Anabolic steroids are amongst the most misused substances in doping control and are intensively metabolized in humans. Adequate screening for misuse of these substances therefore relies on the detection of metabolites in urine samples collected from athletes.
Most of the studies investigating the metabolism of pharmaceutically available steroids were performed in the 1980’s via gas chromatography mass spectrometry (GC-MS). This research resulted in the selection of appropriate metabolites for the detection of steroid misuse. Over the years the selection of metabolites was further elaborated to include several metabolites that can result in prolonged detection times. Over the last decade, liquid chromatography tandem mass spectrometry (LC-MSn) was introduced as a screening technology in doping control laboratories world-wide. As a result several steroids that are difficult to detect via GC-MS became readily detectable. However, it should also be noted that several easily detectable metabolites by GC-MS are virtually undetectable via LC-MS. Indeed, both GC-MS and LC-MS are compatible techniques and the simultaneous application of both technologies is needed to cover the detection of all categories of structures. Recently, it has been shown that precursor ion scanning via LC-MS can be used to detect new steroid metabolites and the use of this technology has resulted in the
detection of previously unreported metabolites. These results have also illustrated
the limitations of GC-MS for the detection of steroid metabolites. Indeed, some
highly abundant metabolites in the LC-MS analysis could hardly be detected by GC-MS. This has highlighted the need for a re-investigation of steroid metabolism to allow for their adequate detection by LC-MS. Preliminary results for several steroids indicate that detection times could be prolonged considerably if more appropriate LC-MS metabolites are selected.