胶体金免疫层析试纸条在猪尿β-兴奋剂多残留检测中的应用  

Investigation of colloidal gold immunochromatographic strip for quantitative determination of variety of β-agonists in swine urine

作　　者：李莎[1] 白瑞樱 曾道平 伍志权 沈玉栋[1] 徐振林[1] 杨金易[1] 

机构地区：[1]华南农业大学食品学院,农业部农产品贮藏保鲜质量安全风险评估实验室,广东省食品质量安全重点实验室,广东广州510642 [2]新乡医学生理学与神经生物学教研室,河南新乡453003 [3]广州万联生物科技有限公司,广东广州510642 [4]广东汇信农产品检验有限公司,广东佛山528200

出　　处：《食品工业科技》2016年第20期53-58,共6页Science and Technology of Food Industry

基　　金：广州市珠江科技新星专项(2013J2200080);国家星火计划项目(2012GA780001;2013GA780035);广东省自然科学基金项目(S2013030013338);NSFC-广东联合基金项目(U1301214);国家科技支持计划课题(2012BAD31B0302);广东省科技计划项目(2012A020100002);广州市科技计划项目(2014J4200015)

摘　　要：本研究采用柠檬酸三钠还原法制备胶体金溶液,并对β-兴奋剂单克隆抗体进行标记。通过棋盘滴定法和单因素实验确定最佳的实验参数,研制出能同时检测多种β-兴奋剂的胶体金免疫层析试纸条。结果表明,本方法对克伦特罗的灵敏度最高,检测线性范围(IC20~IC80)为0.31~4.52μg/L,IC50为1.18μg/L,检出限(LOD,IC10)为0.14μg/L,猪尿样品无需处理,直接检测,单个样品检测时间只需8 min;与沙丁胺醇、马布特罗、溴布特罗、氯丙那林、西马特罗等β-兴奋剂类药物都有交叉,能实现β-兴奋剂的多残留检测。对阴性猪尿的加标回收实验中,试纸条批内实验回收率在83.6%~102.2%之间,试纸条批间实验回收率在84.2%~101.5%之间,且相对标准偏差均在10%以内,与ELISA、HPLC/MS法的对比实验表明,该试纸条能应用于猪尿中β-兴奋剂多残留的定量检测。In this study,the colloidal gold was obtained by reducing gold chloride with sodium citrate.Then, the colloidal gold and beta-agonists monoclonal antibody was conjugated.By optimized experimental parameters with chessboard titration method and single factor experiment, a method for Colloidal gold immunochromatographic quantitative detection of variety of beta-agonists in swine urine was established. The results showed that the method exhibited dynamic linear range（IC2o-ICs0） for the detection of clenbuterol （CL）from 0.3! tjg/L to 4.52 tjg/L, with the median inhibitory concentration（ICso） of 1.18 pg/L,the limit of detection（LOD,IC^o） was 0.14 ~g/L.Samples were detected directly without treatment,the detection time was only 8 min for each sample.The result showed 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.ln the negative swine urine sample standard addition recovery experiment,the recoveries for intra-assay ranged from 83.6% to 102.2% and inter-assay ranged from 84.2% to 101.5% ,and the relative standard deviation of intra-assay and inter-assay were both less than 10%. Ihe comparison experiment Could be used in the multiresidue determination of beta-agonists in swine urine.

关 键 词：胶体金 免疫层析试纸条 Β-兴奋剂 定量检测 多残留检测 

分 类 号：TS207.3[轻工技术与工程—食品科学]