全 文 ： 2012 年 11 月 第 10 卷 第 6 期 Chin J Nat Med Nov. 2012 Vol. 10 No. 6 477

Chinese Journal of Natural Medicines 2012, 10(6): 0477−0480
doi: 10.3724/SP.J.1009.2012.00477
Chinese
Journal of
Natural
Medicines







The structural elucidation and antimicrobial activities of
two new sesquiterpenes from Syringa pinnatifolia Hemsl.
AO Wu-Li-Ji 1, 2, WANG Qing-Hu 2*, Si-Qin 2, Mu-Dan 2, Sa-Ren-Tu-Ya 2,
DAI Na-Yin-Tai 2, Du-Ri-Si-Ha-La-Tu 2
1College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, China;
2College of Traditional Mongolian Medicine, Inner Mongolia University for Nationalities, Tongliao 028000, China
Available online 20 Nov. 2012
[ABSTRACT] AIM: To study the chemical constituents of Syringa pinnatifolia Hemsl. and their antibacterial and antifungal activi-
ties. METHODS: The CHCl3-soluble fraction was separated by chromatography, and the structures of the new compounds were elu-
cidated by spectral experiments. Antimicrobial activity of the compounds was evaluated against various bacteria and fungi. RESULTS:
Two new sesquiterpenes, guai-9-en-4β-ol (1) and 14, 15-dinorguai-1, 11-dien-9, 10-dione (2) were isolated from the stem of Syringa
pinnatifolia Hemsl. var. alashanensis. Compound 1 is more active against Bacillus coagulas, while being less active against Proteus
vulgaris. Compound 2 shows good activity against Escherichia coli, and is less active against Staphylococcus aureus.
CONCLUSIONS: Compounds 1 and 2 are new compounds, and when evaluated against various bacteria showed good results.
[KEY WORDS] Syringa pinnatifolia Hemsl.; Guai-9-en-4β-ol; 14, 15-Dinorguai-1, 11-dien-9,10-dione; Antimicrobial activity
[CLC Number] R284.1 [Document code] A [Article ID] 1672-3651(2012)06-0477-04

1 Introduction
Syringa pinnatifolia Hemsl. var. alashanensis Ma &
S.Q.Zhou (Syringa pinnatifolia Hemsl.) is a member of the
family Oleaceae, genus Syringa, and is found predominantly
in the spinney and scrub in the upland of Helan Mountain,
Inner Mongolia. The stem of Syringa pinnatifolia is one of
the best-known traditional herbal medicines frequently used
to treat cardiovascular symptoms in Mongolian medicine [1-3].
It is widely used in Mongolia as a substitute of the precious
materia medica, Lignum Aquilariae Resinatum., which is
used in treatment of asthma, cardiopalmus and angina pecto-
ris. Lignans [4], the essential oil [5], coumarin [5] and an-
thraquinone [5] have been isolated from this plant. The essen-
tial oil is mainly composed by sesquiterpenes, and more and
more attention has been paid to the pharmaceutical and me-
dicinal role of sesquiterpenes. Many research studies have
showed that sesquiterpenes have various biological activities,
including antibacterial, cytotoxic and antifungal activity [6-7].

[Received on] 19-Jun.-2012
[*Corresponding author] WANG Qing-Hu: Prof., Tel:
86-475-8314242; Fax: 86-475-8314242, E-mail:
WQH196812@163.com
These authors have no any conflict of interest to declare.
In a continuation of an earlier investigation of the plant, the
structural characterization of the new compounds by spectral
analysis and their antimicrobial activities are reported.
2 Apparatus and Reagents
All of the melting points were determined on a thermoe-
lectrically operated melting point apparatus and are uncor-
rected. Optical rotations were measured in CHCl3 at 25 ºС on
a Perkin-Elmer 241 polarimeter. The UV spectra were re-
corded on a Shimadzu UV-2201 spectrometer. The IR spectra
were recorded in KBr discs on a Thermo Nicolet 200 double
beam spectrophotometer. The HR-ESI-MS spectra were
measured on a Bruker Daltonics MicroTOFQ. NMR spectra
were measured on a Bruker ARX-600 NMR spectrometer
with tetramethylsilane (TMS) as the internal reference, and
chemical shifts are expressed in δ. Column chromatography
was performed by using silica gel (200–300 mesh, Marine
Chemical Factory, Qingdao, China) and Sephadex LH-20
(Pharmacia, Uppsala, Sweden). Fractions were monitored by
TLC (silica gel GF254 10–40 μm, Marine Chemical Factory,
Qingdao, China), and spots were visualized by heating silica
gel plates sprayed with 10% H2SO4 in EtOH.
The stems of Syringa pinnatifolia , used as experimental
AO Wuliji, et al. /Chinese Journal of Natural Medicines 2012, 10(6): 477−480
478 Chin J Nat Med Nov. 2012 Vol. 10 No. 6 2012 年 11 月 第 10 卷 第 6 期

material, were collected in Helan Mountain, Inner Mongolia
of China, in July 2010, and identified by Prof. Buhebateer
(Inner Mongolia University for Nationalities). A voucher (No.
20100726) has been deposited in the School of Traditional
Mongolian Medicine of Inner Mongolia University for Na-
tionalities.
3 Extraction and Isolation
The stems of Syringa pinnatifolia (3 Kg) were crushed
and extracted twice under reflux with 95% EtOH. Evapora-
tion of the solvent under reduced pressure afforded the 95%
EtOH extract. The extract was partitioned with petroleum
ether (P.E.), CHCl3, EtOAc and n-BuOH. The CHCl3-soluble
fraction (60.0 g) was separated by column chromatography
on silica gel and gradiently eluted with P.E. - EtOAc (80 : 1
to 5 : 1) to give 10 fractions (Fractions 1–10). Fraction 4 [300
mg, P.E. - EtOAc (40 : 1) eluate ] was loaded onto a column
of silica gel and eluted with P.E. - EtOAc (60 : 1 to 10 : 1) to
give five fractions (Fractions 4-1 to 4-5). Fraction 4-2 (60 mg)
was further purified by column chromatography on silica gel
with P.E. - EtOAc (30 : 1) to give 1 (18 mg). Fraction 4-3 (36
mg) was further chromatographed on Sephadex LH-20 col-
umn eluting with MeOH- CHCl3 (1 : 1), and then separated
by TLC (P.E.-acetone (10 : 1), yielding 2 (18 mg).
4 Antimicrobial Activity
The antimicrobial activity of compounds 1 and 2 were
determined by the filter paper disc diffusion method [8]. The
various bacterial species were first incubated at 45 °C for 48
h. The sterile filter paper discs (6 mm) were soaked with
standard antibacterial agent and various test samples and
were dried at 50 °C.
The discs were then placed on soft nutrient agar (2%)
petri plates previously seeded with suspension of each bacte-
rial species. The diameter of zone of inhibitions were meas-
ured at 37 ± 1 °C after 24 h.
For the antifungal activity, Saborauds broth media [9]
with 4% agar was used for the preparation of plates, and in-
cubated with spores and mycelium suspension of fungi ob-
tained from one week old culture. The diameter of zones of
inhibition were measured at 28 ± 1 °C after 48 h. The results
are recorded in Table 2.
5 Results and Discussion
The 95% ethanol extract of Syringa pinnatifolia was
suspended in water, then partitioned with petroleum ether,
CHCl3, EtOAc and n-BuOH. The CHCl3-soluble fraction was
separated by chromatography and affored two new ses-
quiterpenes, guai-9-en-4β-ol (1) and 14,15-dinorguai-1,11-
dien-9, 10-dione (2). Their structures are shown in Fig. 1, 1H
NMR and 13C NMR spectroscopic data of 1 and 2 are listed
in Table 1.

Fig. 1 Structures of compounds 1 and 2
Table 1 1H and 13C-NMR data (600 MHz for 1H, 150 MHz for 13C, in DMSO-d6) for compounds 1 and 2
Compound 1 Compound 2

δH (J in Hz) δc δH (J in Hz) δc
1 1.70 (m, 1H) 39.8 ＿ 144.5
2 2.00 (m, 1H) 1.22 (m, 1H) 22.6 6.19 (s, 1H) 129.4
3 1.79 (m, 1H) 1.42 (m, 1H) 42.1 2.69 (m, 1H) 54.2
4 ＿ 72.4 2.26 (dd, 1H, J = 13.8 Hz, J = 2.4 Hz) 1.64 (dd, 1H, J = 13.8 Hz, J = 12.6 Hz) 42.0
5 1.21 (m, 1H) 50.0 ＿ 42.8
6 1.64 (m, 1H) 1.59 (m, 1H) 21.9 2.48 (dd, 1H, J = 13.2 Hz, J = 3.0 Hz) 2.31 (dd, 1H, J = 13.2 Hz, J = 12.6 Hz) 45.9
7 1.01 (m, 1H) 46.7 2.21 (m, 1H) 41.1
8 1.97 (m, 2H) 30.9 3.06 (dd, 1H, J = 17.4 Hz, J = 2.4 Hz) 2.61 (dd, 1H, J = 17.4 Hz, J = 4.8 Hz) 31.9
9 5.50 (s, 1H) 122.3 ＿ 208.9
10 ＿ 145.0 ＿ 191.5
11 2.16 (m, 1H) 25.9 ＿ 146.5
12 0.92 (d, 3H, J = 13.2 Hz) 21.5 4.81 (s, 1H) 4.76 (s, 1H) 110.3
13 0.76 (d, 3H, J = 13.2 Hz) 15.1 1.77 (s, 3H) 20.4
14 1.67 (s, 3H) 23.8 1.80 (s, 3H) 10.7
15 1.10 (s, 3H) 20.7 1.40 (s, 3H,) 27.0


AO Wuliji, et al. /Chinese Journal of Natural Medicines 2012, 10(6): 477−480
2012 年 11 月 第 10 卷 第 6 期 Chin J Nat Med Nov. 2012 Vol. 10 No. 6 479

Compound 1 was obtained as colourless crystals. [α] 25
D
= +19° (CHCl3, c 0.10); IR (KBr) υmax: 3 403, 2 963, 2 927, 2
856, 1 548, 1 390, 1 361, 1 128, 1 078 cm–1. The molecular
formula was determined to be C15H26O by HR-ESI-MS at m/z
221.1886 [M – H]– (calcd. for 221.1890). The 13C NMR
spectral data (Table 1) indicated the presence of four methyls,
four methylenes, four methines as well as one trisubstituted
double bond and one hydroxylated quaternary carbon in the
molecule of 1. The signals at δ (C) 39.8, 22.6, 42.1, 72.4,
50.0, 21.9, 46.7, 30.9, 122.3, 145.0, 25.9, 21.5, 15.1, 23.8,
and 20.7, and comparison of these data with those of the
guaiane sesquiterpene, guai-6-en-10-ol [10-11] suggested that 1
had a guaiane skeleton with a double bond at C-9 and a hy-
droxyl group at C-4. The difference between the 13C NMR
spectroscopic data of 1 and those of guai-6-en-10-ol could be
explained by the effects of the hydroxyl at C-4 and the dou-
ble bond at C-9 in the last compound. The proton-bearing
carbon signals were assigned by analysis of the HMQC spec-
trum. In the 1H-1H COSY spectrum of 1, two H-atom systems
were established, the one was from H-1 to H-2, and H-2 to
H-3, the other was from H-5 through H-6, H-7 to H-8. In the
HMBC spectrum (Fig. 2), the methyl proton signal at δ 1.10
(15-CH3, s) was correlated with the carbon signal at δ 41.1
(C-3), 72.4, and 50.0 (C-5), so the oxygenated quaternary
carbon at δ 72.4 was assigned to the C-4 position, and the
methyl proton signal at δ 1.67 (14-CH3, s) was correlated
with the carbon signal at δ 39.8 (C-1), 122.3, and 145.0,
which indicated that the double bond was at C-9. Thus, the
primary structure of 1 was obtained and further confirmed by
the HMQC, 1H-1H COSY, and HMBC spectra. The relative
configuration at the chiral centers in compound 1 was sup-
ported by the ROESY spectrum. The ROESY correlations
were observed from 15-CH3 to H-5, H-1 to H-7. The NOE
interaction from H-1 to H-7 showed that these two protons
were on the same side. When they took a β-orientation,
15-CH3 and H-5 should be in an α-orientation, which was
supported by the NOESY interaction from H-5 to 15-CH3.
Thus, the structure of compound 1 was elucidated and named
as guai-9-en-4β-ol.

Fig. 2 Some key HMBC correlation of 1 and 2
Compound 2 was obtained as colourless crystals. [α] 25
D

+28° (CHCl3, c0.12); IR (KBr) υmax: 3 085, 2 959, 2 924, 2
860, 1 710, 1 692, 1 610, 1 393, 1 364, 1 193, 998, 907 cm–1.
The molecular formula was determined to be C15H20O2 by
HR-ESI-MS at m/z 231.136 4 [M – H]– (Calcd. for 231.138
0). The 13C NMR spectral data (Table 1) indicated the pres-
ence of three methyls, three methylenes, two methines, one
quaternary carbon, and two carbonyls, as well as one trisub-
stituted double bond and one terminal double bond in the
molecule of 2. The 1H NMR spectral data of 2 showed the
presence of two methyl groups linked to two quaternary car-
bons and three olefinic hydrogens (Table 1). The pro-
ton-bearing carbon signals were assigned by analysis of the
HMQC spectra, and the correlations observed in the HMBC
spectral (Fig. 2) confirmed the planar structure of 2, in which
the correlations of 15-CH3 (δH 1.40, s) with C-3 (δc 54.2),
C-2 (δc 129.4) and C-4 (δc 42.0), 14-CH3 (δH 1.80, s) with
C-4 (δc 42.0), C-5 (δc 42.8), C-1 (δc 144.5) and C-6 (δc 45.9)
as well as 13-CH3 (δH 1.77, s) with C-7 (δc 41.1), C-11 (δc
146.5) and C-12 (δc 110.3) indicated the three methyl groups
to be located at C-3, C-5 and C-7. The correlations of H-2
(δH 6.19, s) with C-5 (δc 42.8), C-4 (δc 42.0) and C-10 (δc
191.5), together with H-8 (δH 2.61, 306) with C-9 (δc 208.9),
C-10 (δc 191.5), C-7 (δc 41.1) and C-6 (δc 45.9) indicated
that the two carbonyls were attached to C-9 and C-10, re-
spectively. The relative configuration at the chiral centers in
compound 2 was supported by coupling constants and the
ROESY spectrum. In its 1H NMR spectra, the typical cou-
pling constants, J7, 6α = 12.6 Hz, J7, 6β = 3.0 Hz indicated that
H-7 was in a β-orientation, and in the ROESY spectrum, the
15-CH3 (δ 1.40) and CH3-14 (δ 1.80) showed strong correla-
tion with the protons at δ 1.64 (H-4α) and δ 2.31 (H-6α),
whereas the H-7 (δ 2.21) showed a crosspeak with the proton
at δ 2.48 (H-6β), which was supported by the NOESY inter-
action from 15-CH3 to 14-CH3 and from H-3 to H-7. Thus,
the structure of compound 2 was elucidated and the isolate
named as 14,15-dinorguai-1,11-dien-9,10-dione.
Antimicrobial activity of the above two compounds was
evaluated against various bacteria and fungi. The results re-
ported in Table 2 show that compound 1 is more active
against Bacillus coagulas, while being less active against
Proteus vulgaris. Compound 2 shows good activity against
Escherichia coli, and is less active against Staphylococcus
aureus. In the case of fungicidal activity, compound 1 shows
good activity against Aspergillus niger and is less active
Table 2 Antibacterial and antifungal activity of compounds 1 and 2
Compound Antibacterial activity (zone of inhibition in mm)
Antifungal activity
(zone of inhibition in mm)
E. coli S. aureus B. coagulas P. vulgaris P. digitatum F. oxysporum A. niger
1 11.02 13.41 15.34 9.67 12.56 11.64 13.20
2 15.34 9.45 12.01 14.96 12.34 15.32 11.53
Streptomycin 19.61 18.54 19.69 18.96 Griseofulvin 17.61 20.91 16.02

AO Wuliji, et al. /Chinese Journal of Natural Medicines 2012, 10(6): 477−480
480 Chin J Nat Med Nov. 2012 Vol. 10 No. 6 2012 年 11 月 第 10 卷 第 6 期

against Fusarium oxysporum. Compound 2 was found to be
highly active against Fusarium oxysporum, and less active
against Aspergillus niger.
Acknowledgements
The authors thank SHA Yi and LI Wen for the meas-
urements of the NMR spectra.
References
[1] Nei MG. Standard of traditional Mongolian medicinal materi-
als [M]. Chifeng: Inner Mongolia Science and Technology
Press, 1987: 360.
[2] Ma YQ. Inner Mongolia Flora [M]. Huhehaote: Inner Mongo-
lia People’s Press, 1989: 349.
[3] Qigeqi. Modern Mongolian Medicine [M]. Shenyang: Liaoning
Ethnical Publishing Company, 2002: 201.
[4] Ao WLJ, Bao XH, Wu XL, et al. Lignans from Syringa
pinnatifolia Hemsl. var. alashanensis [J]. J Asian Nat Prod Res,
2012, 14(4): 396-400.
[5] Yan Y, AO WLJ, Zhao XJ, et al. Effect of essential oil of
Syringa pinnatifolia Hemsl. var. alashanensis on ischemi of
myocardium, hypoxia and platelet aggregation [J]. J Ethno-
pharmacol, 2010, 131(6): 248–255.
[6] Zhang SJ, Zhao M, Bai LM, et al. Bioactive guaianolides from
Siyekucai (Ixeris chinensis) [J]. J Nat Prod, 2006, 69(5):
1425-1428.
[7] Seto M, Miyaso T, Umahara K, et al. Sesquiterpene lactones
from Cichorium endivia L. and C. intybus L. and cytotoxic ac-
tivity [J]. Chem Pharm Bull. 1988, 36(7): 2423-2429.
[8] Deohate PP, Berad BN. Synthesis and antimicrobial activity of
1, 3, 5-thiadiazines and their isomerism into 1, 3, 5-thiazines [J].
Indian J Chem, 2005, 44(8): 638-692.
[9] Maddila S, Palakondu L, Chunduri VR. Synthesis, antibacterial,
antifungal and antioxidant activity studies on 2-benzylthio- and
2-benzylsulfonyl-1H-imidazoles [J]. Der Pharm Lett, 2010,
2(6): 393-402.
[10] Lago JHG, Brochini CB, Roque NF. Sesquiterpens from essen-
tial oil fruits of Guarea magrophylla Vahl ssp. tuberculata [J].
J Essen Oil Res, 2002, 17(11): 255.
[11] Mei WL, Chen P, Wang H. Two new sesquiterpenes from
endophytic fungus S49 of Cephalotoxus hainanensis [J]. J
Asian Nat Prod Res, 2010, 12(7): 582-586.

山沉香中两个新倍半萜的结构鉴定和抗菌活性
奥乌力吉 1, 2, 王青虎 2*, 斯 钦 2, 牡 丹 2, 萨仁图雅 2, 代那音台 2, 都日斯哈拉图 2
1华中科技大学生命科学与技术学院, 武汉 430074;
2内蒙古民族大学蒙医药学院, 通辽 028000
【摘 要】 目的：对山沉香中化学成分的分离鉴定和抗菌活性进行研究。方法：运用柱层析等方法对山沉香氯仿层进行分
离纯化, 并通过波谱解析和理化鉴别进行结构鉴定。同时对分离得到化合物的抗菌活性进行了评估。结果：从山沉香分离得到
2个倍半萜类化合物：guai-9-en-4β-ol (1) 和 14, 15-dinorguai-1, 11-dien-9, 10-dione (2)。化合物 1 对 Bacillus coagulas 具有良
好的活性而对 Proteous vulgaris 活性较差; 化合物 2 对 Eschecheria coli 具有良好的活性而对 Staphylococcus aureus 活性较
差。结论：化合物 1 和 2 为新化合物, 具有较好的抗菌活性。
【关键词】 山沉香; Guai-9-en-4β-ol; 14, 15-Dinorguai-1, 11-dien-9, 10-dione; 抗菌活性