title: Determination of 3,4-methylenedioxymethamphetamine and its five main metabolites in rat urine by solid-phase extraction and high performance liquid chromatography with on line mass spectrometry.
abstract: The consumption of psychostimulant amphetamine-like drugs has increased significantly in recent years. Some MDMA metabolites are probably involved in the neurotoxicity and neurodegeneration caused by prolonged use rather than MDMA itself. We recently developed a method to analyze MDMA and its five main metabolites in rat plasma [7]. We have now fully validated this method to the quantification of these drugs in rat urine. We extracted MDMA and its metabolites with Oasis WCX cartridges, separated them on a Nucleodur C18 analytical column and quantified them by ion-trap mass spectrometry. Linearity was excellent: 12.5-1250ng/mL urine for HMA, HMMA, MDA and MDMA, 25-2500ng/mL for HHMA, and 150-7500ng/mL for HHA (r(2)>0.993 for all analytes). The lower limits of quantification were 12.5ng/mL urine for MDMA, MDA, HMA and HMMA, 25ng/mL for HHMA and 150ng/mL for HHA. Reproducibility was good (intra-assay precision=1.7-6.1%; inter-assay precision=0.6-5.7%), as was accuracy (intra-assay deviation=0.1-4.8%; inter-assay deviation=0.7-7.9%). Average recoveries were around 85.0%, except for HHMA (66.2%) and HHA (53.0%) (CV<8.3%). We also checked the stability of stock solutions and the internal standards after freeze-thawing and in the autosampler. Lastly, we measured the MDMA, MDA, HHMA, HHA, HMMA and HMA in urine samples taken over 24h from rats given subcutaneous MDMA.
authors.affiliation: EA 4463 Université Paris Descartes, Laboratoire de Chimie-Toxicologie Analytique et Cellulaire, Faculté de Pharmacie, Avenue de l'Observatoire, F-75006 Paris, France. marie-claude.menet@parisdescartes.fr
authors.forename: Marie-Claude
authors.initials: MC
authors.lastname: Menet
authors.forename: Julien
authors.initials: J
authors.lastname: Fonsart
authors.forename: Françoise
authors.initials: F
authors.lastname: Hervé
authors.forename: Dominique
authors.initials: D
authors.lastname: Fompeydie
authors.forename: Martine
authors.initials: M
authors.lastname: Galliot-Guilley
authors.forename: Florence
authors.initials: F
authors.lastname: Noble
authors.forename: Jean-Michel
authors.initials: JM
authors.lastname: Scherrmann
mesh_terms: D000662:Amphetamines / Q000737:chemistry / Q000378:metabolism / Q000652:urine; D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004298:Dopamine / Q000031:analogs & derivatives / Q000737:chemistry / Q000378:metabolism / Q000652:urine; D004355:Drug Stability; D016014:Linear Models; D008297:Male; D013058:Mass Spectrometry; D018817:N-Methyl-3,4-methylenedioxyamphetamine / Q000737:chemistry / Q000378:metabolism / Q000652:urine*; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D052616:Solid Phase Extraction / Q000379:methods*
chemical_list: D000662:Amphetamines; D018817:N-Methyl-3,4-methylenedioxyamphetamine; D004298:Dopamine