Immunoassay detection of drugs in racing horses. IV. Detection of fentanyl and its congeners in equine blood and urine by a one step ELISA assay.

The researchers have developed an efficient and economical one-step test to detect the presence of fentanyl and its related compounds in horse blood and urine, facilitating their use in pre- and post-race screenings to improve the quality of testing while reducing costs.

Objective of the Research

• The primary goal of this research was to create and assess a one-step enzyme-linked immunosorbent assay (ELISA) for the detection of fentanyl, an opioid used as a pain reliever and a potential performance-enhancing drug in horse racing. The intent was to improve the quality of pre- and post-race narcotics testing for racing horses while simultaneously reducing the expenses associated with the testing.

Methodology

• The test employs an enzyme-linked immunosorbent assay (ELISA) designed to identify fentanyl. ELISA is a common laboratory test that measures the concentration of substances such as peptides, proteins, antibodies, and hormones in biological samples, in this case, horse blood and urine.
• The antibody used in the ELISA test could also identify two other related substances, sufentanil and carfentanil, which are variations of fentanyl.
• The efficacy of the test was evaluated on how well it could detect fentanyl and its analogous compounds in blood and urine samples from horses within a specific timeframe following the administration of doses.

Findings

• The ELISA test developed was found to effectively detect fentanyl with an I-50 (measure of effective concentration) around 100 pg/ml.
• The test could analyze up to ten samples within around twenty minutes, making it quick and efficient for pre-race testing.
• The test was able to detect the presence of fentanyl or its metabolites in a horse’s blood and urine two and twenty-four hours, respectively, after administration of sub-therapeutic doses.
• The antibodies utilized in the test also cross-react with sufentanil and carfentanil, and the test successfully detected these drugs shortly after they were given to the horses.
• In routine screening, the test in combination with another immunoassay identified 10 sufentanil positives.

Conclusion

• The findings of this study demonstrate that this ELISA test for fentanyl and its congeners is capable of improving the quality and reducing the cost of pre- and post-race testing in horse racing.
• With the capability of effectively detecting fentanyl and related substances in horse blood and urine, this test can help maintain the integrity of horse racing by detecting abuse of these medications for performance enhancement.