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MethodsX2020; 7; 100926; doi: 10.1016/j.mex.2020.100926

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

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 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.
© 2020 The Author(s). Published by Elsevier B.V.
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Publication Date: 2020-05-21 PubMed ID: 32547930PubMed Central: PMC7286957DOI: 10.1016/j.mex.2020.100926Google Scholar: Lookup
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Summary

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This research article presents an improved method for detecting commonly abused non-steroidal agents known as selective androgen receptor modulators (SARMs) in urine samples from different species using UHPLC-MS/MS. The method demonstrated enhanced detection rates and fulfillment of legal criteria for doping and food control, suggesting its suitability for routine testing to ensure fair sports practices and contamination-free food.

Research Objectives and Background

  • The primary objective of the research was to incorporate enzymatic hydrolysis using β-glucuronidase/arylsulfatase into a detection protocol for SARMs within urine samples. This was done in response to concern over the misuse of these agents in human and animal sports and potentially as growth-promoting agents in livestock farming.
  • SARMs, or selective androgen receptor modulators, are commonly misused non-steroidal agents. They can potentially contaminate food supply and give unfair advantages to athletes.

Methodology and Findings

  • The researchers used an ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS)-based assay to screen for SARMs. This enhanced the detection of a range of these substances in various species including equines, bovines, humans, canines, and rodents. This method included the use of enzymatic hydrolysis to increase detection efficiency.
  • Validation of the assay showed it to be fit-for-purpose, efficient, and high-throughput, fulfilling the criteria set within relevant doping and food control laws.
  • The deconjugation procedure included in the method boosted the relative abundance of a SARM known as ‘ostarine’ in the urine samples tested, illustrating the necessity for hydrolysis to identify emerging SARMs in their total form.

Potential Applications

  • This improved detection method provides a rapid and efficient screening tool for SARMs in urine samples, making it suitable for routine testing in both human and animal sports to promote fair play.
  • The protocol can also be used to check for contamination of animal-derived food products with SARM residues, guaranteeing that the food supply is safe for consumption.

Cite This Article

APA
Gadaj A, Ventura E, Healy J, Botrè F, Sterk SS, Buckley T, Mooney MH. (2020). Enhanced UHPLC-MS/MS screening of selective androgen receptor modulators following urine hydrolysis. MethodsX, 7, 100926. https://doi.org/10.1016/j.mex.2020.100926

Publication

MethodsX
ISSN: 2215-0161
NlmUniqueID: 101639829
Country: Netherlands
Language: English
Volume: 7
Pages: 100926
PII: 100926

Researcher Affiliations

Gadaj, Anna
  • Institute for Global Food Security, School of Biological Sciences, Queen's University Belfast, BT9 5DL, United Kingdom.
  • Chemical and Immunodiagnostic Sciences Branch, Veterinary Sciences Division, Agri-Food & Biosciences Institute (AFBI), Stoney Road, Belfast BT4 3SD, United Kingdom.
Ventura, Emiliano
  • Institute for Global Food Security, School of Biological Sciences, Queen's University Belfast, BT9 5DL, United Kingdom.
Healy, Jim
  • Laboratory, Irish Greyhound Board, Limerick Greyhound Stadium, Ireland.
  • Applied Science Department, Limerick Institute of Technology, Moylish, Limerick, Ireland.
Botrè, Francesco
  • Laboratorio Antidoping, Federazione Medico Sportiva Italiana, Italy.
Sterk, Saskia S
  • Wageningen Food Safety Research, Wageningen University & Research, European Union Reference Laboratory, Wageningen, the Netherlands.
Buckley, Tom
  • Irish Diagnostic Laboratory Services Ltd., Johnstown, W91 RH93, Co. Kildare, Ireland.
Mooney, Mark H
  • Institute for Global Food Security, School of Biological Sciences, Queen's University Belfast, BT9 5DL, United Kingdom.

Conflict of Interest Statement

There are no conflicts of interest to declare.

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