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The Developemnt Of Immunoassays For Imidaclothiz

Posted on:2017-08-08Degree:MasterType:Thesis
Country:ChinaCandidate:M MaFull Text:PDF
GTID:2321330518480985Subject:Pesticides
Abstract/Summary:PDF Full Text Request
Imidaclothiz,a high efficiently,low toxic and broad-spectrum neonicotinoid insectidide,which developed by Nantong Jangshan Agrochemical and Chemicals Co.,Ltd.,has been widely used for controlling whiteflies,thrips,aphids,leafhoppers and other herbivorous pest.High performance liquid chromatography(HPLC)is the common method for detecting imidaclothiz.But instrument testing methods have some disadvantages,such as expensive equipment,large dose of organic solvents,complex operating procedure and difficult to meet the high-throughput.In this study,the fluorescence poliarizatiion immunoassay(FPIA),time-resolved fluoroimmunoassay(TRFIA),Quantum dots fluoroimmunoassay(FLISA)and enhanced gold immunochromatographic strip(Enhance GICA)based on the monoclonal antibody were developed and used for the rapid detection of imidaclothiz in agricultural and environmental samples.The time-resolved fluoroimmunoassay(TRFIA)was developed based on the monoclonal antibody(MAb),and a variety of conditions including the concentrations of coating antigens and Eu3+-antibody,organic solvent,ionic intension and pH value were determined to optimum assay.Under the optimum conditions,the IC50 and IC10 of the standard curve were 9.37 ng/mL and 0.024 ng/mL,respectively.The cross-reactivities(CRs)of TRFIA with analogues of imidaclothiz were less than 0.2%except imidacloprid(36.05%),which indicated the developed TRFIA had a high specificity for imidaclothiz.The recoveries and relative standard deviations(RSDs)in tomato,pear,rice,cabbage,cucumber,apple and paddy water samples were in the range of 81.3%-117.7%and 1.2%-6.9%,respectively.The detection results of TRFIA for the unknown samples correlated weln with these obtained by HPLC,and the R2 in water samples and pear samples were 0.9920 and 0.9699,respectivily.Compared to the FPIA based on the same antibody,the sensitivity of the TRFIA was improved 9 times,which could be a satisfactory analytical tool for the monitoring of imidaclothiz in agricultural and environmental samples.The quantum dots fluoroimmunoassay(FLISA)was developed based on the monoclonal antibody(MAb),and the concentrations of coating antigens and QDs-antibody,the buffer solution were optimized.Under the optimum conditions,the IC50 and IC10 of the standard curve were 23.63 ng/mL and 0.45 ng/mL,respectively.There were no obvious cross-reactivities with its analogues except for imidacloprid(33.53%),which indicated the developed FLISA had a high specificity for imidaclothiz.The recoveries and relative standard deviations(RSDs)in tomato,pear,rice,cabbage,cucumber,apple and paddy water samples were in the range of 72.7%-117.6%and 2.4%-13.5%,respectively.The detection results of FLISA for the unknown samples showed high correlations with these of HPLC,and the R2 in water samples and pear samples were 0.9806 and 0.9979,respectivily.The proposed FLISA has the advantages of sensitive and accurate,which could be suitable for high-throughput detection of imidaclothiz in agricultural and environmental samples.The fluorescence poliarizatiion immunoassay(FPIA)was developed based on the monoclonal antibody,and the concentrations of tracer and antibody,the buffer solution were optimized.Under the optimum conditions,the IC50 and IC10 of the standard curve were 87.94 ng/mL and of 0.57 ng/mL,respectively.The FPIA had a high cross-reactivity(CR)for imidacloprid(96.87%),and no CR was determined with other analogues,which indicated the developed FPIA had a high specificity for imidaclothiz.The recoveries and relative standard deviations(RSDs)in tomato,pear,rice,cabbage,cucumber,apple and paddy water samples were in the range of 83.3%-117.3%and 5.4%-12.0%,respectively.The detection results of FPIA for the unknown samples were markedly consistent with these of HPLC,and the R2 was 0.9942,which indicated the developed FPIA was reliable,accurate and could be used for the rapid detection of imidaclothiz in agricultural and environmental samples.The enhanced gold immunochromatographic assay(GICA)was developed based on the monoclonal antibody and high affinity between biotin and streptavidin.Under the optimum conditions,the results can be judged by the naked eye within 10 min with the lowest detection limit of 50 ng/mL.The developed GICA has no cross-reactivities(CRs)with analogues of imidaclothiz except imidacloprid.In the study,the matrix effects of different samples were evaluated,and the optimal dilution times to eliminate the matrix effects were achieved.The recoveries of spiked samples were studied by adding different concentrations of imidaclothiz in blank samples,and the results indicated GICA is accurate.The detection results of GICA for the unknown samples were largely consistent with HPLC,which indicated that the developed GICA is simple,rapid and sensitive,which could be used for the qualitative and semi-quantitative detection of imidaclothiz in agricultural and environmental samples.
Keywords/Search Tags:Imidaclothiz, Monoclonal antibody, Time-resolved fluoroimmunoassay, Quantum dots fluorescence immunoassay, Fluorescence polarization immunoassay, Enhanced-gold immunochromatographic assay
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