| The environmental endocrine disrupting Chemicals (EDCs) were also calledenvironmental hormone. Most EDCs have stable chemical properties. They can throughvarious channels to enter the body, and then they can interfere with the natural hormones inthe body. They can influence nerve system, reproductive system and immune system, andwould affect the balance and development of the body oneself, caused variety diseases, toharm human health, and even will harm children’s growth. EDCs and its harm havebecome the focus of the world.Currently, more than80kinds of environmental endocrine disruptors were found andconfirmed in daily life. Its detection method inclued chromatography method,chemiluminescence analysis, electrochemical analysis, and immune analysis, etc. Immuneanalysis method has fast analysis speed, simple, easy, high specificity, high selectivity, etc.And it is especially suitable for the analysis of biological samples, but the sensitivity is stillto be improved.Because of the huge reactive surface area and biocompatible of gold nanoparticles, itcan combine many DNA and protein. The DNA on the nanoparticles can exponentialamplification signal, consequently improve the sensitivity of detection methods. Theresearch and development of DNA-nanoparticle probe technique, which combined withcolor, silver stain and the electrochemical luminescence technology, is rapidly. Sensitivityis obviously better than the commonly detection method. With unique fluorescenceproperties of lanthanides and its chelate as tracer, time-resolved fluorescence immunoassay(TRFIA) has high sensitivity; dual-codified gold nanoparticles (DC-AuNPs) is a kind ofDNA-nanoparticle probe technique. Combined DC-AuNPs and TRFIA, and tested thecontent of environmental endocrine disruptors in biological samples, which will be furtherimprove the detection sensitivity, and own a great importance.The main content of this research is to establish a dual-codified gold nanoparticles(DC-AuNPs) amplification time-resolved fluorescence immunoassay for determination ofenvironmental endocrine disruptors. Using gold nanoparticles and biotin-streptavidin dual signal amplification, we compounded a kind of new fluorescence immunoassay reagentwith high sensitivity, stability, and then establish a dual-codified gold nanoparticlesamplification time-resolved fluorescence immunoassay method.This present thesis consists of four parts as follows:1. Determination of bisphenol A based on dual-codified gold nanoparticles amplifiedfluoroimmunoassayGold nanoparticles was prepared by the citrate reduction of HAuCl4, and modifiedwith antibody and DNA. A fluoroimmunoassay was developed for the determination ofbisphenol A in food packaging material sample based on the dual-codified goldnanoparticles. The method combined the advantage of the dual-codified gold nanoparticlesamplified system with the antigen-immobilization approach. In conclusion, experimentalresults revealed that the linear range of the presented method was from1.0×10-4to10.0ng/mL, the equation of linear regression was ΔI (I0-I)=402.0lg CBPA+4534.5, the limit ofdetection was determined to be0.07pg/mL. The proposed technique is simple, rapid, andsensitive.2. Time-resolved fluoroimmunoassay for bisphenol A based on dual-codified goldnanoparticles amplified techniqueA rapid and sensitive method based on dual-codified gold nanoparticles amplifiedtechnique by time-resolved fluoroimmunoassay for the determination of bisphenol A wasestablished. Streptavidin was labeled with europium ion by means of a simple methodinvolving the anhydride of diethylenetriam inepentaacetia acia. Under the optimalconditions, the linear range of the method was from1.0×10-6to1.0ng/mL, the equation oflinear regression was ΔI (I0-I)=784.0lg CBPA+6572.7, and the limit of detection wasfound to be0.3fg/mL. The method has been applied to determine bisphenol A in watersample with satisfactory results.3. Determination of diethylstilbestrol based on dual-codified gold nanoparticlesamplified time-resolved fluoroimmunoassayA rapid and sensitive method based on dual-codified gold nanoparticles amplified solidphase coated antigen competitive mode time-resolved fluoroimmunoassay for the determination of diethylstilbestrol was established. The linear range of the presentedmethod was from1.0×10-6to10.0ng/mL, the equation of linear regression was ΔI=690.8lg CDES+6198.3, and the limit of detection was found to be0.4fg/mL. The method hasbeen applied to determine diethylstilbestrol in milk sample with satisfactory results. |
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