, the 1H NMR spectrum of
1 (Table 1) indicated two anomeric sugar protons at
δH 5.53 (1H, d, J = 8.0 Hz) and 4.85 (1H, d, J = 7.5
Hz), and one olefinic proton at δH 5.42 (brs, H-12).
The 13C NMR spectrum for the aglycone moiety of 1
exhibited the signals due to one carbonyl group at δC
179.1 (s), three oxymethine groups at δC 68.7 (d), 76.8
(d), and 81.1 (d), one oxymethylene group at δC 64.4 (t),
one C=C group at δC 124.6 (d) and 142.4 (s). The
spectrum also showed two signals at δC 101.5 (d) and
92.7 (d) assignable to anomeric carbons of two sugar
units. The above data suggest that 1 contains two
sugar moieties and a triterpenoid aglycone related to an
oleanane skeleton.

Table 1 1H NMR (500 MHz) and 13C NMR (125 MHz) spectral
data of compound 1 in D2O ( J in Hz). a)overlapped signals
No. δC δH No. δC δH
1 45.8 (t) 1.95, 0.95 (m) 22 31.8 (t) 1.78, 1.71 (m)
2 68.7 (d) 3.81 (m) 23 64.4 (t) 3.44, 3.36 (m)
3 76.8 (d) 3.37 a) 24 12.8 (q) 0.69 (s)
4 42.9 (s) 25 16.7 (q) 0.67 (s)
5 46.8 (d) 1.30 (m) 26 16.3 (q) 1.01 (s)
6 17.8 (t) 1.51, 1.48 (m) 27 24.4 (q) 1.23 (s)
7 31.9 (t) 1.58, 1.25 (m) 28 179.1 (s)
8 39.5 (s) 29 27.6 (q) 0.94 (s)
9 47.4 (d) 1.80 (m) 30 24.1 (q) 0.96 (s)
10 37.7 (s) glc-1 92.7 (d) 5.53 (d, 8.0)
11 23.6 (t) 2.01 (m) 2 70.6 (d) 3.20 (m)
12 124.6 (d) 5.42 (brs) 3 75.8 (d) 3.49 (m)
13 142.4 (s) 4 75.7 (d) 3.93 (m)
14 41.1 (s) 5 76.2 (d) 3.49 (m)
15 28.9 (t) 1.51, 0.99 (m) 6 60.5 (t) 3.88 (d, 11),
3.72 (m)
16 27.6 (t) 1.71, 1.15 (m) glc-1 101.5 (d) 4.85 (d, 7.5)
17 45.7 (s) 2 74.0 (d) 3.25 (m)
18 43.5 (d) 3.00 (brs) 3 76.7 (d) 3.44 (m)
19 81.1 (d) 3.35 (brs) 4 69.2 (d) 3.47 (m)
20 34.2 (s) 5 76.8 (d) 3.37 a)
21 26.5 (t) 2.15, 1.87 (m) 6 61.9 (t) 3.96, 3.66 (m)

On acid hydrolysis of 1, only D-glucose was
detected by GC, suggesting two sugars were all
ZHU Chen-chen, et al: Triterpenes from Callicarpa integerrima Champ · 79 ·

D-glucoses. The β-configuration of the glycosidic
bonds was deduced from the coupling constants of the
anomeric protons and the 13C NMR data of the sugar
units (Table 1). Meanwhile, in the experiment of
positive ESI-MS/MS of 1, ion peaks were observed at
m/z 673 [M+Li−162]+ and 511 [M+Li−162−162]+,
which also confirmed that there were two β-D-glucose
units in 1.
One and two-dimensional NMR techniques
(1H NMR, 13C NMR, 1H-1H COSY, HSQC and HMBC)
permitted assignments of all 1H and 13C signals (Table
1). The presence of a hydroxyl group at C-23 was
deduced from the NMR resonance at δC 64.4 and δH
3.44, 3.36, as well as the long range coupling between
CH3-24 and C-23 in the HMBC experiment. In addition,
the HMBC spectra showed correlations of H-2 (δH
3.81) with C-3 (δC 76.8), and H-3 (δH 3.37) with C-2
(δC 68.7), C-24 (δC 12.8), C-23 (δC 64.4), C-5 (δC
46.8), indicated that two hydroxyl groups were attached
to C-2 and C-3, separately. Moreover, the HMBC
correlations of H-19 (δH 3.35) with C-18 (δC 43.5),
C-29 (δC 27.6), C-20 (δC 34.2), CH3-29 (δH 0.94) with
C-19 (δC 81.1), and CH3-30 (δH 0.96) with C-19 (δC
81.1), showed that a hydroxyl group was linked to C-19.
The 1H-1H COSY (Figure 1) spectrum revealed the
presence of the systems -CH2-CH-CH- (from H-1 to
H-3) and -CH-CH- (from H-18 to H-19), which also
represented that C-2, C-3, and C-19 were oxygenated[6−11].
Therefore, the aglycone of 1 was very similar to
arjungenin (2α, 3β, 19α, 23-tetrahydroxy-olean-12-en-
28-oic acid)[7]. HPLC analysis of the aglycone fraction
of hydrolysis products of 1 and arjungenin (11) under
the same condition demonstrated the aglycone of 1 was
arjungenin. In addition, a cross peak between H-1 of
inner glucose and C-28 of the aglycone indicated that
one glucose was connected to C-28 of the aglycone.
The linkage of terminal glucose at the C-4 of the inner
glucose was indicated by the cross-peak of H-1/C-4.
Hence, the structure of 1 was 2, 3, 19, 23-tetrahydroxy-
olean-12-en-28-oic acid-28-O-β-D-glucopyranosyl-(1→
4)-β-D-glucopyranoside.
The relative stereochemistry of 1 was studied by
NOESY (Figure 2). The NOE correlation between
CH3-25 and CH3-24 indicated the β configuration for
the hydroxymethyl group at C-23, which was further
proved by the chemical shift of C-24 (δC 12.8)[6].
Besides, the configurations of hydroxyl groups on C-3
and C-2 were confirmed as 3β and 2α, according to
the NOE correlations between H-3 and H-5, H-3 and
CH3-23, H-2 and CH3-25, and H-2 and CH3-24 in
the NOESY spectrum. The chemical shifts of C-2 and
C-3 (δC 68.7 and δC 76.8) also provided some evidence
for the configurations of hydroxyl groups on C-3 and
C-2[6]. The α configuration for 19-OH was determined
by comparing the chemical shift of C-19 (δc 81.1)
with that of triterpenoid analogues [6], which was also
proved by the NOE correlations between H-19 and
CH3-30, H-18, H-18 and CH3-26, in the NOESY
spectrum. Thus, the structure of 1 was identified as
2α, 3β, 19α, 23-tetrahydroxy-olean-12-en-28-oic acid-
28-O-β-D-glucopyranosyl-(1→4)-β-D-glucopyranoside.


Figure 2 Key NOESY correlations of compound 1

Experimental
General experimental procedures Melting points
were determined by XT digital melting-point apparatus
with a microscope and uncorrected. Optical rotations
were measured on a JASCO DIP-360 digital polarimeter.
UV spectra were obtained on a CARY 50 Probe UV-Vis
spectrophotometer, and IR spectra on an Avata-330
FT-IR spectrophotometer. NMR spectra were run on a
Bruker AVANCE AV 400 spectrophotometer using TMS
as the internal reference. Precoated silica gel GF254
plates (Qingdao Haiyang Chem. Co.) were employed
for TLC. Spots were visualized under UV light (254
nm) or by spraying with 10% H2SO4 in 95% EtOH
followed with heating. For column chromatography,
silica gel (Qingdao Haiyang Chem. Co.), reversed-
phase C18 silica gel (Merck) and Sephadex LH-20
(Pharmacia) were used. All the reagents were of
analytical grade (Guangzhou Reagent Factory).
Plant materials The leaves and stems of
C. integerrima were collected in Guangdong Province,
China, in October 2007, and authenticated by Prof.
Chao-mei Pan (Department of pharmacognosy, School
of Chinese Materia Medica, Guangzhou University of
· 80 · 药学学报 Acta Pharmaceutica Sinica 2012, 47 (1): 77−83

Chinese Medicine). A voucher specimen (CIL071005)
was deposited in Herbarium of School of Chinese
Materia Medica, Guangzhou University of Chinese
Medicine.
Extraction and isolation
The dried leaves and stems of C. integerrima
(12 kg) were ground and extracted consecutively with
methanol at room temperature. The combined extract
was concentrated under reduced pressure to yield 980 g
of residue, which was suspended in water and extracted
successively with petroleum ether (PE) (60 − 90 ℃),
EtOAc, and n-BuOH. Further purification was done
with silica gel, Sephadex LH-20 chromatography.
The PE extract (60 g) was chromatographed over a
silica gel column eluting with PE-acetone (50∶1−1∶1),
and monitored by TLC analysis to yield nine combined
fractions (Fr. A1-Fr. A9). Fr. A2 (PE-acetone, 40∶1)
gave compound 2 (150 mg). Fr. A4 (PE-acetone,
25∶1) yielded compounds 2 (50 mg) and 3 (29 mg).
Besides, compounds 4 (67 mg), 5 (23 mg), 6 (30 mg),
7 (200 mg), 8 (31 mg) and 9 (102 mg), were isolated
from Fr. A5 (PE-acetone, 20∶1), Fr. A6 (PE-acetone,
15∶1), Fr. A7 (PE-acetone, 10∶1), Fr. A8 (PE-acetone,
5∶1) and Fr. A9 (PE-acetone, 1∶1), respectively.
The EtOAc extract (120 g) was chromatographed
over a silica gel column eluting with CHCl3−MeOH
(50∶1−1∶1), and monitored by TLC analysis to yield
seven combined fractions (Fr. B1−Fr. B7). From Fr.
B1 (CHCl3−MeOH, 50∶1), Fr. B2 (CHCl3−MeOH,
40∶1) and Fr. B4 (CHCl3−MeOH, 25∶1), compounds
10 (80 mg), 11 (25 mg) and 14 (19 mg) were purified
respectively. In addition, Fr. B3 (CHCl3−MeOH, 30∶1)
afforded compounds 12 (30 mg) and 13 (22 mg).
The n-BuOH extract (350 g) was subjected to a
macroporous resin (AB-8) column eluting with water,
10%, 20%, 40% , 60% and 100% methanol, successively.
The 40% MeOH fraction (15 g) was rechromatographed
on a Toyopearl HW-40 column eluting with gradient
mixtures of MeOH-H2O (95∶5 to 60∶40) giving
two fractions. Fr.2 (101 mg) was separated with
Sephadex LH-20 eluting with MeOH−H2O (20%, 40%
respectively) and then purified by ODS (30% aqueous
methanol) to yield compound 1 (26 mg). Compound
15 (29 mg) was isolated from the 60% MeOH fraction.
Structure elucidation
Compound 1 White amorphous powder, mp 260−
262 ℃. [α] 25D −5.2 (c 0.06, MeOH), IR (KBr): 3 409,
2 926, 1 736, 1 457, 1 161, 1 075 cm−1. Positive
ESI-MS m/z 835 [M+Li]+, 673 [M+Li−glc]+, and 511
[M+Li−glc−glc]+. 1H and 13C NMR data were listed
in Table 1.
Acid hydrolysis of compound 1 Compound 1
(2 mg) was hydrolyzed with 9% HCl (1.5 mL) at 90 ℃
for 5 h. After cooling, the reaction mixture was
filtered and the filtrate was freeze-dried. The dried
material was dissolved in pyridine (100 μL), and 200 μL
of 0.1 mol·L−1 L-cysteine methyl ester hydrochloride
was added. The mixture was stirred at 60 ℃ for 1 h,
then 150 μL of HMDS-TMCS (hexamethyldisilazane-
trimethylchlorosilane, 2∶1) was added and the mixture
was stirred at 60 ℃ for another 30 min after centrifu-
gation, the supernatant was analyzed by GC under the
following conditions: capillary column, HP-5 (30.0 m ×
320 μm × 0.25 μm); detection, FID; temperature gradient
system for the oven, 150 ℃ for 1 min and then raised
to 280 ℃ at the rate of 5 ℃·min−1; detector temperature,
250 ℃; carrier, N2 gas. The peak of the hydrolysate
of 1 was detected at 22.02 min (D-glucose) by
comparison with authentic sample[12].
Compound 2 White needle crystal, mp 296−
298 ℃. ESI-MS m/z 456 [M]+; 1H NMR (CDCl3, 500
MHz) δ 5.26 (1H, brs, H-12), 3.32 (1H, m, H-3), 1.26
(3H, s, 27-Me), 1.22, 1.05, 1.00, 0.96, 0.86, 0.78 (3H×6,
s, 23, 24, 25, 26, 29, 30-Me); 13C NMR data were listed
in Table 2. These data were consistent with those of
oleanolic acid in the literature[13].
Compound 3 White crystal, mp 245−246 ℃.
ESI-MS m/z 498 [M]+; 1H NMR (CDCl3, 500 MHz) δ
5.27 (1H, brs, H-12), 4.40 (1H, d, J = 8.0 Hz, H-3),
2.80 (1H, dd, J = 9.6, 3.8 Hz, H-18), 2.14 (3H, s,
CH3CO), 1.35 (3H, s, 27-Me), 1.09, 1.06, 1.03, 0.98,
0.95, 0.90 (3H×6, s, 23, 24, 25, 26, 29, 30-Me);
13C NMR data were listed in Table 2. These data were
consistent with those of 3-acetyl oleanolic acid in the
literature[14].
Compound 4 White powder, mp 276−278 ℃.
ESI-MS m/z 499 [M+H]+; 1H NMR (CDCl3, 500 MHz)
δ 5.55 (1H, brs, H-12), 4.76 (1H, m, H-3), 2.14 (1H, d,
J = 11.0 Hz, H-18), 2.05 (3H, s, CH3CO), 1.35, 1.30,
0.98, 0.95, 0.90 (3H×5, s, 23, 24, 25, 26, 27-Me), 1.07
(3H, d, J = 5.0 Hz, 30-Me), 1.04 (3H, d, J = 5.5 Hz,
29-Me); 13C NMR data were listed in Table 2. These
data were consistent with those of 3β-O-acetyl ursolic
acid in the literature[15].
Compound 5 White powder, mp 254−256 ℃.
1H NMR (C5D5N, 500 MHz) δ 5.42 (1H, brs, H-12),
4.16 (1H, m, H-2), 3.42 (1H, d, J = 9.5 Hz, H-3), 1.28,
ZHU Chen-chen, et al: Triterpenes from Callicarpa integerrima Champ · 81 ·

1.27, 1.21, 1.08, 1.03 (3H×5, s, 23, 24, 25, 26, 27-Me),
1.14 (3H, d, J = 6.5 Hz, 30-Me), 0.93 (3H, d, J = 6.5 Hz,
29-Me); 13C NMR data were listed in Table 2. These
data were consistent with those of 2α-hydroxy-ursolic
acid in the literature[15, 16].

Table 2 13C NMR (125 MHz) spectral data of compounds 2−8
No. 2 3 4 5 6 7 8
1 38.8 39.4 38.0 47.9 47.6 40.2 39.0
2 27.9 22.1 23.1 68.5 68.6 39.0 28.5
3 79.3 79.9 80.6 83.7 78.3 78.9 78.1
4 39.1 37.9 37.9 39.8 44.0 36.9 39.8
5 55.3 54.8 54.8 55.8 48.1 56.5 56.2
6 18.4 16.7 17.7 18.8 18.9 21.3 19.0
7 33.2 32.5 32.5 33.4 33.6 34.7 34.0
8 39.6 38.4 40.2 39.9 40.8 42.4 40.4
9 48.2 46.1 47.2 48.0 48.3 50.8 47.9
10 38.3 37.1 36.1 39.7 38.9 42.0 37.5
11 24.5 31.5 22.9 23.7 24.5 24.6 24.1
12 123.2 121.6 125.6 125.5 128.4 122.3 128.2
13 144.3 143.1 137.6 139.3 140.4 136.5 140.0
14 42.2 40.4 41.8 42.5 42.3 43.6 42.1
15 28.1 26.6 27.6 28.6 29.6 30.3 29.9
16 23.8 21.9 24.1 24.8 26.9 27.0 26.6
17 47.2 44.9 47.5 48.0 48.1 48.5 48.5
18 41.3 40.2 52.2 53.4 55.0 57.2 55.0
19 46.3 44.7 39.2 39.4 73.1 47.3 73.0
20 31.2 29.2 38.7 39.3 42.8 39.3 42.8
21 33.9 36.5 30.3 31.0 26.7 27.9 27.0
22 32.4 35.2 36.7 37.4 38.3 39.7 38.6
23 29.6 15.2 27.4 29.3 66.5 30.3 29.0
24 16.2 26.4 15.3 17.4 14.7 23.4 17.1
25 15.6 14.7 16.6 17.0 17.9 14.8 15.8
26 17.3 15.6 16.8 17.4 17.2 14.9 17.5
27 26.1 25.0 23.2 23.9 24.1 25.9 25.0
28 182.3 183.5 182.7 179.8 181.0 30.5 180.7
29 33.5 33.5 16.8 17.6 27.5 27.7 16.7
30 23.4 22.8 22.0 21.4 16.7 20.0 27.2
31 170.1 170.9
32 20.7 20.6
glc-1 103.1
2 75.8
3 79.4
4 72.0
5 78.9
6 63.0

Compound 6 White powder, mp 283−285 ℃.
ESI-MS m/z 527 [M+Na]+; 1H NMR (C5D5N, 500 MHz)
δ 5.59 (1H, brs, H-12), 3.57 (1H, d, J = 9.5 Hz, H-3),
3.07 (1H, s, H-8), 1.67, 1.42, 1.18, 1.06, 0.94 (3H×5, s,
24, 25, 26, 27, 29-Me), 1.12 (3H, d, J = 6.5 Hz, 30-Me);
13C NMR data were listed in Table 2. These data were
consistent with those of 2α, 3β, 19α, 23-tetrahydroxy-
urs-12-en-28-oic acid in the literature[17].
Compound 7 White amorphous powder. ESI-MS
m/z 589 [M+H]+; 1H NMR (C5D5N, 500 MHz) δ 5.25
(1H, m, H-12), 3.57 (1H, m, H-3), 1.01, 1.00, 0.97,
0.94, 0.90, 0.88, 0.85, 0.69 (3H×8, s, 23, 24, 25, 26, 27,
28, 29, 30-Me); 13C NMR data were listed in Table 2.
These data were consistent with those of α-amyrin-3-
O-β-D-glucopyranoside in the literature[18].
Compound 8 White amorphous powder, mp
276−278 ℃. ESI-MS m/z 473 [M+H]+; 1H NMR
(C5D5N, 500 MHz) δ 5.63 (1H, t, H-12), 3.32 (1H, dd,
J = 11.0, 4.4 Hz, H-3), 2.07 (1H, s, H-18), 1.73, 1.46,
1.24, 1.12, 1.03, 0.94 (3H×6, s, 23, 24, 25, 26, 27,
30-Me), 1.14 (3H, d, J = 6.5 Hz, 29-Me); 13C NMR data
were listed in Table 2. These data were consistent
with those of pomolic acid in the literature[18].
Compound 9 White amorphous powder, mp 285−
287 ℃. ESI-MS m/z 456 [M]+; 1H NMR (C5D5N, 500
MHz) δ 4.96 (1H, s, H-29), 4.79 (1H, s, H-29), 3.24
(1H, m, H-3), 1.88 (3H, s, 30-Me), 1.25, 1.21, 1.08,
0.96, 0.91 (3H×5, s, 23, 24, 25, 26, 27-Me); 13C NMR
data were listed in Table 3. These data were consistent
with those of betulinic acid in the literature[19−21].
Compound 10 White amorphous powder, mp
260−262 ℃. ESI-MS m/z 457 [M+H]+; 1H NMR
(C5D5N, 500 MHz) δ 5.23 (1H, brs, H-12), 3.34 (1H, d,
J = 3.7 Hz, H-3), 1.27, 1.20, 1.05, 1.00, 0.96, 0.80, 0.74
(3H×7, s, 23, 24, 25, 26, 27, 29, 30-Me); 13C NMR data
were listed in Table 3. These data were consistent
with those of ursolic acid in the literature[18, 19, 22].
Compound 11 White power. 1H NMR (C5D5N,
500 MHz) δ 5.26 (1H, brs, H-12), 3.31 (1H, d, J = 3.7
Hz, H-3), 1.27, 1.14, 1.08, 1.07, 1.03, 0.92 (3H×6, s, 24,
25, 26, 27, 29, 30-Me); 13C NMR data were listed in
Table 3. These data were consistent with those of 2α,
3β, 19α, 23-tetrahydroxy-olean-12-en-28-oic acid in the
literature[23].
Compound 12 White power, mp 290−292 ℃.
ESI-MS m/z 472 [M]+; 1H NMR (C5D5N, 500 MHz) δ
5.50 (1H, brs, H-12), 4.11 (1H, m, H-2β), 3.40 (1H, d,
J = 8.0 Hz, H-3α), 3.29 (1H, dd, J = 14.0, 3.5 Hz,
H-18), 1.28, 1.27, 1.08, 1.03, 1.01, 0.99, 0.96 (3H×7, s,
23, 24, 25, 26, 27, 29, 30-Me); 13C NMR data were
listed in Table 3. These data were consistent with those
of 2α-hydroxy-oleanolic acid in the literature[24].
· 82 · 药学学报 Acta Pharmaceutica Sinica 2012, 47 (1): 77−83

Table 3 13C NMR (125 MHz) spectral data of compounds 9−15
No. 9 10 11 12 13 14 15
1 39.8 38.7 47.0 48.1 38.7 42.8 42.3
2 28.8 28.5 68.5 68.4 27.4 66.1 65.9
3 78.2 78.3 77.8 83.3 73.2 79.3 78.8
4 41.7 39.6 43.2 40.2 42.8 38.6 38.2
5 56.0 55.6 48.1 58.9 48.2 48.7 48.1
6 19.0 23.4 18.3 19.2 18.7 18.6 17.9
7 35.0 38.0 33.6 33.1 32.5 33.5 32.9
8 41.2 40.0 39.5 40.2 39.6 40.5 40.2
9 50.1 47.7 48.0 48.5 48.1 47.7 47.0
10 37.6 39.0 38.2 38.8 36.9 38.8 38.1
11 21.4 24.1 28.8 24.0 23.5 24.0 23.4
12 26.3 125.3 123.1 122.1 122.3 127.9 128.2
13 39.4 139.2 144.5 144.6 144.5 139.9 138.9
14 43.0 42.1 41.8 42.6 41.9 42.1 41.4
15 31.3 30.6 29.3 28.6 27.9 29.2 29.1
16 33.0 28.3 23.9 24.1 23.5 26.4 26.0
17 56.7 47.1 46.0 47.0 46.3 48.3 48.4
18 49.9 53.5 44.7 42.3 41.8 54.6 53.9
19 47.9 40.0 81.4 46.8 46.1 72.7 72.4
20 151.2 40.9 35.3 31.3 30.7 42.3 41.7
21 30.4 36.8 28.3 34.6 34.1 27.0 26.2
22 38.7 39.0 32.5 33.5 32.9 38.5 37.5
23 28.5 33.2 66.1 29.7 67.9 29.4 29.0
24 16.6 16.9 13.8 18.0 12.9 16.7 22.0
25 16.6 17.1 17.5 16.0 15.7 16.6 16.3
26 16.6 18.3 16.8 17.6 17.1 17.2 17.3
27 15.1 27.5 24.8 26.5 25.9 24.4 24.4
28 178.7 179.4 180.7 180.1 179.8 180.7 177.4
29 109.9 17.7 28.6 33.6 33.1 26.9 25.8
30 19.6 22.5 24.3 24.3 23.6 22.2 16.5
glc inner/terminal
1 / 1 93.2 / 104.5
2 / 2 79.0 / 75.5
3 / 3 77.7 / 77.7
4 / 4 70.5 / 72.3
5 / 5 78.6 / 77.9
6 / 6 62.1 / 63.5

Compound 13 White needle crystal, mp 331−
333 ℃. ESI-MS m/z 472 [M]+; 1H NMR (C5D5N, 500
MHz) δ 5.46 (1H, brs, H-12), 4.18 (2H, m, H-23), 3.61
(1H, m, H-3), 1.17, 0.98, 0.94, 0.91, 0.82, 0.70 (3H×6,
s, 24, 25, 26, 27, 29, 30-Me); 13C NMR data were listed
in Table 3. These data were consistent with those of
hederagenin in the literature[25, 26].
Compound 14 White powder, mp 333−335 ℃.
1H NMR (C5D5N, 500 MHz) δ 5.60 (1H, brs, H-12),
4.34 (1H, d, J = 9.5 Hz, H-2), 3.18 (1H, d, J = 4.0 Hz,
H-3), 1.14 (3H, d, J = 6.5 Hz, 30-Me), 1.66, 1.43, 1.28,
1.12, 1.00, 0.92 (3H×6, s, 23, 24, 25, 26, 27, 29-Me);
13C NMR data were listed in Table 3. These data were
consistent with those of 2α, 19α-dihydroxy-ursolic acid
in the literature[27].
Compound 15 White powder. ESI-MS m/z 819
[M+Li]+; 1H NMR (C5D5N, 500 MHz) δ 6.15 (1H, d,
J = 6.4 Hz, inner Hglc-1 ), 5.60 (1H, brs, H-12), 5.58
(1H, d, J = 6.0 Hz, terminal Hglc-1 ), 1.02 (3H, d, J =
6.5 Hz, 30-Me), 1.67, 1.42, 1.18, 1.06, 0.94, 0.82
(3H×6, s, 23, 24, 25, 26, 27, 29-Me); 13C NMR data
were listed in Table 3. These data were consistent
with those of pruvuloside A in the literature[28].
References
[1] Pei J, Chen SL. Flora Reipublicae Popularis Sinicae (中国植
物志) [M]. Beijing: Science Press, 2004: 34.
[2] Yu DQ, Yang JS. The Handbook of Analytical Chemistry:
Vol 7 (分析化学手册: 第七分册) [M]. Beijing: Chemical
Industry Press, 1999: 430−434, 796−797.
[3] Tan MX, Li DQ, Tan AZ, et al. Survey on the triterpenoids
from Rubus L and their spectral features [J]. Chem Ind Times
(化工时刊), 2004, 18: 1−4, 8.
[4] Wang ZX, Zhao YY, Chen YY, et al. Triterpenoid compounds
of Prunella genus and their features of 13C NMR spectroscopy
[J]. China J Chin Mater Med (中国中药杂志), 2000, 25:
583−588.
[5] Chen CZ. General methods about triterpenoid saponin structural
elucidation [J]. Chem Res Appl (化学研究与应用), 1999, 2:
119−125.
[6] Mahato SB, Kundu AP. 13C NMR spectra of pentacyclic
triterpenoids―a compilation and some salient features [J].
Phytochemistry, 1994, 37: 1517−1575.
[7] Fumiko A, Tatsuo Y. Glycosides of 19α-hydroxyoleanane-
type triterpenoids from Trachelospermum asiaticum
(Trachelospermum. IV) [J]. Chem Pharm Bull, 1987, 35:
1883−1887.
[8] Ge X, Lin DC, Zhang WD, et al. Triterpenoid saponins and
monoterpenoid glycosides from Incarvillea delavayi [J]. J
Asian Nat Prod Res, 2009, 11: 838−844.
[9] Kim NC, Desjardins AE, Wu CD, et al. Activity of triterpenoid
glycosides from the root bark of Mussaeacla macrophylla
against two oral pathogens [J]. J Nat Prod, 1999, 62: 1379−
1384.
[10] Fumiko A, Tatsuo Y. Trachelosperosides, glycosides of 19α-
hydroxyursane-type triterpenoids from Trachelospermum
asiaticum [J]. Chem Pharm Bull, 1987, 35: 1748−1754.
[11] Zhang ZZ, Li SY. Cytotoxic triterpenoid saponins from the
ZHU Chen-chen, et al: Triterpenes from Callicarpa integerrima Champ · 83 ·

fruits of Aseculus pavia L. [J]. Phytochemistry, 2007, 68:
2075−2086.
[12] Zhao ZZ, Jin J, Ruan JL, et al. Antioxidant flavonoid
glycosides from aerial parts of the fern Abacopteris penangiana
[J]. J Nat Prod, 2007, 70: 1683−1686.
[13] Xu C, Wang ZT. Chemical constituents from roots of
Andrographis paniculata [J]. Acta Pharm Sin (药学学报),
2011, 46: 317−321.
[14] Huang MQ, Ya QK, Chen HY, et al. Chemical constituents
from Rhodomyrtus tomentosa (Ait.) Hassk. [J]. Chin Tradit
Herb Drugs (中草药), 2010, 41: 1079−1080.
[15] Cui Y, Zhang XM, Chen JJ, et al. Chemical constituents
from root of Actinidia chinensis [J]. China J Chin Mater
Med (中国中药杂志), 2007, 32: 1663−1665.
[16] Miao Q, Bao HY, Piao SJ, et al. Study on chemical
constituents of Duchesnea indica Andr. Focke [J]. Acad
J Second Mil Med Univ (第二军医大学学报), 2008, 29:
1366−1370.
[17] Du JW, Yang JZ, Zhang DM. Chemical constituents from
Rubus suavissimus [J]. Chin Tradit Herb Drugs (中草药),
2007, 38: 346−348.
[18] Lin CZ, Zhu CC, Deng GH, et al. Studies on the chemical
constituents of Callicarpa kochiana [J]. Chin Med Mat (中药
材), 2010, 33: 897−900.
[19] Yuan K, Jia A, Zhang AL. Studies on chemical constituents
of Vernonia chunii ( ) [J].Ⅰ Chin Pharm J (中国药学杂志),
2006, 41: 1458−1460.
[20] Sun JM, Yang JS. Studies on chemical constituents of
Jasminum lanceolarium [J]. Chin Pharm J (中国药学杂志),
2007, 42: 489−491.
[21] Zhang Y, Deng ZW, Gao TX, et al. Chemical constituents
from themangrove plant Ceriops tagal [J]. Acta Pharm Sin
(药学学报), 2005, 40: 935−939.
[22] Yang SM, Liu XK, Qing C, et al. Chemical constituents from
the roots of Homonoia riparia [J]. Acta Pharm Sin (药学学
报), 2007, 42: 292−296.
[23] Zhao WQ, Ding LS, Wang MK. Studies on chemical
constituents in radicular part of Rubus swinhoei [J]. Chin
Tradit Herb Drugs (中草药), 2001, 32: 874−876.
[24] Guo QL, Yang JS. Studies on chemical constituents in fruit
of Rubus chingii [J]. China J Chin Mater Med (中国中药杂
志), 2005, 30: 198−200.
[25] Liu GY, Zheng J, Yu ZX, et al. Study on sterols and
triterpenes from the stems of Akebia quinata [J]. Chin Med
Mat (中药材), 2005, 28: 1060−1061.
[26] Zhang QW, Ye WC, Che ZT, et al. Triterpene saponins from
Pulsatilla cernua [J]. Acta Pharm Sin (药学学报), 2000, 35:
756−759.
[27] Wang Q, Ju P, Wang YF, et al. Triterpenoids from Saurauia
napaulensis (Saurauiaceae) [J]. Acta Bot Yunnan (云南植物
研究), 2008, 30: 121−124.
[28] Zhang YJ, Yang CR. Two new ursane glycosides from
Prunella Vulgaris in France [J]. Acta Bot Yunnan (云南植物
研究), 1995, 17: 468−472.