Research On New ELISA Methods For Detection Of Several Typical Environmental Endocrine Disruptors

Environmental endocrine disruptors, also called environmental hormones, act as estrogensin humans and animal bodies. Most of them are discharged into the environment with theproduction and activities of the humanity. It has been found that environmental endocrinedisruptors have adverse effects on endocrine system in humans and animals, once exposed to theenvironment can lead to less secretion of sex hormones and poorer activity in humans and animals,more diseases like azoospermia, hypo genesis of reproductive system and cancer, effecting onimpair fertility, immune system and nervous system, causing gene mutation. Environmentalendocrine disruptors have become the third environmental problem after the global wanning andozonosphere destruction. Now the research on screening and determining the environmentalendocrine disruptors has become one of the hotpots of environmental science.In this study, we researched on new ELISA methods for detecting Naphthalene (NA) , p,p'-Dibromobiphenyl (PBB15) and 3,4 -Dichlorobiphenyl (PCB12) , respectively. Meanwhile,compared the results detected by ELISA and HPLC, made a correlation analysis between them.(â… ) ELISA method was developed for the analysis of NA in the Jingyue Lake, fountain andSuzhou River, with the antibody and coating antigen made by our lab. For the optimum conditionsof experiment,such as pH, sodium chloride and methanol content in PBS, blocking agents,washing solution as well as concentration of goat anti-rabbit HRP-IgG, the linear calibration rangewas 10^-3-10μg/L, the detection limit IC₂₀ was 0.2500 ng/L. Spiked recoveries from three watersamples were 87%-107% for NA from ELISA which was satisfactory. Meanwhile, the linearrelationship between ELISA and HPLC measurements of the dyeing sludge sample was strong inthe combined data, the correlation coefficient reached 0.9867. (â…¡) ELISA method was developed that can used for the detection of PBB15 in the JingyueLake, fountain and Suzhou River with the antibody made by our lab, and the coating antigen madeby me. Under optimized assay conditions, the linear calibration range was 10^-2-10²Î¼g/L, thedetection limit IC₂₀ was 0.02000μg/L. Spiked recoveries from three water samples were97%-147% for PBB15 from ELISA which was satisfactory. Meanwhile, the correlation of theresults detected both by ELISA and HPLC in the dyeing sludge sample was well, both methodswere highly correlated (r = 0.9048).(â…¢) ELISA method was developed for the analysis of PCB12 in the tea with the antibodymade by our lab and the coating antigen made by me. Titer for determination of antibody byindirect ELISA was 100. Under optimized assay conditions, the linear calibration range was 10^-2-10² ug/L, the detection limit IC₂₀was 4.040 ng/L. Spiked recoveries from tea sample were95% -113% for PCB12 from ELISA which was satisfactory. Meanwhile, the correlation of theresults detected by ELISA and HPLC in the tea sample acted well, the correlation coefficient was0.9894.Compared with HPLC, the detection of limit for ELISA was lower, which can meet thetesting requirements of trace materials existing in environment. The line range of HPLC is widerthan ELISA, therefore the analytes have to be enriched, it causes incremental cost andtime-extension.However, the correlation and repeatability of ELISA are not so good as HPLC.