| Beta agonists are a class of benzene amine drugs which chemical structure and physiological functions similar to the norepinephrine and epinephrine. Beta agonists are commonly used in clinical treatment of human, animal bronchial spasm and asthma and other diseases. High dose of beta agonists can affect the reapportion of nutrients in animals, and significantly improve the lean meat percentage. Therefore, beta agonists had been widely used as a feed additive in animal production. The accumulation of beta agonists in the animal visceral is serious, which can enter the human body through the food chain, causing palpitations, dizziness, arrhythmia and other symptoms in human, and even lead to death. Therefore, the detection of beta agonists has been the key issue of food safety.At present, the multi residue detection of beta agonists are mainly GC/MS, HPLC-MS and other instrumental methods or ELISA immunoassay methods. This paper took beta agonists as the research object, combined with nano material technology and applied to immunochromatography technology, established immunochromatographic test strip quantitative detection method for multi residue of beta agonists, and the performance index of the established method is evaluated. The main research contents and results are as follows:(1)Using sodium citrate reduction method prepared the colloidal gold solution, the colloidal gold and beta-agonists monoclonal antibody was conjugated, forming the gold labeled antibody compound.A beta agonists multi residues of GICA detection technology was established by using colloidal gold as marking material, the method exhibited dynamic linear range(IC20-IC80) for the detection of clenbuterol(CL) from 0.26 μg/L to 5.70 μg/L, with the median inhibitory concentration(IC50) of 1.23 μg/L, the limit of detection(LOD, IC10) was 0.11 μg/L, the detection time was only 8 min for each sample; the result show that the developed method had cross reaction rate with salbutamol, mabuterol, brombuterol, cimaterol, clorprenaline and other beta-agonists, which could achieve multiresidue determination of beta-agonists. In the negative swine urine sample standard addition recovery experiment, the recoveries for test strip ranged from 83.6~102.19%, in the negative pork standard addition recovery experiments, the test strip recovery rate between 64.27~87.98%, and the relative standard deviation of intra-assay and inter-assay were both less than 15%, the comparison experiment with ELISA and HPLC-MS tests showed that the test strip can be used in the multi residue determination of beta-agonists.(2) Cd Te/Zn S QDs were used as labeled materials, carboxylated quantum dots and beta-agonists monoclonal antibody was conjugated with EDC/NHS method, forming the QDs-m Abs fluorescent compound. By optimized experimental parameters with chessboard titration method and single factor experiment, a method for fluorescence immunochromatographic quantitative detection of beta agonists was established. The results show that the method exhibited dynamic linear range(IC20-IC80) for the detection of clenbuterol(CL) from 0.14 μg/L to 2.28 μg/L, with the median inhibitory concentration(IC50) of 0.56 μg/L, the limit of detection(LOD, IC10) was 0.06 μg/L, the detection time was only 10 min for each sample; the result show that the developed method had cross reaction rate with salbutamol, mabuterol, brombuterol, cimaterol, clorprenaline and other beta agonists, which could achieve multiresidue determination of beta agonists. In the negative swine urine sample standard addition recovery experiment, the recoveries for intra-assay ranged from 80.4% to 103.7%; in the negative pork standard addition recovery experiments, the test strip recovery rate between 68.67%~82.62%, and the relative standard deviation of intra-assay and inter-assay were both less than 15%, the comparison experiment with ELISA and HPLC-MS tests showed that the test strip can be used in the multiresidue determination of beta agonists. |
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