Quantification of testosterone undecanoate in human hair by liquid chromatography-tandem mass spectrometry

Testosterone undecanoate (T-C11) can be used by athletes in order to improve performance. After oral intake, T-C11 is rapidly metabolized, hampering discrimination between exogenous and endogenous testosterone. A possible alternative is to detect the intact ester in hair. A method based on liquid chromatography-tandem mass spectrometry was developed for the determination of T-C11 in hair. The sample procedure consisted of digestion of 200 mg of pulverized hair with tris(2-carboxyethyl)phosphine hydrochloride and liquid-liquid extraction with n-pentane. Several parameters such as the mobile phase, the ionization source and the washing step were optimized. The method was validated at different spiked levels obtaining satisfactory values for accuracy (between 92 and 102%) with relative standard deviations lower than 7% and a limit of detection of 0.2 ng/g. The applicability of the method was checked by the analysis of three samples from patients using T-C11. A peak for the analyte was detected in all samples with concentrations between 0.4 and 8.4 ng/g.