Detection of testosterone and epitestosterone in human hair using liquid chromatography tandem mass spectrometry

Deshmukh, Nawed, Barker, James, Petroczi, Andrea and Naughton, Declan (2012) Detection of testosterone and epitestosterone in human hair using liquid chromatography tandem mass spectrometry. Journal of Pharmaceutical and Biomedical Analysis, 67-68, pp. 154-158. ISSN (print) 0731-7085

Abstract

The feasibility of using hair analysis as a complimentary test in doping control has received increased attention in the scientific community. The aim of the study was to take a step forward to this goal and develop a method that, for the first time, is able to detect testosterone (T) and epitestosterone (E) in human hair, using liquid chromatography-tandem mass spectrometry (LC-MS/MS) and alkali digestion followed by extraction using pentane. The method was linear within the quantification range of 0.25-100pg/mg for T and 0.5-100pg/mg for E, with determination coefficient (r(2)) values >0.9987. The limits of detection for T and E were 0.1pg/mg and 0.25pg/mg respectively. The accuracy, precision and extraction recovery of the assay were satisfactory for the detection of T and E when ca. 50mg hair was processed. The validated method was successfully applied for the analysis of 75 hair samples collected from healthy volunteers (65.3% males), with the concentration of T between 0.7-11.81pg/mg and 0.33-6.05pg/mg and the concentration of E between 0.63-8.27pg/mg and 0.52-3.88pg/mg in males and in females respectively. In males, the T levels were significantly higher (p=0.020) but there was no difference in the E levels (p=0.359). However, E was not detectable in 34 samples (of which 19 were females). The T and E levels showed linear correlation (r=0.698, p<0.001) with average T/E ratio of 1.32±0.7. The newly developed analytical method was rapid, facile, sensitive, selective, reproducible and reliable for determining the levels of T and E in hair and thus for calculating the T/E ratio in hair.

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