Studies On Immunological Rapid Determination Of Phenobarbital Residues In Animal Food

Phenobarbital(PB) has the effects of pharmacodynamic action such as calmness, hypnosis, anticonvulsion and antiepileptic and is used to treat diseases of anxiety, insomnia, epileptic seizure and hyperbilirubinemia clinically. PB has been usually used as feed additive in animals with its special biological effects of calmness and hypnosis. PB was ruled not to be used in food animals by any way and was not detected in animals and animal foods in every country because it has the toxic effects of carcinogenesis and teratogenesis in humanity. The immunological analytical technique, which has many merits of sensitiveness, specificity, rapidity and convenience, have been generally used in determination of drug residues in animal food. Immunological analytical products for detection of PB residues in animal food have been developed in USA and Germany, but the products have not to be developed in this country.This study aimed at immunological rapid determination of PB residues in animal food, including the immunogenicity of PB, synthesis of artificial antigen, preparation and special properties of PB polyclonal antibody(PB pAb), PB monoclonal antibody(PB mAb) and reagent kits, the preparation and quality of immunological gold-labeled strip by GC-MS.The main contents and experimental results of this study were as follows:Basis on analysis of molecular structure and immunogenicity of PB, the active group amino was introduced to phenyl of PB and formed p-amidophenobarbital(pAPB) which has hapten structure by chemical modification of nitration and deoxidization. Diazotization was used to conjugate pAPB to bovine serum albumin(BSA) and obtained artificial immunogen (BSA-pAPB), the coating antigen OVA-pAPB was obtained in the same way. BSA-pAPB was synthesized successfully and its conjugation ratio of pAPB to BSA was about 19∶1 by identification with infrared, ultraviolet and SDS-PAGE, the high-titer, sensitive and specific PB antiserum of Balb/C mice immunized with BSA-pAPB had been produced by identification with indirect ELISA.Five New Zealand white rabbits were immunized with BSA-pAPB and PB pAb had been...