Validation and application of a screening method for beta2-agonists, anti-estrogenic substances and mesocarb in human urine using liquid chromatography/tandem mass spectrometry

Electrospray ionization (ESI) mass spectra of 15 anti-estrogenic substances, beta2-agonists and mesocarb were investigated in terms of fragmentation patterns. On the basis of this product ion information, a simultaneous screening method for anti-estrogenic substances, beta2-agonists and mesocarb was developed for doping control purposes. After hydrolysis, liquid-liquid extraction was adopted for the sample preparation. The recoveries for all compounds were 30 and 96%. A single liquid chromatography/tandem mass spectrometry (LC/MS/MS) analysis could be performed in 13 min for the analysis of 15 anti-estrogenic substances, beta2-agonists and mesocarb. A quantitative analysis was also validated. Inaccuracies were below +/-12% and precisions varied from 0 to 15.8%. The limit of detection was below 10 ng/mL except formestane (300 ng/mL) and aminoglutethimide (100 ng/mL). The validated method was applied for the analysis of excretion samples.