Detection and mass spectrometric characterization of novel long-term dehydrochloromethyltestosterone metabolites in human urine

The biotransformation of dehydrochloromethyltestosterone (DHCMT, 4-chloro-17 beta-hydroxy, 17 alpha-methylandrosta-1,4-dien-3-one) in man was studied with the aim to discover long-term metabolites valuable for the antidoping analysis. Having applied a high performance liquid chromatography for the fractionation of urinary extract obtained from the pool of several DHCMT positive urines, about 50 metabolites were found. Most of these metabolites were included in the GC-MS/MS screening method, which was subsequently applied to analyze the post-administration and routine doping control samples. As a result of this study, 6 new long-term metabolites were identified tentatively characterized using GC-MS and GC-MS/MS as 4-chloro-17 alpha-methyl-5 beta-androstan-3 alpha, 16, 17 beta-triol (M1), 4-chloro-18-nor-17 beta-hydroxymethyl,17 alpha-methyl-5 beta-androsta-1,13-dien-3 alpha-ol (M2), 4-chloro-18-nor-17 beta-hydroxymethyl, 17 alpha-methyl-5 beta-androst-13-en-3 alpha-ol (M3), its epimer 4-chloro-18-nor-17 alpha-hydroxymethyl, 17 beta-methyl-5 beta-androst-13-en-3 alpha-ol, 4-chloro-18-nor-17 beta-hydroxymethyl, 17 alpha-methylandrosta-4,13-dien-3 alpha-ol (M4) and its epimer 4-chloro-18-nor-17 alpha-hydroxymethyl, 17 beta-methylandrosta-4,13-dien-3 alpha-ol. The most long-term metabolite M3 was shown to be superior in the majority of cases to the other known DHCMT metabolites, such as 4-chloro-18-nor-17 beta-hydroxymethyl, 17 alpha-methylandrosta-1,4,13-trien-3-one and 4-chloro-3 alpha,6 beta,17 beta-trihydroxy-17 alpha-methyl-5 beta-androst-1-en-16-one. (C) 2011 Elsevier Ltd. All rights reserved.