Determination of Steroid Hormones in Human Saliva by High-Performance Liquid Chromatography with Tandem Mass Spectrometry Detection

A method is developed for the simultaneous determination of steroid hormones of various classes (androgens, progestins, and glucocorticoids) in human saliva using liquid-liquid extraction and detection by ultrahigh-performance liquid chromatography-tandem mass spectrometry. It was found that the most suitable solvent for the extraction of the target analytes is methyl tert-butyl ether. The volume of the extractant and other factors affecting the extraction of analytes from the analyzed matrix were optimized using multivariate analysis (Box-Behnken design). Under optimal conditions, the recoveries of analytes are more than 90%, and the limits of detection are in the range of 50-250 pg/mL, which indicates the high sensitivity of the proposed procedure.