The use of ELISA tests and immunoaffinity chromatography combined with reversed-phase high-performance liquid chromatography for dexamethasone detection in equine urine.
Abstract: Dexamethasone is a corticosteroid drug widely used in racehorses because of its anti-inflammatory effect. It is, therefore, frequently detected in antidoping tests. A method for the antidoping control of dexamethasone in equine urine using screening by ELISA and confirmation by immunoaffinity chromatography combined with reversed-phase high-performance liquid chromatography-diode array detection (HPLC-DAD) is described. The ELISA test is frequently used in antidoping tests for its sensitivity, relative speed, and low cost. The test showed linearity in the range of 4-500 ng/mL of urine, and the intra-assay and interassay imprecision were 9.4 and 9.7%, respectively. The confirmation method showed a limit of detection of 4 ng/mL for dexamethasone. The intra-assay and interassay imprecisions were 10.3 and 14.4%, respectively. The HPLC-DAD showed a limit of detection of 5 ng and linearity in the range of 25-500 ng of dexamethasone. The absolute method recovery was 56.4%. The proposed method detected dexamethasone up to 52 h after administration and proved to be adequate for the antidoping control.
Publication Date: 1997-09-01 PubMed ID: 9288594DOI: 10.1093/jat/21.5.393Google Scholar: Lookup
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- Journal Article
- Anti-inflammatory
- Clinical Study
- Corticosteroids
- Dexamethasone
- Diagnostic Technique
- Doping
- Drug
- Enzyme-Linked Immunosorbent Assay (ELISA)
- Equine Diseases
- Equine Health
- High-performance Liquid Chromatography (HPLC)
- Horse Management
- Horse Racing
- Laboratory Methods
- Pharmaceuticals
- Urine Analysis
- Veterinary Care
- Veterinary Medicine
- Veterinary Research
Summary
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The research work explores a method for detecting Dexamethasone – a controversial anti-inflammatory drug used in racehorses, using screening by ELISA, and then confirming with immunoaffinity chromatography and high-performance liquid chromatography-diode array detection (HPLC-DAD).
Elaborate Explanation
This particular research revolves around the propensity of Dexamethasone, a corticosteroid drug, to be detected in antidoping tests for racehorses. Dexamethasone is commonly used due to its anti-inflammatory effects. However, its abuse can lead to unfair advantages in racing competitions and health issues for horses.
- The study describes a two-part method for the antidoping control of dexamethasone in equine urine. The first stage uses the enzime-linked immunosorbent assay (ELISA) for screening purposes, and the results are then confirmed through immunoaffinity chromatography combined with reversed-phase high-performance liquid chromatography-diode array detection (HPLC-DAD).
- The ELISA test has been chosen here for its sensitivity, cost-effectiveness, and rapidity. The research found its linearity to be in the range of 4–500 ng/mL of urine. It also calculated the intra-assay and interassay imprecision rates to be 9.4% and 9.7% respectively.
- Post the ELISA test, the researchers made use of immunoaffinity chromatography and HPLC-DAD for re-confirmation. The detection limit using this method was found to be 4 ng/mL for dexamethasone, with intra-assay and interassay imprecision rates of 10.3% and 14.4% respectively. The linearity range of this method was 25–500 ng, with 56.4% absolute method recovery.
- The study concluded that the proposed method could successfully detect dexamethasone up to 52 hours post administration, making this an adequate method for antidoping control.
Given the potential misuse of Dexamethasone in racehorses, this research proves invaluable. It provides a robust, sensitive, and precise method to detect any instances of drug use, ensuring the fair treatment of animals and the integrity of horse racing competitions.
Cite This Article
APA
Ribeiro Neto LM, Spinosa HS, Salvadori MC.
(1997).
The use of ELISA tests and immunoaffinity chromatography combined with reversed-phase high-performance liquid chromatography for dexamethasone detection in equine urine.
J Anal Toxicol, 21(5), 393-396.
https://doi.org/10.1093/jat/21.5.393 Publication
Researcher Affiliations
- Departamento de Controle e Pesquisas Antidopagem, Jockey Club de São Paulo Rua Bento Frias, Brazil.
MeSH Terms
- Animals
- Anti-Inflammatory Agents / urine
- Calibration
- Chromatography, Affinity / methods
- Chromatography, High Pressure Liquid / methods
- Dexamethasone / urine
- Enzyme-Linked Immunosorbent Assay
- Horses / urine
- Linear Models
Citations
This article has been cited 3 times.- Ribeiro Neto LM, Salvadori MC, Spinosa HS. Hydrocortisone concentrations in post-race urine from horses. Vet Res Commun 2000 May;24(4):239-44.
- Albeitawi S, Bani-Mousa SU, Jarrar B, Aloqaily I, Al-Shlool N, Alsheyab G, Kassab A, Qawasmi B, Awaisheh A. Associations Between Follicular Fluid Biomarkers and IVF/ICSI Outcomes in Normo-Ovulatory Women-A Systematic Review. Biomolecules 2025 Mar 20;15(3).
- Xue Y, Liu H, Zhang Y, Yang W, Li H, Gong Y, Zhang Y, Li B, Liu C, Li Y. Label-Free and Ultra-Sensitive Detection of Dexamethasone Using a FRET Aptasensor Utilizing Cationic Conjugated Polymers. Biosensors (Basel) 2024 Jul 26;14(8).
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