Validation of a GC-MS screening method for anabolizing agents in solid nutritional supplements

A sensitive and selective method for the screening of 28 different compounds including testosterone and prohormones, nandrolone and prohormones, stanozolol and metandienone in solid nutritional supplements is described and validated. The different substances are extracted from the solid nutritional supplements by liquid-liquid extraction with a mixture of pentane and freshly distilled diethylether (9/1) after dissolving the supplement in NaOH (1 N). The anabolizing agents are derivatized with a mixture of MSTFA/NH(4)I/ethanethiol (320/1/2), routinely used for the derivatization of anabolic steroids extracted from urine. The TMS-derivatives are analysed by GC-MS in the SIM mode. The limits of detection were in the range from 2 to 40 ng/g. One supplement was analysed with this method and was found to contain several forbidden substances according to IOC doping regulations. All detected compounds, except dihydrotestosterone, could be confirmed with GC-MS(2), proving that the proposed method is suitable for the screening of anabolizing agents in solid nutritional supplements.