全 文 ：· 1258 · 药学学报 Acta Pharmaceutica Sinica 2009, 44 (11): 1258−1261



A new erythrodiol triterpene fatty ester from Scorzonera mongolica
WANG Bin1*, LI Guo-qiang2*, GUAN Hua-shi2, YANG Li-ye1, TONG Guo-zhong1
(1. School of Food and Pharmacy, Zhejiang Ocean University, Zhoushan 316000, China; 2. Key Laboratory of Marine Drugs,
Chinese Ministry of Education, School of Medicine and Pharmacy, Ocean University of China, Qingdao 266003, China)
Abstract: Two erythrodiol triterpene fatty esters, 3β-dodecanoyl erythrodiol (1) and 3β-tetradecanoyl
erythrodiol (2), were isolated from Scorzonera mongolica. Their structures were elucidated on the basis of IR,
MS and extensive 2D NMR spectroscopic analysis. Compound 1 was identified to be a new compound and 2
was confirmed to be a new natural compound. Their antitumor effects in vitro were evaluated with MTT and
SRB assays, but compounds 1 and 2 only showed moderate cytotoxicities on A-549 cell line.
Key words: Scorzonera mongolica; erythrodiol; triterpene fatty ester
CLC number: R284.1 Document code: A Article ID: 0513-4870 (2009) 11-1258-04
蒙古鸦葱中的一个新三萜脂肪酸酯
王 斌 1*, 李国强 2*, 管华诗 2, 杨立业 1, 童国忠 1
(1. 浙江海洋学院食品与药学学院, 浙江 舟山 316000;
2. 中国海洋大学医药学院教育部海洋药物重点实验室, 山东 青岛 266003)

摘要: 利用硅胶、Sephadex LH-20 及半制备 HPLC 等柱色谱方法从菊科鸦葱属植物蒙古鸦葱 Scorzonera
mongolica Maxim.的甲醇提取物中分离得到 2 个化合物, 根据理化性质和光谱数据鉴定其结构分别为 3β-十二酰
高根二醇 (1) 和 3β-十四酰高根二醇 (2)。其中, 化合物 1 为新化合物, 2 为新天然产物。体外细胞毒活性显示化
合物 1 和 2 对人肺癌细胞 A-549 显示一定的细胞毒活性。
关键词: 蒙古鸦葱; 高根二醇; 三萜脂肪酸酯

Scorzonera mongolica Maxim., one of the 32 known
species varieties of the genus Scorzonera found in
China, is mainly distributed in the north and north-
western regions of China[1]. Its tender stem is one of
the vegetables for Chinese people and the root is used
as traditional medicine drugs for the treatment of fever,
carbuncle mastitis and cancers[2]. These functional
characteristics of the plant are mainly due to the active
chemical composition. However, no previous chemical

Received 2009-07-03.
Project supported by the National 863 project (2007AA091701); Natural
Science Foundation of Zhejiang (Y2080579) and
Important Project of Zhejiang Ocean University
(21135030107).
*Corresponding author Tel: 86-580-2555005, Fax: 86-580-2554781,
E-mail: binwang4159@hotmail.com
Tel: 86-532-82032323, Fax: 86-532-82033054,
E-mail: liguoqiang@ouc.edu.cn
works on the species have been recorded, and only a
few articles on other members of the genus have re-
vealed the presence of sesquiterpenoids[3, 4], triterpe-
noids[5−7], steroids[5−7], flavonoids[7], tyrolobibenzyls[8−10]
and dihydroisocoumarins[11]. In the continuing research
for the bioactive components, the chemical constituents
of S. mongolica were studied systematically and two
new moradiol triterpene fatty esters ware reported[12].
Our detailed investigation has led to the discovery of
two erythrodiol triterpene fatty esters, 3β-dodecanoyl
erythrodiol 1 and 3β-tetradecanoyl erythrodiol 2
(Figure 1). In this paper, we report the isolation and
structure elucidation of these two compounds.

Results and discussion
Compound 1, a white amorphous powder, showed
DOI:10.16438/j.0513-4870.2009.11.004
WANG Bin, et al: A new erythrodiol triterpene fatty ester from Scorzonera mongolica · 1259 ·

light pink color upon treatment with sulfuric acid on a
TLC plate followed by heating. Its molecular formula
was established as C42H72O3Na by HR-SI-MS (m/z
647.537 0 [M+Na]+, calcd. for C42H72O3Na, 647.537 9).
The IR spectrum indicated the presence of ester
carbonyl group (1 705 cm−1) in its structure.


Figure 1 Structures of compounds 1, 1a and 2

The 1H NMR spectrum of compound 1 (Table 1)
exhibited eight methyl proton signals at δ 0.87 (9H, s,
CH3-24, 29, 30), 1.16 (3H, s, CH3-27), 0.96 (3H, s,
CH3-26), 0.94 (3H, s, CH3-25), 0.89 (3H, s, CH3-23)
and 0.88 (3H, t, J = 7.4 Hz, CH3-12), one oxygenated
methine proton signal at δ 4.48 (dd, 1H, J = 11.5, 5.5 Hz),
one olefinic proton signal at δ 5.19 (t, 1H, J = 3.5 Hz),
two hydroxylmethyl proton signals at δ 3.55 and 3.21 (d,
each 1H, J = 10.6 Hz), respectively. With analysis of
the 1H NMR in detail, the triplet of olefinic proton at δ
5.19 (t, J = 3.5 Hz) indicated that the olefinic carbon is
connected with a secondary carbon, so the double bond
could only locate at C-5/C-6, C-9/C-11 or C-12/C-13.
By comparing experimental data with those described
in the literature[13, 14], compound 1 was identified as
erythrodiol-type triterpenoids. A careful analysis of
DEPT, 1H-1H COSY and HMQC spectra permitted to
assigning all proton and carbon signals in the molecule
(Table 1). The double bond at C-12/C-13 was
confirmed by the key HMBC correlations between
δ 5.19 (1H, t, H-12) and δC 47.5 (C-9), 23.5 (C-11),
144.2 (C-13), 41.7 (C-14) and the hydroxylmethyl
group at C-28 was also confirmed by the key
HMBC correlations between δH 3.55, 3.21 (each 1 H, d,
H-28) and 22.0 (C-16), 42.3 (C-18), 31.0 (C-22). The
presence of a dodecanoyl ester was deduced from the
characteristic fragment ion peak in the mass spectrum at
m/z 425 (M-C12H23O2−), due to the loss of a dodecanoyl
saturated fatty acid; and the location of dodecanoyl
group was determined to link at C-3 according to the
HMBC correlations between at H-3 (δH 4.48) and C-1
(δC 173.7). So according to 1D, 2D NMR data and the
data in literature[13], especially the NOESY correlations
between H-24 and H-25, H-25 and H-26, H-26 and
H-12, the relative stereochemistry of 1 was as shown in
Figure 2 and the structure of 1 was determined as
3β-dodecanoyl-28-hydroxyl-olean-12-ene.

Table 1 The 1H (600 MHz) and 13C (150 MHz) NMR data of
compounds 1, 1a and 2 in CDCl3 (δ in Hz)
No. δC (1) δH (1) δC (1a)[13] δC (2)
1 38.2 1.57 (m, 1H), 1.05 (m, 1H) 38.2 38.2
2 23.5 1.90 (m, 1H), 1.25 (m, 1H) 25.5 23.5
3 80.5 4.48 (dd, 1H, 11.5, 5.5) 80.9 80.5
4 37.7 − 37.7 37.7
5 55.2 0.83 (1H, br d, 9.6) 55.2 55.2
6 18.2 1.52 (m, 1H), 1.34 (m, 1H) 18.2 18.2
7 32.5 1.50 (m, 1H), 1.36 (m, 1H) 32.5 32.5
8 39.8 − 39.8 39.8
9 47.5 1.58 (m, 1H) 47.5 47.5
10 36.8 − 36.8 36.8
11 23.5 1.25 (m, 1H), 1.16 (m, 1H) 23.5 23.5
12 122.3 5.19 (t, 1H, 3.5) 122.2 122.3
13 144.2 − 144.2 144.2
14 41.7 − 41.7 41.7
15 25.5 1.62 (m, 1H), 1.25 (m, 1H) 25.5 25.5
16 22.0 1.63 (m, 1H), 1.32 (m, 1H) 22.0 22.0
17 36.9 1.89 (m, 2H) 36.9 36.9
18 42.3 2.30 (m, 1H) 42.3 42.3
19 46.4 1.19 (m, 1H), 1.88 (m, 1H) 46.4 46.4
20 31.0 − 30.9 31.0
21 34.1 1.25 (m, 1H), 1.29 (m, 1H) 34.1 34.1
22 31.0 1.25 (m, 1H), 1.54 (m, 1H) 31.0 31.0
23 28.0 0.89 (s, 3H) 28.0 28.0
24 15.6 0.87 (s, 3H) 15.5 15.6
25 16.7 0.94 (s, 3H) 15.5 16.7
26 16.8 0.96 (s, 3H) 16.7 16.8
27 25.9 1.16 (s, 3H) 25.9 25.9
28 69.8 3.55 (d, 1H, 10.6), 3.21 (d, 1H, 10.6) 69.7 69.8
29 34.1 0.87 (s, 3H) 33.2 34.1
30 23.6 0.87 (s, 3H) 23.6 23.6
1 173.7 − 171.0 173.7
2 34.9 2.30 (t ,2H, 7.2) 21.3 34.9
3 25.2 1.62 (m, 2H) 25.2
4−7 29.7 1.27 (m, 24H) 29.7
8 29.7 1.27 (m, 2H) 29.7
9 29.7 1.27 (m, 2H) 29.7
10 31.9 1.27 (m, 2H) 29.7
11 22.7 1.27 (m, 2H) 29.7
12 14.1 0.88 (t, 3H, 7.4) 31.9
13 22.7
14 14.1
· 1260 · 药学学报 Acta Pharmaceutica Sinica 2009, 44 (11): 1258−1261


Figure 2 The key NOESY (H↔H) and HMBC (H→C) correla-
tions of compound 1

Compound 2 was obtained as white amorphous
powder. The 1H and 13C NMR spectral data of 2
were almost identical with those of 1 (Table 1), but the
obvious difference was observed from the retention
time in HPLC, which were respectively 120 minutes
and 170 minutes under HPLC (Kromasil C18, 10 μm,
250 mm × 10 mm, methanol as mobile phase, flow rate
1.5 mL·min−1). Its molecular formula was determined
as C44H76O3Na according to HR-SI-MS (m/z 675.568 6
[M+Na]+, calcd. for C44H76O3Na, 675.569 2), and the
molecular weight of 2 was 28.031 6 amu which is more
heavy than that of 1. Based on the above analysis, the
structure of 2 was identified as 3β-tetradecanoyl-28-
hydroxyl-olean-12-ene (Figure 1).
Using the MTT and SRB dye reduction assays, the
cytotoxicity of crude extract and fractions A to E at the
concentration of 50 µg·mL−1 against P-388, A-549 and
Bel-7402 human hepatoma cell lines was tested.
Higher activity of fraction B and C was observed
(Table 2).

Table 2 Cytotoxicity of different fractions of S. mongolica
Maxim. on three tumor cell lines
Inhibitory rate / %
Sample Mouse leukemic
cell line P-388
Human lung cancer
cell line A-549
Human hepatoma
cell line Bel-7402
Crude extract 34.6 ± 0.24 66.4 ± 0.53 53.9 ± 0.42
Fraction A 5.8 ± 0. 44 6.8 ± 0.10 3.7 ± 0.23
Fraction B 57.2 ± 0.29 83.7 ± 0.37 78.6 ± 0.15
Fraction C 49.2 ± 0.26 86.7 ± 0.23 83.0 ± 0.28
Fraction D 10.3 ± 0.35 12.8 ± 0.42 4.2 ± 0.53
Fraction E 6.9 ± 0.54 6.2 ± 0.38 4.0 ± 0.27
Inhibition ratio (%) = mean ± SD, n = 3

Cytotoxic activity was evaluated by using the
P-388, Bel-7402 cell lines with the MTT method and
the A-549 cell line by the SRB method, the inhibition
ratio (%) of the compounds on each cell lines is
presented in Table 3. The results indicated that
compounds 1 and 2 only showed moderate cytotoxicity
on A-549 cell line with the inhabitation rate 66.8% and
69.8% at the concentration of 50 µg·mL−1, respectively.

Table 3 Effects of compounds 1 and 2 on three tumor cell lines
at the concentration of 50 µg·mL−1
Inhibitory rate / %
Compound Mouse leukemic
cell line P-388
Human lung cancer
cell line A-549
Human hepatoma
cell line Bel-7402
1 21.0 ± 0.21 66.8 ± 0.03 22.5 ± 0.11
2 13.6 ± 0.33 69.8 ± 0.17 15.3 ± 0.48
Inhibition ratio (%) = mean ± SD, n = 3

Experimental
General experimental procedures Optical
rotations were obtained on a JASCO P-1020 digital po-
larimeter. IR spectra were taken on a Nicolet NEXUS
470 spectrophotometer in KBr disks. 1H, 13C NMR
and DEPT and 2D NMR were recorded on a JEOL
JNM-ECP-600 spectrometer using TMS as internal
standard, and chemical shifts were recorded as δ values.
NOESY experiments were carried out using a mixing
time of 0.5 s. 1D NOE spectra were obtained on a
Varian INOVA-400 spectrometer. HR-SI-MS were
performed with an AEI-MS-50 mass spectrometer, and
ESI-MS were recorded in Q-STAR ESI-TOFMS/ MS
mass spectrometer. Column chromatography (CC)
was carried out with silica gel (200–300 mesh), and
HF254 silica gel for TLC was provided by Qingdao
Marine Chemistry Company. Higher pressure liquid
chromatography (HPLC) was performed on a Alltech
426 apparatus using Kromasil prepack column (ODS,
250 mm × 10 mm, for reverse phase) and monitored by
UV detector.
Plant material The original plant S. mongolica
was collected in Dongying (Shandong province) in
October, 2005, and authenticated by Prof. ZHOU
Feng-qin, College of Traditional Chinese Medicine,
Shandong University of Traditional Chinese Medicine.
The voucher specimen has been deposited in Institute of
Marine Drugs and Food, Ocean University of China
under reference DY20051007.
Extraction and isolation The air-dried plant
(1.2 kg) was pulverized and extracted with methanol
three times (7 days each time) at room temperature.
The solvent was evaporated under reduced pressure,
and the residue (100 g) was subjected to silica gel
column chromatography, eluted with increasing polarity
from petroleum ether to acetone to give five fractions
(A-E). Fraction C was purified by silica gel CC with a
WANG Bin, et al: A new erythrodiol triterpene fatty ester from Scorzonera mongolica · 1261 ·

gradient of petroleum ether–acetone (10∶1) and semi-
preparative HPLC employing isocratic elution with
methanol to give compound 1 (3.4 mg) and 2 (2.8 mg).
Cytotoxicity determination Cytotoxic activity
was evaluated using the P-388, Bel-7402 cell lines by
the MTT method and the A-549 cell line by the SRB
method[15].
In the MTT assay, the cell line was grown in
RPMI-1640 supplemented with 10% FBS under a
humidified atmosphere of 5% CO2 and 95% air at 37 ℃.
Cell suspensions (200 μL) at a density of 5 × 104 cells/
mL were plated in 96-well microtiter plates and
incubated for 24 h. The test compound solutions
(2 μL in MeOH) at different concentrations were added
to each well and further incubated for 72 h under the
same condition. MTT solution (20 μL of a 5 mg·mL−1
solution in RPMI-1640 medium) was added to each
well and incubated for 4 h. Old medium (150 μL)
containing MTT was gently replaced by DMSO and
pipetted to dissolve any formazan crystals formed.
Absorbance was then determined on a Spectra Max Plus
plate reader at 540 nm.
In the SRB assay, cell suspensions (200 μL) were
plated in 96-cell plates at a density of 2 × 105 cell/mL.
Then the test compound solutions (2 μL in MeOH) at
the concentration of 50 µg·mL−1 were added to each
well and further incubated for 24 h. Following drug
exposure, the cells were fixed with 12% trichloroacetic
acid and the cell layer was stained with 0.4% SRB.
The absorbance of SRB solution was measured at 515
nm.
Inhibition ratio (%) = ((Acontrol − Asample) / Acontrol) × 100
Where the Acontrol is the absorbance of the control, the
Asample is the absorbance of the test sample.

Identification
Compound 1 White amorphous powder. [α] 20D
+14.4 (c 0.25, CHCl3), IR (ν
KBr
max cm
−1) 3 463, 2 923,
2 852, 1 705, 1 464, 1 358, 1 245, 905, 845; HR-SI-
MS: m/z 647.537 0 [M+Na]+ (calcd. For C42H72O3Na,
647.537 9). 1H NMR and 13C NMR data see Table 1.
Compound 2 White amorphous powder. [α] 20D
+14.8 (c 0.25, CHCl3), IR (ν
KBr
max cm
−1) 3 463, 2 923,
2 852, 1 705, 1 464, 1 358, 1 245, 905, 845; HR-SI-
MS: m/z 675.568 6 [M+Na]+ (calcd. For C44H76O3Na,
675.569 2). 1H NMR and 13C NMR data see Table 1.
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