Devel Opment Of Fluorescence Immunoassay For Dibutyl Phthalate And Bisphenol A In Environments

Dibutyl phthalate（DBP）and Bisphenol A（BPA）are two typical emerging organic pollutants,which have attracted wide attention due to their terrible endocrine disrupting effects.The two compounds go into the environment by a variety of ways during the process of daily production and use,and accumulate residue in multi-media environment,causing serious harm to human body.At present,the rapid analytical method for the two pollutants is mainly enzyme-linked immunosorbent assay（ELISA）based on antigen-antibody,and not suitable for analysis of sample in complex matrix,which is limited due to the bad stability of natural enzymes.However,peroxisase-like nanomaterials show unique advantages.The aim of this study was to prepare novel peroxidase-like nanomaterials and establish enzyme-free fluorescence immunoassay（FELISA）,which can provide a reliable,rapid and sensitive method for DBP and BPA pollution investigation and risk assessment.AuPtCo nanopolyhedron was synthesized and its catalytic activity was identified by TMB-H₂O₂ color system.Based on the nanomaterial,we established FELISA analysis method for DBP detection,and optimized factors such as H₂O₂ concentration,p H,reaction time and substrate OPD concentration.The detection limit of the method was 2.26 ng/m L,IC₅₀ was 13.85 ng/m L,and the detection range was 6.38～65.25ng/m L;the recoveries ranged from 88.60 to 108.40%,and the coefficients of variation（CV）ranged from 3.41 to 9.45%.The optimized method was used to detect the real samples,the results showed a good correlation with the results of traditional ELISA,R²=0.95.Silver nanoparticles（Ag NPs）were synthesized and its catalytic activity was identified by TMB-H₂O₂ color system.Based on the core reagent Ag NPs,we established ratio FELISA analysis method for DBP detection,and optimized factors such as H₂O₂ concentration,substrate concentration,p H and reaction time.The LOD of the method was 0.86 ng/m L,the IC₅₀ was 12.70 ng/m L,and the detection range was2.48～57.27 ng/m L;the recoveries ranged from 87.2 to 106.62%,and the coefficients of variation was less than 6.54%.The results showed a good correlation with the results of ELISA and LC-MS/MS in environmental water samples,R²=0.97.RhB@ZIF-67 was synthesized,and its catalytic activity was identified by TMB-H₂O₂ acid system.Based on the composite material,we established a dual-mode FELISA method for BPA detection,and optimized factors such as reaction time,p H,H₂O₂ concentration and RhB@ZIF-67 concentration.The detection limit of the method was 1.52 ng/m L,IC₅₀ was 13.23 ng/m L,and the detection range was 3.38～51.79ng/m L;the recoveries ranged from 94.20 to 106.88%,and the coefficients of variation ranged from 2.37 to 6.61%.In environmental water samples,the results showed a good correlation with the results of ELISA,R²=0.96.In conclusion,three novel enzyme-like nanomaterials were successfully prepared in this study.Based on it,the enzyme-free FELISA analysis method was established can be applied for the detection of DBP and BPA in environmental water.The method was sensitive,rapid and reliable,which can meet the needs of pollution investigation and environmental risk assessment of these two pollutants.