After a single oral dose of MDMA (ecstasy), the drug and its metabolites are excreted in urine over an extended period, with the metabolite HMMA detectable longer than MDMA itself. In a double-blind study, healthy adult MDMA users received placebo, 1.0 mg/kg, or 1.6 mg/kg doses. From 916 urine specimens provided by 16 participants, median peak concentrations after the higher dose were 21,470 ng/mL for MDMA and 20,793 ng/mL for HMMA, with HMMA's last detection exceeding MDMA's by over 33 hours. In the first 24 hours, 30.2-34.3% of total urinary excretion occurred. Including HMMA in urine testing improves detection of MDMA exposure but requires hydrolysis of the sample.
A sensitive gas chromatography-mass spectrometry method was developed and validated to simultaneously measure MDEA, MDMA, and its metabolites (HMA, MDA, HMMA) in human urine. The method involves acid hydrolysis of 1 mL urine, solid-phase extraction, and derivatization before analysis. Limits of quantification were 25 μg/L for all analytes, with linear calibration up to 5000 μg/L. Extraction efficiencies exceeded 85.5%, and imprecision (coefficient of variation) was below 15% at tested concentrations. The assay meets federal workplace drug testing guidelines for these substances.
Acid hydrolysis recovers more MDMA urinary metabolites than enzymatic hydrolysis with beta-glucuronidase from either E. coli or Helix pomatia. Acid hydrolysis yielded 40.0% and 39.3% higher HMA recovery compared to E. coli and H. pomatia hydrolysis, respectively. E. coli beta-glucuronidase gave 17.1% and 26.5% greater MDA recovery than acid hydrolysis and H. pomatia hydrolysis. HMMA recovery by acid hydrolysis was 336.1% and 159.8% greater than E. coli and H. pomatia beta-glucuronidase. Optimal conditions were 100 µL hydrochloric acid per 1 mL urine incubated at 120°C for 40 minutes. Based on recovery, time, availability, and cost, acid hydrolysis is preferred.