title: Identification of 20(S)-protopanaxatriol metabolites in rats by ultra-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry and nuclear magnetic resonance spectroscopy.
abstract: 20(S)-Protopanaxatriol (PPT), one of the aglycones of ginsenosides, has been shown to exert cardioprotective effects against myocardial ischemic injury. However, studies on PPT metabolism have rarely been reported. This study is the first to investigate the in vivo metabolism of PPT following oral administration by ultra-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry (UPLC-Q/TOF-MS) and nuclear magnetic resonance (NMR) spectroscopy. The structures of the metabolites were identified based on the characteristics of their MS data, MS(2) data, and chromatographic retention times. A total of 22 metabolites, including 17 phase I and 5 phase II metabolites, were found and tentatively identified by comparing their mass spectrometry profiles with those of PPT. Two new monooxygenation metabolites, (20S,24S)-epoxy-dammarane-3,6,12,25-tetraol and (20S,24R)-epoxy-dammarane-3,6,12,25-tetraol, were chemicallly synthesized and unambiguously characterized according to the NMR spectroscopic data. The metabolic pathways of PPT were proposed accordingly for the first time. Results revealed that oxidation of (1) double bonds at Δ((24,25)) to form 24,25-epoxides, followed by rearrangement to yield 20,24-oxide forms; and (2) vinyl-methyl at C-26/27 to form corresponding carboxylic acid were the predominant metabolic pathways. Phase II metabolic pathways were proven for the first time to consist of glucuronidation and cysteine conjugation. This study provides valuable and new information on the metabolism of PPT, which is indispensable for understanding the safety and efficacy of PPT, as well as its corresponding ginsenosides.
authors.affiliation: Department of Pharmacognosy, China Pharmaceutical University, Nanjing 210038, China.
authors.forename: Chunyong
authors.initials: C
authors.lastname: He
authors.forename: Dandan
authors.initials: D
authors.lastname: Zhou
authors.forename: Jia
authors.initials: J
authors.lastname: Li
authors.forename: Han
authors.initials: H
authors.lastname: Han
authors.forename: Guang
authors.initials: G
authors.lastname: Ji
authors.forename: Li
authors.initials: L
authors.lastname: Yang
authors.forename: Zhengtao
authors.initials: Z
authors.lastname: Wang
mesh_terms: D000284:Administration, Oral; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D003545:Cysteine / Q000737:chemistry; D004852:Epoxy Compounds / Q000737:chemistry; D005243:Feces; D036145:Ginsenosides / Q000652:urine; D009682:Magnetic Resonance Spectroscopy; D008297:Male; D015394:Molecular Structure; D010100:Oxygen / Q000737:chemistry; D051381:Rats; D017207:Rats, Sprague-Dawley; D012502:Sapogenins / Q000493:pharmacokinetics / Q000652:urine*; D012984:Software; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry
chemical_list: D004852:Epoxy Compounds; D036145:Ginsenosides; D012502:Sapogenins; C081552:protopanaxatriol; D003545:Cysteine; D010100:Oxygen
keywords: 20(S)-Protopanaxatriol; Metabolite identification; NMR; Rats; UPLC–Q/TOF-MS