Lipidomics investigations for improved enquine anti-doping using LC-HRMS
- Publication Type:
- Thesis
- Issue Date:
- 2025
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Lipidomics is a novel analysis tool for the equine anti-doping field. The development of analytical methods for the analysis of lipids (e.g. eicosanoids) has been necessary due to the integrity and ethical issues that arise from the use of legitimate therapeutics and mis-use of non-approved anti-inflammatory substances. With the discovery of novel biomarkers for detecting doping practices, the potential exists for a larger number of drugs to be indirectly detected over extended periods of time. With these additional novel biomarkers, these can be included into the equine biological passport, an intelligence based anti-doping program allowing for longitudinal profiling of individual horses.
The arachidonic acid cascade (i.e. eicosanoids) are involved in the inflammatory pathways with the ability to decrease in the presence of anti-inflammatory drugs. A method was developed to monitor 20 lipids and 5 corticosteroids. From these targeted biomarkers, 2 lipids (Oleoyl ethanolamide (OEA) and Arachidonoyl ethanolamide (AEA)) and 2 corticosteroids (hydrocortisone (HC) and cortisone (C)) were quantified using equine plasma from 750 pre-race blood samples to determine basal concentrations with upper and lower limits proposed.
With an understanding of the basal levels for specific biomarkers, this knowledge was applied to four separate administration studies: 2 corticosteroids and 2 bisphosphonates. Exogenous corticosteroids have long been used in the equine racing industry to treat inflammation and whilst many are legitimate therapeutics, these are prohibited during competition. The 2 corticosteroids studied in this project were triamcinolone acetonide and fluticasone propionate. Administration of these corticosteroids caused endogenous fluctuations within each individual horse allowing for HC, HC/C and OEA to give extended indirect detection compared to the targeted parent compound.
The bisphosphonate administrations of tiludronic acid and zoledronic acid is of significance due to the growing use of this class of compounds to treat bone disorders in the horse. At present, there are only two non-nitrogenous bisphosphates approved for the treatment of Navicular syndrome in the horse with no nitrogenous bisphosphonates approved for use. From the bisphosphonate administration studies, specific biomarkers were able to provide evidence of a non-nitrogenous bisphosphonate whilst others provided evidence of a nitrogenous bisphosphonate.
Further work translating the research method using the routine analysis method was necessary to allow for a better understanding of these biomarkers using an accredited plasma method at the Australian Racing Forensic Laboratory. The monitoring of newly discovered biomarkers could provide greater confidence for interpretations of a misuse of a doping agent compared to natural variation.
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