Abstract

Nowadays, testosterone (T) remains a highly abused anabolic androgenic steroid for the practice of doping. There are many causes that make its detection particularly complicated, since it is an endogenous compound and its concentration in spot urine samples is highly variable. Many approaches have been proposed for the detection of T misuse. From the classical T/E (reference population) to personalize the references, the individual became its own reference. To improve the urinary steroid profile one way is to extend its evaluation with the inclusion of new biomarkers, as proposed by Renterghem et al. and Pozo et al. But little is known about the origin of these metabolites and on their stability in the different conditions of extraction. In the first step of the study we have studied the stability of 25 hydroxylated steroids in different conditions: usual screening condition, soft hydrolysis and two aggressive hydrolysis conditions (acid hydrolysis with H2SO4/organic solvent and basic hydrolysis with KOH). Their behavior and their relationship with the new possible compounds formed were analyzed using two techniques GC-MS and LC-MS (TOF). We could clearly demonstrate that, following different hydrolytic procedures, the hydroxylated steroid are stable as expected in screening or soft hydrolysis conditions, while in aggressive hydrolysis conditions the changes are very pronounced. New compounds are formed, and after a detailed characterization, it was found that the hydroxy group position on the steroid skeleton is important for the hydroxylated steroid stability and type of product formed. Verf.-Referat