title: A validated liquid chromatography tandem mass spectrometry method for simultaneous determination of pioglitazone, hydroxypioglitazone, and ketopioglitazone in human plasma and its application to a clinical study.
abstract: A simple and rapid high performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for the simultaneous determination of pioglitazone and its active metabolites hydroxypioglitazone and ketopioglitazone in human plasma. Samples were processed by protein precipitation with acetonitrile and selective phospholipid depletion in a 96-well plate format. The method used deuterated internal standards for each analyte. Chromatographic separation was achieved with gradient elution on a Hypersil GOLD C18 column. The mass spectrometer was operated in electrospray positive ion mode with detection by selected reaction monitoring using the transitions m/z 357.1>134.0 for pioglitazone, m/z 373.1>150.0 for hydroxypioglitazone, and m/z 371.0>148.0 for ketopioglitazone. A linear standard curve was established for the range of 10-1800ng/mL for all three analytes. Intra-run and inter-run precision and accuracy (relative error) were less than 15%, and the mean extraction recoveries of all analytes were more than 87.8%. The validated method is sensitive and selective and was successfully applied to analyze clinical samples obtained from patients with nonalcoholic fatty liver disease taking pioglitazone.
authors.affiliation: Department of Pharmacotherapy and Translational Research, College of Pharmacy, University of Florida, Gainesville, FL, USA.
authors.forename: Marina
authors.initials: M
authors.lastname: Kawaguchi-Suzuki
authors.affiliation: Division of Endocrinology, Diabetes and Metabolism, College of Medicine, University of Florida, Gainesville, FL, USA.
authors.forename: Fernando
authors.initials: F
authors.lastname: Bril
authors.affiliation: Division of Endocrinology, Diabetes and Metabolism, College of Medicine, University of Florida, Gainesville, FL, USA.
authors.forename: Paola Portillo
authors.initials: PP
authors.lastname: Sanchez
authors.affiliation: Division of Endocrinology, Diabetes and Metabolism, College of Medicine, University of Florida, Gainesville, FL, USA.
authors.forename: Kenneth
authors.initials: K
authors.lastname: Cusi
authors.affiliation: Department of Pharmacotherapy and Translational Research, College of Pharmacy, University of Florida, Gainesville, FL, USA. Electronic address: frye@cop.ufl.edu.
authors.forename: Reginald F
authors.initials: RF
authors.lastname: Frye
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D007004:Hypoglycemic Agents / Q000097:blood / Q000737:chemistry / Q000627:therapeutic use; D016014:Linear Models; D065626:Non-alcoholic Fatty Liver Disease / Q000097:blood / Q000188:drug therapy; D000077205:Pioglitazone; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry / Q000379:methods*; D045162:Thiazolidinediones / Q000097:blood* / Q000737:chemistry* / Q000627:therapeutic use
chemical_list: D007004:Hypoglycemic Agents; D045162:Thiazolidinediones; D000077205:Pioglitazone
keywords: Human plasma; LC–MS/MS; Liquid chromatography; Mass spectrometry; Metabolites; Pioglitazone