Urinary Excretion Kinetics of 3,4-Methylenedioxymethamphetamine (MDMA, Ecstasy) and Its Phase I and Phase II Metabolites in Humans following Controlled MDMA Administration

Abstract

BACKGROUND 3,4-Methylendioxymethamphetamine (MDMA) is excreted in human urine as unchanged drug and phase I and II metabolites. Previous urinary excretion studies after controlled oral MDMA administration have been performed only after conjugate cleavage. Therefore, we investigated intact MDMA glucuronide and sulfate metabolite excretion. METHODS We used LC–high-resolution MS and GC-MS to reanalyze blind urine samples from 10 participants receiving 1.0 or 1.6 mg/kg MDMA orally. We determined median Cmax, tmax, first and last detection times, and total urinary recovery; calculated ratios of sulfates and glucuronides; and performed in vitro–in vivo correlations. RESULTS Phase II metabolites of 3,4-dihydroxymeth-amphetamine (DHMA), 4-hydroxy-3-methoxymeth-amphetamine (HMMA), 3,4-dihydroxyamphetamine (DHA), and 4-hydroxy-3-methoxyamphetamine were identified, although only DHMA sulfates, HMMA sulfate, and HMMA glucuronide had substantial abundance. Good correlation was observed for HMMA measured after acid hydrolysis and the sum of unconjugated HMMA, HMMA glucuronide, and HMMA sulfate (R2 = 0.87). More than 90% of total DHMA and HMMA were excreted as conjugates. The analyte with the longest detection time was HMMA sulfate. Median HMMA sulfate/glucuronide and DHMA 3-sulfate/4-sulfate ratios for the first 24 h were 2.0 and 5.3, respectively, in accordance with previous in vitro calculations from human liver microsomes and cytosol experiments. CONCLUSIONS Human MDMA urinary metabolites are primarily sulfates and glucuronides, with sulfates present in higher concentrations than glucuronides. This new knowledge may lead to improvements in urine MDMA and metabolite analysis in clinical and forensic toxicology, particularly for the performance of direct urine analysis.

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

“…In vivo, sulfation initially predominated, changing to comparable amounts of both conjugates after approximately 3 hours, explainable by differences in enzymatic affinity and capacity observed in vitro. While SULTs show much higher affinity for HMMA, they are easily saturated, and the higher capacity UGTs can take over (Schwaninger et al, 2009(Schwaninger et al, , 2011a(Schwaninger et al, , 2011c. Consequently, the phase II metabolites should be the main metabolites in plasma.…”

“…A minor pathway includes the formation of 3,4-methylenedioxyamphetamine (MDA) by N-demethylation followed again by O-demethylation, O-methylation, and conjugation (Maurer, 1996;Maurer et al, 2000;de la Torre et al, 2004). Analysis of urine samples after recreational MDMA consumption revealed MDMA and primarily phase II metabolites (DHMA 3-sulfate, HMMA 4-sulfate, and HMMA 4-glucuronide) as major excretion products (Schwaninger et al, 2011a). Other metabolites could only be detected at negligible concentrations.…”

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

“…The S-enantiomer of MDMA is eliminated at a higher rate than the R-enantiomer The analytes included in the method were chosen based on the main metabolites identified in urine [20] and on the measurement of five authentic MDMA blood samples checking for all possible metabolites. In these samples mainly MDMA, HMMA 4-sulfate, HMMA 4-glucuronide, and DHMA 3-sulfate could be detected.…”

“…A minor pathway includes demethylation to 3,4-methylendioxyamphetamine (MDA) followed by demethylenation to 3,4-dihydroxyamphetamine, O-methylation to 4-hydroxy-3-methoxyamphetamine (HMA) and conjugation [17][18][19]. In urine samples of recreational MDMA users following a controlled single dose of MDMA, DHMA 3-sulfate, HMMA 4-sulfate and HMMA 4-glucuronide were detected as major metabolites next to unchanged MDMA [20].…”

Targeting misuse of 2‐amino‐N‐ethyl‐1‐phenylbutane in urine samples: in vitro–in vivo correlation of metabolic profiles and development of LC‐TOF‐MS method

“…In general, this metabolite was predominantly conjugated but interestingly, the main fraction was glucuronide-conjugated in Cases 2 and 6, sulpho-conjugated in Cases 1 and 3, and somewhat of an equal distribution in Cases 4 and 5. [14] Although number of cases in our study is small, and no background information was available on the dosing and timing of 2-AEPB, these observations underlines the variability of the routine samples that the laboratory should be able to handle in order to maintain effective drug-of-abuse testing. Furthermore, taking into account the origin of the samples and potentially high degree of drug abuse, the effect of enzyme induction or inhibition cannot be ruled out.…”