Development and validation of an LC‐MS/MS method after chiral derivatization for the simultaneous stereoselective determination of methylenedioxy‐methamphetamine (MDMA) and its phase I and II metabolites in human blood plasma

Abstract

This is the peer reviewed version of the following article: Steuer, A. E., Schmidhauser, C., Liechti, M. E., and Kraemer, T. (2015), Development and validation of an LC-MS/MS method after chiral derivatization for the simultaneous stereoselective determination of methylenedioxy-methamphetamine (MDMA) and its phase I and II metabolites in human blood plasma. Drug Test. Analysis, 7, 592-602, which has been published in final form at http://dx.doi.org/10.1002/dta.1740. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving. resulting in the formation of diastereomers which were easily separable on standard reverse phase stationary phases. This simple and fast method was validated according to international guidelines including specificity, recovery, matrix effects, accuracy and precision, stabilities, and limits of quantification. The method proved to be selective, sensitive, accurate and precise for all tested analytes except for DHMA. The method was applied to the analysis of more than 400 samples from a controlled study after application of MDMA.

Interactions between Bupropion and 3,4-Methylenedioxymethamphetamine in Healthy Subjects

“…It was important, however, to use a DA transporter inhibitor with no psychoactive effects, as shown for bupropion in the present study and previously (Peck and Hamilton, 1983;Oliveto et al, 2001) and in contrast to methylphenidate Schmid et al, 2014). Third, both MDMA and bupropion exhibit stereoselective metabolism (Kharasch et al, 2008;Steuer et al, 2014). The analytical method used in the present study was not stereoselective.…”

Chiral Plasma Pharmacokinetics of 3,4-Methylenedioxymethamphetamine and its Phase I and II Metabolites following Controlled Administration to Humans

“…Data on stability of MDMA and its phase I metabolites were previously published and no instability could be observed up to 6 months (Clauwaert et al, 2001). Sufficient stability of phase II metabolites under these storage conditions for 24 months and over at least three freeze/thaw cycles were shown during method validation (Steuer et al, 2015). Aliquots of plasma analyzed in the present pharmacokinetic study were thawed at a maximum of two times.…”

“…Owing to rather high LOQs (Steuer et al, 2015) presence of low DHMA amounts cannot be excluded. However, free HMMA also was not detectable (LOQ 0.0025 mM per enantiomer).…”

“…Blood plasma samples were analyzed using a Thermo Fisher Ultimate 3000 UHPLC system coupled to an ABSciex QTRAP 5500 in positive electrospray ionization (ESI) mode. Chromatography was performed on a Phenomenex Kinetex C18 column after chiral derivatization with Marfey's reagent (N-(2,4-dinitro-5-fluorophenyl) L-valinamide) as described in detail by Steuer et al (2015). Briefly, to 200 ml plasma mixed with 20 ml of the internal standard (IS) mixture (MDMA-d5, MDA-d5, pholedrine, dihydroxybenzylamine, 2.5 mM each), 1000 ml of acetonitrile were added; the mixture was shaken and centrifuged (10,000g, 5 minutes).…”

“…Full derivatization was observed with no peaks for underivatized analytes left and the formed diastereomers were baselineseparated from all analytes except for DHMA. The method was fully validated, including selectivity, recovery, matrix effects, bias and imprecision, stabilities, and limit of quantification (LOQ) and criteria were fulfilled and LOQs sufficient for the expected plasma concentrations of all analytes except for DHMA (Steuer et al, 2015). All blood plasma samples of a previous study (Schmid et al, 2015) were stored at -20°C prior to analysis.…”