In Vitro Synthesis of Metabolites of three Anabolic Androgenic Steroids, by Human Liver Microsomes

Abstract

Anabolic androgenic steroids are substances frequently misused to improve physicalperformance in sports. To reveal substances misused as doping, athlete urinesamples are collected and tested. To identify the steroid and/or its metabolitesin urine, reference compounds must exist for comparison. The time-consumingand ethical concerns about in vivo excretion studies for the examination of thesecompounds, make the use of liver fragment microsomes an attractive alternative.The aim of this thesis was to synthesize and identify metabolites from known andrare anabolic androgenic steroids, by the use of human liver microsomes. Liveris an important organ in steroid metabolism. By incubating AAS with humanliver microsomes and co factors, an in vitro simulation of the liver metabolism wascarried out. A conrmation of metabolites was performed by gas chromatographytandem mass spectrometry in full scan, MRM mode, or both. 6beta-hydroxymethylmetandienon, epimetandienon and 17,17-dimethylmetandienon were successfullysynthesized from metandienon, and the 17beta-hydroxymetandienon was producedfrom the 17,17-dimethyl metabolite. Respectively three and one metabolite(s)were found for the "designer steroids" methylnortestosteron and madol. Metabolitevariations were observed regarding the optimal time of incubation, and enzymaticrequirements of formation.