Screening for gestagens in kidney fat using accelerated solvent extraction and liquid chromatography electrospray tandem mass spectrometry Original Research Article

A screening method has been developed for the determination of various anabolic steroids in kidney fat. Fat samples are extracted and steroids are trapped “on-line” during accelerated solvent extraction (ASE). Following this initial extraction samples are further purified with C18 solid-phase extraction (SPE). Compounds were finally analyzed by liquid chromatography (LC) coupled with tandem mass spectrometry (MS). The method was validated using blank kidney fat and fat samples fortified at 2 ng g−1. The detection capability (CCβ) was <2 ng g−1 and the decision limit (CCα) varied from 0.3 to 0.9 ng g−1. Recoveries were in the range 17–58%, the accuracy using isotopically-labeled internal standards varied from 100 to 135%, and the method was successfully applied to screening of the gestagens flurogestone acetate (FGA), delmadinone acetate (DMA), megestrol acetate (MEGA), chloromadinone acetate (CMA), melengestrol acetate (MELA) and medroxyprogesterone acetate (MPA), and the androgen chlorotestosterone acetate (CTA) in kidney fat.