title: Simultaneous determination of glipizide and its four hydroxylated metabolites in human urine using LC-MS/MS and its application in urinary phenotype study.
abstract: Cytochrome P450 (CYP) 2C9 and CYP2C19 genetic mutant could influence the plasma concentration of glipizide in human subjects, which refers to glipizide safety and adverse effects in clinic practice. A further study to investigate the relationship of the concentrations between glipizide and its metabolites in human with different CYP mutants was valuable. We firstly develop a validated liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for simultaneous quantification of glipizide and its hydroxylated metabolites in human urine. After simple protein precipitation with methanol including 4'-OH-tolbutamide and gliclazide (both are internal standards), the analytes were chromatographed on a reversed-phased column with a mobile phase of 0.1% formic acid in acetonitrile and 0.1% formic acid in water by a gradient elution. The ion transitions of the precursor to the product ion were principally protonated ions [M+H]+at m/z 446.4→m/z 321.1 for glipizide, m/z 462.2→m/z 321.1 for the four hydroxylated forms of glipizide, m/z 287.2→m/z 188.0 for 4'-OH-tolbutamide, and m/z 324.1→m/z 127.1 for gliclazide. The method was linear over a concentration range of 0.02-20.0ng/mL. The intraday and inter-day variances were less than 9.9%, and accuracy was within ±6.8%. The method was successfully applied to the urinary phenotyping study in volunteers after a single oral administration of 5-mg glipizide tablet, and two new hydroxycyclohexyl metabolites of glipizide (OH-gp), 4-cis-OH-gp and 3-trans-OH-gp, were found in this study.
authors.affiliation: Clinical Pharmacokinetic Laboratory, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, 528 Zhangheng Road, Shanghai 201203, China; Department of Clinical Pharmacology, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, 528 Zhangheng Road, Shanghai 201203, China.
authors.forename: Bo
authors.initials: B
authors.lastname: Tan
authors.affiliation: Department of Febrile Disease of Basic Medical College, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China.
authors.forename: Aidong
authors.initials: A
authors.lastname: Yang
authors.affiliation: Department of Clinical Pharmacology, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, 528 Zhangheng Road, Shanghai 201203, China.
authors.forename: Weian
authors.initials: W
authors.lastname: Yuan
authors.affiliation: Clinical Pharmacokinetic Laboratory, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, 528 Zhangheng Road, Shanghai 201203, China; Department of Clinical Pharmacology, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, 528 Zhangheng Road, Shanghai 201203, China.
authors.forename: Yue
authors.initials: Y
authors.lastname: Li
authors.affiliation: Department of Febrile Disease of Basic Medical College, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Jiang
authors.affiliation: Clinical Pharmacokinetic Laboratory, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, 528 Zhangheng Road, Shanghai 201203, China; Department of Clinical Pharmacology, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, 528 Zhangheng Road, Shanghai 201203, China. Electronic address: jiangjiansg@126.com.
authors.forename: Jian
authors.initials: J
authors.lastname: Jiang
authors.affiliation: Clinical Pharmacokinetic Laboratory, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, 528 Zhangheng Road, Shanghai 201203, China; Department of Clinical Pharmacology, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, 528 Zhangheng Road, Shanghai 201203, China. Electronic address: furong_qiu@126.com.
authors.forename: Furong
authors.initials: F
authors.lastname: Qiu
mesh_terms: D000328:Adult; D002853:Chromatography, Liquid / Q000379:methods; D005913:Glipizide / Q000378:metabolism / Q000652:urine*; D006801:Humans; D006900:Hydroxylation; D008297:Male; D010641:Phenotype; D053719:Tandem Mass Spectrometry / Q000379:methods*; D055815:Young Adult
chemical_list: D005913:Glipizide
keywords: Glipizide; Hydroxylated metabolite; LC–MS/MS; Urine