Quantification of horse plasma proteins altered by xylazine using the fluorogenic derivatization-liquid chromatography-tandem mass spectrometry.
Abstract: In the doping tests currently used in horse racing, prohibited substances or their metabolites are usually directly detected in urine or blood samples. However, despite their lasting pharmaceutical effects, some prohibited substances are rapidly eliminated from horse urine and blood, making them difficult to detect. Therefore, new indirect biomarkers for doping, such as plasma proteins that are increased by the prohibited substances, have recently attracted much attention. Here, a fluorogenic derivatization-liquid chromatography-tandem mass spectrometry (FD-LC-MS/MS) method was adopted for horse plasma proteomics analysis, in order to identify plasma proteins whose concentrations were altered in response to xylazine in Thoroughbred horses. Xylazine, which is rapidly absorbed and eliminated and has possibility of the change in the levels of plasma proteins, was selected as a model drug. Of the ten plasma proteins identified, four proteins, including three acute phase proteins (haptoglobin, ceruloplasmin, and α-2-macroglobulin-like), were significantly increased after xylazine administration. Therefore, our present approach might be useful in identifying indirect biomarkers of drug administration.
Publication Date: 2016-02-03 PubMed ID: 26858580PubMed Central: PMC4739145DOI: 10.1294/jes.26.141Google Scholar: Lookup
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- Journal Article
Summary
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The study uses advanced analytical methods to identify potential plasma proteins in horse blood that change in response to a specific drug called xylazine. This effort is aimed at improving the accuracy and sensitivity of drug doping tests in horse racing.
Background and Purpose
- The research was conducted to address an ongoing challenge in horse racing, where prohibited substances or their metabolites are often hard to detect through conventional urine or blood tests due to their rapid elimination from the body.
- The paper proposes the possibility of using altered plasma proteins as indirect biomarkers to detect the use of certain substances. This could provide more accurate results than direct testing methods.
- Xylazine, a drug that is swiftly absorbed and eliminated from a horse’s body, was chosen as a model drug for testing this method. This suggests that the aim was to prove the concept with substances known to pose detection challenges.
Research Methodology
- The researchers employed a fluorogenic derivatization-liquid chromatography-tandem mass spectrometry (FD-LC-MS/MS) method, a technique commonly used in bioanalysis, to study plasma proteins in horse blood.
- The use of the FD-LC-MS/MS method enabled the researchers to analyze changes at the molecular level, specifically the change in plasma protein concentrations post xylazine administration.
Results and Insights
- Out of ten plasma proteins identified during the FD-LC-MS/MS analysis, four proteins, including three acute phase proteins (haptoglobin, ceruloplasmin, and α-2-macroglobulin-like), showed a significant increase after xylazine administration.
- This increase in specific plasma proteins post xylazine administration illustrates the potential of using altered plasma proteins as indirect biomarkers for drug administration. This approach could provide an essential tool for bolstering drug testing in horse racing.
Conclusion
- The paper concluded that the technique used might help in identifying indirect biomarkers for drug administration. While the test was conducted with xylazine, the approach could potentially be applied to other substances used to dope horses in competitions.
- The success of the research could lead to the development of more sensitive, effective antidoping controls, thereby ensuring fair competition and the welfare of horses in racing events.
Cite This Article
APA
Mori M, Ichibangase T, Yamashita S, Kijima-Suda I, Kawahara M, Imai K.
(2016).
Quantification of horse plasma proteins altered by xylazine using the fluorogenic derivatization-liquid chromatography-tandem mass spectrometry.
J Equine Sci, 26(4), 141-146.
https://doi.org/10.1294/jes.26.141 Publication
Researcher Affiliations
- Laboratory of Bio-analytical Chemistry, Research Institute of Pharmaceutical Sciences, Musashino University, Tokyo 202-8585, Japan; Laboratory of Proteomics Analysis, Research Institute of Pharmaceutical Sciences, Musashino University, Tokyo 202-8585, Japan; Laboratory of Racing Chemistry, Tochigi 320-0851, Japan.
- Laboratory of Proteomics Analysis, Research Institute of Pharmaceutical Sciences, Musashino University, Tokyo 202-8585, Japan.
- Laboratory of Racing Chemistry, Tochigi 320-0851, Japan.
- Laboratory of Racing Chemistry, Tochigi 320-0851, Japan.
- Laboratory of Bio-analytical Chemistry, Research Institute of Pharmaceutical Sciences, Musashino University, Tokyo 202-8585, Japan.
- Laboratory of Proteomics Analysis, Research Institute of Pharmaceutical Sciences, Musashino University, Tokyo 202-8585, Japan.
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Citations
This article has been cited 1 times.- Tozaki T, Kikuchi M, Kakoi H, Hirota KI, Mukai K, Aida H, Nakamura S, Nagata SI. Profiling of exercise-induced transcripts in the peripheral blood cells of Thoroughbred horses.. J Equine Sci 2016;27(4):157-164.
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