Chinese

Journal of
Natural
Chinese Journal of Natural Medicines 2012, 10(2): 0088−0091 Medicines
doi: 10.3724/SP.J.1009.2012.00088

A new C30 sterol glycoside from the fresh fruits of

Momordica charantia
LIU Peng1, LU Jian-Feng1, 2, KANG Li-Ping1, 2, YU He-Shui1, 2, ZHANG Li-Juan 1,
SONG Xin-Bo1*, MA Bai-Ping2*
1
Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China;
2
Beijing Institute of Radiation Medicine, Beijing 100850, China
Available online 20 Mar. 2012

[ABSTRACT] AIM: To investigate the chemical constituents from the fresh fruits of Momordica charantia L. METHODS: Chemical
constituents were isolated and purified by chromatography on silica gel and octadecylsilanized (ODS) silica gel, and the structures were
determined by spectral data and chemical evidence. RESULTS: A new C30 sterol glycoside, named 25ξ-isopropenylchole-5,
(6)-ene-3-O-β-D-glucopyranoside (1), was isolated from the fresh fruits of M. charantia L. CONCLUSION: Compound 1 is a new
C30 sterol glycoside.
[KEY WORDS] Momordica charantia L.; Sterol glycoside; 25ξ-isopropenylchole-5, (6)-ene-3-O-β-D-glucopyranoside
[CLC Number] R284.1 [Document code] A [Article ID] 1672-3651(2012)02-0088-04

chromatography. TLC was performed on precoated silica gel

1 Introduction GF254 plates (Qingdao marine Chemical, China). HPLC was
The Cucurbitaceae vegetable plant M. charantia L. is performed on Waters 2695 (Waters, USA), YMC-Pack
widely cultivated in Asian countries and the fruits of this ODS-A C18 (YMC, 4.6 mm × 250 mm, 5 μm, Japan). The
plant have been used as a bitter stomachic, a laxative, an HR-ESI-MS was recorded on 9.4 T Q-FT-MS Apex Qe
antidiabetic, and an anthelmintic for children in China [1]. In (Bruker Co.). FAB-MS: Micromass Zabspec. Optical rota-
addition, several cucurbitane-type triterpene glycosides were tions were measured with Perkin-Elmer 343 polarimeter. The
reported as chemical constituents of the fruits [2-10]. However, NMR spectra were recorded with Varian UNITY Inova 600
chemical and pharmacological studies [11-13] on the fruits of M. (599.8 MHz for 1H NMR and 150.8 MHz for 13C NMR), and
charantia were yet left uncharacterised. In our studies of the the chemical shifts were given on δ scale with tetramethyl-
chemical constituents of the fresh fruits of M. charantia, a silane as an internal standard. Gas chromatographic analysis
new C30 sterol glycoside, named 25ξ-isopropenylchole-5, was performed with an Agilent 6890, gas chromatographer
(6)-ene-3-O-β-D-glucopyranoside, was isolated, together equipped with a 5973 mass spectrographer. The column was
with momordicoside L, a known compound (Fig. 1). The new an HP-5 capillary column (30 m × 0.25 mm × 0.25 μm)
compound was determined by analysis of 1D and 2D NMR (Agilent, USA).
data, HR-ESI-MS and chemical methods. 2.2 Plant Material
The fresh fruits of M. charantia L. were collected from
2 Experimental
the local market in May 2007 in Tianjin. The plant was iden-
2.1 General tified by Prof. ZHANG Li-Juan (Tianjin University of Tradi-
Silica-gel (Qingdao Haiyang Chemical Co., Ltd., China) tional Chinese Medicine) and a voucher specimen (No.
and ODS silica-gel (50 μm, YMC, Japan) were used for 070403) has been deposited in the Herbarium of Beijing In-
stitute of Radiation Medicine, Beijing, China.
2.3 Extraction and Isolation
[Received on] 02-Aug.-2011
The fresh fruits of M. charantia (3.0 Kg) were refluxed
[*Corresponding author] SONG Xin-Bo: Tel: 86-22-27494976,
E-mail: [email protected]; MA Bai-Ping: Tel: 86-10-6821-
thrice with 90% EtOH (3 × 24 L, each for 1.5 h). The com-
0077 Ext. 930265, E-mail: [email protected] bined extract was concentrated under reduced pressure. The
These authors have no any conflict of interest to declare. extract was separated chromatographically on silica-gel col-

88 Chin J Nat Med Mar. 2012 Vol. 10 No. 2 2012 年 3 月 第 10 卷 第 2 期

 LIU Peng, et al. /Chinese Journal of Natural Medicines 2012, 10(2): 88−91

umn and eluted with a gradient mixture of CHCl3− (C-24), 30.1 (C-23), 112.0 (C-28) and 141.0 (C-5) were ob-
MeOH−H2O (20 : 1: 0.01−80 : 10 : 1) to give five fractions served in the HMBC spectrum. Thus, the aglycone of 1 was
A1-A5. Fraction A4 was purified by silica-gel column with identified as 25ξ-isopropenylchole-5, (6)-ene-3β-hydroxyl.
gradient mixture of CHCl3−MeOH−H2O (10 : 1 : 0.01−40 : Acid hydrolysis of 1 was dissolved in 2 mol·L−1
10 : 1) as eluent. Fractions A4-70-74 were recrystallised in CF3COOH―H2O (5 mL) and heated for 6 h at 95 °C. After
methanol and gave compound 1 (20 mg). Fraction A5 was extracted with CH2Cl2 (5 mL) three times. The aqueous layer
purified by ODS silica-gel column with gradient mixture of was repeatedly evaporated to dryness with EtOH for getting
MeOH−H2O (50 : 50−76 : 24) as eluent and gave compound rid of CF3COOH. Then, in monosaccharide mixture and glu-
2 (9 mg). cose were detected by TLC over silica gel (CHCl3−
MeOH−H2O, 8 : 5 : 1) by comparison with standard sample:
glucose (Rf 0.45) Furthermore, the residue of sugars was dis-
solved in anhydrous pyridine (2 mL), and added L-cysteine
methyl ester hydrochloride (2.4 mg) and the mixture was
stirred at 60 °C for 1 h. After that, HMDS-TMCS (hexame-
thyldisilazane−trimethylchlorosilane 2 : 1) (6.0 mL) was
added and kept at 60 °C for 0.5 h [14]. Finally, the supernatant
was analyzed by GC as following condition: Agilent 6890
system coupled with mass spectrographer 5973; HP-5 capil-
lary column (30 m × 0.25 mm × 0.25 μm); column tempera-
ture: 180.0−250.0 °C, programmed increase, 15.0 °C·min−1;
carrier gas: N2 (1 mL·min-1); injection temperature: 250.0 °C;
injection volume: 1.0 μL, split ratio: 1/50. Consequently, the
D-configuration of glucose was established by comparison of
Fig. 1 Structures of compounds 1 and 2 retention times with the authentic sample, tR (min): D-glucose
(17.92, 19.67). The large coupling constant (JGlc1 = 7.8) indi-
3 Results and Discussion
cated the β configuration of the sugar [15−16]. The HMBC
Compound 1 was obtained as a white amorphous powder. spectrum of 1 showed the long range correlations between δ
The molecular formula was assigned as C36H60O6 on the 5.05 (C1'-Glc-H) and δ 74.8 (C-3), indicated the sugar linkage
basis of the positive-ion HR-ESI-MS at m/z 611.411 8 [M + site to the aglycone moiety. Thus, the structure of 1 was de-
Na]+ (Calcd. 611.413 1), together with its 1H- and 13C NMR termined to be 25ξ-isopropenylchole-5, (6)-ene-3-O-β-D-
(pyridine-d5) spectral data (Table 1). FAB-MS showed the ion glucopyranoside.
peaks at m/z 589.2 [M + H]+, 427.2 [M + H – 162]+ and 409.2 Compound 2 was obtained as a white amorphous powder.
[M + H – 162 – H2O]+, attributable to the sequential losses of which gave positive Libermann-Burchard and Molish reagent
a hexose and a moleculer of water residues. The 1H NMR tests. Its molecular formula was assigned as C36H58O9 on the
spectrum of 1 showed four methyl signals at δ 0.93 (3H, s, basis of the high-resolution electrospray ionization mass
C18-CH3), 0.66 (3H, s, C19-CH3), 0.96 (3H, d, J = 6.6, spectrometry (FAB-MS) m/z 657.1 [M + Na]+, 635.1 [M +
C21-CH3) and 0.86 (t, J = 7.8 Hz, C27-CH3), one anomeric H]+ and the 1H- and 13C NMR data. The FAB-MS showed the
proton signal at δ 5.05 (1H, d, J = 7.8 Hz), and three olefinic significant fragment peaks at m/z 455.1 [M + H – 162 –
proton signals at δ 5.34 (1H, br s, C6-H), 4.87 (1H, br s, H2O]+, 437.1 [M + H – 162 – H2O – H2O]+, 419.1 [M + H –
C29-Ha), and 4.80 (1H, br s, C29-Hb). The 13C NMR spectrum 162 – H2O – H2O – H2O]+. The 1H NMR spectrum showed
showed 36 carbon signals, in which the characteristic carbon signals for seven Me groups at δ 0.72 (3H, s, C18-CH3), 0.85
signals at δ 122.0 (C-6), 112.0 (C-28), 141.0 (C-5) and 147.8 (3H, s, C30-CH3), 0.94 (3H, d, J = 6.0, C21-CH3), 1.12 (3H, s,
(C-29) were assigned readily. C29-CH3), 1.43 (3H, s, C28-CH3), 1.53 (3H, s, C26-CH3) and
The combined use of 1H- and 13C NMR, 1H-1H COSY, 1.54 (3H, s, C27-CH3), two anomeric proton signals at δ 6.16
HSQC and HMBC spectroscopy allowed the sequential as- (1H, d, J = 4.2 Hz, H-6) and δ 4.59 (1H, d, J = 5.4 Hz, H-23,
signments of all resonances for saponin (Table 1). In the H-24), and δ 4.97 (1H, d, J = 7.8 Hz, H-7). The 13C NMR
HMBC spectrum, the long-range correlations between δ 0.93 spectrum of 2 showed two groups olefinic carbon signals at δ
(C19-CH3) with δ 37.0 (C-10), 37.5 (C-1), 50.4 (C-9) and 147.6 and 135.2, δ 141.8 and 122.3, in addition, one ano-
141.0 (C-5), and between δ 1.61 (C30-CH3) with δ 49.8 meric carbon signals at δ 101.8. Analysis of the 1H- and 13C
(C-25), 112.0 (C-28) and 147.8 (C-29), indicated the double NMR spectral data of 2 in comparison with those of mo-
bonds were at C5-C6 and C26-C28. On the other hand, the mordicoside L in the literature [9] implied that compound 2
long-range correlations between δ 0.96 (C21-CH3) with δ 35.8 was the same as 3β, 7β, 25-trihydroxy-cucurbita-5, 23- di-
(C-20), 34.1 (C-22) and 56.3 (C-17), and between δ 1.90 ene-19-al-7-O-β-D-glucopyranoside. Thus, the structure of 2
(C25-H) with δ 12.3 (C-30), 17.9 (C-27), 26.8 (C-26), 29.7 was determined to be momordicoside L.

2012 年 3 月 第 10 卷 第2期 Chin J Nat Med Mar. 2012 Vol. 10 No. 2 89

 LIU Peng, et al. /Chinese Journal of Natural Medicines 2012, 10(2): 88−91

Acknowledgments
MS and NMR spectra were recorded at the National
Center of Biomedical Analysis. The assistance of the staff is
gratefully acknowledged.

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新鲜苦瓜中的一个新 C30 甾醇苷

刘 芃 1，陆剑锋 1, 2，康利平 1, 2，余河水 1, 2，张丽娟 1，宋新波 1*，马百平 2*
1
天津中医药大学，天津 300193;
2
北京放射与辐射医学研究所，北京 100850

【摘 要】 目的：研究新鲜苦瓜中的化学成分。方法：采用硅胶柱色谱和 ODS 柱色谱进行化学成分分离纯化，并利用波

谱数据分析和理化性质进行结构鉴定。结果：从新鲜苦瓜中分离得到 1 个新 C30 甾醇苷，结构鉴定为 25ξ-异丙烯-胆甾-5, (6)-
烯-3-O-β-D-吡喃葡萄糖苷(1)。结论：化合物 1 为新的 C30 甾醇苷。
【关键词】 苦瓜; 甾醇苷; 25ξ-异丙烯-胆甾-5, (6)-烯-3-O-β-D-吡喃葡萄糖苷

2012 年 3 月 第 10 卷 第2期 Chin J Nat Med Mar. 2012 Vol. 10 No. 2 91