Development an automated and high -throughput analysis methods for detecting 64 anabolic androgenic steroids in urine using gas chromatography-mass spectrometry

Abstract

A gas chromatography tandem mass spectrometry (GC–MS/MS) assay was developed for the simultaneous detection of 64 anabolic androgenic steroids (AAS). Urine is extracted with tertbutyl methyl ether under alkaline conditions, purified with C18, and derived through silanization, Detection is carried out in dynamic multiple reaction monitoring (dMRM) mode, The overall recoveries ranged from 74.31 % to 119.66 %, The relative standard deviation is <10 %, with a linear range of 1–200 ng/ml, The linear range of endogenous androsterone (A) and etiocholanolone (Etio) is 1.7–10,000 ng/ml, with a correlation coefficient 0.9991– 0.9999. The limit of detection (LOD) range of this experiment was 0.2 ng/ml-1.2 ng/ml, and limit of quantification (LOQ) was 0.7 ng/ml-4 ng/ml, The results of testing 100 actual samples show that this method is simple, effective and feasible, and can be used for the detection of AAS, meeting the requirements of the World Anti Doping Agency (WADA)