Control of anabolic steroids misuse in sportPotential of direct detection of phase ii metabolites

Resumen

Anabolic androgenic steroids (AAS) are widely used by athletes to enhance sports performance. In this thesis, different studies were performed to evaluate the potential of the direct detection of phase II metabolites by LC-MS/MS to improve the detection of AAS misuse in sport. First, the direct detection of glucuronide metabolites was investigated using metandienone, allowing the detection of thirteen glucuronides, one of them resistant to enzymatic hydrolysis. The proposed analytical strategy provides a comprehensive approach and it is recommended to be used for metabolic studies of other AAS. Second, the detection of bisglucuronide conjugates of AAS was investigated. The mass spectrometric behavior was studied using synthesized AAS bisglucuronides to develop strategies for their detection. The formation of bisglucuronides was demonstrated in samples collected after norandrostenediol administration. Finally, the role of sulfate metabolites to detect endogenous AAS administration was evaluated. A LC-MS/MS method was developed for the direct quantitation of fourteen endogenous steroid sulfates. The use of some ratios between steroid sulfates improved the retrospectivity after oral and intramuscular testosterone administration. Steroid sulfates should be included in the steroid profile to complement the current markers. The results of the thesis demonstrate the increased role of the direct analysis of phase II metabolites of AAS in doping control.