FREE SHIPPING over $40
OVER 9000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Home / Equine Research Bank / Rapid Commun Mass Spectrom / Metabolic study of GW1516 in equine urine using liquid chrom...
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.
Get Full Text
Publication Date: 2020-12-16 PubMed ID: 33319421DOI: 10.1002/rcm.9028Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
LinkedInCopy
  • Evaluation Study
  • Journal Article

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 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

References

This article includes 27 references
  1. Pokrywka A, Cholbinski P, Kaliszewski P, Kowalczyk K, Konczak D, Zembron-Lacny A. Metabolic modulators of the exercise response: Doping control analysis of an agonist of the peroxisome proliferator-activated receptor δ (GW501516) and 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR).. J Physiol Pharmacol 2014;65(4):469-476.
  2. Narkar VA, Downes M, Yu RT. AMPK and PPARdelta agonists are exercise mimetics.. Cell 2008;134(3):405-415.
    doi: 10.1016/j.cell.2008.06.051google scholar: lookup
  3. Nishimura K, Nakano N, Chowdhury VS. Effect of PPARβ/δ agonist on the placentation and embryo-fetal development in rats.. Birth Defects Res B Dev Reprod Toxicol 2013;98(2):164-169.
    doi: 10.1002/bdrb.21052google scholar: lookup
  4. Sahebkar A, Chew GT, Watts GF. New peroxisome proliferator-activated receptor agonists: Potential treatments for atherogenic dyslipidemia and non-alcoholic fatty liver disease.. Expert Opin Pharmacother 2014;15(4):493-503.
    doi: 10.1517/14656566.2014.876992google scholar: lookup
  5. Chen W, Gao R, Xie X. A metabolomic study of the PPARδ agonist GW501516 for enhancing running endurance in Kunming mice.. Sci Rep 2015;5(1):9884.
    doi: 10.1038/srep09884google scholar: lookup
  6. International Federation of Horseracing Authorities. International Agreement.. .
  7. Fédération Équestre Internationale. 2020 Equine Prohibited Substances List.. .
  8. Thevis M, Möller I, Thomas A. Characterization of two major urinary metabolites of the PPARdelta-agonist GW1516 and implementation of the drug in routine doping controls.. Anal Bioanal Chem 2010;396(7):2479-2491.
    doi: 10.1007/s00216-009-3283-xgoogle scholar: lookup
  9. Thevis M, Möller I, Beuck S, Schänzer W. Synthesis, mass spectrometric characterization, and analysis of the PPARδ agonist GW1516 and its major human metabolites: Targets in sports drug testing.. Methods Mol Biol 2013;952:301-312.
    doi: 10.1007/978-1-62703-155-4_22google scholar: lookup
  10. Sobolevsky T, Dikunets M, Sukhanova I, Virus E, Rodchenkov G. Detection of PPARδ agonists GW1516 and GW0742 and their metabolites in human urine.. Drug Test Anal 2012;4(10):754-760.
    doi: 10.1002/dta.1413google scholar: lookup
  11. Gray B, Viljanto M, Menzies E, Vanhaecke L. Detection of prohibited substances in equine hair by ultra-high performance liquid chromatography-triple quadrupole mass spectrometry - application to doping control samples.. Drug Test Anal 2018;10(7):1050-1060.
    doi: 10.1002/dta.2367google scholar: lookup
  12. Ishii H, Leung GNW, Yamashita S. Doping control analysis of GW1516 in equine plasma using liquid chromatography/electrospray ionization Q-Exactive high-resolution mass spectrometry.. Rapid Commun Mass Spectrom 2020;34:e8920.
    doi: 10.1002/rcm.8920google scholar: lookup
  13. Ishii H, Shimada M, Yamaguchi H, Mano N. A simultaneous determination method for 5-fluorouracil and its metabolites in human plasma with linear range adjusted by in-source collision-induced dissociation using hydrophilic interaction liquid chromatography-electrospray ionization-tandem mass spectrometry.. Biomed Chromatogr 2016;30(11):1882-1886.
    doi: 10.1002/bmc.3743google scholar: lookup
  14. Takeda A, Nakata M, Kijima-Suda I, Tanaka H. Trial for ionisation enhancement in negative ion electrospray ionisation and rapid screening of acid drugs by liquid chromatography/mass spectrometry.. Proceedings of the 16th International Conference of Racing Analysts and Veterinarians Tokyo, Japan, 2006. PA: R & W Communications, 2007:239-245.
  15. Thevis M, Schmickler MH, Schänzert W. Mass spectrometric behavior of thiazide-based diuretics after electrospray ionization and collision-induced dissociation.. Anal Chem 2002;74(15):3802-3808.
    doi: 10.1021/ac020020egoogle scholar: lookup
  16. Garcia P, Popot MA, Fournier F, Bonnaire Y, Tabet JC. Gas-phase behaviour of negative ions produced from thiazidic diuretics under electrospray conditions.. J Mass Spectrom 2002;37(9):940-953.
    doi: 10.1002/jms.353google scholar: lookup
  17. Kuuranne T, Vahermo M, Leinonen A, Kostianen R. Electrospray and atmospheric pressure chemical ionization tandem mass spectrometric behavior of eight anabolic steroid glucuronides.. J Am Soc Mass Spectrom 2000;11(8):722-730.
    doi: 10.1016/s1044-0305(00)00135-5google scholar: lookup
  18. Lu AY, Wang RW, Lin JH. Cytochrome P450 in vitro reaction phenotyping: A re-evaluation of approaches used for P450 isoform identification.. Drug Metab Dispos 2003;31(4):345-350.
    doi: 10.1124/dmd.31.4.345google scholar: lookup
  19. Cashman JR. Some distinctions between flavin-containing and cytochrome P450 monooxygenases.. Biochem Biophys Res Commun 2005;338(1):599-604.
    doi: 10.1016/j.bbrc.2005.08.009google scholar: lookup
  20. Hines RN, Cashman JR, Philpot RM, Williams DE, Ziegler DM. The mammalian flavin-containing monooxygenases: Molecular characterization and regulation of expression.. Toxicol Appl Pharmacol 1994;125(1):1-6.
    doi: 10.1006/taap.1994.1042google scholar: lookup
  21. Hollenberg PF. Mechanisms of cytochrome P450 and peroxidase-catalyzed xenobiotic metabolism.. FASEB J 1992;6(2):686-694.
    doi: 10.1096/fasebj.6.2.1537457google scholar: lookup
  22. Zhu BT. Catechol-O-Methyltransferase (COMT)-mediated methylation metabolism of endogenous bioactive catechols and modulation by endobiotics and xenobiotics: Importance in pathophysiology and pathogenesis.. Curr Drug Metab 2002;3(3):321-349.
    doi: 10.2174/1389200023337586google scholar: lookup
  23. Scarth JP, Teale P, Kuuranne T. Drug metabolism in the horse: A review.. Drug Test Anal 2011;3(1):19-53.
    doi: 10.1002/dta.174google scholar: lookup
  24. Lan W, Bian L, Zhao X. Liquid chromatography/quadrupole time-of-flight mass spectrometry for identification of in vitro and in vivo metabolites of bornyl gallate in rats.. J Anal Methods Chem 2013;2013:473649.
    doi: 10.1155/2013/473649google scholar: lookup
  25. Oda S, Fukami T, Yokoi T, Nakajima M. a comprehensive review of UDP-glucuronosyltransferase and esterases for drug development.. Drug Metab Pharmacokinet 2015;30(1):30-51.
    doi: 10.1016/j.dmpk.2014.12.001google scholar: lookup
  26. Thevis M, Walpurgis K, Thomas A. Analytical approaches in human sports drug testing: Recent advances, challenges, and solutions.. Anal Chem 2020;92(1):506-523.
    doi: 10.1021/acs.analchem.9b04639google scholar: lookup
  27. Walpurgis K, Rubio A, Wagener F. Elimination profiles of microdosed ostarine mimicking contaminated products ingestion.. Drug Test Anal 2020;12:1570-1580.
    doi: 10.1002/dta.2933google scholar: lookup

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
Subscribe to our newsletter
Join 99,357 horse owners like you who receive our email updates on horse health and nutrition.