Study On A Rapid Detection Technology For Nitrofuran Metabolites With Magnetic Particle-based Chemiluminescence Enzyme Immunoassay

In this research, a rapid detection technology of nitrofuran metabolites with magnetic particle-based chemiluminescence enzyme immunoassay was studied. The magnetic particle-based chemiluminescence reagent kit to determine the nitrofuran metabolite was established, and the processing conditions of samples were optimized.At first, the standard curve of magnetic chemiluminescence enzyme immunoassay for nitrofuran metabolites antigen and antibody for was established by checkerboard method through screening the concentration of antigen and antibody, and then the performance of the kit was analyzed. The standard curve for nitrofurazone metabolite was with a linear range between 0.05 and 4 ng/mL,the IC50 was 0.272 μg/kg,R2 was 0.999. Different testing routes of magnetic particle-based chemiluminescence enzyme immunoassay were compared by sensitivity and stability to choose the reagent kit for furaltadone metabolites. As a result, the MB-BSA-AMOZ was choosed, the linear range was 0.05 to 4 ng/m L and IC50 and correlation coefficient was 0.2277 μg/kg and 0.999 respectively..The process of sample pretreatment for nitrofuran metabolites was optimized. The results showed that the main factors were as follows: the volume of acid used for hydrolysis and derivatization, the neutralization pH of sample extraction, heating temperature and the time of water bath. Optimising the sample pretreatment conditions with incurred residues samples of crucian carp,the optimization results of sample pretreatment showed that: compared with the traditional 37 ℃overnight for sample pretreatment methods, the pretreatment with 300 μL of HCl and bath in 75 ℃ water for 30 min for hydrolysis and derivatization, and then neutralize the sample to pH 6 for the ethyl acetate extraction, can effectively shorten the pretreatment time and improve the efficiency of extraction of the sample.Based on the optimized conditions of the sample processing, the samples were spiked with 0.5, 1, 2 μg/kg of SEM,the sample recovery is 96 ~ 113%, coefficient of variation ≤9%, the detection limit ≤0.08 μg/kg. The results of detection for nitrofurazone metabolite with incurred residues samples by magnetical particle chemiluminescence and ELISA kit were with good correlation, indicating that this method is effective and feasible. Samples spiked with 0.5, 1, 2 μg/kg of AMOZ, recovery was 36~43%, coefficient of variation ≤7%, the detection limit ≤0.02 μg/kg. Analysis the reason may be the optimal sample processing method of different nitrofuran metabolites samples were different.Above all, the rapid detection technology of nitrofuran metabolites with magnetic particle-based chemiluminescence enzyme immunoassay have high sensitivity, specificity, rapidity and reproducibility, so it is suitable for the rapid screening of large quantities of samples,And ti will have broad application prospects in the field of rapid detection for food safety.