Equine Doping Controls of Thymosin β $$ beta $$ 4: A Population Study and Strategy for Misuse Detection.
Abstract: Thymosin 4 (TB4) is a ubiquitous, highly conserved and abundant peptide among mammals with a critical role in cytoskeleton organization. In spite of its yet non-authorized use as a medicine and being forbidden by the IFHA, the FEI, and the WADA, intelligence and doping control laboratories reported numerous products available online claiming to contain a synthetic acetylated fragment of TB4 or TB4 itself, promoted as a growth factor with regenerative properties. In this article, the first estimation of the endogenous TB4 concentration in racing horses' blood samples was performed through a population study. We reveal that this concentration does not significantly depend on gender, age, nor horse breed. We highlight that the TB4 concentration increases significantly and rapidly in plasma stored at 4°C when not separated from blood cells due to cell lysis. Finally, we also demonstrate that the detection of a non-natural synthesis impurity is possible in equine plasma after a single dose administration of a TB4 containing product to a horse.
© 2024 John Wiley & Sons Ltd.
Publication Date: 2024-09-24 PubMed ID: 39314109DOI: 10.1002/dta.3806Google Scholar: Lookup
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Summary
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The research study focuses on controlling the misuse of Thymosin β4 (TB4), a protein often claimed to promote growth and regeneration in horses, despite its unauthorized use in veterinary medicine. Through a population study, the research establishes base TB4 concentration levels in horse blood samples, showing these are not significantly influenced by gender, age, or breed, but notably increase when blood cells undergo lysis in stored plasma. The study also shows it is possible to detect synthetic impurities of TB4 in horse plasma following a single-dose administration.
Objective and Background of the Study
- The study was commissioned due to the presence of numerous TB4 products online, promoted as substances for growth and regeneration, despite TB4’s non-licensed use as a medical substance and restrictions imposed by major equine and anti-doping authorities.
- The researchers aimed to establish baseline TB4 levels in horses, which can serve as a point of reference in detecting possible misuse of this substance.
Approach of the Study
- A population study was conducted to emphasise the endogenous TB4 concentration in the blood samples of racing horses.
- The researchers examined if the levels of TB4 were influenced by the horse’s age, sex, and breed. They also investigated the change in TB4 levels during cell lysis (cell breakdown), which happens when plasma is stored at 4°C and not separated from the blood cells.
Key Findings
- The study identified that the TB4 concentration remained consistent, irrespective of the age, gender, or breed of the horse, thereby implying these factors don’t contribute significantly as variables during testing procedures.
- It was discovered that when blood cell lysis occurs during plasma storage at 4°C, the level of TB4 increases significantly. This finding is critical in ensuring appropriate handling and storage of samples for accurate results.
- The research also revealed that a synthetic impurity resulting from non-natural synthesis of TB4 can be detected in horse plasma after a single dose of a TB4 product is administered, establishing a method to detect potential misuse.
Cite This Article
APA
Delcourt V, Garcia P, Chabot B, Aber N, Pescher M, Cacault M, Scholtes P, Loup B, Barnabé A, Popot MA, Bailly-Chouriberry L.
(2024).
Equine Doping Controls of Thymosin β $$ beta $$ 4: A Population Study and Strategy for Misuse Detection.
Drug Test Anal.
https://doi.org/10.1002/dta.3806 Publication
Researcher Affiliations
- GIE LCH, Laboratoire des Courses Hippiques, Verrières-le-Buisson, France.
- GIE LCH, Laboratoire des Courses Hippiques, Verrières-le-Buisson, France.
- GIE LCH, Laboratoire des Courses Hippiques, Verrières-le-Buisson, France.
- GIE LCH, Laboratoire des Courses Hippiques, Verrières-le-Buisson, France.
- GIE LCH, Laboratoire des Courses Hippiques, Verrières-le-Buisson, France.
- GIE LCH, Laboratoire des Courses Hippiques, Verrières-le-Buisson, France.
- GIE LCH, Laboratoire des Courses Hippiques, Verrières-le-Buisson, France.
- GIE LCH, Laboratoire des Courses Hippiques, Verrières-le-Buisson, France.
- GIE LCH, Laboratoire des Courses Hippiques, Verrières-le-Buisson, France.
- GIE LCH, Laboratoire des Courses Hippiques, Verrières-le-Buisson, France.
- GIE LCH, Laboratoire des Courses Hippiques, Verrières-le-Buisson, France.
Grant Funding
- Institut Franu00e7ais du Cheval et de l'Equitation
References
This article includes 23 references
- Safer D, Elzinga M, Nachmias VT. Thymosin β$$ eta $$4 and fx, an Actin‐Sequestering Peptide, Are Indistinguishable. Journal of Biological Chemistry 266, no. 7 (1991): 4029–4032.
- Low TL, Hu SK, Goldstein AL. Complete Amino Acid Sequence of Bovine Thymosin Beta 4: A Thymic Hormone That Induces Terminal Deoxynucleotidyl Transferase Activity in Thymocyte Populations. Proceedings of the National Academy of Sciences 78, no. 2 (1981): 1162–1166.
- . Uniprot: The Universal Protein Knowledgebase. Nucleic Acids Research 45, no. D1 (2017): D158–D169.
- Sanders MC, Goldstein AL, Wang Y‐L. Thymosin Beta 4 (fx Peptide) Is a Potent Regulator of Actin Polymerization in Living Cells. Proceedings of the National Academy of Sciences 89, no. 10 (1992): 4678–4682.
- Wishart DS, Feunang YD, Guo AC. Drugbank 5.0: A Major Update to the Drugbank Database for 2018. Nucleic Acids Research 46, no. D1 (2018): D1074–D1082.
- Delcourt V, Garcia P, Chabot B. Tb500/tb1000 and sgf1000: A Scientific Approach for a Better Understanding of Misbranded and Adulterated Drugs. Drug Testing and Analysis 15, no. 4 (2023): 458–464.
- Esposito S, Deventer K, Goeman J, Van der Eycken J, Van Eenoo P. Synthesis and Characterization of the n‐Terminal Acetylated 17‐23 Fragment of Thymosin Beta 4 Identified in TB‐500, a Product Suspected to Possess Doping Potential. Drug Testing and Analysis 4, no. 9 (2012): 733–738.
- Ho ENM, Kwok WH, Lau MY. Doping Control Analysis of TB‐500, a Synthetic Version of an Active Region of Thymosin β$$ eta $$4, in Equine Urine and Plasma by Liquid Chromatography–Mass Spectrometry. Journal of Chromatography A 1265 (2012): 57–69.
- Krug O, Thomas A, Walpurgis K. Identification of Black Market Products and Potential Doping Agents in Germany 2010–2013. European Journal of Clinical Pharmacology 70 (2014): 1303–1311.
- Hannapel E, van Kampen M. Determination of Thymosin β$$ eta $$4 in Human Blood Cells and Serum. Journal of Chromatography A 397 (1987): 279–285.
- Hannappel E, Xu G‐J, Morgan J, Hempstead J, Horecker BL. Thymosin Beta 4: A Ubiquitous Peptide in Rat and Mouse Tissues. Proceedings of the National Academy of Sciences 79, no. 7 (1982): 2172–2175.
- MacLean B, Tomazela DM, Shulman N. Skyline: An Open Source Document Editor for Creating and Analyzing Targeted Proteomics Experiments. Bioinformatics 26, no. 7 (2010): 966–968.
- Pino LK, Searle BC, Bollinger JG, Nunn B, MacLean B, MacCoss MJ. The Skyline Ecosystem: Informatics for Quantitative Mass Spectrometry Proteomics. Mass Spectrometry Reviews 39, no. 3 (2020): 229–244.
- R Core Team. R: A Language and Environment for Statistical Computing. Vienna, Austria: R Foundation for Statistical Computing, 2022.
- Dowle M, Srinivasan A. data.table: Extension of ‘data.frame’. R package version 1.14.8, (2023).
- Firke S. janitor: Simple Tools for Examining and Cleaning Dirty Data. R package version 2.2.0, (2023).
- Wickham H, Averick M, Bryan J. Welcome to the Tidyverse. Journal of Open Source Software 4, no. 43 (2019): 1686.
- Wilke CO. cowplot: Streamlined Plot Theme and Plot Annotations for ‘ggplot2’. R package version 1.1.1, (2020).
- Wan KX, Vidavsky I, Gross ML. Comparing Similar Spectra: From Similarity Index to Spectral Contrast Angle. Journal of the American Society for Mass Spectrometry 13, no. 1 (2002): 85–88.
- Nanjappa V, Thomas JK, Marimuthu A. Plasma Proteome Database as a Resource for Proteomics Research: 2014 Update. Nucleic Acids Research 42, no. D1 (2014): D959–D965.
- Farrah T, Deutsch EW, Omenn GS. A High‐Confidence Human Plasma Proteome Reference Set With Estimated Concentrations in PeptideAtlas. Molecular & cellular proteomics 10, no. 9 (2011): M110.006353.
- AORC. AORC Guidelines for the Minimum Criteria for Identification by Chromatography and Mass Spectrometry. (2016).
- Rahaman KA, Muresan AR, Hasan ML. Detection and Quantification of the Metabolite Ac‐Tβ$$ eta $$1–14 in In Vitro Experiments and Urine of Rats Treated With Ac‐Tβ$$ eta $$4: A Potential Biomarker of Ac‐Tβ$$ eta $$4 for Doping Tests. Drug Testing and Analysis 15, no. 11‐12 (2023): 1454–1467.
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