Direct determination of p-hydroxymethamphetamine glucuronide in human urine by high-performance liquid chromatography

High-performance liquid chromatography (HPLC) with UV detection for the simultaneous determination of the free form of p-hydroxymethamphetamine (p-OHMA) and its metabolite, glucuronide (p-OHMAG) was accomplished for the first time. We achieved this by employing 1) an ion pair reagent for retention of sample to a solid-phase extraction (SPE) cartridge, Sep-Pak Light C18 and 2) a simple two-step stepwise elution technique for subsequent ion pair RP-HPLC. The proposed method was optimized for resolution of p-OHMAG, p-OHMA and MA. The method was successfully applied to urine samples collected from MA abusers.