On the Metabolism and the Toxicological Analysis of Methylenedioxyphenylalkylamine Designer Drugs by Gas Chromatography-Mass Spectrometry

Designer drugs of the methylenedioxyphenylalkylamine type, such as MDA, MDMA, MDE, BDB, and MBDB, are increasingly abused. Their metabolism in humans involves two overlapping pathways: O-dealkylation of the methylenedioxy group to dihydroxy derivatives followed by methylation of one hydroxy group, and successive side-chain degradation to N-dealkyl and deaminooxo metabolites. MDA, MDMA, and MDE are further metabolized to glycine conjugates of 3,4-disubstituted benzoic acids. A gas chromatography–mass spectrometry (GC-MS) screening procedure was developed to detect these drugs and their metabolites in urine after acid hydrolysis, isolation at pH 8-9, and acetylation. Using mass chromatography with characteristic fragment ions, the method can detect abuse or intoxication at 5-50 ng/ml.

Study at a glance

Abstract

Designer drugs of the methylenedioxyphenylalkylamine type are increasingly abused. Studies on their metabolism in humans are necessary to develop a reliable gas chromatography--mass spectrometry (GC-MS) screening procedure. Such a method must allow their detection in urine for drug testing in clinical and forensic toxicology. Studies on racemic methylenedioxyamphetamine (MDA), methylenedioxymetamphetamine (MDMA), methylenedioxyethylamphetamine (MDE), benzodioxazolylbutanamine (BDB), and N-methylbenzodioxazolylbutanamine (MBDB) are presented. The metabolites were identified by GC-MS after enzymatic hydrolysis, isolation (pH 4.5 and 8-9), and derivatization (acetylation followed by methylation). The drugs undergo two overlapping metabolic pathways: O-dealkylation of the methylenedioxy group to dihydroxy derivatives followed by methylation of one of the hydroxy groups and successive degradation of the side chain to N-dealkyl and deaminooxo metabolites. MDA, MDMA, and MDE are subsequently metabolized to glycine conjugates of the corresponding 3,4-disubstituted benzoic acids. The hydroxy metabolites are excreted in a conjugated form. Based on these results, a GC-MS procedure was developed for simultaneous screening and identification of these designer drugs and/or their metabolites in urine after acid hydrolysis, isolation at pH 8-9, and acetylation. With use of mass chromatography with the most characteristic fragment ions m/z 58, 72, 86, 150, 162, 164, 176, and 178, the presence of the designer drugs was indicated and the peak underlying spectra could be identified by computerized comparison with reference spectra recorded during the presented studies. The procedure was suitable to detect an abuse of or an intoxication with the studied designer drugs (detection limit 5-50 ng/ml).