Determination of Ketosteroids in Human Urine Using Dispersive Liquid-Liquid Microextraction and Ultra High-Performance Liquid Chromatography-High Resolution Mass Spectrometry

Abstract

A method is proposed for the determination of some ketosteroids in human urine, including enzymatic hydrolysis using β-glucuronidase from E. coli followed by dispersive liquid-liquid microextraction, derivatization of analytes with hydroxylamine, and detection by reversed-phase ultra-HPLC–quadrupole time-of-flight mass spectrometry. Optimization of extraction and derivatization conditions of the studied compounds made it possible to find that the highest recoveries were achieved using an acetone–chloroform mixture as a dispersant and an extractant, and the completeness of the derivatization reaction was achieved by thermostating the sample at 70°C for 90 min. The proposed method has high sensitivity (limits of detection in the range of 0.1–0.25 ng/mL) and a wide linearity range.

Cyanoacetohydrazide as a Novel Derivatization Agent for the Determination of UHPLC-HRMS Steroids in Urine

“…Regardless of the form of steroids determined during the analysis, the quantification of steroid hormones at trace levels necessitates preliminary sample concentration. The most commonly employed techniques for this purpose include liquid-liquid extraction [ 19 , 20 , 21 ], solid-phase extraction [ 22 , 23 , 24 ], and dispersive liquid-liquid microextraction [ 25 , 26 , 27 ]. In addition, it is imperative to consider that performing non-targeted steroid screening, encompassing estrogens, androgens, progestins, and corticosteroids simultaneously, poses significant challenges due to the switching the polarity required during analysis.…”