Study On Immunological Detection Methods Of Small Molecular Environmental Pollutants PAHs And PAEs

Small molecular pollutants of various kinds are frequently reported in the environment along with the development of industrialization.Polycyclic aromatic hydrocarbons(PAHs)and phthalate esters(PAEs)are two of the most abundant small molecular pollutants found in various environmental matrices.In this thesis,naphthalene and diethyl phthalate were selected from PAHs and PAEs groups for further immunological studies to meet the requirements of rapid detection for such environmental pollutants in on-site water samples.According to the principle of hapten design,2-naphthoic acid and monoethyl phthalate were selected as the hapten of naphthalene and diethyl phthalate to prepare 2-naphthoic acid-keyhole limpet hemocyanin(KLH)and monoethyl phthalate-bovine serum albumin(BSA)artificial immune antigen,respectively.The carbodiimide method and the mixed acid anhydride ratio scheme were optimized to achieve a coupling ratio between 10-20.NAP and DEP monoclonal antibodies were prepared from the artificial immune antigen.The consequential NAP and DEP monoclonal antibody titers were up to 1:15625 and 1:30000 respectively.The stable DEP gold standard antibody solution was obtained under the optimal conditions for the monoclonal antibody-labeled colloidal gold particles at p H=7.2 and the antibody labeling amount of 6 ?L/m L.The coated antigen,gold standard antibody and goat anti-mouse Ig G were bound to the blank test strip via colloidal gold enzyme-linked immunochromatography to obtain DEP enzyme-linked immunochromatographic test strip.The sensitivity against DEP can reach 20?g/L,the cross-reaction rate of 11 structural analogs is less than 2%,and the test strip can be stored for at least 90 days at room temperature under drying conditions.The test strip was validated with the national standard method(GB5009.271-2016)using the simulated DEP-contaminated seawater samples.The results showed that when the test strip coloration result is ‘-',the corresponding concentration measured by GC-MS is below 7.32 ?g/L;when the color rendering result is ‘+',the corresponding concentration ranges from 38.93 to 156.72 ?g/L;for ‘++',the responding concentration is 156.72-628.69 ?g/L;for ‘+++',the concentration is above 628.69 ?g/L.In the application to the actual water body detection,the DEP in the water samples from 36 sampling sites in the Taihu Lake Basin and the East China Sea region were analyzed and the concentrations were all below 7.32 ?g/L.