title: Simultaneous enantiomeric determination of MDMA and its phase I and phase II metabolites in urine by liquid chromatography-tandem mass spectrometry with chiral derivatization.
abstract: A liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) procedure was developed for the simultaneous determination of enantiomers of the prevalent designer drug 3,4-methylenedioxymethamphetamine (MDMA) and its phase I and phase II metabolites in urine with chiral derivatization. The analytes in urine were directly derivatized with chiral Marfey's reagent, N(α)-(5-fluoro-2,4-dinitrophenyl)-D-leucinamide, without extraction. The diastereomers of the N(α)-(2,4-dinitrophenyl)-D-leucinamide derivatives generated were determined by LC-MS/MS. Satisfactory chromatographic separation was achieved for the enantiomers of MDMA and its metabolites 3,4-methylenedioxyamphetamine, 4-hydroxy-3-methoxymethamphetamine (HMMA), HMMA glucuronide, and HMMA sulfate on a semimicro octadecylsilane column using linear gradient elution. With use of multiple reaction monitoring mode, the limits of detection of these analytes ranged from 0.01 to 0.03 μg/mL. Linear calibration curves were obtained for all enantiomers from 0.1 to 20 μg/mL in urine. The method showed sufficient reproducibility and quantitative ability. This is the first report of a simple LC-MS/MS-based analytical procedure with direct chiral derivatization in aqueous media that allows simultaneous enantiomeric determination of drugs and their metabolites, including glucuronide and sulfate derivatives.
authors.affiliation: Forensic Science Laboratory, Osaka Prefectural Police Headquarters, Chuo-ku, Osaka, Japan. keikonakanishi@office.zaq.jp
authors.forename: Keiko
authors.initials: K
authors.lastname: Nakanishi
authors.forename: Munehiro
authors.initials: M
authors.lastname: Katagi
authors.forename: Kei
authors.initials: K
authors.lastname: Zaitsu
authors.forename: Noriaki
authors.initials: N
authors.lastname: Shima
authors.forename: Hiroe
authors.initials: H
authors.lastname: Kamata
authors.forename: Akihiro
authors.initials: A
authors.lastname: Miki
authors.forename: Hajime
authors.initials: H
authors.lastname: Kato
authors.forename: Ken-Ichi
authors.initials: K
authors.lastname: Harada
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Tsuchihashi
authors.forename: Koichi
authors.initials: K
authors.lastname: Suzuki
mesh_terms: D015104:3,4-Methylenedioxyamphetamine / Q000378:metabolism / Q000592:standards / Q000652:urine*; D002853:Chromatography, Liquid / Q000592:standards*; D006801:Humans; D015394:Molecular Structure; D011786:Quality Control; D013237:Stereoisomerism; D053719:Tandem Mass Spectrometry / Q000592:standards*; D016482:Urinalysis / Q000379:methods*
chemical_list: D015104:3,4-Methylenedioxyamphetamine