Research On Immunoassay Techniques For Carbofuran And The Metabolite Residues In Agricultural Products And Water

Carbofuran is a highly effective pesticide with strong toxicity.It has an effective killing effect on a variety of pests.It has a wide range of applications in the prevention and control of pests of cotton,sugar,beans and other crops.Due to the strong toxicity of carbofuran,it may cause a series of hazards such as dizziness,headache and nerve damage.It has been banned in many countries and regions.Carbofuran will be partially converted into 3-hydroxy carbofuran in the environment.Therefore,the detection of carbofuran residues should also consider the detection of 3-hydroxy carbofuran.In this study,based on the new carbofuran hapten,a broad-spectrum monoclonal antibody against carbofuran and 3-hydroxy carbofuran was prepared.Based on the principle of immunoassay,three different immunoassay technologies for carbofuran and 3-hydroxy carbofuran had been developed respectively.And for different application scenarios to optimize detection methods to meet different detection needs.Use vegetables,fruits,water and other samples for actual sample detection and method evaluation to achieve simultaneous rapid detection of carbofuran and 3-hydroxy carbofuran.The main findings are as follows:(1)A new carbofuran hapten was chemically synthesized,and a broad-spectrum monoclonal antibody against carbofuran and 3-hydroxy carbofuran was prepared.Based on the principle of immunoassay,an indirect competitive enzyme-linked immunoassay(ic ELISA)for simultaneous detection of carbofuran and 3-hydroxy carbofuran was established.The average recovery rate of carbofuran and 3-hydroxy carbofuran was 94.9.Between-111.6% and 95.2-108.8 %,IC50 values were 0.76ng/m L and 0.69 ng/m L,and the detection linear range was 0.09-13.6 and 0.04-18.9ng/m L,respectively.(2)Preparation of colloidal gold-monoclonal antibody conjugates,based on immunochromatographic detection technology,developed colloidal gold immunochromatographic test strips for simultaneous detection of carbofuran and 3-hydroxy carbofuran.The detection limit of carbofuran and 3-hydroxy carbofuran is 7-10 ng/m L.And for a variety of water samples for application testing,eliminating matrix interference,suitable for testing a variety of water samples.The operation is simple,the detection is rapid,and the single detection time is within 10 minutes,which is suitable for field detection without laboratory conditions and qualitative screening of large quantities of samples.(3)Based on the evanescent wave theory,planar waveguide fluorescent immunosensor and the principle of competitive immune detection,a fluorescent immunosensor detection method that can simultaneously detect carbofuran and 3-hydroxy carbofuran has been established.A series of related sensor parameters were optimized to realize the automatic detection of carbofuran and 3-hydroxy carbofuran in actual samples.The average recovery rates of carbofuran and 3-hydroxy carbofuran were 98.7%-109.6% and 94.3%-104.9%,respectively,IC50 values were 0.89 ng/m L and 0.75 ng/m L,and the detection range was 0.29 –2.69 ng/m L and 0.12–4.59 ng/m L.The detection cost is low,the operation is simple,and the degree of automation is high.The single detection time is about 30 minutes,which is suitable for the quantitative detection of large quantities of samples and automatic online monitoring.(4)Three types of immunoassay technologies are used,and typical horticultural products in Hainan Province are selected for testing to verify the application of the established immunoassay technology in actual samples.A comprehensive analysis of the residual effects of carbofuran and related metabolites in agricultural products in Hainan Province provides feasible methods for the detection of carbofuran residues in actual samples and the monitoring of illegal addition of pesticide preparations.