Development of a rapid chemiluminescent ciELISA for simultaneous determination of florfenicol and its metabolite florfenicol amine in animal meat products




A rapid one-step chemiluminescent competitive indirect enzyme-linked immunosorbent assay (CL-ciELISA) for florfenicol (FF) and its major metabolite florfenicol amine (FFA) residues in animal meat products has been developed.


The 50% binding inhibition (IC50) values of the method were 0.195 µg kg−1 for FFA and 0.24 µg kg−1 for FF under optimum conditions. The cross-reactive rates for FF and FFA were 100.0% and 81.2%, respectively. FF and FFA were easily extracted from animal meat product with an FF/FFA extraction buffer, obtaining recoveries of 81.8–92.0% (FF) and 77.2–100% (FFA). The whole one-step CL-ciELISA test can be accomplished within 40 min in theory. The detection limits (LODs) of the assay were 0.98 µg kg−1 for FF and 0.80 µg kg−1 for FFA in animal meat samples. Finally, field animal meat samples were analyzed with the CL-ciELISA method, and the results correlated well with those obtained using traditional ELISA and a previously reported liquid chromatographic–tandem mass spectrometric method.


The combined results confirmed the utility of this faster one-step CL-ciELISA for simultaneous trace analysis of FF and FFA. To date, this is the most rapid developed ELISA and CL-ELISA method for detection of FF and FFA. © 2013 Society of Chemical Industry