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Drug testing and analysis2020; 12(6); 752-762; doi: 10.1002/dta.2769

In vivo metabolism of the designer anabolic steroid hemapolin in the thoroughbred horse.

Abstract: Hemapolin (2α,3α-epithio-17α-methyl-5α-androstan-17β-ol) is a designer steroid that is an ingredient in several "dietary" and "nutritional" supplements available online. As an unusual chemical modification to the steroid A-ring could allow this compound to pass through antidoping screens undetected, the metabolism of hemapolin was investigated by an in vivo equine drug administration study coupled with GC-MS analysis. Following administration of synthetically prepared hemapolin to a thoroughbred horse, madol (17α-methyl-5α-androst-2-en-17β-ol), reduced and dihydroxylated madol (17α-methyl-5α-androstane-2β,3α,17β-triol), and the isomeric enone metabolites 17β-hydroxy-17α-methyl-5α-androst-3-en-2-one and 17β-hydroxy-17α-methyl-5α-androst-2-en-4-one, were detected and confirmed in equine urine extracts by comparison with a library of synthetically derived reference materials. A number of additional madol derivatives derived from hydroxylation, dihydroxylation, and trihydroxylation were also detected but not fully identified by this approach. A yeast cell-based androgen receptor bioassay of available reference materials showed that hemapolin and many of the metabolites identified by this study were potent activators of the equine androgen receptor. This study reveals the metabolites resulting from the equine administration of the androgen hemapolin that can be incorporated into routine GC-MS antidoping screening and confirmation protocols to detect the illicit use of this agent in equine sports.
© 2020 John Wiley & Sons, Ltd.
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Publication Date: 2020-02-04 PubMed ID: 31950580DOI: 10.1002/dta.2769Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research explores the metabolism of Hemapolin, a designer steroid often used in dietary supplements, in thoroughbred horses. The study reveals the metabolites that result from the equine administration of Hemapolin, which can be leveraged for GC-MS antidoping tests to detect its illicit use in horse racing.

About Hemapolin

  • Hemapolin (2α,3α-epithio-17α-methyl-5α-androstan-17β-ol) is a designer steroid found in several dietary and nutritional supplements sold online.
  • Due to its unusual chemical modification to the steroid A-ring, this compound can slip through doping detection screens unnoticed.

Investigating Hemapolin Metabolism

  • The research team performed an in vivo drug administration study on a thoroughbred horse to investigate Hemapolin metabolism followed by Gas Chromatography-Mass Spectrometry (GC-MS) analysis.
  • Synthetically prepared Hemapolin was administered to a thoroughbred horse for this study.

The Metabolites

  • After administration, several metabolites including madol, reduced and dihydroxylated madol, and the isomeric enone metabolites were detected in the equine urine extracts.
  • The detection was confirmed by comparison with a library of synthetically derived reference materials.
  • A number of additional madol derivatives derived from hydroxylation, dihydroxylation, and trihydroxylation were also detected, but their identities were not fully established in this study.

Endocrine Activity

  • A yeast cell-based androgen receptor bioassay of available reference materials was conducted.
  • The results showed that Hemapolin and many of the metabolites identified by the study were potent activators of the equine androgen receptor.

Impact of the Study

  • The study helps expose the metabolites resulting from the equine administration of the androgen Hemapolin.
  • These findings can be incorporated into regular GC-MS antidoping screening and confirmation protocols to detect the illicit use of Hemapolin in equine sports.

Cite This Article

APA
Waller CC, Weththasinghe SA, McClure L, Cawley AT, Suann C, Suann E, Sutherland E, Cooper E, Heather A, McLeod MD. (2020). In vivo metabolism of the designer anabolic steroid hemapolin in the thoroughbred horse. Drug Test Anal, 12(6), 752-762. https://doi.org/10.1002/dta.2769

Publication

Drug testing and analysis
ISSN: 1942-7611
NlmUniqueID: 101483449
Country: England
Language: English
Volume: 12
Issue: 6
Pages: 752-762

Researcher Affiliations

Waller, Christopher C
  • Research School of Chemistry, Australian National University, Canberra, Australia.
Weththasinghe, Sumudu A
  • Research School of Chemistry, Australian National University, Canberra, Australia.
McClure, Lauren
  • Australian Racing Forensic Laboratory, Racing NSW, Sydney, Australia.
Cawley, Adam T
  • Australian Racing Forensic Laboratory, Racing NSW, Sydney, Australia.
Suann, Craig
  • Australian Racing Forensic Laboratory, Racing NSW, Sydney, Australia.
Suann, Emily
  • Australian Racing Forensic Laboratory, Racing NSW, Sydney, Australia.
Sutherland, Emma
  • Department of Physiology, School of Biomedical Sciences, University of Otago, Dunedin, New Zealand.
Cooper, Elliot
  • Department of Physiology, School of Biomedical Sciences, University of Otago, Dunedin, New Zealand.
Heather, Alison
  • Department of Physiology, School of Biomedical Sciences, University of Otago, Dunedin, New Zealand.
McLeod, Malcolm D
  • Research School of Chemistry, Australian National University, Canberra, Australia.

MeSH Terms

  • Animals
  • Biotransformation
  • Designer Drugs / metabolism
  • Doping in Sports / methods
  • Gas Chromatography-Mass Spectrometry
  • Horses / metabolism
  • Receptors, Androgen / metabolism
  • Reference Standards
  • Steroids / metabolism
  • Steroids / urine
  • Substance Abuse Detection / methods
  • Testosterone Congeners / metabolism
  • Testosterone Congeners / urine

Grant Funding

  • LP120200444 / Australian Research Council

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Citations

This article has been cited 2 times.
  1. Pounina TA, Gloriozova TA, Savidov N, Dembitsky VM. Sulfated and Sulfur-Containing Steroids and Their Pharmacological Profile. Mar Drugs 2021 Apr 24;19(5).
    doi: 10.3390/md19050240pubmed: 33923288google scholar: lookup
  2. Klöppner L, Harps LC, Parr MK. Sample Preparation Techniques for Growth-Promoting Agents in Various Mammalian Specimen Preceding MS-Analytics. Molecules 2024 Jan 9;29(2).
    doi: 10.3390/molecules29020330pubmed: 38257243google scholar: lookup
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