Doping control analysis of GW1516 in equine plasma using liquid chromatography/electrospray ionization Q-Exactive high-resolution mass spectrometry.
Abstract: GW1516 is a peroxisome proliferator-activated receptor-δ agonist in the class of hormones and metabolic modulators. The use of GW1516 is banned in both horseracing and equestrian competitions. To the best of our knowledge, this is the first metabolic study of GW1516 in horses. Methods: After protein precipitation of pre- and post-administration plasma GW1516 samples, the supernatants were analyzed using liquid chromatography/electrospray ionization Q-Exactive high-resolution mass spectrometry to detect GW1516 and its metabolites. Monoisotopic ions of GW1516 and its metabolites were monitored from the full-scan mass spectral data of pre- and post-administration samples. Quantification methods were developed and validated to establish the elimination profiles of GW1516, its sulfoxide, and its sulfone in equine plasma. Results: GW1516 and its four metabolites GW1516 sulfoxide, GW1516 sulfone, 5-(hydroxymethyl)-4-methyl-2-(4-trifluoromethylphenyl)thiazole (HMTT), and M1 were detected in post-administration plasma samples. GW1516 sulfoxide, GW1516 sulfone, and HMTT were identified by comparison with their respective reference standards whereas M1 was tentatively identified as 4-methyl-2-[4-(trifluoromethyl)phenyl]-1,3-thiazole-5-carboxylic acid by mass spectral interpretation. GW1516 had the longest detection time in post-administration plasma. The elimination profiles of GW1516, its sulfoxide, and its sulfone in plasma were established. Conclusions: For the purpose of doping control, GW1516 is recommended as the target analyte to be monitored in equine plasma due to its long detection time (around 1 week) and the ready availability of its reference material.
© 2020 John Wiley & Sons, Ltd.
Publication Date: 2020-08-11 PubMed ID: 32776613DOI: 10.1002/rcm.8920Google Scholar: Lookup
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- Journal Article
- Analytical Methods
- Doping
- Equestrian Sports
- Equine Health
- Equine Science
- Experimental Methods
- High-performance Liquid Chromatography (HPLC)
- Hormones
- Horse Management
- Horse Training
- Laboratory Methods
- Metabolism
- Metabolites
- Performance Horses
- Pharmaceuticals
- Plasma
- Veterinary Medicine
- Veterinary Research
- Veterinary Science
Summary
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The research presents a method to detect GW1516, a banned substance in horse racing and equestrian competitions, in horse plasma. The study utilized liquid chromatography and high-resolution mass spectrometry to identify GW1516 and its metabolites. The detection method was developed and validated, presenting the elimination profiles of the substance and its metabolites in horse plasma.
Objective of the Study
- This research intended to develop a method for monitoring and detection of GW1516, a substance prohibited in equestrian sports, in horse plasma.
- The objective also included determining GW1516’s metabolites and their respective elimination profiles in horse plasma after adminstration.
Methodology
- Pre and post-administration plasma samples were treated with protein precipitation. Afterward, the supernatants were analyzed using liquid chromatography / electrospray ionization Q-Exactive high-resolution mass spectrometry to identify GW1516 and its subsequent metabolites.
- Monoisotopic ions of GW1516 and its metabolites were monitored from the full-scan mass spectral data of the samples taken before and after administration.
- A quantification method was developed and validated to establish elimination profiles of GW1516 and its two specific metabolites (sulfoxide and sulfone) in horse plasma.
Results
- The research successfully detected GW1516 and its four metabolites in the plasma samples taken after administration.
- Three of the metabolites (sulfoxide, sulfone, and HMTT) were identified through comparison with their reference standards. The fourth metabolite, M1, was tentatively identified as 4-methyl-2-[4-(trifluoromethyl)phenyl]-1,3-thiazole-5-carboxylic acid by mass spectral interpretation.
- The study found that GW1516 remained detectable for the longest period in horse plasma among its metabolites.
- Additionally, the elimination profiles of GW1516, its sulfoxide, and sulfone were established.
Conclusions
- The research concluded that, for doping control in equine sports, GW1516 should be the primary target monitored. This is because it has a long detection time of around one week, and its reference material is readily available.
Cite This Article
APA
Ishii H, Leung GN, Yamashita S, Yamada M, Kushiro A, Kasashima Y, Okada J, Kawasaki K, Kijima-Suda I.
(2020).
Doping control analysis of GW1516 in equine plasma using liquid chromatography/electrospray ionization Q-Exactive high-resolution mass spectrometry.
Rapid Commun Mass Spectrom, 34(23), e8920.
https://doi.org/10.1002/rcm.8920 Publication
Researcher Affiliations
- 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.
- Drug Analysis Department, Laboratory of Racing Chemistry, 1731-2 Tsuruta-machi, Utsunomiya, Tochigi, 320-0851, Japan.
- Drug Analysis Department, Laboratory of Racing Chemistry, 1731-2 Tsuruta-machi, Utsunomiya, Tochigi, 320-0851, Japan.
- Drug Analysis Department, Laboratory of Racing Chemistry, 1731-2 Tsuruta-machi, Utsunomiya, Tochigi, 320-0851, Japan.
- Equine Research Institute, Research Planning & Coordination Division, JRA, 1400-4 Shiba, Shimotsuke, Tochigi, 329-0412, Japan.
- Equine Research Institute, Research Planning & Coordination Division, JRA, 1400-4 Shiba, Shimotsuke, Tochigi, 329-0412, Japan.
- Veterinarian Section, Equine Department, JRA, 6-11-1 Roppongi, Minato-ku, Tokyo, 105-0003, Japan.
- Veterinarian Section, Equine Department, JRA, 6-11-1 Roppongi, Minato-ku, Tokyo, 105-0003, Japan.
- Drug Analysis Department, Laboratory of Racing Chemistry, 1731-2 Tsuruta-machi, Utsunomiya, Tochigi, 320-0851, Japan.
MeSH Terms
- Administration, Intranasal
- Animals
- Chromatography, Liquid / methods
- Doping in Sports
- Female
- Horses / blood
- Limit of Detection
- Linear Models
- Reproducibility of Results
- Spectrometry, Mass, Electrospray Ionization / methods
- Thiazoles / administration & dosage
- Thiazoles / blood
- Thiazoles / chemistry
- Thiazoles / pharmacokinetics
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Citations
This article has been cited 2 times.- 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.
- 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.
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