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.
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.