title: Identification of chemical constituents in two traditional Chinese medicine formulae by liquid chromatography-mass spectrometry and off-line nuclear magnetic resonance.
abstract: There have been increasing works on identification of the chemical constituents in traditional Chinese medicines (TCMs) by liquid chromatography-mass spectrometry (LC-MS). However, isomers cannot be distinguished generally only by MS data, and the structures of unknown compounds cannot be confirmed. In this study, semi-preparative LC guided by LC-MS was used to prepare the isomers in microscale followed by off-line NMR analysis to confirm their structures. This approach was applied to identifying the constituents in two TCM formulae, Zhi-Zi-Gan-Cao-Chi-Tang (ZZGCCT) and Zhi-Zi-Bai-Pi-Tang (ZZBPT). A total number of 119 constituents were identified tentatively or unambiguously by LC-IT-MS, LC-Q-TOF-MS, and NMR. Among them, 20 constituents were characterized unambiguously by comparing with the reference substances. In addition, 21 constituents without reference substances were prepared for the following 1D-NMR and/or 2D-NMR analysis, and their structures were unambiguously identified by MS, 1D-NMR, and 2D-NMR. Two triterpenoid glycosides (compounds 134 and 140) and one flavonoid glycoside (compound 62a or 62b) were showed to be novel compounds. Compounds 125 and 129, as well as 62a,b, were epimers.
authors.affiliation: Pharmaceutical Informatics Institute, College of Pharmaceutical Sciences, Zhejiang University, Zijingang Campus, No. 866 Yuhangtang Road, Hangzhou 310058, PR China. Electronic address: wangsf@zju.edu.cn.
authors.forename: Shufang
authors.initials: S
authors.lastname: Wang
authors.affiliation: Pharmaceutical Informatics Institute, College of Pharmaceutical Sciences, Zhejiang University, Zijingang Campus, No. 866 Yuhangtang Road, Hangzhou 310058, PR China.
authors.forename: Yunxiang
authors.initials: Y
authors.lastname: Zhu
authors.affiliation: Pharmaceutical Informatics Institute, College of Pharmaceutical Sciences, Zhejiang University, Zijingang Campus, No. 866 Yuhangtang Road, Hangzhou 310058, PR China.
authors.forename: Qing
authors.initials: Q
authors.lastname: Shao
authors.affiliation: Pharmaceutical Informatics Institute, College of Pharmaceutical Sciences, Zhejiang University, Zijingang Campus, No. 866 Yuhangtang Road, Hangzhou 310058, PR China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Pharmaceutical Informatics Institute, College of Pharmaceutical Sciences, Zhejiang University, Zijingang Campus, No. 866 Yuhangtang Road, Hangzhou 310058, PR China.
authors.forename: Xiaohui
authors.initials: X
authors.lastname: Fan
authors.affiliation: Pharmaceutical Informatics Institute, College of Pharmaceutical Sciences, Zhejiang University, Zijingang Campus, No. 866 Yuhangtang Road, Hangzhou 310058, PR China.
authors.forename: Yiyu
authors.initials: Y
authors.lastname: Cheng
mesh_terms: D002853:Chromatography, Liquid / Q000379:methods; D004365:Drugs, Chinese Herbal / Q000032:analysis* / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D012503:Saponins / Q000032:analysis / Q000737:chemistry; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D004365:Drugs, Chinese Herbal; D012503:Saponins
keywords: Liquid chromatography–mass spectrometry (LC–MS); Nuclear magnetic resonance (NMR); Semi-preparative liquid chromatography; Zhi-Zi-Bai-Pi-Tang; Zhi-Zi-Gan-Cao-Chi-Tang