Screening and confirmation of recombinant human follistatin in equine plasma for doping control purposes.
Abstract: Recombinant human follistatin (rhFST) is a potential performance-enhancing agent owing to its stimulating effect on muscle growth. Administration of rhFST to athletes is prohibited in human sports by the World Anti-Doping Agency (WADA) and in horseracing according to Article 6 of the International Agreement on Breeding, Racing and Wagering published by the International Federation of Horseracing Authorities (IFHA). For effective control of the potential misuse of rhFST in flat racing, methods for screening and confirmatory analysis are required. This paper describes the development and validation of a complete solution for detecting rhFST and confirming its presence in plasma samples collected from racehorses. A high-throughput analysis of rhFST with a commercially available enzyme-linked immunosorbent assay (ELISA) was evaluated for the screening of equine plasma samples. Any suspicious finding would then be subjected to a confirmatory analysis using immunocapture, followed by nano-liquid chromatography/high-resolution tandem mass spectrometry (nanoLC-MS/HRMS). The confirmation of rhFST by nanoLC-MS/HRMS was achieved by comparing the retention times and relative abundances of three characteristic product-ions with those from the reference standard in accordance with the industry criteria published by the Association of Official Racing Chemists. The two methods achieved comparable limit of detection (~2.5-5 ng/mL) and limit of confirmation (2.5 ng/mL or below), as well as adequate specificity, precision and reproducibility. To our knowledge, this is the first report of the screening and confirmation methods for rhFST in equine samples.
© 2023 John Wiley & Sons Ltd.
Publication Date: 2023-07-04 PubMed ID: 37401514DOI: 10.1002/dta.3540Google Scholar: Lookup
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- Journal Article
Summary
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The research paper presents a method for detecting and confirming the presence of recombinant human follistatin (a potential performance-enhancing agent) in horses for anti-doping purposes.
Objectives of the Research
- The study aimed at the development and validation of a method to detect and confirm the presence of recombinant human follistatin (rhFST), a substance that can enhance performance by stimulating muscle growth, in horse plasma samples.
- This work was done since the administration of rhFST is prohibited in human sports by the World Anti-Doping Agency (WADA) and is also forbidden in horse racing.
- There was a need for an effective control method to curb potential misuse of rhFST in horse racing.
Methodology Employed
- The authors first used a commercially available enzyme-linked immunosorbent assay (ELISA), which is a common lab test, for a high-throughput analysis of rhFST in horse plasma samples.
- If the ELISA test exhibited suspicious findings, the samples would then go through confirmatory analysis. This entailed using immunocapture to isolate the rhFST, followed by nano-liquid chromatography/high-resolution tandem mass spectrometry (nanoLC-MS/HRMS) to confirm its presence.
- The confirmation step involved comparing the retention times and relative abundances of three characteristic product-ions from the test sample with those from the reference standard in compliance with the industry criteria published by the Association of Official Racing Chemists.
Findings of the Study
- The proposed method achieved a comparable detection limit (~2.5-5 ng/mL) and confirmation limit (2.5 ng/mL or below). It also had adequate specificity, precision, and reproducibility.
- The authors claim that this is the first report of such screening and confirmation methods for rhFST in equine samples.
Cite This Article
APA
Wong KS, Cheung HW, Choi YC, To NS, Wan TSM, Ho ENM.
(2023).
Screening and confirmation of recombinant human follistatin in equine plasma for doping control purposes.
Drug Test Anal.
https://doi.org/10.1002/dta.3540 Publication
Researcher Affiliations
- Racing Laboratory, The Hong Kong Jockey Club, Sha Tin Racecourse, Sha Tin, N. T., Hong Kong, China.
- Racing Laboratory, The Hong Kong Jockey Club, Sha Tin Racecourse, Sha Tin, N. T., Hong Kong, China.
- Racing Laboratory, The Hong Kong Jockey Club, Sha Tin Racecourse, Sha Tin, N. T., Hong Kong, China.
- Racing Laboratory, The Hong Kong Jockey Club, Sha Tin Racecourse, Sha Tin, N. T., Hong Kong, China.
- Racing Laboratory, The Hong Kong Jockey Club, Sha Tin Racecourse, Sha Tin, N. T., Hong Kong, China.
- Racing Laboratory, The Hong Kong Jockey Club, Sha Tin Racecourse, Sha Tin, N. T., Hong Kong, China.
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