Simultaneous enantiomeric determination of MDMA and its phase I and phase II metabolites in urine by liquid chromatography–tandem mass spectrometry with chiral derivatization

Abstract

A liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) procedure was developed for the simultaneous determination of enantiomers of the prevalent designer drug 3,4-methylenedioxymethamphetamine (MDMA) and its phase I and phase II metabolites in urine with chiral derivatization. The analytes in urine were directly derivatized with chiral Marfey's reagent, N(α)-(5-fluoro-2,4-dinitrophenyl)-D-leucinamide, without extraction. The diastereomers of the N(α)-(2,4-dinitrophenyl)-D-leucinamide derivatives generated were determined by LC-MS/MS. Satisfactory chromatographic separation was achieved for the enantiomers of MDMA and its metabolites 3,4-methylenedioxyamphetamine, 4-hydroxy-3-methoxymethamphetamine (HMMA), HMMA glucuronide, and HMMA sulfate on a semimicro octadecylsilane column using linear gradient elution. With use of multiple reaction monitoring mode, the limits of detection of these analytes ranged from 0.01 to 0.03 μg/mL. Linear calibration curves were obtained for all enantiomers from 0.1 to 20 μg/mL in urine. The method showed sufficient reproducibility and quantitative ability. This is the first report of a simple LC-MS/MS-based analytical procedure with direct chiral derivatization in aqueous media that allows simultaneous enantiomeric determination of drugs and their metabolites, including glucuronide and sulfate derivatives.

Development and validation of an LC‐MS/MS method after chiral derivatization for the simultaneous stereoselective determination of methylenedioxy‐methamphetamine (MDMA) and its phase I and II metabolites in human blood plasma

“…Recently, Nakanishi et al described a new LC-MS method after chiral derivatization with N-(2,4-dinitrophenyl)-D-leucinamide for stereoselective analysis of MDMA as well as of HMMA sulfate and HMMA glucuronide in urine, but unfortunately this method did not cover DHMA sulfates. [37] To the best of our knowledge no stereoselective methods for detection and quantification of MDMA and all relevant phase II metabolites in blood plasma are available at the moment. Therefore, the aim of the presented study was the development and validation of a stereoselective method for simultaneous analysis of MDMA and all relevant phase I and phase II metabolites in human blood plasma.…”

Rapid quantitative chiral amphetamines liquid chromatography–tandem mass spectrometry: Method in plasma and oral fluid with a cost-effective chiral derivatizing reagent

“…Foster et al implemented Marfey's reagent for MAMP chiral resolution and AMP in urine by HPLC-UV/VIS [15]. Recently, Nakanishi et al utilized a Marfey's reagent analogue, N(α)-(5-fluoro-2,4-dinitrophenyl)-d-leucinamide (d-FDLA), replacing the alanine moiety with leucine, for the resolution of MDMA and its phase I and II metabolites in urine via LC-MS/MS [13]; sample preparation was minimal and did not include a liquid-liquid or solid-phase extraction, and all enantiomers were resolved with an L-column2 semimicro octadecylsilane column. Our derivatization procedure was similar to Marfey's and Foster's.…”

“…A MAMP and AMP oral fluid (OF) method described chiral resolution by gas chromatography negative-ion chemical ionization mass spectrometry (GC-NICI-MS) [10]. Only a few methods utilized LC-MS for AMPs chiral analysis, performed in urine [6,13], hair [7] and human liver microsomes [8], but none in OF. OF is an increasingly popular alternative matrix for drugs of abuse testing in workplace, forensic, pain management, treatment, and clinical settings because its collection is easy compared to urine or blood, does not require a same-sex collector, and minimizes the opportunity for adulteration because of direct observation during collection [14].…”

Review of in situ derivatization techniques for enhanced bioanalysis using liquid chromatography with mass spectrometry

“…D‐FDLA enabled direct derivatization of amino groups of target analytes in aqueous media, such as urine, without prior hydrolysis or extraction. The limits of detection of these analytes ranged from 0.01 to 0.03 μg/mL .…”

“…Multiple reaction monitoring (MRM) chromatograms of DDLA derivatives of 3,4‐methylenedioxymethamphetamine (MDMA) and its metabolites added to drug‐free human urine at 1 μg/mL for each one (2 μg/mL as the racemic mixture) Peaks: a ( S )‐MDMA, b ( R )‐MDMA, c ( R )‐MDA, d ( S )‐MDA, e ( S )‐HMMA, f ( R )‐HMMA, g ( S )‐HMMA‐sul, h ( R )‐HMMA‐sul, i ( S )‐HMMA‐glu, j ( R )‐HMMA‐glu. Reproduced with permission from Springer (copyright 2012) .…”