Study Of Enzyme Immunoassay For Florfenicol Residues

Since FFC residues in Animal derived food are harmful to human’s health,the residue limit is explicitly stipulated by our government.They have some advantages,like a low false positive rate and accurate detection.However,there are some disadvantages to be reckoned with.For example,sample pretreatment procedure is complicated,and professionals are needed to operate expensive instruments.What’s more,these methods are time-consuming and unable to be used to detect veterinary drug residuess in a large scale.Facing a growing amount in veterinary drugs and increasingly serious problems of veterinary drug residuess,these disadvantages restrict field detection of FFC residuess,leaving administrative law enforcement officers unable to detect FFC effectively.The thesis aims at establishing the EMIA method in rapid detection of animal derived FFC residuess,a quantitative determination of FFC residuess with accuracy.First,in the experiment,mixed anhydride method is used to prepare complete antigen of FFC.The complete antigen proves a success after qualification tests of UV spectrophotometry and infrared spectrophotometry.As the immune animals,New Zealand white rabbits are immunized five times to get serum with high potency.After purification of serum by proteinA-sepharuse affinity chromatography and SDS-PAGE identification,the antibody is prepared successfully.Second,after optimizing multiple experiment conditions,a complete IC-ELISA method is established.In accordance with the linear equation of standard curve(y=-0.19943+1.2313),the IC50 is 4.5μg/mL,and the LOD is 0.53ng/mL.Third,after optimizing multiple experiment conditions,a complete BA-ELISA method is established.The linear equation is y=-0.32216+1.5559,the IC50 is 1.9μg/mg,and the LOD is 0.15ng/mg.Both of IC-ELISA and BA-ELISA methods have cross reaction to chloramphenicol,a structural analog of FFC.However,neither has cross reaction to its functional analogs.The experiment result shows that both have a high specificity and sensitivity.Compared to IC-ELISA,BA-ELISA has a lower LOD.