Studies on the metabolism of the α-pyrrolidinophenone designer drug methylenedioxy-pyrovalerone (MDPV) in rat and human urine and human liver microsomes using GC-MS and LC-high-resolution MS and its detectability in urine by GC-MS.
Since the late 1990s, many derivatives of the α-pyrrolidinophenone (PPP) drug class appeared on the drugs of abuse market. The latest compound was described in 2009 to be a classic PPP carrying a methylenedioxy moiety remembering the classic entactogens (ecstasy). Besides Germany, 3,4-methylene-dioxypyrovalerone (MDPV) has appeared in many countries in Europe and Asia, indicating its worldwide importance for forensic and clinical toxicology. The aim of the presented work was to identify the phase I and II metabolites of MDPV and the human cytochrome-P450 (CYP) isoenzymes responsible for its main metabolic step(s). Finally, the detectability of MDPV in urine by the authors' systematic toxicological analysis (STA) should be studied. The urine samples were extracted after and without enzymatic cleavage of conjugates. The metabolites were separated and identified after work-up by GC-MS and liquid chromatography (LC)-high-resolution MS (LC-HR-MS). The studies revealed the following phase I main metabolic steps in rat and human: demethylenation followed by methylation, aromatic and side chain hydroxylation and oxidation of the pyrrolidine ring to the corresponding lactam as well as ring opening to the corresponding carboxylic acid. Using LC-HR-MS, most metabolite structures postulated according to GC-MS fragmentation could be confirmed and the phase II metabolites were identified. Finally, the formation of the initial metabolite demethylenyl-MDPV could be confirmed using incubation of human liver microsomes. Using recombinant human CYPs, CYP 2C19, CYP 2D6 and CYP 1A2 were found to catalyze this initial step. Finally, the STA allowed the detection of MDPV metabolites in the human urine samples.
Department of Experimental and Clinical Toxicology, Institute of Experimental and Clinical Pharmacology and Toxicology, Saarland University, D-66421 Homburg (Saar), Germany. markus.meyer@uks.eu
Markus R
MR
Meyer
Peng
P
Du
Frank
F
Schuster
Hans H
HH
Maurer
D000818:Animals; D001189:Aryl Hydrocarbon Hydroxylases / Q000378:metabolism; D052117:Benzodioxoles / Q000032:analysis / Q000737:chemistry / Q000378:metabolism* / Q000652:urine; D002853:Chromatography, Liquid / Q000379:methods*; D015198:Designer Drugs / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D006801:Humans; D053858:Metabolic Networks and Pathways; D008862:Microsomes, Liver / Q000378:metabolism*; D011759:Pyrrolidines / Q000032:analysis / Q000737:chemistry / Q000378:metabolism* / Q000652:urine; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism; D000094982:Synthetic Cathinone
D052117:Benzodioxoles; D015198:Designer Drugs; D011759:Pyrrolidines; D011994:Recombinant Proteins; D001189:Aryl Hydrocarbon Hydroxylases; D000094982:Synthetic Cathinone