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Drug testing and analysis2022; 14(7); 1244-1254; doi: 10.1002/dta.3247

Long-term monitoring of IOX4 in horse hair and its longitudinal distribution with segmental analysis using liquid chromatography/electrospray ionization Q Exactive high-resolution mass spectrometry for the purpose of doping control.

Abstract: IOX4, a hypoxia-inducible factor stabilizer, is classified as a banned substance for horses in both horse racing and equestrian sports. We recently reported the pharmacokinetic profiles of IOX4 in horse plasma and urine and also identified potential monitoring targets for the doping control purpose. In this study, a long-term longitudinal analysis of IOX4 in horse hair after a nasoesophageal administration of IOX4 (500 mg/day for 3 days) to three thoroughbred mares is presented for the first time for controlling the abuse/misuse of IOX4. Six bunches of mane hair were collected at 0 (pre), 1, 2, 3, and 6 month(s) postadministration. Our results showed that the presence of IOX4 was identified in all postadministration horse hair samples, but no metabolite could be detected. The detection window for IOX4 could achieve up to 6-month postadministration (last sampling point) by monitoring IOX4 in hair. In order to evaluate the longitudinal distribution of IOX4 over 6 months, a validated quantification method of IOX4 in hair was developed for the analysis of the postadministration samples. Segmental analysis of 2-cm cut hair across the entire length of postadministration hair showed that IOX4 could be quantified up to the level of 1.84 pg/mg. In addition, it was found that the movement of the incorporated IOX4 band in the hair shaft over 6 months varied among the three horses due to individual variation and a significant diffusion of IOX4 band up to 10 cm width was also observed in the 6-month postadministration hair samples.
© 2022 John Wiley & Sons, Ltd.
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Publication Date: 2022-03-07 PubMed ID: 35195358DOI: 10.1002/dta.3247Google 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.

The research study is about the monitoring of a banned substance called IOX4 in horse hair over a six-month period using high-resolution mass spectrometry. This substance is forbidden in horse racing and equestrian sports. The researchers were able to detect IOX4 in horse hair samples up to six months after its administration and found that its detection was possible without identifying any metabolites.

Research Objectives and Methodology

  • The study aimed to track the presence and distribution of IOX4, a hypoxia-inducible factor stabilizer, in the hair of horses over a period of six months. This substance, which can potentially be abused or misused in horse racing and equestrian sports, is prohibited.
  • Three thoroughbred mares were administrated IOX4 nasoesophageally at a dose of 500 mg per day for three days. Six bunches of mane hair were collected from each horse at specific intervals — before administration, and at 1, 2, 3 and 6 months post-administration.
  • The research team used liquid chromatography/electrospray ionization Q Exactive high-resolution mass spectrometry to gauge the presence of IOX4 and its metabolites in the hair samples.

Findings

  • IOX4 was detected in all hair samples taken after the administration of the substance, however, no metabolites were identified.
  • The ability to detect IOX4 continued up to six months (the last sampling point). This indicates that IOX4 could potentially be long-term tracking marker for doping in horses.
  • To evaluate the distribution of IOX4 over the six months, the researchers developed a quantification method to monitor IOX4 in hair. Segmental analysis of 2-cm long hair strands demonstrated that IOX4 could be quantified up to the level of 1.84 pg/mg.
  • Note was made of the fact that the movement of the incorporated IOX4 band in the hair varied among the individual horses. A significant diffusion of IOX4 band up to 10 cm width was observable in the hair samples six months after administration.

Conclusion

  • This research demonstrated a viable method for the long-term monitoring of banned substances in equestrian sports, specifically focusing on the detection of IOX4 in horse hair. The findings may contribute to more effective measures for doping control in horse racing and other equestrian sports.

Cite This Article

APA
Ishii H, Shibuya M, So YM, Wong JKY, Ho ENM, Kusano K, Sone Y, Kamiya T, Wakuno A, Ito H, Miyata K, Yamada M, Leung GN. (2022). Long-term monitoring of IOX4 in horse hair and its longitudinal distribution with segmental analysis using liquid chromatography/electrospray ionization Q Exactive high-resolution mass spectrometry for the purpose of doping control. Drug Test Anal, 14(7), 1244-1254. https://doi.org/10.1002/dta.3247

Publication

Drug testing and analysis
ISSN: 1942-7611
NlmUniqueID: 101483449
Country: England
Language: English
Volume: 14
Issue: 7
Pages: 1244-1254

Researcher Affiliations

Ishii, Hideaki
  • Drug Analysis Department, Laboratory of Racing Chemistry, Utsunomiya, Tochigi, Japan.
  • Department of Pharmaceutical Sciences, Tohoku University Hospital, Sendai, Miyagi, Japan.
Shibuya, Mariko
  • Drug Analysis Department, Laboratory of Racing Chemistry, Utsunomiya, Tochigi, Japan.
So, Yat-Ming
  • Racing Laboratory, The Hong Kong Jockey Club, Sha Tin, N.T., Hong Kong, China.
Wong, Jenny K Y
  • Racing Laboratory, The Hong Kong Jockey Club, Sha Tin, N.T., Hong Kong, China.
Ho, Emmie N M
  • Racing Laboratory, The Hong Kong Jockey Club, Sha Tin, N.T., Hong Kong, China.
Kusano, Kanichi
  • Veterinarian Section ∣ Equine Department, Japan Racing Association, Minato-ku, Tokyo, Japan.
Sone, Yu
  • Veterinarian Section ∣ Equine Department, Japan Racing Association, Minato-ku, Tokyo, Japan.
Kamiya, Takahiro
  • Equine Veterinary Clinic ∣ Horse Racing School, Japan Racing Association, Shiroi, Chiba, Japan.
Wakuno, Ai
  • Equine Veterinary Clinic ∣ Horse Racing School, Japan Racing Association, Shiroi, Chiba, Japan.
Ito, Hideki
  • Equine Veterinary Clinic ∣ Horse Racing School, Japan Racing Association, Shiroi, Chiba, Japan.
Miyata, Kenji
  • JRA Equestrian Park Utsunomiya Office, Japan Racing Association, Utsunomiya, Tochigi, Japan.
Yamada, Masayuki
  • Drug Analysis Department, Laboratory of Racing Chemistry, Utsunomiya, Tochigi, Japan.
Leung, Gary Ngai-Wa
  • Drug Analysis Department, Laboratory of Racing Chemistry, Utsunomiya, Tochigi, Japan.

MeSH Terms

  • Animals
  • Chromatography, Liquid / methods
  • Doping in Sports / prevention & control
  • Female
  • Hair / chemistry
  • Horses
  • Spectrometry, Mass, Electrospray Ionization
  • Substance Abuse Detection / methods
  • Tandem Mass Spectrometry / methods

Grant Funding

  • 2021 / Japan Racing Association (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. 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
  2. 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.
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  3. Checkouri A, Gheddar L, Arbouche N, Raul JS, Kintz P. Simultaneous detection of three hypoxia-inducible factor stabilizers-molidustat, roxadustat, and vadadustat-in multiple keratinized matrices and its application in a doping context. Drug Test Anal 2025 May;17(5):647-654.
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