Approaches for the determination of florfenicol and thiamphenicol in pork using a chemiluminescent ELISA

Abstract

A chemiluminescent competitive indirect enzyme-linked immunosorbent assay (CL-ciELISA) for detection of florfenicol (FF) and thiamphenicol (TAP) residues in pork has been developed for the first time. The 50% binding inhibition (IC₅₀) value of the CL-ciELISA was 0.15 μg kg⁻¹ for FF with a cross-reactivity of 48.4% for TAP. FF and TAP were co-extracted from pork with ethyl acetate, obtaining recoveries of 80.0–93.3% (FF) and 80.0–88.3% (TAP) above the limit of detection (LOD). The LODs were 0.015 μg kg⁻¹ for FF and 0.030 μg kg⁻¹ for TAP, respectively. Moreover, 20 field pork samples were analyzed with the developed CL-ciELISA and the results correlated well with those obtained using traditional ELISA and a previously reported liquid chromatography-tandem mass spectrometry (LC-MS/MS), confirming the utility of CL-ciELISA for quantitation of FF and TAP in pork with a good accuracy and reliability. Furthermore, the CL-ciELISA method has been established for the first time for detection of FF and TAP.