5 - Nitro-2 - Furan Aldehydes, Chlorpromazine And Salbutamol Enzyme-linked Immunosorbent Assay Of Study

With the improvement of the life, the demand of animal derived food was increased. Because of the economic interest, veterinary drugs were used more and more widely. But most of these drugs were banned because of their side effects, such as carcinogenicity, teratogenicity and drug resistance. In order to ensure the food safety, many methods for detection drug residues were developed, such as HPLC, LC-MS and GC-MS. These methods were expensive and time consuming, therefore, they were hard to spread. The enzyme-linked immunosorbent assay (ELISA) technique has been known as a rapid, sensitive, specific, and costeffective analytical method and has been used for detection drug residue for many years. Chemiluminescent enzyme-linked immunosorbent assay (CL-ELISA) is more popular because of its improved performance in range of sensitivity and specificity compared with the conventional colorimetric ELISA. This paper aimed to develop ELISA for detection nitrofuran aldehydes and chlorpromazine and establish CL-ELISA for analysis albuterol.Nitrofuran aldehydes owed similar structure to nitrofuranes. Its coating antigen and immunogen were synthesized via EDC and mixed acid anhydride. Firstly, nitrofuran aldehydes was coupled with p-aminobenzoic acid and then the products were coupled with OVA and cBSA. The antibody was obtained with immuned rabbits. The value of IC50 was 6.8 ng.mL-1 and the limit of determination (LOD) was 0.2 ng·mL-1.Chlorpromazine, a kind of transquilizer, was used in breeding industry. Its coating antigen was synthesized via DCC and immunogen was obtained with mixed acid anhydride. The monoclonal antibody was collected from the immuned mouse. IC50 was 0.73 ng.mL-1 toward chlorpromazine in buffer and the antibody showed high specificity to chlorpromazine. The recovery rates were in range of 86.2-95.6% and 88.9-94.1% in pork meat and chicken meat, respectively. And the intra-assay and inter-assay coeffients of variation were all less than 16%.Salbutamol is a kind ofβ-exhilarants. The immunogen was conjugated with para aminobenzoic acid while the coating antigen was obtained by coupling the clenbuterol with OVA. The CL-ELISA gave an IC50 of 0.45 ng.mL-1 whereas IC50 obtained by traditional ELISA was 8.5 ng.mL-1. The recovery rate was in range of 82-112% and the coeffients of variation were all less than 25%.In this paper, we obtained three antibodies and developed the detection methods. Moreover, monoclonal anti-chlorpromazine antibody was obtained for the first time. Based on our work, the ELISA kits can be made with the antibodies we obtained.