Detection of the anabolic steroid methyltestosterone in hair by HPLC-EIA

An analytical method to detect the illegal application of the anabolic steroid methyltestosterone (MT) in cattle by hair analysis was developed. The time course of the incorporation of this orally active xenobiotic steroid into growing hair and the duration of its possible detection by hair analysis were measured. Female veal calves were fed with 35 mu g MT per kg body weight, twice daily, for 10 days. Before, during and after the treatment, hair samples were obtained and analyzed for MT residues. An appropriate method to extract MT from hair was developed. The extraction procedure consisted of liquid-liquid and solid-phase extraction and was followed by an essential high performance liquid chromatography (HPLC) step for further purification of the extracts. Final quantification was done with a specific enzyme immunoassay (EIA). MT residues could be detected in hair from day 4 (approximately 5 ng MT/g hair) of the experiment up to day 94 (approximately 0.5 ng MT/g). Our results suggest that hair analysis may be a powerful means to detect and track the illegal use of anabolic steroids.