title: A practical strategy for characterization of the metabolic profile of chiral drugs using combinatory liquid chromatography-mass spectrometric techniques: application to tetrahydropalmatine enantiomers and their metabolites in rat urine.
abstract: The characterization and quantification of the metabolites of chiral drugs still remain a great challenge due to the complexity of the metabolites and most of them are not commercially available. In this study, a practical approach based on the combinatory liquid chromatography-mass spectrometric techniques has been proposed for the evaluation of metabolism profiles and urinary excretion kinetics of chiral drugs and their metabolites. Racemic tetrahydropalmatine (rac-THP), a biologically active ingredient isolated from a traditional Chinese herb Rhizoma Corydalis, was chosen as the model chiral drug. Ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QTOF/MS) was applied to characterize the metabolites of THP enantiomers in rat urine after administration of (+)-THP or (-)-THP. Accurate mass measurement was used to determine the elemental composition of metabolites and thus to confirm the proposed structures of these metabolites. More than 30 potential metabolites were found in rat urine, most of which were identified for the first time, and the metabolic pathways in vivo were involved in demethylation, oxidation, glucuronide conjugation and sulfation, etc. And the tridesmethlyzed metabolite and didesmethlyzed coupled with oxidation metabolite were found only in (+)-THP treated rats. Afterwards, a liquid chromatography tandem mass spectrometry (LC-QqQ/MS) assay was developed and validated for the determination of the urine level of THP enantiomers and their metabolites. Semi-quantification of three phase I metabolites and two phase II metabolites were performed. Enantiomeric (-/+) cumulative urinary excretion ratios of THP and its five metabolites were obtained, which indicated the stereoselective aspects of metabolites of THP enantiomers in vivo. The study demonstrated the enormous potential of this strategy for the qualitative characterization, quantitative assay and the stereoselectivity of chiral drugs and their metabolites.
authors.affiliation: School of Pharmacy, Second Military Medical University, Shanghai Key Laboratory for Pharmaceutical Metabolites Research, No. 325 Guohe Road, Shanghai, PR China.
authors.forename: Yinying
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: School of Pharmacy, Second Military Medical University, Shanghai Key Laboratory for Pharmaceutical Metabolites Research, No. 325 Guohe Road, Shanghai, PR China.
authors.forename: Xin
authors.initials: X
authors.lastname: Dong
authors.affiliation: Shanghai Institute for Food and Drug Control, Shanghai 201203, PR China.
authors.forename: Jian
authors.initials: J
authors.lastname: Le
authors.affiliation: School of Pharmacy, Second Military Medical University, Shanghai Key Laboratory for Pharmaceutical Metabolites Research, No. 325 Guohe Road, Shanghai, PR China.
authors.forename: Jun
authors.initials: J
authors.lastname: Wen
authors.affiliation: School of Pharmacy, Second Military Medical University, Shanghai Key Laboratory for Pharmaceutical Metabolites Research, No. 325 Guohe Road, Shanghai, PR China.
authors.forename: Zebin
authors.initials: Z
authors.lastname: Lin
authors.affiliation: School of Pharmacy, Second Military Medical University, Shanghai Key Laboratory for Pharmaceutical Metabolites Research, No. 325 Guohe Road, Shanghai, PR China.
authors.forename: Yinli
authors.initials: Y
authors.lastname: Liu
authors.affiliation: School of Pharmacy, Second Military Medical University, Shanghai Key Laboratory for Pharmaceutical Metabolites Research, No. 325 Guohe Road, Shanghai, PR China.
authors.forename: Ziyang
authors.initials: Z
authors.lastname: Lou
authors.affiliation: School of Pharmacy, Second Military Medical University, Shanghai Key Laboratory for Pharmaceutical Metabolites Research, No. 325 Guohe Road, Shanghai, PR China.
authors.forename: Yifeng
authors.initials: Y
authors.lastname: Chai
authors.affiliation: School of Pharmacy, Second Military Medical University, Shanghai Key Laboratory for Pharmaceutical Metabolites Research, No. 325 Guohe Road, Shanghai, PR China. Electronic address: hongzhy001@163.com.
authors.forename: Zhanying
authors.initials: Z
authors.lastname: Hong
mesh_terms: D000818:Animals; D001600:Berberine Alkaloids / Q000737:chemistry* / Q000378:metabolism* / Q000652:urine; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D031311:Corydalis / Q000737:chemistry; D004365:Drugs, Chinese Herbal / Q000737:chemistry; D008297:Male; D050216:Metabolic Detoxication, Phase I / Q000502:physiology; D050217:Metabolic Detoxication, Phase II / Q000502:physiology; D051381:Rats; D017207:Rats, Sprague-Dawley; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D001600:Berberine Alkaloids; D004365:Drugs, Chinese Herbal; C014215:tetrahydropalmatine
keywords: LC-QqQ/MS; Metabolites; Rat urine; Tetrahydropalmatine; UPLC-QTOF/MS