2. Academic Validation
  3. Enhanced UHPLC-MS/MS screening of selective androgen receptor modulators following urine hydrolysis

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

  • MethodsX. 2020 May 21;7:100926. doi: 10.1016/j.mex.2020.100926.
Anna Gadaj 1 2,  Emiliano Ventura 1,  Jim Healy 3 4,  Francesco Botrè 5,  Saskia S Sterk 6,  Tom Buckley 7,  Mark H Mooney 1
Affiliations

Affiliations

  • 1 Institute for Global Food Security, School of Biological Sciences, Queen's University Belfast, BT9 5DL, United Kingdom.
  • 2 Chemical and Immunodiagnostic Sciences Branch, Veterinary Sciences Division, Agri-Food & Biosciences Institute (AFBI), Stoney Road, Belfast BT4 3SD, United Kingdom.
  • 3 Laboratory, Irish Greyhound Board, Limerick Greyhound Stadium, Ireland.
  • 4 Applied Science Department, Limerick Institute of Technology, Moylish, Limerick, Ireland.
  • 5 Laboratorio Antidoping, Federazione Medico Sportiva Italiana, Italy.
  • 6 Wageningen Food Safety Research, Wageningen University & Research, European Union Reference Laboratory, Wageningen, the Netherlands.
  • 7 Irish Diagnostic Laboratory Services Ltd., Johnstown, W91 RH93, Co. Kildare, Ireland.
PMID: 32547930 DOI: 10.1016/j.mex.2020.100926
Abstract

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 livestock-based 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 (β-glucuronidase/arylsulfatase) into a previously established protocol while maintaining the minimal volume (200 µL) 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.•CCβ 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 animal-derived food is free from contamination with SARM residues.

Keywords

Doping analysis; Food safety; Hydrolysis; SARMs; UHPLC-MS/MS; Urine.

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