DEVELOPMENT OF AN ELISA USING A UNIVERSAL METHOD OF ENZYME-LABELING DRUG-SPECIFIC ANTIBODIES .1. DETECTION OF DEXAMETHASONE IN EQUINE URINE

The development, validation, and application of an ELISA for dexamethasone in equine urine is described. The drug-protein conjugate was immobilised in microtitre plate wells and antiserum raised against the same drug-protein conjugate was allowed to compete with sample or standard drug and the immobilised drug-protein conjugate. The proportion of antiserum binding to the immobilised drug-protein conjugate was detected using a biotinylated protein G/extravidin-alkaline phosphatase complex in situ and measurement of the substrate product. The method was used to detect the presence of drug-derived material in unextracted diluted urine after the administration of a single i.m. dose of dexamethasone at approximately 0.04 mg/kg to a thoroughbred horse. Validation of the method was carried out against a radioimmunoassay and GC/MS analysis.