胶体金免疫层析法快速检测蔬菜中3种农药残留  

Rapid Detection of Three Pesticide Residues in Vegetables Using Colloidal Gold Immunoassay

作　　者：魏茂琼 王丽[1] 兰珊珊[1,2] 刘振环 沙凌杰[1,2] 林昕[1,2] 李俊 普娅丽[3] 刘宏程 WEI Maoqiong;WANG Li;LAN Shanshan;LIU Zhenhuan;SHA Lingjie;LIN Xin;LI Jun;PU Yali;LIU Hongcheng(Agri‑food Quality Standard and Testing Technology Institute,Yunnan Academy of Agricultural Sciences,Kunming 650222,China;Laboratory of Quality and Safety Risk Assessment for Agro‑product,Ministry of Agriculture and Rural Affairs(Kunming),Kunming 650223,China;Yuanjiang County Agricultural Products Quality and Safety Detecting Test Station,Yuanjiang 653300,China)

机构地区：[1]云南省农业科学院质量标准与检测技术研究所,云南昆明650222 [2]农业农村部农产品质量安全风险评估实验室(昆明),云南昆明650223 [3]元江县农产品质量安全检测站,云南元江653300

出　　处：《河南农业科学》2024年第6期79-90,共12页Journal of Henan Agricultural Sciences

基　　金：云南省院士(专家)工作站项目(202305AF150015)。

摘　　要：基于胶体金免疫层析原理研制吡虫啉、丙溴磷及克百威快速检测试纸条,评价其在大蒜、白菜检测中的应用效果。采用柠檬酸三钠还原法制备胶体金,用胶体金颗粒标记吡虫啉、丙溴磷及克百威单克隆抗体作为检测探针,以硝酸纤维素膜为固相载体,包被吡虫啉-BSA、丙溴磷-BSA及克百威-BSA偶联物为检测线,羊抗鼠IgG抗体为质控线,制备胶体金免疫层析试纸条,并对金标抗体用量、标记抗体浓度、检测线包被浓度等参数条件进行优化,评价试纸条的灵敏度、特异性、稳定性和准确性。以大蒜、白菜为对象,优化了样品前处理方法,并对这2种蔬菜样品进行阴性检测及添加标准品农药检测。结果表明,优化后的工艺参数中吡虫啉、丙溴磷、克百威的最适标记抗体质量浓度分别为9.6、8.4、9.6μg/mL,抗原的包被质量浓度均是0.3 mg/mL,检测用时约15 min。吡虫啉、丙溴磷及克百威的检出限分别为20、3000、20 ng/mL,符合国标对2种蔬菜的限量规定。检测与吡虫啉、丙溴磷及克百威功能和结构相似的其他药物时无交叉反应,特异性较好。克百威和吡虫啉农药的最适提取剂分别为4 mL乙酸乙酯和4 mL甲醇,样品用量均为2 g,上清液3 mL;丙溴磷农药的最适提取剂是9 mL乙酸乙酯,样品用量3 g,上清液8 mL,上清液用氮气吹干。50份蔬菜阴性样品和加标样品的检测结果显示,假阳性率为6%,假阴性率为0。可见,研制的试纸条适用于大蒜、白菜现场快速检测以及基层实验室中大量样本的筛查,可为大蒜、白菜中农药残留控制提供有效监管手段。Rapid test strips for the detection of imidacloprid,profenofos and carbofuran were developed based on colloidal gold immunochromatography,and its application effect in the detection of garlic and cabbage was evaluated.The test strips for rapid detection of imidacloprid,profenofos and carbofuran were established by preparation of colloidal gold by reduction with trisodium citrate,with the monoclonal antibodies of imidacloprid,profenofos and carbofuran labeled with colloidal gold particles as the detection probes,nitrocellulose membrane as the solid‑phase carrier,imidacloprid‑BSA,profenofos‑BSA and carbofuran‑BSA conjugate as the detection lines,and goat anti‑mouse IgG antibody as the control line.The parameter conditions,including gold‑labeled antibody usage,concentrations of antibody for labeling,and concentration of the coating antigen were optimized.Under the optimal condition,the sensitivity,specificity and stability,as well as the accuracy of the test strips were evaluated.With optimization of sample extraction,the test strips were employed for the detection of pesticides in two vegetable samples by negative detection and addition of standardized pesticide products.The results showed that in the optimized process parameters,the concentrations of colloidal gold labeled antibody of imidacloprid,profenofos and carbofuran were 9.6,8.4,9.6μg/mL,respectively,the concentrations of the antigens coated were all 0.3 mg/mL,and the detection time was about 15 min.The limits of detection of imidacloprid,profenofos and carbofuran were 20,3000,and 20 ng/mL,which were in compliance with the limits of the national standard for the two kinds of vegetables.There was no cross‑reactivity and good specificity in the detection of other drugs with similar function and structure as imidacloprid,profenofos and carbofuran.The most suitable extractants for carbofuran and imidacloprid pesticides were 4 mL of ethyl acetate and 4 mL of methanol,respectively,with a sample size of 2 g and a supernatant of 3 mL;for profenofos p

关 键 词：大蒜 白菜 胶体金快速检测 吡虫啉 丙溴磷 克百威 

分 类 号：S481.8[农业科学—农药学]