伏安免疫法检测牛奶中氯霉素残留  

Immuno-voltammetric Determination of Chloramphenicol Residues in Milk

作　　者：宋巍巍[1] 丁明星[1] 张挪威[1] 刘海峰[2] 徐明刚[2] 刘国艳[2] 柴春彦[2] 

机构地区：[1]华中农业大学动物医学院,武汉430070 [2]上海交通大学农业与生物学院,上海201101

出　　处：《分析化学》2007年第12期1731-1735,共5页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金资助项目(No.30571368);上海市科委攻关计划项目(No.06dz05040)资助

摘　　要：为探索用于现场检测牛乳中氯霉素（CAP）残留的高灵敏度及特异性强的免疫传感器方法，本实验在制备CAP单克隆抗体的基础上，以卵清蛋白-氯霉素（OVA-CAP）偶联物为包被抗原，并将其包被到聚苯乙烯反应板上；在孵育反应中，样品中的CAP与OVA-CAP竞争结合CAP单克隆抗体，洗涤后加入碱性磷酸酶（ALP）标记的二抗，经再次孵育及洗涤后加入对硝基苯磷酸（pNPP）底物液；反应终止后用线性导数伏安法记录pNPP水解产物的氧化峰电流。实验结果表明，用免疫传感法测试CAP的灵敏度高于传统的间接竞争酶联免疫吸附法（ELISA）。该方法检测CAP的检出限为0．064μg／L；检测线性范围为0．15～600μg／L，测试牛奶样品的平均回收率为89．8％。另外，由于免疫电化学传感器体积较小，便于携带，操作简单，可实现牛乳样品中CAP残留的现场检测。To explore the immuno-vohammetric techniques with high sensitivity and specificity for the determination of chloramphenicol （CAP） residues in milk, the conjugation of oalbumin （OVA）-CAP was immobilized onto the micro-reaction plate and the monoclonal anti-chloramphenicol antibody prepared to set up competitive enzyme-linked immunoabsorbent assays （ELISA）. After the procedure of competitive combination of OVA-CAP and CAP in sample with anti-chloramphenicol monoclonal antibody during the incubation period, the plate was rinsed and alkaline phosphatase （ALP）-labeled goat anti-mouse IgG added into the micro-cuvettes, p-Nitrophenylphate was applied as substrates and the oxidative peak currents were recorded by differential pulse volta- mmetry after the reaction on the plate had been stopped by 2 mol/L NaOH. The results showed that the sensitivity of immuno-voltammetry was better than that of ELISA in the determination of chloramphenicol residues. This method allows the detection limit as low as to 0.064 μg/L with the linear range of 0.15 -600 μg/L and the average recovery rate reaches 89.8% in milk sample. Furthermore, the immuno-voltammetric apparatus are portable and can be used for the determination of chloramphenicol residues in milk on the sites due to its small size and easy operating.

关 键 词：氯霉素 微分脉冲伏安法 免疫传感方法 

分 类 号：TS252.7[轻工技术与工程—农产品加工及贮藏工程]