Detection of Δ6-methyltestosterone in a dietary supplement and GC-MS/MS investigations on its urinary metabolism

Since a few years more and more products have appeared on the market for dietary supplements containing steroids that had never been marketed as approved drugs, mostly without proper labeling of the contents. Syntheses and few data on pharmacological effects are available dated back mainly to the 1950s or 1960s. Only little knowledge exists about effects and side effects of these steroids in humans. The present study reports the identification of Delta 6-methyltestosterone in a product named Jungle Warfare, which was obtained from a web-based supplement store. The main urinary metabolites, 17 alpha-hydroxy-17 beta-methylandrosta-4,6-dien-3-one (Delta 6-epimethyltestosterone), 17 alpha-methyl-5 beta-androstane-3 alpha,17 beta-diol (3 alpha,5 beta-THMT), and 17 beta-methyl-5 beta-androstane-3 alpha,17 alpha-diol, as well as the parent compound excreted after a single oral administration were monitored by GC-MS/MS. Delta 6-Epimethyltestosterone and 3 alpha,5 beta-THMT served for long-term detection (still present in the 181-189h urine). 17 alpha-Methyltestosterone and its 17-epimer were not detected in the urines (LOD 0.3 ng/mL). The highest concentrations were found in the 14-20.5 h urine for Delta 6-epimethyltestosterone (600 ng/mL), and 3 alpha,5 beta-THMT (240 ng/mL) and in the 36-44.5 h urine for 17 beta-methyl-5 beta-androstane-3 alpha,17 alpha-diol (7 ng/mL). For reference methyltestosterone and epimethyltestosterone were dehydrogenated with chloranil. The characterization of the products was performed by GC-MS(/MS) and NMR (C) 2010 Elsevier Ireland Ltd. All rights reserved.