Immunoassay detection of drugs in horses. I. Particle concentration fluoroimmunoassay detection of fentanyl and its congeners.

We investigated the use of particle concentration fluorescence immunoassay (PCFIA) as a technique for drug detection in racing horses. The test was constructed from an antiserum to a carboxyfentanyl-BSA conjugate and carboxyfentanyl linked to b-Phycoerythrin. Using these reagents and a PCFIA apparatus levels of fentanyl as low as 0.1 ng/ml could be detected by the assay. In addition, cross-reactivity studies on this assay showed that the anti-serum cross-reacted well with carfentanil, sufentanil and the methylated analogs of fentanyl. We therefore evaluated the ability of these agents to produce pharmacological effects in the horse and the ability of this test to detect pharmacologically significant doses of this drug in racing horses. All of these agents produced good locomotor responses in horses at doses of between 0.1 and 10 micrograms/kg. Of these agents, carfentanyl was the most potent followed by 3-methylfentanyl, sufentanyl, alpha-methylfentanyl, and fentanyl. Similarly, when these agents were administered to horses at doses sufficient to produce a pharmacological response, all produced sufficient inhibition of fluorescence in the PCFIA system to enable their detection in post-race urines from these horses. Since PCFIA is a much faster technique than radioimmunoassay, is of approximately similar sensitivity, and requires much less instrumentation we concluded that this technique holds considerable promise as an equine drug testing technique.