全 文 ： 2009 年 11 月 第 7 卷 第 6 期 Chin J Nat Med Nov. 2009 Vol. 7 No. 6 436

Chinese Journal of Natural Medicines 2009, 7(6): 0436−0439
doi: 10.3724/SP.J.1009.2009.00439
Chinese
Journal of
Natural
Medicines








Analysis of Secoiridoid Glucosides in Jasminum lanceo-
larium Roxb. by HPLC-MS
SUN Jia-Ming1,2, ZHANG Hui1*, YANG Jun-Shan2*
1Development Center of Traditional Chinese Medicine and Bioengineering, Changchun University of Traditional Chinese Medicine,
Changchun 130117;
2Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing
100193,China
Available online 20 Nov. 2009
[ABSTRACT] AIM: To develop high performance liquid chromatography-electrospray ionization-mass spectrometry (HPLC-ESI-
MS) for the analysis and identification of secoiridoid glucosides in Jasminum lanceolarium Roxb. METHOD: The ultraviolet (UV)
chromatogram, total ion current chromatogram (TIC) and extracted ion chromatogram (EIC) of the sample and ESI/MS/MS spectra of
the peaks in the chromatograms were obtained in order to identify the structure of secoiridoid glucosides. RESULT: The six identified
secoiridoid glucosides were Jasminoside, Jaslanceoside A, Jaslanceoside B, Jaslanceoside C, Jaslanceoside D and Jaslanceoside E. The
structures of another four unknown secoiridoid glucosides were speculated. CONCLUSION: The developed simple and rapid method
was useful to isolate the new secoiridoid glycoside from Jasminum lanceolarium quickly and to authenticate the species of the herb.
[KEY WORDS] Jasminum lanceolarium Roxb.; HPLC-ESI-MS; Secoiridoid glucosides
[CLC Number] R917 [Document code] A [Article ID]1672-3651(2009)06-0436-04

1 Introduction
Jasminum lanceolarium is a climbing shrub distributed
over thickets at low altitudes from south-eastern China to
India[1]. Its stems and roots are used in Chinese medicine for
the treatment of fever and rheumatic pain. The leaves are also
used as an antiinflammatory agent for releasing eye pain[2].
Previous phytochemical investigation resulted in the isolation
of hundreds of secoiridoid glucosides in Jasminum. Nine
secoiridoid glucosides have been isolated from the stems and
leaves of J. lanceolarium[2,3]. Therefore quantitative evalua-
tion of the secoiridoid glucosides in J. lanceolarium is sig-
nificant for guiding the isolation of new compounds and the
quality control of this medicinal herb.
Liquid chromatography combined with tandem mass
spectrometry (LC-MS-MS), a relatively new technique with
rapidly growing popularity, has been successfully applied to
elucidate the structures of the active components in medicinal
plants. Recently, it was also employed to determine the con-

[Received on] 24-Mar.-2009
[*Corresponding author] ZHANG Hui: Tel: 86-431-86172080, Fax:
86-431-86172080, E-mail: zhanghui_8080@163.com; YANG Jun-
Shan: Tel: 86-10-62899707, Fax: 86-10-62898425, e-mail: junshan
yang@hotmail.com
tents of bioactive components, such as saponins and flavon-
oids, in herbal medicines. The qualitative research of the
secoiridoid glucosides in J. lanceolarium by LC-MS-MS has
not been reported yet in the literature so far.
In this study, a HPLC-MS-MS method has been devel-
oped and validated for the quantitative determination of six
known secoiridoid glucosides in J. lanceolarium. At the same
time the structures of four unknown secoiridoid glu- cosides
were inferred.
2 Experimental
2.1 Instrumentation and reagents
An Agilent Technology 1100 Series HPLC system
equipped with a quaternary pump, a degasser, a thermostatic
auto-sampler and a photodiode array detector (DAD), was
used for analysis (Agilent, America). A SK3200LH ultraso-
nic cleaning instrument (Shanghai Kudos Ultrasonic Instru-
ment Co., Shanghai, China) was used for extraction. Acetoni-
trile is of chromatographic grade and was purchased from
Fisher (America). The water used was treated with a Milli-Q
water purification system (Millipore, Molsheim, France). The
MS instrument used to perform the studies is a 6320 ion trap
LC/MS from Agilent.
SUN Jia-Ming, et al. /Chinese Journal of Natural Medicines 2009, 7(6): 436−439
2009 年 11 月 第 7 卷 第 6 期 Chin J Nat Med Nov. 2009 Vol. 7 No. 6 437

2.2 Materials
The stems and leaves of J. lanceolarium were collected
in Jiujiang, Jiangxi Province, China, in September of 2002.
The identity of the plant material was verified by Professor
Ceming Tan, and a voucher specimen (XC-02-0917) was
deposited at the Institute of Medicinal Plant Development,
Chinese Academy of Medical Sciences and Peking Union
Medical College. The samples for analysis were stored in the
regfrigerator at -20oC.
Jaslanceoside A, B, C, D, E and Jasminoside were iso-
lated from J. lanceolarium. Their structures were elucidated
by their spectral data (MS, 1H NMR and 13C NMR). The
purity of each compound was determined to be higher than
95% by HPLC.
2.3 Sample preparation
The stems and leaves (1.0 g) of J. lanceolarium were
shade-dried, ground, and extracted with refluxing 95% EtOH
successively (20 mL, 2 h, 2 times). The EtOH extract was
evaporated in vacuo to yield a semisolid, which was dis-
solved in MeOH (10 mL). MeOH solution (1 mL) was puri-
fied successively with 100% MeOH over Sephadex LH-20
yielding fractions 1-10. Fr. 5 was filtered through a 0.45 µm
filter membrane before analysis. Twenty microliters of the
sample solution was injected to HPLC column and separated
under below chromatographic conditions.
2.4 HPLC procedures
Chromatographic separations were carried out on a C18
analytical column (Agilent Zorbax XDB-C18 4.6 mm × 150
mm, I.D. 5 µm) supplied by Agilent, America. The mobile
phase consisted of methanol (A)-acetonitrile (B)-0.025%
trifluo-roacetic acid aqueous (C); A∶B∶C was as follows: 0
min, 19∶10∶71; 25 min, 19∶10∶71; 26 min, 29∶ 0∶71;
the flow-rate was 0.3 ml·min-1 and the column temperature
was maintained at 30°C. The chromatogram was recorded at
320 nm. All solvents were filtered through a 0.45 µm filter
and were then degassed by sonication in an ultrasonic bath
before use.
2.5 ESI-MS parameter
The HPLC analytical conditions for the LC-MS were the
same as those used for the HPLC-DAD analysis. Agilent
1100 HPLC/MSD Trap mass spectrometer (Agilent, America)
equipped with an electrospray ionization source was used in
positive ion mode. An HPLC system coupled with DAD was
controlled by an HPLC-MSD ChemStation software system.
Auto MS2 mode of mass spectrometer was chosen to analyze
the sample. The following operation parameters were used:
Capillary voltage: 4 000 V; nebulizer pressure: 35 psi; drying
gas: 9.0 L·min-1; gas temperature: 350°C; fragmentor voltage:
200 V; skimmer voltage: 60 V. LC-ESI-MS accurate mass
spectra were recorded across the range from 50 to 1 000 m/z.
The data recorded was processed with the Applied HPLC-
MSD ChemStation software system.
3 Results and discussion
3.1 Optimization of HPLC–DAD–ESI-MS conditions
Photodiode array detector (DAD) was used in HPLC
analysis, and full scan runs were made initially to select the
optimum wavelength that provided the best result in chroma-
tographic analysis. Chromatogram at 320 nm showed the
most abundant component. The gradient was applied in order
to ensure the good repeatability without reducing their reso-
lutions. Satisfactory results were obtained within 60 min for
the HPLC separation. The effect of the composition of mo-
bile phase on the chromatographic separation of the extracts
was investigated in this study. The results indicated that a
mobile phase composed of methanol-acetonitrile- 0.025%
trifluoroacetic acid aqueous was suitable for the separation of
the extracts.
The secoiridoid glucosides could be analyzed in both
positive and negative ionization. However, the signals of
them were obvious in the positive ionmode. Therefore, posi-
tive ion mode had to be employed to identify the corre-
sponding signals.
3.2 Identification of secoiridoid glucosides in Jasminum
lanceolarium by HPLC–DAD–ESI-MS2
The dominant fragmentation pathways of authentic
compounds were studied. All authentic compounds exhibited
[M + Na]+ ions of sufficient abundance that could be sub-
jected to MS2 analysis. MS2 data were obtained by colli-
sion-induced dissociation (CID), and utilized for the struc-
tural identification of compounds with similar fragmen- ta-
tion patterns.
Jaslanceoside A showed [M + Na]+ at m/z 619 in posi-
tive mode. The ion m/z 457 given by MS2 spectrum suggest-
ing the loss of one hexose (162 Da) from the precursor ion
[M + Na]+. A series of ions at m/z 439 [M + Na − 162 −
H2O]+, 413 [M + Na − 162 − CO2]+ and 263 [M + Na − 162 −
H2O − CO2 − 194]+ were also observed, which strongly sug-
gest the structure of a C–OH, a C–COOH and a ferulic acid.
Fig. 1 shows the proposed fragmentation pattern of
Jaslanceoside A in positive mode.
Six peaks in the HPLC–DAD and HPLC–MS(TIC)
chromatograms were unequivocally identified by compari-
son of their retention times, MS data and UV spectra with
those of authentic compounds. The other four peaks were
presumed by comparing their UV spectra, molecular weight
and structural information from MS2 spectra with authentic
compounds and the literature [4-9]. The TIC ESI-MS mass
spectrum of the sample are shown in Fig. 2. The mass data
and the tentatively identified compound names of the peaks
are given in Table 1 and Fig. 3.
4 Conclusion
An HPLC-MS method has been developed and validated
for the quantitative determination of six known and four un-

SUN Jia-Ming, et al. /Chinese Journal of Natural Medicines 2009, 7(6): 436−439
438 Chin J Nat Med Nov. 2009 Vol. 7 No. 6 2009 年 11 月 第 7 卷 第 6 期



Fig. 1 Fragment pathway of Jaslanceoside A



Fig. 2 TIC chromatogram (A- used in positive ion mode) and HPLC chromatogram (B- recorded at 320 nm) of secoiridoid glu-
cosides in Jasminum lanceolarium



Unknown substance 1 R1 = H, R2 = H, R3 = OH
Jaslanceoside E R1 = CH3, R2 = OH , R3 = OH
Jaslanceoside B R1 = CH3, R2 = H , R3 = OH
Jaslanceoside A R1 = CH3, R2 = OCH3 , R3 = OH
Jasminoside R1 = CH3, R2 = H , R3 = H
Unknown substance 4 R1 = CH3, R2 = OCH3 , R3 = OCH3


Unknown substance 2 R1=CH3, R2=OH , R3=OH
Jaslanceoside D R1=CH3, R2=OH , R3=H
Jaslanceoside C R1=CH3, R2=OH , R3=OCH3
Unknown substance 3 R1=CH3, R2=H , R3=H
Fig. 3 Chemical structures of secoiridoid glucosides identified in extract in Jasminum lanceolarium
Table 2 HPLC-DAD-ESI-MS2 data of secoiridoid glucosides in Jasminum lanceolarium
No. tR (min) UVλmax (nm) [M + Na]+ (m/z) MS2 characteristic ions (m/z) Compound presumed
1 9.3 230, 315 575 531, 413, 395, 369, 249 Unknown substance 1
2 10.3 220, 235, 295, 320 605 443, 425, 399, 263 Jaslanceoside E
3 11.4 220, 235, 290, 320 605 443, 425, 399, 263 Unknown substance 2
4 18.2 230, 310 589 427, 383, 263 Jaslanceoside B
5 19.3 230, 310 589 427, 383, 263 Jaslanceoside D
6 20.8 220, 240, 290, 320 619 457, 439, 413, 263 Jaslanceoside A
7 21.8 220, 240, 290, 320 619 457, 439, 413, 263 Jaslanceoside C
8 43.3 225, 280 573 411, 393, 367, 263 Jasminoside
9 43.8 225, 280 573 411, 393, 367, 263 Unknown substance 3
10 46.2 220, 240, 290, 320 633 471, 439, 263 Unknown substance 4

SUN Jia-Ming, et al. /Chinese Journal of Natural Medicines 2009, 7(6): 436−439
2009 年 11 月 第 7 卷 第 6 期 Chin J Nat Med Nov. 2009 Vol. 7 No. 6 439

known secoiridoid glucosides in J. lanceolarium. The method
was used to instruct us to isolate the new secoiridoid glyco-
side from J. lanceolarium quickly and to authenticate the
species of the herb.
References
[1] Jiangsu New Medicinal College. Cyclopediaof Pharmacy
[M]. Shanghai Science and Technology Press, 1977: 1825-
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[2] Shen YC, Lin SL. New secoiridoid glucosides from Jas-
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[3] Shen YC, Lin SL. Three secoiridoid glucosides from Jas-
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[4] Shen YC, Hsieh PW, Kuo YH. Neolignan glucosides from
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[5] Shen YC, Lin SL, Chein CC, et al. A secoiridoid glucosides
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[6] Tanahashi T, Takenaka Y, Nagakura N. Two dimeric se-
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[7] Trujillo JM, Hernandaez JM, Perez JA, et al. A secoiridoid
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[8] Chen HY. Jasmultiside. A new secoiridoid glucoside from
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2905-2908

高效液相色谱-串联质谱法分析破骨风中
裂环环烯醚萜苷类成分

孙佳明 1, 2, 张 辉 1* 杨峻山 2*

1长春中医药大学中医药与生物工程研究开发中心, 长春 130117
2中国医学科学院-中国协和医科大学药用植物研究所, 北京 100193

【摘 要】 目的：利用高效液相色谱-电喷雾串联质谱法(HPLC-DAD-ESI-MS2)分析了破骨风中裂环环烯醚萜苷类成
分。方法：采用该方法得到了破骨风中裂环环烯醚萜苷类成分的紫外(UV)色谱图、总离子流(TIC)色谱图和萃取离子(EIC)
色谱图, 以及相应色谱峰的紫外光谱图和 EIC/MS2的质谱图, 结合文献对其进行解析。结果：鉴别出破骨风中 6 个裂环环
烯醚萜苷类成分, 分别为 Jasminoside, Jaslanceoside A, Jaslanceoside B, Jaslanceoside C, Jaslanceoside D 和 Jaslanceoside E;
同时发现还有 4 个未知结构裂环环烯醚萜苷类化合物存在, 并对它们的结构进行了推测。结论：高效液相色谱-电喷雾串
联质谱法可以用于分析破骨风中裂环环烯醚萜苷类成分。
【关键词】 破骨风; 高效液相色谱; 电喷雾串联质谱; 裂环环烯醚萜苷

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