| Benzo[a]pyrene, abbreviated as Bap, is one of the most potent carcinogenic polycyclic aromatic hydrocarbons. Polycyclic aromatic hydrocarbons are formed as a result of incomplete combustion of organic compounds. The distribution of Benzopyrene is very wide and mainly found in soil, water and air. The primary routes of human exposure are inhalation of polluted air, as well as ingestion of contaminated food and water.Carbonyl derivatives of benzo(a)pyrene were synthesized through two-step chemical reactions of iodination and Heck reaction, and then transformation into carboxylic acids was achieved by oximation reaction. The carboxyl haptens were conjugat with the carrier proteins,bovine serum albumin (BSA), using activated ester method for preparation of artificial antigen of benzo(a)pyrene. In this work, the intermediate products, including benzo(a)pyren-6-yl-substituted carboxylic acids (carboxyl hapten) and6-iodobenzo[a]pyrene, were characterized by IR, MS and NMR methods, and the artificial antigen was qualitatively analyzed by UV spectrophotometry. The results indicated that the carboxyl haptens and the artificial antigen were successfully synthesized. Then, the suitable working dilution of coatigen, monoclonal antibody and second antibody was explored by indirect ELISA. Meantime, the effects of pH, ionic strength and methanol concentration on sensitivity of ELISA were investigated and optimized by using indirect compete ELISA (ciELISA), and then the method of ciELISA for detection of Bap was initially established. The results indicated that the imitation degree of standard curve in linear regression model was high. The linear equation is:y=-28.105Ln(x)+123.66, R2is0.9937, and IC50is13.74ng/mL. |
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