The Establishment Of The Enzyme-linked Imumunosorbent Method To Detect The Residue Of Nitroimidazole, Ractopamine And Diazepam

In livestock production, the use of antibiotics additives and growth promoters will undoubtedly improve production efficiency and bring huge economic benefits, but veterinary medicine in animal products not only engender the harm to the health of consumers,such as cancer, mutation and the increase of bacteria resistant, but also cause effect on our country's livestock and poultry products export trade. At present many countries and area all carry out a variety of ways to control or reduce the elixir residues animal in order to protect consumers' safety. However, breeding illegally added veterinary phenomenon is on the rise in pursuit of the economic interests. In order to protect the people's health, the safety of animal products must be made in all links, the effective method made to control the source. So the research of the methods for drug residue detection has the vital significance.At present, there are the three main methods to detect the drug residue in food: chromatography, enzyme-linked immunosorbent assay and microbial assay. Although the method of chromatography has precise determination, but it needs expensive instrument and require professional operation; Microbial method is simple, but with only qualitative, not quantitative analysis. The enzyme-linked immunosorbent assay what we study can not only make large quantities of qualitative analysis, but also quantitative analysis. In this thesis, I made the detection of nitroimidazole drugs, lake dopamine and anti-psychotic by enzyme-linked immunosorbent assay.When it comes to the drug testing for nitroimidazole medicament, firstly half antigen was retained by los dalai thiazole hydrolysis product with glutaraldehyde. By using mixed anhydride method blocking antigen was gotten by half antigen and ovalbumin (OVA), half antigen and Keyhole Limpet Hemocyanin (KLH) reacting with EDC will generate immune antigen. Use of indirect colorimetric enzyme-linked immunosorbent assay(ELISA) to detect antibody, the results:IC50=0.2 ng/mL, the limitation of the drug in food is 0.1 ng/mL, crossover rate of the antibody with other drugs which are similar in structure or function is small, coefficient of variation is less than 14% and recovery ratio is between 84% and 114%. When ractopamine were detected, blocking antigen was produced by half antigen and bovine serum albumin with EDC. Using the chemical luminescence enzyme-linked immunosorbent assay (CLEIA) to detect produced antibody, the results:IC50=0.35 ng/mL, the limitation of the drug in food is 0.1 ng/mL, crossover rate of the antibody with other drugs which are similar in structure or function is small, coefficient of variation is less than 10% and recovery ratio is between 92% and 110%.For the diazepam's detecting, firstly half antigen was generated by hydroxyl methyl hydroxyl amine reacting with diazepam. Then, the blocking antigen was gotten by the reactions of half antigen and thyroid carrier protein with hybrid anhydride, while the immune antigen was produced by the half antigen and the human serum albumin (HSA) with EDC. Using the chemical luminescence enzyme-linked immunosorbent assay (ELISA) to detect produced antibody, the results:IC50=0.145 ng/mL, the limitation of the drug in food is 0.1 ng/mL, crossover rate of the antibody with other drugs which are similar in structure or function is small, coefficient of variation is less than 20% and recovery ratio is between 68% and 92%.Through the above research, we established successfully the kit for detection of nitroimidazole medicament, ractopamine and diazepam.