Separation and Characterization of Triterpenoid Saponins in Gleditsia sinensis by Comprehensive Two-Dimensional Liquid Chromatography Coupled with Mass Spectrometry

Abstract

A comprehensive two-dimensional liquid chromatography coupled with mass spectrometry method was established to separate and characterize triterpenoid saponins in Gleditsia sinensis, the Chinese herbal medicine Zhu-Ya-Zao used for the treatment of apoplexy. The saponins were separated on an Agilent Zorbax Eclipse plus C₁₈ column (2.1 mm × 150 mm, 1.8 µm) for the first dimension, and a Poroshell 120 phenyl-hexyl column (3.0 mm × 50 mm, 2.7 µm) for the second dimension. Methanol and acetonitrile were used as the organic mobile phase for ¹D and ²D, respectively. The theoretical peak capacity was 640, and the orthogonality was 57 %. Particularly, saponins with different numbers of monoterpene groups could be well separated on the second dimension. The structures were characterized by electrospray ionization tandem mass spectrometry in both positive and negative ion modes. In the MS/MS spectra, the relative abundances for [B _α + Na]⁺ and [Y _0α + Na]⁺ were closely correlated with the number of sugar residues of the α-chain and β-chain, respectively, which facilitated the differentiation of isomers. Finally, a total of 72 saponins with molecular weights greater than 1500 Da were characterized. Among them, 49 compounds, including 2 acetylated saponins, were detected from G. sinensis for the first time.

Key words

Gleditsia sinensis - Leguminosae - comprehensive two-dimensional liquid chromatography - mass spectrometry - triterpenoid saponins

• References

• 1 Chinese Pharmacopoeia Commission. Pharmacopoeia of the Peopleʼs Republic of China, Vol 1. Beijing: Chinese Medical Science and Technology Press; 2015
• 2 Xia Y, Gao Z, Dai Y, Wang Q. Analysis of bioactive saponins in Fructus Gleditsiae abnormalis and Fructus Gleditsiae sinensis by LC-ELSD. Chromatographia 2009; 70: 1361-1366
• 3 Feng Y, Jia YY, Li GH, Liang RX, Zhang XJ. The study on antineoplastic effective part of the stings of Gleditsia sinensis Lam. Lishizhen Med Mat Medica Res 2006; 17: 319-320
• 4 Dai Y, Chan YP, Chu LM, But PPH. Antiallergic and anti-inflammatory properties of the ethanolic extract from Gleditsia sinensis . Biol Pharm Bull 2002; 25: 1179-1182
• 5 Zhang ZZ, Koike K, Jia ZH, Nikaido T, Guo D, Zheng JH. Four new triterpenoidal saponins acylated with one monoterpenic acid from Gleditsia sinensis . J Nat Prod 1999; 62: 740-745
• 6 Zhang ZZ, Koike K, Jia ZH, Nikaido T, Guo D, Zheng JH. Gleditsiosides N–Q, new triterpenoid saponins from Gleditsia sinensis . J Nat Prod 1999; 62: 877-881
• 7 Zhang ZZ, Koike K, Jia ZH, Nikaido T, Guo D, Zheng JH. Triterpenoidal saponins acylated with two monoterpenic acids from Gleditsia sinensis . Chem Pharm Bull 1999; 47: 388-393
• 8 Zhang ZZ, Koike K, Jia ZH, Nikaido T, Guo D, Zheng JH. Triterpenoidal saponins from Gleditsia sinensis . Phytochemistry 1999; 52: 715-722
• 9 Zhong L, Li P, Han J, Qu GQ, Guo D. Structure-activity relationships of saponins from Gleditsia sinensis in cytotoxicity and induction of apoptosis. Planta Med 2004; 70: 797-802
• 10 Chen J, Li Z, Zheng KYZ, Guo AJY, Zhu KY, Zhang WL, Zhan JYX, Dong TTX, Su Z, Tsim KWK. Chemical fingerprinting and quantitative analysis of two common Gleditsia sinensis fruits using HPLC-DAD. Acta Pharm 2013; 63: 505-515
• 11 Lian XY, Zhang Z. Quantitive analysis of gleditsia saponins in the fruits of Gleditsia sinensis Lam. by high performance liquid chromatography. J Pharm Biomed Anal 2013; 75: 41-46
• 12 Bo T, Zhong L, Li M, Luo YQ, Li KA, Liu HW, Guo DA. Separation of isoquinoline alkaloids and saponins by microemulsion electrokinetic chromatography with anionic and cationic surfactants. Chromatographia 2002; 56: 709-716
• 13 Han L, Pan G, Wang Y, Song X, Gao X, Ma B, Kang L. Rapid profiling and identification of triterpenoid saponins in crude extracts from Albizia julibrissin Durazz. by ultra high-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry. J Pharm Biomed Anal 2011; 55: 996-1009
• 14 Yang WZ, Ye M, Qiao X, Liu CF, Miao WJ, Bo T, Tao HY, Guo DA. A strategy for efficient discovery of new natural compounds by integrating orthogonal column chromatography and liquid chromatography/mass spectrometry analysis: Its application in Panax ginseng, Panax quinquefolium and Panax notoginseng to characterize 437 potential new ginsenosides. Anal Chim Acta 2012; 739: 56-66
• 15 Wang Y, Lu X, Xu G. Development of a comprehensive two-dimensional hydrophilic interaction chromatography/quadrupole time-of-flight mass spectrometry system and its application in separation and identification of saponins from Quillaja saponaria . J Chromatogr A 2008; 1181: 51-59
• 16 Jeong EK, Cha HJ, Ha YW, Kim YS, Ha IJ, Na YC. Development and optimization of a method for the separation of platycosides in Platycodi Radix by comprehensive two-dimensional liquid chromatography with mass spectrometric detection. J Chromatogr A 2010; 1217: 4375-4382
• 17 Yang WZ, Bo T, Ji S, Qiao X, Guo DA, Ye M. Rapid chemical profiling of saponins in the flower buds of Panax notoginseng by intergrating MCI gel column chromatography and liquid chromatography/mass spectrometry analysis. Food Chem 2013; 139: 762-769
• 18 Qiao X, Song W, Ji S, Li YJ, Wang Y, Li R, An R, Guo DA, Ye M. Separation and detection of minor constituents in herbal medicines using a combination of heart-cutting and comprehensive two-dimensional liquid chromatography. J Chromatogr A 2014; 1362: 157-167
• 19 Konoshima T, Umegaki Y, Sawada T. Legume saponins of Gleditsia japonica MIQUEL. III. Further desmonoterpenyl glycosides of echinocystic acid. Chem Pharm Bull 1981; 29: 2695-2699