| Imidacloprid(IMI)is a kind of the new nicotine insecticides,which has become the fastest growing insecticide in the world and is widely used in the process of crop pest control.The water,soil,air and agricultural products would be contaminated,further,damaging the ecosystem seriously,if the pesticides were used exceedingly or improperly.Therefore,it is of great significance to explore the establishment of pesticide imidacloprid residues in agricultural food,environmental samples,and the sustainable development of agriculture,animal husbandry and the environment.IMI-BSA and IMI-OVA were synthesized by chemical method.The results showed that IMI-BSA and IMI-OVA were successfully coupled.The hybridized cell line secreting anti imidacloprid antibody was obtained by cell fusion and subcloning.The cell supernatant was injected intraperitoneally into mice,and the final antibody secreted was κ light chain IgG3 subtype after purified by octanoic acid ammonium sulfate method,the concentration of coated antigen was optimized by chessboard titration,and the indirect competitive enzyme-linked immunosorbent assay(ELISA)was established to detect IMI.The IC50 of the competitive inhibition curve of ELISA was 1.405 ng/mL,the lowest detection limit was 0.05 ng/mL,and the correlation coefficient was R2=0.9853,moreover,the antibody had stronger specificity,and the cross reaction rate with thiamethoxamine was 0.46%in five structural analogues;there was no cross reaction with thiamethoxamine,thiametholine,nitenpyram and acetamiprid.On this basis,a chemiluminescent enzyme-linked immunosorbent assay(CLEIA)method was further established.Its IC50 was 1.741 ng/mL,R2=0.9871,and its linear range was 0.5～50 ng/mL.The established HPLC standard curve detection method,R2=0.9988,has a good linear detection range,which lays a good foundation for the later detection of imidacloprid residues.In this paper,ELISA,CLEIA and HPLC were used to detect IMI method aims to establish a rapid,stable,sensitive and high-throughput detection system for the detection of Imidacloprid in environmental samples and agricultural products,in order to provide reference for the detection of Imidacloprid in actual samples. |
