Optimization of enantioselective high-performance liquid chromatography-tandem mass spectrometry method for the quantitative determination of 3,4-methylenedioxy-methamphetamine (MDMA) and its phase-1 metabolites in human biological fluids.
Giorgia Sprega, Giorgi Kobidze, Alfredo Fabrizio Lo Faro, Simona Pichini, Tivadar Farkas, Anastasio Tini, Antonina Mskhiladze, Francesco Paolo Busardò, Bezhan Chankvetadze
Journal of pharmaceutical and biomedical analysis June 15, 2024 DOI: 10.1016/j.jpba.2024.116076 via PubMed
Summary
AI-generated from the abstractAn improved chiral LC-MS/MS method separates all four pairs of enantiomers of MDMA and its major phase-1 metabolites (HMA, HMMA, MDA) on a single Lux AMP column within six minutes, using an optimized mobile phase and column dimensions. The method was applied to human plasma, oral fluid, and urine. In urine, hydrolysis of glucuronides with hydrochloric acid or glucuronidase was tested to evaluate effects on the concentration and enantiomeric distribution of the hydroxy metabolites HMA and HMMA.
Study at a glance
| Characteristics | Method development and validation Peer reviewed |
|---|---|
| Population | Human biological fluids (plasma, oral fluid, urine) |
| Keywords | Chiral hplc Enantioselective metabolism Enantioseparation Hydrolysis of glucuronides MDMA And Metabolites |
| Citations | 4 |
| Key finding | Baseline separation of all four pairs of enantiomers of MDMA and its major phase-1 metabolites was achieved on a single Lux AMP column within six minutes. |
Abstract
Recently we published in this journal an enantioselective high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the quantitative determination of 3,4-methylenedioxymethamphetamine (MDMA) and its major phase-1 metabolites, 4-hydroxy-3-methoxyamphetamine (HMA), 4-hydroxy-3-methoxymethamphetamine (HMMA) and 3,4-methylenedioxyamphetamine (MDA) in human plasma, sweat, oral fluid and urine. Since we did not achieve simultaneous enantioseparation of all 4 compounds with a single chiral column, two amylose-based chiral columns were used alternatively. Further optimization of the mobile phase in the present study enabled baseline separation of all four pairs of enantiomers on a single Lux AMP column. In addition, by optimization of the column dimension and applied flow-rate it became possible to complete the separation within 6 minutes. These new methods were applied to the analysis of human plasma, oral fluid and urine. While results on the concentration of MDMA and its metabolites in various biological fluids were reported in our recent publication, in the present study an attempt was made to hydrolyze glucuronides in urine samples by using alternatively, hydrochloric acid or glucuronidase and to evaluate the effect of hydrolysis on the concentration and enantiomeric distribution of hydroxy metabolites of MDMA such as HMA and HMMA.