title: Analysis of quazepam and its metabolites in human urine by gas chromatography-mass spectrometry: application to a forensic case.
abstract: A sensitive method for the simultaneous determination of quazepam and two of its metabolites, 2-oxoquazepam and 3-hydroxy-2-oxoquazepam, in human urine was developed using gas chromatography-mass spectrometry (GC/MS) with an Rtx-5MS capillary column. The quazepam and its metabolites were extracted from human urine using a simple solid-phase extraction Oasis(®) HLB cartridge column, and the 3-hydroxy-2-oxoquazepam was derivatised using BSTFA/1%TMCS and pyridine at 60 °C for 30 min. The mass spectrometric detection of the analytes was performed in the full scan mode, m/z 60-480, and selected ion monitoring (SIM) mode, m/z 386, for quazepam; m/z 342, for 2-oxoquazepam; m/z 429, for 3-hydroxy-2-oxoquazepam-TMS; and m/z 284, for alprazolam-d5 (internal standard), by electron ionization. The calibration curves of quazepam and its metabolites in urine showed good linearity in the concentration range of 2.5-500 ng/0.2 ml of urine. The average recoveries of quazepam and its metabolites from 0.2 ml of urine containing 500 ng and 50 ng of each drug were 71-83% and 88-90%, respectively. The limits of detection of quazepam, 2-oxoquazepam and 3-hydroxy-2-quazepam in urine by the selected ion monitoring mode were 0.096-0.37 ng/ml. This method would be applicable to other forensic biological materials containing low concentrations of quazepam and its metabolites.
authors.affiliation: Department of Legal Medicine, School of Medicine, Toho University, 5-21-16 Oomorinishi, Tokyo 143-8540, Japan. mterada@med.toho-u.ac.jp
authors.forename: Masaru
authors.initials: M
authors.lastname: Terada
authors.forename: Tatsuo
authors.initials: T
authors.lastname: Shinozuka
authors.forename: Chika
authors.initials: C
authors.lastname: Hasegawa
authors.forename: Einosuke
authors.initials: E
authors.lastname: Tanaka
authors.forename: Makiko
authors.initials: M
authors.lastname: Hayashida
authors.forename: Youkichi
authors.initials: Y
authors.lastname: Ohno
authors.forename: Kunihiko
authors.initials: K
authors.lastname: Kurosaki
mesh_terms: D001569:Benzodiazepines / Q000652:urine*; D001570:Benzodiazepinones / Q000652:urine; D053593:Forensic Toxicology; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods; D006801:Humans; D006993:Hypnotics and Sedatives / Q000652:urine*; D008297:Male; D052616:Solid Phase Extraction; D015813:Substance Abuse Detection / Q000295:instrumentation; D014229:Triazolam / Q000031:analogs & derivatives / Q000652:urine
chemical_list: C584129:3-hydroxy-2-oxoquazepam; D001570:Benzodiazepinones; D006993:Hypnotics and Sedatives; D001569:Benzodiazepines; D014229:Triazolam; C028638:alpha-hydroxytriazolam; C039103:2-oxoquazepam; C025130:quazepam