Research And Application Of Rapid Detection Technology For Endocrine Disruptors In Water

Endocrine disruptors are common substances in social activities.In recent years,endocrine disruptors have been detected in natural water environment frequently.It poses a great threat to human health and ecological safety.Based on the principle of immunoassay of antigen-antibody specific reaction,the enzyme-linked immunosorbent assay method and the planar waveguide biosensor for endocrine disruptors were developed.The research aims to provide technical support for the quick detection of endocrine disruptors in water environment.Firstly,based on the principle of indirect competitive immune response,the optimized concentration for coating antigen and primary antibody of bisphenol A,estradiol and nonylphenol were determined.The enzyme-labeled antibody concentration and reaction time for ELISA detection were studied,the best reaction conditions for endocrine disruptors were obtained.Indirect competition ELISA detection methods for bisphenol A,estradiol and nonylphenol were established.The results showed that the detection limit of bisphenol A was 3.60 ?g/L,the quantitative detection interval was 8.15?64.70 ?g/L,the detection limit of estradiol was 3.18 ?g/L,the quantitative detection interval was 4.18?11.98 ?g/L,and the detection limit of nonylphenol was 24.78 ?g/L,the quantitative detection interval was 36.51 ?363.43?g/L.Secondly,based on the planar waveguide immunosensor developed by Tsinghua University research group,the preparation of immunochips and the fluorescent labeling of antibodies were studied.The effects of antibody concentration,p H and heavy metal ion concentration on the detection method of the sensor were studied.The planar waveguide immunosensor detection method of bisphenol A,estradiol and nonylphenol were established.The research results showed that the detection limit of bisphenol A was 0.04 ?g/L,the quantitative detection interval was 0.30?3.68 ?g/L,the detection limit of estradiol was 0.12 ?g/L,the quantitative detection interval was0.10?0.59 ?g/L,and the detection limit of nonylphenol was 2.10 ?g/L,the quantitative detection interval was 6.28?56.12 ?g/L.Finally,using the established indirect competitive ELISA and planar waveguide immunsensor,bisphenol A,estradiol and nonylphenol were spiked and recovered inactual water samples.The relative standard deviations of the above two methods were less than 20%,and spiked recoveries were between 80% and 120%,indicating that the precision and accuracy of the two detection methods are good,which is suitable for the detection of endocrine disruptors in actual water samples.