ABSTRACT
A long-term metabolite of the doping agent oxymetholone (OXM-M2, 17ß-hydroxymethyl-2,17α-methyl-18-norandrost-13-en-3-one) which has been identified by GC-MS/MS was synthesized from commercially available materials. Two efficient synthetic routes to access both C-17 epimers of tentative metabolites were developed. The identity and molecular configuration of the in vivo metabolite: 17ß-hydroxymethyl-2α,17α-methyl-18-norandrost-13-en-3-one was confirmed by single crystal X-ray diffraction.
Subject(s)
Oxymetholone/chemical synthesis , Oxymetholone/metabolism , Crystallography, X-Ray , Humans , Models, Molecular , Molecular Conformation , Oxymetholone/chemistryABSTRACT
The metabolism of two anabolic steroids - oxymetholone and desoxymethyltestosterone - was reinvestigated to identify new targets potentially valuable for the antidoping analysis. Excretion urine samples from the laboratory reference collection were used in this study. Following fractionation of the urinary extract by means of high performance liquid chromatography (HPLC), each fraction was subjected to gas chromatography-mass spectrometry (GC-MS) and gas chromatography-tandem mass spectrometry (GC-MS/MS) analysis after trimethylsilylation. About 20 metabolites were found for desoxymethyltestosterone and more than 40 for oxymetholone, with many of them being isomeric compounds. In addition to the well-known reduced and hydroxylated metabolites, 18-nor-17,17-dimethyl and 18-nor-17-hydroxymethyl-17-methyl steroids were also identified. Having evaluated all the metabolites in terms of how long they could be detected, we suggest that 18-nor-2ξ,17ß-hydroxymethyl-17α-methyl-5α-androst-13-en-3α-ol is an important marker of oxymetholone abuse. In case of desoxymethyltestosterone, better detectability could be achieved if 18-nor-17,17-dimethyl-5α-androst-13-en-2ξ,3α-diol is monitored. These novel metabolites could be detected using GC-MS/MS at least for 14 days after administration of these anabolic steroids compared to 5-7 days for previously reported metabolites.
Subject(s)
Anabolic Agents/urine , Androgens/urine , Androstenols/urine , Oxymetholone/urine , Substance Abuse Detection/methods , Anabolic Agents/metabolism , Androgens/metabolism , Androstenols/metabolism , Chromatography, High Pressure Liquid/methods , Doping in Sports , Gas Chromatography-Mass Spectrometry/methods , Humans , Male , Oxymetholone/metabolism , Tandem Mass Spectrometry/methodsABSTRACT
An acidic metabolite, 2alpha-carboxy-5alpha-androstane-3alpha,16alpha,17alpha-triol and two neutral metabolites, 2alpha-hydroxymethyl-5alpha-androstane-3alpha,17alpha-diol, and 2alpha-hydroxymethyl-5alpha-androstane-3alpha,16alpha,17alpha-triol have been identified in the urine of rabbits orally dosed with 17beta-hydroxy-2-hydroxymethylene-5alpha-androstan-3-one. 2alpha-Hydroxymethyl-5alpha-androstane-3alpha,16alpha,17alpha-triol was previously obtained from the urine of rabbits dosed with 17beta-hydroxy-2alpha-methyl-5alpha-androstan-3-one. The acidic metabolite was the major urinary excretion product.
