Development of an enzyme-linked immunosorbent assay for the determination of florfenicol and florfenicol amine in eggs

A monoclonal antibody against florfenicol amine (FFA) was produced, and a competitive enzyme-linked immunosorbent assay (ELISA) was developed for detecting florfenicol (FF) and FFA in eggs. This proposed method exhibited the 50% inhibiting concentrations (IC50) of 0.120 ng/mL for FF and 0.118 ng/mL for FFA under the optimization of experimental parameters. At the fortified levels of 0.4, 0.8, 2.4, and 30.0 μg/kg, the inter-assay recoveries of the target compounds ranged from 78.3 to 103.2%, and the intra-assay recoveries ranged from 81.5 to 104.1%, respectively. The coefficients of variation were all below 12%. Results for the determination of spiked samples by this ELISA showed a good relationship with the high-performance liquid chromatography (HPLC) technique. This newly developed method confirmed to be rapid, user-friendly, and sensitivity for simultaneous detecting FF and FFA in eggs.