全 文 ： 药学学报 Acta Pharmaceutica Sinica 2009, 44 (8): 891−894 · 891 ·



Triterpenoid from Lycopodium obscurum L.
DENG Tie-zhong, AI Yong, CHEN Yu, YANG Guang-zhong*
(Laboratory for Natural Product Chemistry, College of Pharmacy, South Central University for Nationalities, Wuhan 430074, China)
Abstract: To study the triterpenoid constituents from air-dried whole plants of Lycopodium obscurum L., the
constituents were isolated by normal-phase and reverse-phase silica gel column chromatorgraphy from the
EtOAc extract. Their structures were elucidated on the basis of spectral analysis. Five triterpenoids were
purified and identified as 3β, 19α-dihydroxy-20β-acetate-serrat-14-en-21β-ol (1), serratenediol (2), α-onocerin
(3), 26-nor-8-oxo-α-onocerin (4), (3β, 8β, 14α, 21α)-26, 27-dinoronocerane-3, 8, 14, 21-tetrol (5). Compound 1
is a new serratane-type triterpene and compound 5 is isolated from this plant for the first time.
Key words: Lycopodium; Lycopodium obscurum L.; serratane triterpenoid
CLC number: R284.1 Document code: A Article ID: 0513-4870 (2009) 08-0891-04
玉柏石松三萜成分研究
邓铁忠, 艾 勇, 陈 玉, 杨光忠*
(中南民族大学药学院天然产物化学研究室, 湖北 武汉 430074)

摘要: 为了研究玉柏石松全草的三萜类成分, 运用正相和反相硅胶柱色谱技术对玉柏石松的乙酸乙酯提
取物进行分离纯化, 并运用波谱技术对其结构进行鉴定。共分离得到 5 个三萜类化合物, 其结构分别鉴定为 3β,
19α-dihydroxy-20β-acetate-serrat-14-en-21β-ol (1), serratenediol (2), α-onocerin (3), 26-nor-8-oxo-α-onocerin (4), (3β,
8β, 14α, 21α)-26, 27-dinoronocerane-3, 8, 14, 21-tetrol (5)。其中化合物 1 为新的石松型三萜化合物, 化合物 5 为首
次从该植物中分离得到。
关键词: 石松属; 玉柏石松; 石松三萜

Lycopodium obscurum L. is widely used as traditional
Chinese herbal medicine for the treatment of arthritic
pain, quadriplegia, dysmenorrhea, and contusion[1].
Previous phytochemical studies of this plant had
showed that serratane-type triterpenoids[2] or deriva-
tives of its biogenetic precursor α-onocerin[3] were
well-known characteristic constituents. Serratane is
an unique family of pentacyclic triterpenoids possessing
seven tertiary methyls and a central seven-member
C-ring, usually with a C=C bond between C (14) and
C (15) and O-functionalities at C (3) and C (21)[4].
Pharmacological studies have shown that serratane-type

Received 2009-02-16.
Project supported by National Natural Science Foundation of China
(No.30670215).
*Corresponding author Tel / Fax: 86-27-67841196,
E-mail: yanggz888@126.com
triterpenoids have pharmacological action, including
cancer chemopreventive activity[5] and inhibitory effects
against Candida albicans secreted aspartic proteases[6].
As part of research projects on the study of ethnomedi-
cinal plants, we have investigated L. obscurum and
reported herein the isolation and structure elucidation
of a new serratane-type triterpenoid together with four
known compounds: serratenediol (2), α-onocerin (3),
26-nor-8-oxo-α-onocerin (4), and (3β, 8β, 14α, 21α)-26,
27-dinoronocerane-3, 8, 14, 21-tetrol (5) (Figure 1).
Compound 1 was obtained as white amorphous
powder, its molecular formula was deduced as
C32H52O5 by HR-EI-MS (found [M]+ m/z 516.380 7,
calcd. 516.381 4), requiring seven degrees of unsatura-
tion. The NMR spectral showed the presence of an
acetyl group at δC 171.4, δC 21.3, and δH 1.93 (3H, s),
which was supported by the ion fragment at m/z
DOI:10.16438/j.0513-4870.2009.08.007
· 892 · 药学学报 Acta Pharmaceutica Sinica 2009, 44 (8): 891−894


Figure 1 Structures of compound 1−5

456.359 2 [M-CH3COOH]+. With the exception of an
acetyl group signal, its 1H NMR (Table 1) showed
seven tertiary methyls (δ 0.83 (3H, s), 0.87 (3H, s), 1.06
(3H, s), 1.17 (3H, s), 1.20 (3H, s), 1.24 (3H, s) and 1.26
(3H, s)), three hydroxy groups (δ 6.30 (1H, d, J = 4.8
Hz), 6.70 (1H, d, J = 4.2 Hz), 5.80 (1H, s)), and four
oxygenated methines (δ 3.52 (1H, dd, J = 14.8, 8.4 Hz),
4.08 (1H, br s), 4.74 (1H, dd, J = 10.0, 4.8 Hz), 5.71
(1H, dd, J = 10.0, 2.4 Hz)), and an olefinic proton
δ 5.54 (1H, br s). The 13C NMR (Table 1) and DEPT
spectrum of 1 displayed 32 carbon signals, including
a C=C bond at δC 139.8 (s), δC 122.8 (d), four
oxygenated methines, seven methyls, eight methylenes,
four methines, as well as five quarternary carbons.
All the above data suggested that 1 belongs to a
serratane-type triterpenoid with an acetyl group as ester
substituent[7]. Comparison of the 1H and 13C NMR
spectroscopic data of 1 with those of the known com-
pounds serratenediol[8], showed that the two compounds
were very similar. Compound 1 and serratenediol
were mainly different in the ring E. With the help of
1H-1H COSY and HSQC spectrum, the partial structure
-CH(O)-CH(O)-CH(O)- was deduced in ring E. Thus,
two CH2 (19) and CH2 (20) signals in serratenediol
were replaced by two oxygenated CH in 1 which was
supported by HMBC correlations H-19/C-13 (δC 58.9),
C-18 (δC 43.1), C-20 (δC 76.7), and H-20/ C-19 (δC
74.6), and H-21/C-17 (δC 43.3), C-19 (δC 74.6), C-20
(δC 76.7), C-22 (δC 38.5). The ester unit was positioned
at C-20 on the basis of HMBC correlation H-20/δC
171.4. The relative configuration of 1 was derived
from the ROESY spectrum. The β-orientation of the
3-OH was deduced from the correlations of H-3/ H3-23,
H-5. The ROSEY correlations H-19/H-13 and H-13/
H3-26 suggested that 19-OH was α-oriented. The
20-OAc and 21-OH was deduced to be in β-position
which was based on the correlations H3-29/H-20, H-21
and H-20/H-21 (Figure 2). Therefore, compound 1
was elucidated as 3β, 19α-dihydroxy-20β-acetate-serrat-
14-en-21β-ol.

Table 1 1H NMR, 13C NMR and HMBC data of compound 1
(in C5D5N)
Position 1H NMR 13C NMR HMBC
1

0.95 (1H, m)
1.71 (1H, m)
38.8


2 1.88 (2H, m) 28.1
3

3.52 (1H, dd, J = 14.8,
8.4 Hz)
77.8

C (2), C (4), C (23), C (24)

4 39.4
5 0.85 (1H, m) 55.8 C (4), C (10)
6 1.54 (2H, m) 19.2
7 1.43 (2H, m) 45.3 C (5), C (8)
8 37.2
9 0.81 (1H, m) 62.8 C (8), C (10)
10 38.1
11 1.85 (2H, m) 27.5
12

1.91 (1H, m)
3.45 (1H, t, J = 7.8 Hz)
26.9

C (9), C (13), C (14)

13

2.48 (1H, br d, J =
11.4 Hz)
58.9


14 139.8
15 5.54 (1H, br s) 122.8
16 2.10 (2H, m) 24.7
17

2.40 (1H, m)

43.3

C (13), C (16), C (18),
C (22), C (29)
18 43.1
19

4.74 (1H, dd, J = 10.0,
4.8 Hz)
74.6

C (13), C (18), C (20)

20

5.71 (1H, dd, J = 10.0,
2.4 Hz)
76.7

C (19), C (31)

21 4.08 (1H, br s) 77.2 C (17), C (19), C (20), C (22)
22 38.5
23 1.20 (3H, s) 28.5 C (3), C (4), C (5), C (24)
24 1.06 (3H, s) 16.2 C (3), C (4), C (5), C (23)
25 0.83 (3H, s) 15.9 C (1), C (5), C (9), C (10)
26 0.87 (3H, s) 20.0 C (7), C (8), C (9), C (27)
27

1.88 (1H, s)
2.45 (1H, m)
56.3

C (7), C (8), C (9), C (13),
C (14), C (15), C (26)
28 1.26 (3H, s) 9.8 C (13), C (18), C (17), C (19)
29 1.17 (3H, s) 21.3 C (17), C (21), C (22), C (30)
30 1.24 (3H, s) 28.4 C (17), C (21), C (22), C (29)
31 171.4
32 1.93 (3H, s) 21.3 C (31)
OH 6.70 (1H, d, J = 4.2 Hz)
OH 6.30 (1H, d, J = 4.8 Hz)
OH 5.80 (1H, d, br s)

DENG Tie-zhong, et al: Triterpenoid from Lycopodium obscurum L. · 893 ·


Figure 2 Key ROESY of compound 1

Table 2 13C NMR data of compound 2−5
Position 2a 3a 4a 5b
1 38.0 37.4 37.7 36.3
2 28.8 28.6 29.1 27.8
3 78.3 77.9 78.1 79.7
4 40.0 39.6 40.0 39.0
5 56.4 54.8 55.3 56.0
6 19.8 24.4 24.7 17.6
7 45.9 38.5 42.8 41.1
8 37.8 149.1 211.9 67.7
9 63.3 57.7 65.1 56.1
10 39.4 39.6 42.9 38.7
11 25.9 23.0 22.0 23.5
12 27.9 23.0 24.2 23.5
13 58.0 57.7 58.0 56.1
14 139.1 149.1 148.8 67.2
15 123.1 38.5 39.0 41.1
16 25.0 24.4 24.5 17.6
17 50.4 54.8 53.9 56.6
18 38.8 39.6 40.0 38.7
19 36.8 37.4 37.9 36.3
20 29.1 28.6 29.1 27.8
21 78.6 77.9 78.4 79.7
22 39.8 39.6 40.1 39.0
23 28.7 28.8 29.2 29.0
24 16.5 16.2 16.6 16.6
25 16.8 14.7 15.2 16.3
26 20.5 106.7
27 56.9 106.7 108.3
28 14.2 14.7 15.3 16.3
29 15.9 16.2 16.4 16.6
30 29.0 28.8 29.2 29.0
ain C5D5N; bin CD3OD

Experimental
General experimental procedures NMR spectra
were run on Bruker AM-300 and 600 spectrometers
with TMS as internal standard. EI-MS and HR-EI-MS
were measured with a Finnigan MAT 95 instrument.
Thin-layer chromatography (TLC) was performed on
silica gel 60 GF254, and column chromatography was
carried using silica gel (200−300 mesh) from Qingdao
Haiyang Chemical Group Co., China and C18 reversed-
phase silica gel from YMC CO., LTD., Japan.
Plant material The whole plant of Lycopodium
obscurum L. was collected from Jianshi county, Hubei
province, China and identified by Professor WAN
Ding-rong, College of Pharmacy, South Central
University for Nationalities.
Extraction and isolation The air-dried whole
plant of Lycopodium obscurum L. (12.3 kg) was
powdered and then extracted with MeOH three times at
room temperature. The MeOH extract (1.65 kg) was
suspended in 3% tartaric acid/H2O (pH 3) and then par-
titioned with EtOAc. The EtOAc extract (890 g) was
suspended in 90% H2O/MeOH and then successively
partitioned with petroleum ether (PE), EtOAc and
n-BuOH. The EtOAc extract (324 g) was subjected to
CC (silica gel, PE:acetone 9∶1, 8∶2, 7∶3, 1∶1,
3∶7, 0∶1, v/v) to give 9 fractions (Fr.1−Fr.9). Fr.5
was subjected to CC (silica gel, CHCl3-acetone 1∶0 →
1∶1) to give 5 subfractions (Fr.5.1−Fr.5.5). Fr.5.4
was subjected to CC (ODS, H2O-MeOH 9∶1 → 1∶9)
to give compound 1 (4 mg). Compound 3 (40 mg)
was recrystallized from Fr.5.2. Fr.5.3 was subjected
to CC (silica gel, cyclohexane-EtOAc 9∶1 → 1∶1) to
give 4 subfractions (Fr.5.3.1−Fr.5.3.4), Fr.5.3.3 was
subjected to CC (ODS, H2O-MeOH 7∶3 → 3∶7) to
give compound 4 (10 mg). Fr.2 was subjected to CC
(silica gel, cyclohexane-acetone 99∶1 → 7∶3) to give
5 subfractions (Fr.2.1−Fr.2.5). Fr.2.3 was subjected to
CC (silica gel, cyclohexane-EtOAc 98∶2 → 9∶1) to
obtain compound 2 (29 mg). Fr.7 was subjected to CC
(ODS, H2O-MeOH 7∶3 → 1∶0) to give 7 subfractions
(Fr.7.1−Fr.7.7). Fr.7.4 was subjected to CC (silica gel,
CHCl3-EtOAc 9∶1 → 1∶1) to give 4 subfractions
(Fr.7.4.1−Fr.7.4.4). Fr.7.4.2 was subjected to CC
(ODS, H2O-MeOH 7∶3 → 3∶7) to give compound 5
(10 mg).
Identification of compounds 1−5
Compound 1: white amorphous powder, C32H52O5,
EI-MS (70 eV) m/z 516 [M]+ (32), 456 (19), 438 (20),
294 (23), 207(95), 190 (100), 189 (74), 135 (42),
121(51), 107(42); HR-EI-MS m/z 516.380 7 (calcd. for
C32H52O5, 516.381 4). 1H NMR and 13C NMR data
were listed in Table 1.
Compound 2: white amorphous powder, C30H50O2,
1H NMR (600 MHz, C5D5N) δ: 0.81 (3H, s, CH3), 0.89
(3H, s, CH3), 0.96 (3H, s, CH3), 1.08 (3H, s, CH3), 1.13
· 894 · 药学学报 Acta Pharmaceutica Sinica 2009, 44 (8): 891−894

(3H, s, CH3), 1.21 (3H, s, CH3), 1.24 (3H, s, CH3), 3.54
(1H, dd, J = 10.2, 5.4 Hz, H-3), 3.49 (1H, dd, J = 8.4,
7.8 Hz, H-21), 5.49 (1H, s, H-15), 5.85(1H, br s, OH),
5.95 (1H, br s, OH); 13C NMR data were listed in Table
2. These data were identical to the literature values of
serratenediol[8].
Compound 3: white amorphous powder, C30H50O2,
1H NMR (600 MHz, C5D5N) δ: 0.78 (6H, s, H3-25, 28),
1.05 (6H, s, H3-24, 30), 1.23 (6H, s, H3-23, 29), 3.52
(2H, dd, J = 11.4, 4.8 Hz, H-3, 21), 4.80 (2H, s, H2-26,
27), 5.05 (2H, s, H2-26, 27), 5.90 (2H, s, 3, 21-OH);
13C NMR data were listed in Table 2. These data
coincide with the literature values of α-onocerin[3].
Compound 4: white amorphous powder, C29H48O3,
1H NMR (600 MHz, C5D5N) δ: 0.73 (3H, s, CH3), 0.74
(3H, s, CH3), 0.98 (3H, s, CH3), 1.00 (3H, s, CH3), 1.20
(3H, s, CH3), 1.25 (3H, s, CH3), 3.50 (2H, m, H-3, 21),
5.07 (1H, s, H2-27), 5.29 (1H, s, H2-27); 13C NMR data
were listed in Table 2. These data agree with the
literature values of 26-nor-8-oxo-α-onocerin[3].
Compound 5: white amorphous powder, C28H50O4,
1H NMR (300 MHz, CD3OD) δ: 0.79 (6H, s, 2×CH3),
0.97 (6H, s, 2×CH3), 0.98 (6H, s, 2×CH3), 3.12 (2H, dd,
J = 11.7, 6.0 Hz, H-3, 21), 3.94 (2H, br s, H-8, 21);
13C NMR data were listed in Table 2. These data
correspond to the literature values of (3β, 8β, 14α,
21α)-26, 27-dinoronocerane-3, 8, 14, 21-tetrol[9].
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