Doping Control Analysis of Total Carbon Dioxide (TCO2) in Equine Plasma by Headspace Gas Chromatography-Mass Spectrometry (HS-GC/MS).
Abstract: The use of alkalinising agents prior to racing for manipulating performance in the horse has been identified since the 1990s. To mitigate the risk, an international threshold for available carbon dioxide in equine plasma based on analyses using the Beckman Synchron EL-ISE analyser was adopted in 1994 by the International Federation of Horseracing Authorities (IFHA) and revised from 37 to 36 mM in 2004. In 2009, the technical support for the above instrument was discontinued by its manufacturer. Based on the same measurement principle (i.e., ion selective electrode), the Beckman DxC600 analyser was selected as an alternative and validated against the protocol developed by the Association of Official Racing Chemists (AORC). Recently, the DxC600 analyser is also no longer supported by Beckman. Various alternative methods for measuring total carbon dioxide (TCO) in plasma have been explored. Among these, a headspace gas chromatography-mass spectrometry (HS-GC/MS) method was first reported by the Analytical Forensic Testing Laboratory (AFTL) in 2017. Methods based on the same measurement principle were later developed by different horseracing laboratories. With the objective of cross-validating the new HS-GC/MS methods and to establish an absolute (rather than instrument-dependent or empirical) threshold, an international research collaboration was initiated among different racing laboratories. This paper describes the results of cross-validation studies conducted in November 2019 and December 2022 using horse administration samples from Canada and France, respectively, the determination of a threshold based on population data, and some technical insights on the HS-GC/MS methods.
© 2024 John Wiley & Sons, Ltd.
Publication Date: 2024-11-25 PubMed ID: 39586582DOI: 10.1002/dta.3832Google Scholar: Lookup
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Summary
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The research conducted cross-validation tests on new methods of conducting gas chromatography-mass spectrometry to measure the total carbon dioxide in horse plasma samples. The new method was compared to previous ways, and it also aimed at creating an absolute threshold for total carbon dioxide measurements that were not tied to a specific instrument or empirical data.
Background
- In horse racing, performance-enhancing substances have been an issue, with alkalinising agents identified as a form of doping where the horses’ performance could be manipulated.
- Due to this, international standards set limits for total carbon dioxide in horse plasma, initially with a threshold of 37mM that was later revised to 36mM in 2004. These limits were determined using a Beckman Synchron EL-ISE analyzer.
- This analyzer’s technical support was discontinued in 2009, leading to the use of an alternative, the Beckman DxC600 analyzer, which was validated against the original protocol.
- This Beckman DxC600 analyzer also lost support, necessitating the exploration of new methods for measurement of total carbon dioxide in horse plasma.
New Method
- Since 2017, headspace gas chromatography-mass spectrometry (HS-GC/MS) method has been explored as a potential alternative and various horse racing laboratories developed this method.
- This research aimed to cross-validate the new methods and establish an absolute threshold, not just relying on instrument-dependent or empirical thresholds.
- International research collaboration was initiated involving various racing laboratories to further validate the findings.
Methodology and Results
- Cross-validation studies were conducted in November 2019 and December 2022 using horse administration samples from Canada and France to further validate the HS-GC/MS method.
- An absolute threshold for total carbon dioxide in horse plasma was determined based on population data.
- The researchers also provided technical insights into the HS-GC/MS methods.
Conclusion
- This study effectively employed a new and valid method for measuring total carbon dioxide in horse plasma.
- Not only can this method offer accurate readings, but it also presents a new absolute threshold of total carbon dioxide that stands regardless of the instrument used or empirical data – a likely significant improvement in efforts towards fair and clean horse racing events.
Cite This Article
APA
Kwok KY, Kwok WH, Wan TSM, Brooks L, Popot MA, Jaubert M, Bailly-Chouriberry L, Heffron BT, Choo CK, Tso J, Tso R, McKenzie B, Selvadurai N, Batty D, Gray B, Hudson A, Ragazzoni S, Claudio M, Ho ENM.
(2024).
Doping Control Analysis of Total Carbon Dioxide (TCO2) in Equine Plasma by Headspace Gas Chromatography-Mass Spectrometry (HS-GC/MS).
Drug Test Anal.
https://doi.org/10.1002/dta.3832 Publication
Researcher Affiliations
- Racing Laboratory, The Hong Kong Jockey Club, Sha Tin, Hong Kong, China.
- Racing Laboratory, The Hong Kong Jockey Club, Sha Tin, Hong Kong, China.
- Racing Laboratory, The Hong Kong Jockey Club, Sha Tin, Hong Kong, China.
- Canadian Pari-Mutuel Agency, Ottawa, Ontario, Canada.
- Laboratoire des Courses Hippiques (GIE-LCH), Verrieres le Buisson, Paris, France.
- Laboratoire des Courses Hippiques (GIE-LCH), Verrieres le Buisson, Paris, France.
- Laboratoire des Courses Hippiques (GIE-LCH), Verrieres le Buisson, Paris, France.
- Department of Biopharmaceutical Sciences, College of Pharmacy, Analytical Forensic Testing Laboratory, University of Illinois at Chicago, Chicago, Illinois, USA.
- Racing Forensics Inc., Toronto, Ontario, Canada.
- Racing Forensics Inc., Toronto, Ontario, Canada.
- Racing Forensics Inc., Toronto, Ontario, Canada.
- Racing Forensics Inc., Toronto, Ontario, Canada.
- Racing Analytical Services Limited, Flemington, Victoria, Australia.
- Racing Analytical Services Limited, Flemington, Victoria, Australia.
- LGC, Fordham, London, UK.
- LGC, Fordham, London, UK.
- Unirelab, Settimo Milanese, Milan, Italy.
- Unirelab, Settimo Milanese, Milan, Italy.
- Racing Laboratory, The Hong Kong Jockey Club, Sha Tin, Hong Kong, China.
References
This article includes 17 references
- Lloyd DR, Rose RJ. The Detection and Performance Effects of Sodium Bicarbonate Administration in the Racehorses. Proceedings of 9th International Conference of Racing Analysts and Veterinarians New Orleans, 1992, 131–135.
- Lloyd DR, Rose RJ. Effects of Several Alkalising Agents on Acid‐Base Balance in Horses. Proceedings of 11th International Conference of Racing Analysts and Veterinarians Queensland, 1996, 104–110.
- Lloyd DR, Rose RJ. Effects of Sodium Bicarbonate on Fluid, Electrolyte and Acid‐Base Balance in Racehorses. British Veterinary Journal 151, no. 5 (1995): 523–545.
- Wilkes D, Gledhill N, Smyth R. Effect of Acute Induced Metabolic Alkalosis on 800‐M Racing Time. Medicine & Science in Sports & Exercise 15, no. 4 (1983): 277–280.
- Auer DE, Skelton K, Tay S, Baldock FC. Detection of Bicarbonate Administration (Milkshake) in Standardbred Horses. Australian Veterinary Journal 70, no. 9 (1993): 336–340.
- Reilly P, Duffield A, Suann CJ. The Analysis of the Plasma Total Carbon Dioxide in Racehorses. Proceedings of 11th International Conference of Racing Analysts and Veterinarians Queensland, 1996, 142–147.
- Auer DE, Hughes K, Vine J. Measurement of Total Plasma Carbon Dioxide Concentrations in Thoroughbreds Using the ASE Linearity Set. Proceedings of 13th International Conference of Racing Analysts and Veterinarians Cambridge, 2000, 142–147.
- Tang PW, Crone DL. Validation of Australia's Data for Plasma Bicarbonate. Proceedings of 10th International Conference of Racing Analysts and Veterinarians Stockholm, 1994, 327–328.
- Scarth J, Davis M, Houghton R. Plasma Total Carbon Dioxide in Thoroughbred Horses in Training and Prior to Racing in the UK. Proceedings of 13th International Conference of Racing Analysts and Veterinarians Cambridge, 2000, 364–367.
- Hibbert DB, Stenhouse A, Murphy C. A GUM‐Compliant Uncertainty Budget for the Analysis of Total Carbon Dioxide (TCO2) in Equine Plasma. Accreditation and Quality Assurance 13 (2008): 523–530.
- Jarrett M, Hibbert DB, Osbourne R, Young EB. Alternative Instrumentation for the Analysis of Total Carbon Dioxide (TCO2) in Equine Plasma. Analytical and Bioanalytical Chemistry 397, no. 2 (2010): 717–722.
- . Association of Racing Chemists AORC Protocol for Cross‐Validating TCO2‐Measuring Instruments Against the Beckman Synchron EL‐ISETM™ (First Edition). 2011.
- Heffron B, Bash J, Larsen AK. Cross Validation of HS‐GC/MS to Quantify Total Carbon Dioxide in Horse Plasma. Journal of Analytical Toxicology 41, no. 3 (2016): 230–235.
- Zahra PW, Jayasinghe NS, Selvadurai NS. A Fully Automated Method to Quantitate Total Carbon Dioxide in Equine Plasma by Headspace GC‐MS. Drug Testing and Analysis 14, no. 10 (2022): 1724–1731.
- Dirikolu L, Waller P, Waguespack ML, Andrews FM, Keowen ML, Guant SD. The Effect of Sodium Bicarbonate and Validation of Beckman Coulter AU680 Analyzers for Measuring Total Carbon Dioxide (TCO2) Concentrations in Horse Serum. Veterinary Science and Medicine 3, no. 4 (2017): 263–269.
- Waller A, Lindinger MI. The Effect of Oral Sodium Acetate Administration on Plasma Acetate Concentration and Acid‐Base State in Horses. Acta Veterinaria Scandinavica 49, no. 1 (2007): 38.
- Segers A, Faict D, Duponchelle A, Hartman J-P, Peluso F. Method for Manufacturing and Storing Stable Bicarbonate Solutions. US Patent Patent Number: 5,383,324 1995.
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