氯霉素残留的胶体金免疫层析法的研究

发布时间：2018-02-25 09:01

本文关键词： 氯霉素检测胶体金免疫层析技术　出处：《中国农业科学院》2015年硕士论文　论文类型：学位论文

【摘要】：氯霉素曾作为广谱抗菌素应用于畜牧业和水产养殖业，由于其严重的毒副作用而被美国FDA、欧盟、中国等相关标准规定为不得检出。因其稳定的抗菌性好、价格低廉，违法添加的现象屡见不鲜。胶体金技术是利用抗原抗体的特异性结合，放大反应结果，使结果通过直接观察即可判定的检测技术。通过对胶体金免疫层析法制备及性能，超高效液相色谱-串联质谱仪的验证等进行研究。主要研究结果如下： 1、确定以柠檬酸三钠为还原剂，制备出直径约为20nm的胶体金颗粒。用氯霉素多抗进行标记，最佳标记量为300μg/mL。 2、成功研制出氯霉素胶体金免疫层析试纸条，优化工艺条件，选择Millipore135s的硝酸纤维素膜为反应载体，用以0.5%吐温的磷酸盐缓冲液处理的全血滤膜为样品垫，使本试纸条的灵敏度达到1ppb。 3、确定了试纸条检测生牛乳、蜂蜜和鸡蛋的检测限分别是2ng/mL，，5ng/g，3ng/g，并用超高效液相色谱-串联质谱仪验证了试纸条的测定结果，达到了用一种试纸条检测不同种类样品的研究目的。
[Abstract]:Chloramphenicol has been used in animal husbandry and aquaculture as a broad-spectrum antibiotic, and has been banned from detection in the FDAs, EU, China and other related standards because of its serious toxic side effects. Colloidal gold technique is a kind of detection technique which can be determined by direct observation by using the specific binding of antigen and antibody and amplifying the reaction result. The preparation and properties of colloidal gold immunochromatography and the verification of ultra-high performance liquid chromatography-tandem mass spectrometer were studied. The main results are as follows:. 1. The colloidal gold particles with diameter of 20 nm were prepared with trisodium citrate as reducing agent, and the best labeling amount was 300 渭 g / mL with chloramphenicol polyclonal antibody. 2. The test strip of chloramphenicol colloidal gold immunochromatography was successfully prepared. The optimum technological conditions were optimized. The nitrocellulose membrane of Millipore135s was selected as the reaction carrier, and the whole blood filter membrane treated with 0.5% tween phosphate buffer was used as the sample pad. The sensitivity of the test strip is 1 ppb. 3. The detection limits of milk, honey and egg were determined to be 2ng / mLL ~ 5ng / g / g respectively. The results were verified by ultra-high performance liquid chromatography-tandem mass spectrometer, and the purpose of detecting different kinds of samples with a kind of test strip was achieved.
【学位授予单位】：中国农业科学院
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R446.6

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