A Sensitive And Specific ELISA Method To Determine Medroxy Progesterone Acetate And Ciprofloxacin In Animal Food

The veterinary drug residues in food has become a weighty and disputed problem in recent years since it is close to public health and has greatly affected the economic benefits of the animal husbandry. The residue level of the veterinary drug residues is nearly 1 μg ~1mg in the food or lower, and there are many interference factors while measuring, so it is extremely necessary and urgent to develop convenient and high-efficient previous treatment method , and the high sensitivity and alternative detection method.This dissertation consists of three parts. The first chapter summarized the progress of veterinary drug residues analysis in animal tissue. The regulatory analysis methods are briefly summarized and the purpose and meaning of the research in this paper are given finally. Chapter two is the Synthesis and identification of artificial antigen , as a half-antigen can't lead to immune reaction and has to be conjugated with a carrier, Medroxy Progesterone Acetate(MPA) and Ciprofloxacin(CIP) were covalently conjugated to bovine serum albumin(BSA) or ovalbumin (OVA) to prepare immunogen and coating antigen .The conjugates and their molecular ration of hapten to carrier protein were identified by UV spectrophotometry and SDS electrophoresis , and animal immunization test.In the third chapters ELISA detect system were established to detect the residues in actual samples,the titers of the antisera were 1: 512000 , 1: 1024000, the linear detection of MPA was 2μg/ml~50μg/ml,and the detection limit was 1μg/ml,the relative standard deviation was less than 10.03%,the average recovery of MPA added to sample was above 50.66% . respectively, the cross reactives of anti-MPA-BSA serum to some other hormones were less than 0.1%. The validity of kits was six months or more.The linear detection of CIP was 0.1μg/ml~5μg/ml ,and the detection limit was 0.05μg/ml, the relative standard deviation was less than 9.736% ,the average recovery of CIP added to sample were above 57.06% . respectively , the cross reactives of anti-CIP-BSA serum to some other antibiotic were less than 0.1%.The validity of kits was six months or more.In the fourth chapters a method for analysis of MPA and CIP in actual samples by HPLC was proposed. The result of ELISA detect system is accurate proved by HPLC and can be regarded as screening detection method for MPA and CIP in animal food.