Detection of the misuse of steroids in doping control

The list of prohibited substances of the World Anti-Doping Agency (WADA) classifies the administration of several steroids in sports as doping. Their analysis is generally performed using urine specimen as matrix. Lots of the steroids are extensively metabolised in the human body. Thus, knowledge of urinary excretion is extremely important for the sensitive detection of steroid misuse in doping control. The methods routinely used in steroid screening mainly focus on substances, that are excreted unconjugated or as glucuronides. Common procedures include deconjugation using a beta-glucuronidase enzyme. Following extraction and concentration the analytes are submitted to LC-MS(/MS) analysis and/or GC-MS(/MS) analyses. Besides the classical steroids, more and more products appear on the market for "dietary supplements" containing steroids that have never been marketed as approved drugs, mostly without proper labelling of the contents. To cover the whole range of potential products comprehensive screening tools have to be utilised in addition to the classical methods. Endogenous steroids, e.g. testosterone, represent a special group of compounds. As classical chemical methodology is incapable of discriminating synthetic hormones from the biosynthesised congeners, the method of steroid profiling is used for screening purpose. Additionally, based on isotope signatures a discrimination of synthetic and natural hormones can be achieved.