【作者】 于秋香；

【导师】 翟志席； 王保民；

【作者基本信息】 中国农业大学 ， 作物栽培学与耕作学， 2003， 硕士

【摘要】 本研究通过制备对硫磷的特异性抗体，建立了蔬菜中对硫磷残留的酶联免疫吸附分析法(enzyme-linked immunosorbent assay, ELISA)。先将对硫磷苯环上的硝基还原为氨基，然后采用重氮化法将氨基-对硫磷与牛血清白蛋白(BSA)或卵清白蛋白(OVA)相偶联，分别合成了免疫原和包被抗原：氨基对硫磷-BSA、氨基对硫磷-OVA。对合成的抗原进行紫外可见扫描，确证偶联成功。用免疫原免疫新西兰大耳白兔，制得对硫磷抗血清，测得其效价为1：2000，用饱和硫酸铵沉淀法得到纯化抗体，与其它类似物的交叉反应为甲基对硫磷：1.22％，毒死蜱：0.09％，杀螟松：0.06％，标准曲线的线性范围为5-1000ng／mL。用此血清建立的ELISA方法检测蔬菜样品(小白菜、豆角、辣椒)中添加的对硫磷标准样品，平均回收率为96.52％，与GC方法的相关性很高，准确性大大高于酶抑制法，整个测定大约在(不包括包被时间)1.5h内完成。更多还原

【Abstract】 Enzyme-linked immunosorbent assay (ELISA) was developed for the analysis of parathion in vegetable samples. A diazotization reaction was employed to conjugate parathion-amino to large carrier protein-BSA or OVA, and parathion-amino-BSA, parathion-amino-OVA were been synthesized. The antigen was identification by the method of ultraviolet and visible light scanning, the result demonstrated that the conjugates were synthesized successfully. The parathion was obtained after three months by using the immunogen in rabbits.The cross-reaction tests indicated most analogues and the compounds structurally related to the parathion not showed cross-reactions to the antibodies. The antibodies to parathion were purified. The effects of pH on affinity of the antibody and antigen for parathion were investigated. The tests showed the affinity of the antibody to antigens between pH higher than 6.64 and lower 8.04. The ELISA was shown to be very accurate and precise for measuring the concentrations of parathion. Results from the analysis of vegetable samples were validated by GC. The preliminary results demonstrated that the high sensitivity and selectivity of our assay systems held promise for the monitoring of parathion in the environment. The whole procedure could be completed in about 1.5h(except coating).更多还原