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Rapid communications in mass spectrometry : RCM2021; 35(8); e9050; doi: 10.1002/rcm.9050

Detection and longitudinal distribution of GW1516 and its metabolites in equine hair for doping control using liquid chromatography/high-resolution mass spectrometry.

Abstract: GW1516 is a peroxisome proliferator-activated receptor-δ (PPAR-δ) agonist that is banned in horseracing and equestrian sports. Long-term detection and longitudinal distribution of GW1516 in the mane of a horse are reported for the first time and this hair analysis could prolong the detection window of GW1516 for doping control. Methods: Mane hairs were divided into three segments (0-7, 7-15, and >15 cm from the cut end) and completely pulverized and homogenized for analysis. The pulverized hair samples were extracted with methanol followed by further purification and the extracts were analyzed by liquid chromatography/electrospray ionization high-resolution mass spectrometry (LC/ESI-HRMS) using a Q-Exactive instrument. This method was successfully validated and applied to post-administration samples to confirm the presence of GW1516 and its metabolites and estimate the uptake amounts of GW1516. Results: After administration of 150 mg of GW1516 to a thoroughbred mare, GW1516 was detected in one of two segments of all mane hairs, and four metabolites, namely GW1516 sulfoxide, GW1516 sulfone, 5-(hydroxymethyl)-4-methyl-2-(4-trifluoromethylphenyl)thiazole (HMTT), and 4-methyl-2-[4-(trifluoromethyl)phenyl]-1,3-thiazole-5-carboxylic acid (MTTC), were also identified. The longitudinal distribution analysis results showed that the maximum uptake of GW1516 into hair (approximately 0.05 pg/mg) was observed at around 13 weeks post-administration and GW1516 could be detected and confirmed up to 6 months post-administration. Conclusions: The parent drug GW1516 was identified as the most appropriate monitoring target in equine hair for controlling its misuse in horses. The use of hair analysis could extend the detection time of GW1516 to at least 6 months after the administration of 150 mg of GW1516 to a thoroughbred mare.
© 2021 John Wiley & Sons Ltd.
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Publication Date: 2021-01-21 PubMed ID: 33470485DOI: 10.1002/rcm.9050Google 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 demonstrates the ability to detect a banned performance-enhancing drug, GW1516, and its metabolites in horse mane hair. The study also highlights that the drug and its metabolites can be detected up to 6 months after administration, potentially giving authorities a more prolonged window to discover doping infractions in horse-racing and equestrian sports.

Research Methodology

  • Mane hairs of a horse were utilized in this study and segmented into three parts: 0-7, 7-15, and more than 15 cm from the cut end. These segments were then pulverized and homogenized for analysis.
  • The pulverized hair samples underwent extraction with methanol. This process allowed drugs or other substances within the hair samples to be isolated for further analysis.
  • Theextracts were then analysed using a liquid chromatography/high-resolution mass spectrometry (LC/ESI-HRMS) tool, specifically, a Q-Exactive instrument. Liquid chromatography separates the compounds in the extract, while the mass spectrometry identifies and quantifies the compounds. This combination allows the researchers to detect the presence of GW1516 and its metabolites.

Results

  • 150 mg of GW1516 was administered to a thoroughbred mare for testing. After administration, GW1516 was found in one of the two segments of all mane hairs. It was also discovered that four metabolites of GW1516 (GW1516 sulfoxide, GW1516 sulfone, HMTT, and MTTC) could be identified in mane hair.
  • The longitudinal distribution analysis showed that GW1516 intake into the hair recorded the maximum (approximately 0.05 pg/mg) at about 13 weeks post-administration. Detection was possible six months after administration.

Conclusions

  • From the research, the parent drug GW1516 was identified as the most suitable target to monitor in equine hair for controlling its misuse in horses.
  • The use of hair analysis method could extend the detection time of GW1516 in samples to at least 6 months after the drug was administered. This extended window of detection could have powerful implications for doping control in high-level equestrian sports.

Cite This Article

APA
Ishii H, Shibuya M, Leung GN, Nozawa S, Yamashita S, Yamada M, Kushiro A, Kasashima Y, Okada J, Kawasaki K, Kijima-Suda I. (2021). Detection and longitudinal distribution of GW1516 and its metabolites in equine hair for doping control using liquid chromatography/high-resolution mass spectrometry. Rapid Commun Mass Spectrom, 35(8), e9050. https://doi.org/10.1002/rcm.9050

Publication

Rapid communications in mass spectrometry : RCM
ISSN: 1097-0231
NlmUniqueID: 8802365
Country: England
Language: English
Volume: 35
Issue: 8
Pages: e9050

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.
Nozawa, Satoshi
  • 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
  • Research Planning & Coordination Division, JRA, Equine Research Institute, 1400-4, Shiba, Shimotsuke, Tochigi, 329-0412, Japan.
Kasashima, Yoshinori
  • Research Planning & Coordination Division, JRA, Equine Research Institute, 1400-4, Shiba, Shimotsuke, Tochigi, 329-0412, Japan.
Okada, Jun
  • Veterinarian Section, Equine Department, Japan Racing Association, 6-11-1 Roppongi, Minato-ku, Tokyo, 105-0003, Japan.
Kawasaki, Kazumi
  • Veterinarian Section, Equine Department, Japan Racing Association, 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, Liquid / methods
  • Doping in Sports
  • Female
  • Hair / chemistry
  • Horses
  • Performance-Enhancing Substances / analysis
  • Reproducibility of Results
  • Spectrometry, Mass, Electrospray Ionization / methods
  • Thiazoles / administration & dosage
  • Thiazoles / analysis
  • Thiazoles / isolation & purification
  • Thiazoles / metabolism
  • Time Factors

References

This article includes 25 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. 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
  4. International Federation of Horseracing Authorities. International Agreement.. .
  5. Fédération Équestre Internationale. 2020 Equine Prohibited Substances List.. .
  6. 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
  7. 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
  8. 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
  9. Ishii H, Shibuya M, Leung GN-W. Metabolic study of GW1516 in equine urine using liquid chromatography/electrospray ionization Q-Exactive high-resolution mass spectrometyr for doping control.. Rapid Commun Mass Spectrom 2021;35(5):e9028.
    doi: 10.1002/rcm.9028google scholar: lookup
  10. Ishii H, Leung GN-W, 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(23):e8920.
    doi: 10.1002/rcm.8920google scholar: lookup
  11. Dunnett M. The diagnostic potential of equine hair: a comparative review of hair analysis for assessing nutritional status, environmental poisoning, and drug use and abuse.. .
  12. Popot MA, Boyer S, Maciejewski P, Garcia P, Dehennin L, Bonnaire Y. Approaches to the detection of drugs in horse hair.. In: Proceedings of the 13th International Conference of Racing Analysts and Veterinarians Cambridge, UK 2000. PA: R & W Publications (Newmarket) Limited; 2001:115-120.
  13. Kuwayama K, Miyaguchi H, Iwata YT. Three-step drug extraction from a single sub-millimeter segment of hair and nail to determine the exact day of drug intake.. Anal Chim Acta 2016;948:40-47.
    doi: 10.1016/j.aca.2016.10.029google scholar: lookup
  14. Madry MM, Spycher BS, Kupper J. Long-term monitoring of opioid, sedative and anti-inflammatory drugs in horse hair using a selective and sensitive LC-MS/MS procedure.. BMC Vet Res 2016;12(84):1-10.
    doi: 10.1186/s12917-016-0709-5google scholar: lookup
  15. Popot MA, Stojiljkovic N, Garcia P, Richard CA, Bonnaire Y, Tabet JC. Additional studies on the detection of drugs in horse hair samples.. In: Proceedings of the 14th International Conference of Racing Analysts and Veterinarians Florida, USA, 2002. PA: R & W Publications (Newmarket) Limited; 2003:224-233.
  16. Boyer S, Garcia P, Popot MA, Steiner V, Lesieur M. Detection of testosterone propionate administration in horse hair samples.. J Chromatogr B Analyt Technol Biomed Life Sci 2007;852(1-2):684-688.
    doi: 10.1016/j.jchromb.2007.02.046google scholar: lookup
  17. 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
  18. Kintz P, Ameline A, Gheddar L, Raul JS. Testing for GW501516 (cardarine) in human hair using LC/MS-MS and confirmation by LC/HRMS.. Drug Test Anal 2020;12(7):980-986.
    doi: 10.1002/dta.2802google scholar: lookup
  19. Kaufmann A, Widmer M, Maden K. Post-interface signal suppression, a phenomenon observed in a single-stage Orbitrap mass spectrometer coupled to an electrospray interfaced liquid chromatograph.. Rapid Commun Mass Spectrom 2010;24(14):2162-2170.
    doi: 10.1002/rcm.4615google scholar: lookup
  20. Kaufmann A, Walker S. Improved performance of multiplexed targeted tandem mass spectrometry scans using customized Q Orbitrap data acquisition.. Rapid Commun Mass Spectrom 2016;30(9):1131-1138.
    doi: 10.1002/rcm.7532google scholar: lookup
  21. Kaufmann A, Walker S. Comparison of linear intrascan and interscan dynamic ranges of Orbitrap and ion-mobility time-of-flight mass spectrometers.. Rapid Commun Mass Spectrom 2017;31(22):1915-1926.
    doi: 10.1002/rcm.7981google scholar: lookup
  22. 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
  23. 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.. In: Proceedings of the 16th International Conference of Racing Analysts and Veterinarians Tokyo, Japan, Vol. 2007. 2006:239-245.
  24. Association of Official Racing Chemists. AORC Guidelines for the Minimum Criteria for Identification by Chromatography and Mass Spectrometry.. .
  25. Stenn KS, Paus R. Controls of hair follicle cycling.. Physiol Rev 2001;81(1):449-494.
    doi: 10.1152/physrev.2001.81.1.449google 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
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