title: Development and validation of LC-HRMS and GC-NICI-MS methods for stereoselective determination of MDMA and its phase I and II metabolites in human urine.
abstract: 3,4-Methylenedioxymethamphetamine (MDMA) is a racemic drug of abuse and its R- and S-enantiomers are known to differ in their dose-response curve. The S-enantiomer was shown to be eliminated at a higher rate than the R-enantiomer most likely explained by stereoselective metabolism that was observed in various in vitro experiments. The aim of this work was the development and validation of methods for evaluating the stereoselective elimination of phase I and particularly phase II metabolites of MDMA in human urine. Urine samples were divided into three different methods. Method A allowed stereoselective determination of the 4-hydroxy-3-methoxymethamphetamine (HMMA) glucuronides and only achiral determination of the intact sulfate conjugates of HMMA and 3,4-dihydroxymethamphetamine (DHMA) after C18 solid-phase extraction by liquid chromatography-high-resolution mass spectrometry with electrospray ionization. Method B allowed the determination of the enantiomer ratios of DHMA and HMMA sulfate conjugates after selective enzymatic cleavage and chiral analysis of the corresponding deconjugated metabolites after chiral derivatization with S-heptafluorobutyrylprolyl chloride using gas chromatography-mass spectrometry with negative-ion chemical ionization. Method C allowed the chiral determination of MDMA and its unconjugated metabolites using method B without sulfate cleavage. The validation process including specificity, recovery, matrix effects, process efficiency, accuracy and precision, stabilities and limits of quantification and detection showed that all methods were selective, sensitive, accurate and precise for all tested analytes.
authors.affiliation: Department of Experimental and Clinical Toxicology, Institute of Experimental and Clinical Pharmacology and Toxicology, Saarland University, D-66421 Homburg (Saar), Germany.
authors.forename: Andrea E
authors.initials: AE
authors.lastname: Schwaninger
authors.forename: Markus R
authors.initials: MR
authors.lastname: Meyer
authors.forename: Marilyn A
authors.initials: MA
authors.lastname: Huestis
authors.forename: Hans H
authors.initials: HH
authors.lastname: Maurer
mesh_terms: D002853:Chromatography, Liquid; D003846:Deoxyepinephrine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000652:urine; D004355:Drug Stability; D008401:Gas Chromatography-Mass Spectrometry; D020719:Glucuronides / Q000737:chemistry / Q000652:urine; D006801:Humans; D016018:Least-Squares Analysis; D013058:Mass Spectrometry / Q000379:methods*; D008694:Methamphetamine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000652:urine; D018817:N-Methyl-3,4-methylenedioxyamphetamine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000652:urine*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013237:Stereoisomerism; D013431:Sulfates / Q000737:chemistry / Q000652:urine
chemical_list: D020719:Glucuronides; D013431:Sulfates; C079646:4-hydroxy-3-methoxymethamphetamine; C069512:alpha-methylepinine; D008694:Methamphetamine; D018817:N-Methyl-3,4-methylenedioxyamphetamine; D003846:Deoxyepinephrine