Enhanced UHPLC-MS/MS screening of selective androgen receptor modulators following urine hydrolysis

Selective androgen receptor modulators (SARMs) represent non-steroidal agents commonly abused in human and animal (i.e. equine, canine) sports, with potential for further misuse as growth promoting agents in livestockbased farming. As a direct response to the real and possible implications of illicit application in both sport as well as food production systems, this study incorporated enzymatic hydrolysis (beta-glucuronidase/arylsulfatase) into a previously established protocol while maintaining the minimal volume (200 mu L) of urine sample required to detect SARMs encompassing various pharmacophores in urine from a range of species (i.e. equine, bovine, human, canine and rodent). The newly presented semi-quantitative UHPLC-MS/MS-based assay is shown to be fit-for-purpose, being rapid and offering high-throughput, with validation findings fulfilling criteria stipulated within relevant doping and food control legislation. CC beta values determined at 1 ng mL(-1) for majority of analytes. Deconjugation step included in the method led to significantly increased relative abundance of ostarine in analysed incurred urine samples demonstrating the requirement for hydrolysis to detect a total form of emerging SARMs. Assay amenable for use within routine testing to ensure fair play in animal and human sports and that animalderived food is free from contamination with SARM residues. (C) 2020 The Author(s). Published by Elsevier B.V.