Analysis of MDMA and its Metabolites in Urine and Plasma Following a Neurotoxic Dose of MDMA

After a single neurotoxic dose (20 mg/kg) of MDMA given to male Dark Agouti rats, the drug and three of its metabolites—MDA, HMMA, and HMA—appeared in urine at different peak times: MDMA at 4 hours, MDA at 8 hours, HMMA at 12 hours, and HMA at 16 hours. All four compounds remained detectable in urine for up to 168 hours (detection limit 25 ng/mL). In plasma, MDMA and MDA peaked at 2 hours, HMMA at 4 hours, and HMA at 24 hours, but with high variability at that time point. No MDMA or metabolites were detectable in plasma at 96 or 168 hours (detection limit 10 ng/mL).

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Abstract

3,4-Methylenedioxymethamphetamine (MDMA), a commonly encountered drug of abuse, has been shown in a variety of studies to cause neurotoxic effects. Because MDMA itself is not neurotoxic, identifying the potential neurotoxic metabolite(s) was of significant importance. Evaluation of urine and plasma concentrations of MDMA and three of its main metabolites, 3,4-methylenedioxyamphetamine (MDA), 4-hydroxy-3-methoxyamphetamine (HMA), and 4-hydroxy-3-methoxymethamphetamine (HMMA), following administration of a neurotoxic dose (20 mg/kg) to male Dark Agouti rats was accomplished. Currently there are no data available describing urine and plasma concentrations of MDMA and these metabolites over a period of 7 days. The rats received a single 20 mg/kg i.p. dose of MDMA. Blood and urine samples were collected prior to administration and at 2, 4, 8, 12, 16, 20, 24, 48, 96, and 168 h following drug administration. Plasma and urine samples were extracted using solid-phase extraction, derivatized with N-methyl-bis(trifluoroacetamide), then analyzed using gas chromatography-mass spectrometry. Urine samples showed peak concentrations of MDMA at 4 h, MDA at 8 h, HMMA at 12 h, and HMA at 16 h post dose. MDMA and its metabolites were detectable (limit of detection 25 ng/mL) in the urine for up to 168 h post dose. Plasma samples showed mean peak concentrations of MDMA and MDA at 2 h post dose and HMMA at 4 h. Although the highest mean concentration of HMA was seen at 24 h post dose, variability between sample results for this time point was significant. No detectable levels of MDMA, MDA, HMA, and HMMA (LOD 10 ng/mL) were found in plasma at 96 and 168 h post dose.