Gas chromatography–mass spectrometric confirmation of anabolic compounds in injection sites

Abstract

Intramuscularly administered, many anabolic residues are present in injection sites in an esterified form. Routine high-performance thin-layer chromatography (HPTLC) results show different spots recognizable as the esters of a certain anabolic compound, e.g., testosterone or estradiol, without giving the exact identity of the ester. Hydrolysis of the extract and respotting on HPTLC plates can confirm these spots to be esters owing to the absence of such spots on the plate after hydrolysis. The residues are then seen in their unesterified form. Identification is possible after respotting on reversed phase plates. As this method is complex and time consuming, an alternative method was tried out using gas chromatography–mass spectrometry (GC–MS). GC–MS analysis of injection site extracts is not only an independent method to confirm HPTLC results, but also provides the opportunity to identify the specific ester of the anabolic steroid. Identification is only possible when a standard of the ester is available for comparing the respective mass spectra. This report presents the mass spectra of some esters of testosterone, estradiol and nortestosterone found during the confirmation analysis of 35 injection sites by using GC–MS.