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Rapid communications in mass spectrometry : RCM2020; 35(5); e9028; doi: 10.1002/rcm.9028

Metabolic study of GW1516 in equine urine using liquid chromatography/electrospray ionization Q-Exactive high-resolution mass spectrometry for doping control.

Abstract: The use of GW1516, a peroxisome proliferator-activated receptor δ (PPAR δ) agonist, is strictly prohibited in both horseracing and equestrian competitions. However, little is known about its metabolic fate in horses. To the best of our knowledge, this is the first reported metabolic study of GW1516 in equine urine. Methods: Urine samples obtained from a thoroughbred after nasoesophageal administration with GW1516 were protein-precipitated and the supernatants were subsequently analyzed by liquid chromatography/electrospray ionization high-resolution mass spectrometry (LC/ESI-HRMS) with a Q-Exactive mass spectrometer. Monoisotopic ions of GW1516 and its metabolites were monitored from the full-scan mass spectral data of pre- and post-administration samples. A quantification method was developed and validated to establish the excretion profiles of GW1516, its sulfoxide, and its sulfone in equine urine. Results: GW1516 and its nine metabolites [including GW1516 sulfoxide, GW1516 sulfone, 5-(hydroxymethyl)-4-methyl-2-(4-trifluoromethylphenyl)thiazole (HMTT), methyl 4-methyl-2-[4-(trifluoromethyl)phenyl]-1,3-thiazole-5-carboxylate (MMTC), 4-methyl-2-[4-(trifluoromethyl)phenyl]-1,3-thiazole-5-carboxylic acid (MTTC), and M1 to M4] were detected in post-administration urine samples. GW1516 sulfoxide and GW1516 sulfone showed the longest detection times in post-administration urine samples and were therefore recommended as potential screening targets for doping control purposes. Quantitative analysis was also conducted to establish the excretion profiles of GW1516 sulfoxide and GW1516 sulfone in urine. Conclusions: For the purposes of doping control of GW1516, the GW1516 sulfoxide and GW1516 sulfone metabolites are recommended as the target analytes to be monitored in equine urine due to their high specificities, long detection times (1 and 4 weeks, respectively), and the ready availability of their reference materials.
© 2020 John Wiley & Sons Ltd.
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Publication Date: 2020-12-16 PubMed ID: 33319421DOI: 10.1002/rcm.9028Google Scholar: Lookup
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Summary

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This research investigates the metabolic fate of the banned substance GW1516 in horses. Utilizing liquid chromatography and mass spectrometry, the study identifies and quantifies its metabolites in horse urine, suggesting two metabolites, GW1516 sulfoxide and GW1516 sulfone, as potential screening targets for doping detection due to their long detection times.

Introduction and Background

  • This research is centered around GW1516, a substance strictly banned in horseracing and horse competitions due to its performance-enhancing properties.
  • Yet, there was a lack of knowledge about the metabolic implications of this substance in horses.

Methods

  • The study used equine urine samples taken after the administration of GW1516 to study its metabolites.
  • High-resolution mass spectrometry was used to analyze the samples and monitor the ions of GW1516 and its metabolites.
  • The study established a method to measure the excretion profiles of GW1516 and its sulfoxide and sulfone metabolites.

Results

  • The researchers found GW1516 and nine of its metabolites in the urine samples taken after administration of the substance.
  • Of these, GW1516 sulfoxide and GW1516 sulfone showed the longest detection times and were therefore recommended as potential screening targets for doping control.
  • Quantitative analysis was conducted to study the excretion profiles of these two metabolites.

Conclusions

  • The study recommends that, for doping control purposes, testing for the presence of GW1516 sulfoxide and GW1516 sulfone in horse urine is ideal due to their high specificities and long detection times (1 and 4 weeks respectively).
  • The availability of reference materials also facilitates their use as target analytes for anti-doping checks.

Cite This Article

APA
Ishii H, Shibuya M, Leung GN, Yamashita S, Yamada M, Kushiro A, Kasashima Y, Okada J, Kawasaki K, Kijima-Suda I. (2020). Metabolic study of GW1516 in equine urine using liquid chromatography/electrospray ionization Q-Exactive high-resolution mass spectrometry for doping control. Rapid Commun Mass Spectrom, 35(5), e9028. https://doi.org/10.1002/rcm.9028

Publication

Rapid communications in mass spectrometry : RCM
ISSN: 1097-0231
NlmUniqueID: 8802365
Country: England
Language: English
Volume: 35
Issue: 5
Pages: e9028

Researcher Affiliations

Ishii, Hideaki
  • Drug Analysis Department, Laboratory of Racing Chemistry, 1731-2 Tsuruta-machi, Utsunomiya, Tochigi, 320-0851, Japan.
  • Department of Pharmaceutical Sciences, Tohoku University Hospital, 1-1 Seiryo-machi, Aoba-ku, Sendai, Miyagi, 980-8574, Japan.
Shibuya, Mariko
  • Drug Analysis Department, Laboratory of Racing Chemistry, 1731-2 Tsuruta-machi, Utsunomiya, Tochigi, 320-0851, Japan.
Leung, Gary Ngai-Wa
  • Drug Analysis Department, Laboratory of Racing Chemistry, 1731-2 Tsuruta-machi, Utsunomiya, Tochigi, 320-0851, Japan.
Yamashita, Shozo
  • Drug Analysis Department, Laboratory of Racing Chemistry, 1731-2 Tsuruta-machi, Utsunomiya, Tochigi, 320-0851, Japan.
Yamada, Masayuki
  • Drug Analysis Department, Laboratory of Racing Chemistry, 1731-2 Tsuruta-machi, Utsunomiya, Tochigi, 320-0851, Japan.
Kushiro, Asuka
  • Equine Research Institute, Research Planning & Coordination Division, JRA, 1400-4, Shiba, Shimotsuke, Tochigi, 329-0412, Japan.
Kasashima, Yoshinori
  • Equine Research Institute, Research Planning & Coordination Division, JRA, 1400-4, Shiba, Shimotsuke, Tochigi, 329-0412, Japan.
Okada, Jun
  • Veterinarian Section, Equine Department, JRA, 6-11-1 Roppongi, Minato-ku, Tokyo, 105-0003, Japan.
Kawasaki, Kazumi
  • Veterinarian Section, Equine Department, JRA, 6-11-1 Roppongi, Minato-ku, Tokyo, 105-0003, Japan.
Kijima-Suda, Isao
  • Drug Analysis Department, Laboratory of Racing Chemistry, 1731-2 Tsuruta-machi, Utsunomiya, Tochigi, 320-0851, Japan.

MeSH Terms

  • Animals
  • Chromatography, High Pressure Liquid / methods
  • Doping in Sports / prevention & control
  • Horses / metabolism
  • Horses / urine
  • Spectrometry, Mass, Electrospray Ionization / methods
  • Substance Abuse Detection / methods
  • Substance Abuse Detection / veterinary
  • Thiazoles / metabolism
  • Thiazoles / urine
  • Urine / chemistry

Grant Funding

  • Grant-in-Aid from the JRA

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Citations

This article has been cited 3 times.
  1. Ishii H, Shibuya M, Kusano K, Sone Y, Kamiya T, Wakuno A, Ito H, Miyata K, Sato F, Kuroda T, Yamada M, Leung GN. Generic approach for the discovery of drug metabolites in horses based on data-dependent acquisition by liquid chromatography high-resolution mass spectrometry and its applications to pharmacokinetic study of daprodustat. Anal Bioanal Chem 2022 Nov;414(28):8125-8142.
    doi: 10.1007/s00216-022-04347-2pubmed: 36181513google scholar: lookup
  2. Ishii H, Shibuya M, Kusano K, Sone Y, Kamiya T, Wakuno A, Ito H, Miyata K, Sato F, Kuroda T, Yamada M, Leung GN. Pharmacokinetic Study of Vadadustat and High-Resolution Mass Spectrometric Characterization of its Novel Metabolites in Equines for the Purpose of Doping Control. Curr Drug Metab 2022;23(10):850-865.
    doi: 10.2174/1389200223666220825093945pubmed: 36017833google scholar: lookup
  3. Ishii H, Shigematsu R, Takemoto S, Ishikawa Y, Mizobe F, Nomura M, Arima D, Kunii H, Yuasa R, Yamanaka T, Tanabe S, Nagata SI, Yamada M, Leung GN. Quantification of osilodrostat in horse urine using LC/ESI-HRMS to establish an elimination profile for doping control. Bioanalysis 2024;16(17-18):947-958.
    doi: 10.1080/17576180.2024.2385848pubmed: 39235065google scholar: lookup
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