全 文 ： 366 Chin J Nat Med Sep. 2012 Vol. 10 No. 5 2012 年 9 月 第 10 卷 第 5 期

Chinese Journal of Natural Medicines 2012, 10(5): 0366−0369
doi: 10.3724/SP.J.1009.2012.00366
Chinese
Journal of
Natural
Medicines







Chemical constituents from the aerial parts of
Codonopsis nervosa
AGA Er-Bu, LI Hui-Jun, CHEN Jun*, LI Ping*
State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing 210009, China
Available online Sep. 2012
[ABSTRACT] AIM: To study the chemical constituents of the aerial parts of Codonopsis nervosa (Chipp.) Nannf. (Campanulaceae), a
commonly used Tibetan herb. METHODS: The constituents were isolated by D-101 porous polymer resin, silica gel, ODS and
Sephadex LH-20 column chromatography, and their structures were elucidated on the basis of spectroscopic analysis. RESULTS: 15
compounds were isolated and identified as chrysoeriol (1), tricin (2), apigenin (3), succinic acid (4), β-sitosterol-3-O-β-D-glu-
copyranoside (5), luteolin (6), luteolin-7-O-β-D-glucopyranoside (7), ethylsyringin (8), apigenin-7-O-β-D-glucopyranoside (9), luteo-
lin-7-O-β-D-glucopyranosyl-(1→6)-[6-O-caffeoyl]-β-D-glucopyranoside (10), luteolin-7-O-β-D-gentiobioside (11), syringin (12),
3-O-caffeoylquinic acid (13), 5-O-caffeoylquinic acid (14), and 4-O-(β-D-glucopyranosyl)-benzoic acid (15), respectively.
CONCLUSION: Compounds 1-5, 8, 9, 12, 14, 15 were reported from C. nervosa for the first time.
[KEY WORDS] Codonopsis nervosa; Campanulaceae; Tibetan medicine; Flavones; Phenylpropanoids
[CLC Number] R284.1 [Document code] A [Article ID] 1672-3651(2012)05-0366-04

1 Introduction
Codonopsis nervosa Nannf. (Campanulaceae), known as
luduiduoji in Tibetan, is a plateau plant endemic to China
which is distributed in grass, forest edge or slope shade at an
altitude of 3 500−4 500 m [1]. The aerial parts of C. nervosa
are utilized to treat arthritis, neuralgia, leprosy, beriberi and
hysteria in Tibetan medicine [2], while the roots of some other
species of this genus are used to treat appetite loss, diarrhea,
and vomiting in traditional Chinese medicine. Previously, the
chemical compositions of C. pilosula has been investigated in
detail but few studies on the aerial parts of C. nervosa have
been reported [3]. To investigate the chemical constituents of
the aerial parts of C. nervosa, we systematically examined
the aerial parts of C. noversa and 15 compounds were iso-
lated, whose structures were elucidated as chrysoeriol (1),
tricin (2), apigenin (3), succinic acid (4), β-sitosterol-3-O-β-D

[Received on] 20-Jan.-2012
[Research funding] The project was supported by the National New
Drug Innovation Major Project of China (2011ZX09307-002-02) and
the Program for New Century Excellent Talents in University (No.
NCET-09-0769).
[*Corresponding author] LI Ping: Prof., Tel: 86-25-83271379,
E-mail: liping2004@126.com; chenj2002@gmail.com (CHEN Jun)
These authors have no any conflict of interest to declare.
-glucopyranoside (5), luteolin (6), luteolin-7-O-β-D-glucopy-
ranoside (7), ethylsyringin (8), apigenin-7-O-β-D-glucopy-
ranoside (9), luteolin-7-O-β-D-glucopyranosyl-(1→6)-[(6-
O-caffeoyl)]-β-D-glucopyranoside (10), luteolin-7-O-β-D-
gentiobioside (11), syringin (12), 3-O-caffeoylquinic acid
(13), 5-O-caffeoylquinic acid (14), 4-(β-D-glucopyranosyl)-
benzoic acid (15). Compounds 1-5, 8, 9, 12, 14, and 15 were
reported from C. nervosa for the first time.
2 Experimental
2.1 General
NMR experiments were recorded with Bruker AV-500 or
Bruker AV-300 spectrometers, using TMS as internal stan-
dard. The ESI-MS spectra were recorded on an Agilent 1100
LC/MSD TOF (ESI mode) instrument. Column chroma-
tographic separations were carried out using D-101 porous
polymer resin (Hebei Bonherb Technology Corporation, He-
bei Province, China), Silica gel (Qingdao Marine Chemistry
Company, China), Rp-C18 (40−60 μm, Sigma, USA) and
Sephadex LH-20 (20−100 μm, Pharmacia, Sweden). Silica
gel GF254 for TLC was purchased from Qingdao Marine
Chemical Inc.
2.2 Plant material
The aerial parts of C. nervosa were collected in August
2010 in Tibet, China, and authenticated by Prof. Gesang
Bazhu, Tibet Institute of Tibetan Medicine, Lhasa, China.
AGA Er-bu, et al. /Chinese Journal of Natural Medicines 2012, 10(5): 366−369
2012 年 9 月 第 10 卷 第 5 期 Chin J Nat Med Sep. 2012 Vol. 10 No. 5 367

The voucher specimen (No. 100046) was deposited in the
State Key Laboratory of Natural Medicines, China Pharma-
ceutical University, Nanjing, China.
2.3 Extraction and isolation
The air-dried aerial parts of C. nervosa (10 kg) were ex-
tracted three times with 70% aqueous ethanol (V/V) (100 L ×
3 h) at reflux. The combined extracts were concentrated in
vacuo to afford the residue, which was suspended in H2O and
partitioned successively with petroleum ether, EtOAc and
n-BuOH. The EtOAc fraction was applied to silica-gel col-
umn chromatography eluting with petroleum ether-acetone
(20 : 1−1 : 1) to obtain fractions 1−19 (Frs. 1−19). Repeated
chromatography of Fr. 11 with silica gel (MeOH/CHCl3, 90 :
1−10 : 1) and Sephadex LH-20 (CHCl3/MeOH, 1 : 1) gave
compounds 1 (4 mg), 2 (20 mg), 3 (10 mg) and 4 (50 mg).
From Frs. 12 and 14, compounds 5 (40 mg) and 6 (300 mg)
were obtained by recrystallization, respectively. A portion of
the n-BuOH extract (500 g) was applied on D101 macropor-
ous resin column chromatography eluted with H2O, 20%,
70% and 95% ethanol respectively. Then the extract (60 g)
from the 70% ethanol elution was stepwise eluted on silica
gel column (100−200 mesh) with MeOH/CHCl3 (50 : 1−1 : 1)
to afford ten fractions (Frs. A−J). Meanwhile, compound 7
(900 mg) was precipitated from Fr. I. Fr. C was further
purified by Sephadex LH-20 with CHCl3/CH3OH (1 : 1) to
yield compounds 8 (35 mg) and 9 (20 mg). Fr. J was further
subjected to Rp-C18 column with CH3OH/H2O (20 : 80−60 :
40) obtained compound 10 (200 mg). The 20% ethanol ex-
tract was subjected to silica gel column (200−300 mesh) and
stepwise eluted with MeOH/CHCl3/H2O (9 : 1 : 0.1−6 : 4 : 1),
followed by recrystallization in MeOH to afford compounds
11 (300 mg), 12 (100 mg) and 13 (200 mg). Compounds 14
(50 mg) and 15 (30 mg) were obtained from the water extract
by recrystallization respectively.

Fig. 1 Structures of compounds 1-3, 6-15
3 Structural Identification
Compound 1 Yellow powder (MeOH), C16H12O6.
ESI-MS m/z 299 [M − H]−. 1H NMR (300 MHz, DMSO-d6) δ:
12.95 (1H, br s, 5-OH), 10.78 (1H, br s, 7-OH), 9.19 (1H, br
s, 4-OH), 7.56 (2H, m, H-2 , 6), 6.93 (1H, d, J = 8.4 Hz,
H-5), 6.88 (1H, s, H-3), 6.50 (1H, d, J = 1.2 Hz, H-8), 6.19
(1H, d, J = 1.2 Hz, H-6), 3.89 (3H, s, OCH3). Compound 1
was characterized as chrysoeriol by comparing the physical
and spectral data with the literature of Zhang et al [4].
Compound 2 Light yellow needles (MeOH), C17H14O7.
ESI-MS m/z 329 [M − H]−. 1H NMR (300 MHz, DMSO-d6) δ:
12.96 (1H, s, 5-OH), 10.77 (1H, s, 7-OH), 9.29 (1H, s,
4-OH), 7.32 (2H, s, H-2, 6), 6.96 (1H, s, H-3), 6.55 (1H, d,
J = 1.9 Hz, H-8), 6.20 (1H, d, J = 1.9 Hz, H-6), 3.88 (6H, s, 2
× OCH3). Compound 2 was characterized as tricin by com-
paring the physical and spectral data with the literature [5].
Compound 8 White plate crystal (MeOH). The mole-
cular formula of compound 8 C19H28O9 was deduced from
HR-ESI-MS (observed m/z 445.172 5 [M + HCOOH], calcd
m/z 445.171 5). The 1H NMR spectrum resembled that of
syringin (compound 12) except for the signals corresponding
to a ethoxy group: 3.58 (2H, q, J = 7.0 Hz, OCH2-CH3) and
1.22 (3H, t, J = 7.0 Hz, -CH2CH3). By comparing the NMR
spectrum data of compound 8 and compound 12 (Table 1),
compound 8 was characterized as ethylsyringin.
Compound 9 Yellow block crystals (MeOH), C21H20O10.
ESI-MS m/z 431 [M − H]−. 1H NMR (300 MHz, DMSO-d6) δ:
12.96 (1H, br s, 5-OH), 10.4 (1H, br s, 4-OH), 7.95 (2H, d, J
= 8.5 Hz, H-2, 6), 6.94 (2H, d, J = 8.5 Hz, H-3, 5), 6.86
(1H, s, H-3), 6.82 (1H, d, J = 1.8 Hz, H-8), 6.45 (1H, d, J =
1.9 Hz, H-6), 5.09 (1H, d, J = 8.5 Hz, H-1). These data were
identical with those of apigenin-7-O-β-D-glucopyranoside [6].
Compound 10 Pale yellow block crystal (MeOH),
C36H36O19. ESI-MS m/z 771 [M − H]−. 1H NMR (500 MHz,
DMSO-d6) δ: 7.44 (1H, dd, J = 8.4, 2.3 Hz, H-6), 7.41 (1H,
AGA Er-bu, et al. /Chinese Journal of Natural Medicines 2012, 10(5): 366−369
368 Chin J Nat Med Sep. 2012 Vol. 10 No. 5 2012 年 9 月 第 10 卷 第 5 期

d, J = 2.3 Hz, H-2), 6.97 (1H, d, J = 2.0 Hz, H-2), 6.94
(1H, d, J = 8.4 Hz, H-5), 6.91 (1H, dd, J = 8.2, 2.3 Hz,
H-6), 6.75 (1H, d, J = 2.2 Hz, H-6), 6.71 (1H, d, J = 8.2 Hz,
H-5), 6.68 (1H, s, H-3), 6.53 (1H, d, J = 2.2 Hz, H-8), 5.02
(1H, d, J = 7.2 Hz, H-1), 4.28 (1H, d, J = 7.8 Hz, H-1).
13C NMR data was given in Table 2. Compound 10 was
characterized as luteolin-7-O-β-D-glucopyranosyl-(1→6)-
[6-O-caffeoyl]-β-D-glucopyranoside by comparing the
physical and spectral data with the results reported by Fan et
al [7].
Table 1 1H and 13C NMR data (δ) of compounds 8 and 12 (CD3OD, 300 MHz, 75 MHz)
8 12
Position
δH (J in Hz) δC δH (J in Hz) δC
1 4.13 (2H, dd, J = 5.5, 1.3 Hz) 66.7 4.24 (2H, dd, J = 5.5, 1.3 Hz) 63.6
2 6.28 (1H, dt, J = 15.6, 6.0 Hz) 127.2 6.33 (1H, dt, J = 15.8, 5.6Hz) 130.1
3 6.60 (1H, d, J = 15.8 Hz, ) 133.2 6.56 (1H, d, J = 15.8 Hz,) 131.3
4 ─ 135.0 ─ 135.3
5 6.76 (1H, s) 105.6 6.76 (1H, s) 105.6
6 ─ 154.4 ─ 154.4
7 ─ 135.8 ─ 136.0
8 ─ 154.4 ─ 154.4
9 6.76 (1H, s) 105.6 6.76 (1H, s) 105.6
1 4.87 (1H, d, J = 7.2 Hz) 105.3 4.88 (1H, d, J = 7.2 Hz) 105.4
2 3.39 (1H, m) 75.7 3.39 (1H, m) 75.7
3 3.22 (1H, m) 77.8 3.22 (1H, m) 77.8
4 3.42 (1H, m) 71.4 3.42 (1H, m) 71.4
5 3.31 (1H, m) 78.4 3.31 (1H, m) 78.4
6 3.79 (1H, dd, J = 12.0 Hz, 2.5 Hz) 62.6 3.80 (1H, dd, J = 12.0 , 2.5 Hz) 62.6
3.67 (1H, dd, J = 12.0, 5.1 Hz) 3.68 (1H, dd, J = 12.0, 5.1 Hz)
-OCH3 3.86 (6H, s) 57.0 3.87 (6H, s) 57.0
-OCH2-CH3 3.58 (2H, q, J = 7.0 Hz) 48.8 ─ ─
-CH2CH3 1.22 (3H, t, J = 7.0 Hz) 15.4 ─ ─
Table 2 13C NMR (125 MHz) data of compounds 10 and 11 in DMSO-d6.
δC δC δC
Position
10 11
Position
10 11
Position
10 11
2 164.4 164.5 4 149.8 149.9 4 69.7 69.4
3 103.0 103.1 5 116.0 116.0 5 73.9 76.7
4 181.9 181.8 6 119.0 119.1 6 63.3 61.0
5 161.2 161.1 1 100.0 99.8 1 125.4 ─
6 99.6 99.6 2 73.1 73.1 2 115.0 ─
7 162.8 162.8 3 76.3 76.2 3 145.3 ─
8 94.8 94.7 4 69.8 70.0 4 148.2 ─
9 156.8 156.9 5 75.4 75.4 5 115.6 ─
10 105.3 105.3 6 69.4 69.4 6 121.4 ─
1 121.2 121.3 1 103.8 103.5 C-α 113.6 ─
2 113.5 113.5 2 73.6 73.4 C-β 145.2 ─
3 145.6 145.7 3 76.5 75.4 C=O 166.5 ─

Compound 11 Yellow powder (MeOH), C27H30O16.
ESI-MS m/z 609 [M − H]−. 1H NMR (300 MHz, DMSO-d6) δ:
7.48 (1H, dd, J = 8.3, 2.1 Hz, H-6), 7.43 (1H, d, J = 2.1 Hz,
H-2), 6.92 (1H, d, J = 8.3 Hz, H-5), 6.81 (1H, d, J = 2.1 Hz,
H-8), 6.73 (1H, s, H-3), 6.51 (1H, d, J = 2.1 Hz, H-6), 5.10
(1H, d, J = 7.0 Hz, H-1), 4.45 (1H, H-1), 4.19 (1H, d, J =
AGA Er-bu, et al. /Chinese Journal of Natural Medicines 2012, 10(5): 366−369
2012 年 9 月 第 10 卷 第 5 期 Chin J Nat Med Sep. 2012 Vol. 10 No. 5 369

7.6 Hz, H-6a), 4.00 (1H, d, J = 10.2 Hz, H-6b), 3.71 (1H,
dd, J = 11.1 Hz, H-6a), 3.64 (1H, d, J = 10.7 Hz, H-6b).
13C NMR data was given in Table 2. Compound 11 was
characterized as luteolin-7-O-β-D-gentiobioside by compar-
ing the physical and spectral data with those reported by Sun
et al [8].
Compound 12 White plate crystals (MeOH), C17H24O9.
ESI-MS m/z 417 [M + HCOOH]−. The 1H and 13C NMR data
were given in table 1. By comparing the physical and spectral
data with the data reported by Lee et al [9], compound 12 was
characterized as syringin .
Compound 15 Colorless needles (MeOH), C13H16O8.
ESI-MS m/z 299 [M − H]−. 1H NMR (300 MHz, CD3OD) δ:
7.97 (2H, m, H-2, 6), 7.15 (2H, m, H-3, 5), 5.01 (1H, m,
H-1); 13C NMR (75 MHz, CD3OD) δ: 169.56 (COOH),
162.79 (C-1), 125.65 (C-2, 6), 132.65 (C-3, 5), 117.15 (C-4),
101.67 (C-1), 74.82 (C-2), 78.26 (C-3), 71.30 (C-4), 77.94
(C-5), 62.46 (C-6). Compound 15 was characterized as
4-O-(β-D-glucopyranosyl)-benzoic acid by comparing of the
physical and spectral data with the literature of Qin et al [10].
Compounds 3-7, 13 and 14 were identified by comparing
with authentic samples and their structures were elucidated as
apigenin, succinic acid, β-sitosterol-3-O-β-D-glucopyrano-
side, luteolin, luteolin-7-O-β-D-glucopyranoside, 3-O-caf-
feoylquinic acid and 5-O-caffeoylquinic acid, respectively.
Acknowledgements
We appreciate the help from Prof. WANG Ju-Le and
ZHUOMA Dong-Zhi (Medical College of Tibet University,
China) for the collection of the plant material, and are grate-
ful to Prof. GESANG Ba-Zhu (Tibet Institute of Tibetan
Medicine, China) for the identification of the original plant
material.
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藏药脉花党参的化学成分
阿呷尔布, 李会军, 陈 君*, 李 萍*
天然药物活性组分与药效国家重点实验室(中国药科大学), 南京 210009
【摘 要】 目的：研究藏药脉花党参 Codonopsis nervosa ( Chipp.) Nannf.地上部分的化学成分。方法：采用 D101 大孔树脂
柱、硅胶柱、ODS 柱、凝胶柱等色谱手段进行化学分离和纯化, 并利用各种波谱技术进行结构鉴定。结果：分离并鉴定了 15 个
化合物, 分别为柯伊利素(1), 苜蓿素(2), 芹菜素(3), 琥珀酸(4), β-胡萝卜苷(5), 木犀草素(6), 木犀草素-7-O-β-D-葡萄糖苷(7),
ethylsyringin (8), 芹菜素-7-O-β-D-葡萄糖苷(9), 木犀草素-7-O-β-D-葡萄糖基-(1→6)-[(6-O-咖啡酸)-β-D-葡萄糖苷](10), 木犀草
素-7-O-β-D-龙胆二糖苷(11), 丁香苷(12), 绿原酸(13), 新绿原酸(14), 对羟基苯甲酸葡萄糖苷(15)。结论：化合物 1−5, 8, 9, 12, 14,
15 均首次从该植物中分离得到。
【关键词】 脉花党参; 桔梗科；藏药; 黄酮; 苯丙素

【基金项目】 国家“重大新药创制”科技重大专项(2011ZX09307-002-02)和“教育部新世纪优秀人才计划”(NCET-09-0769)资
助项目