Investigation of colloidal gold immunochromatographic strip for quantitative determination of variety of β-agonists in swine urine
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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（IC20IC80）为0.314.52μg/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。Abstract: 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( IC_(20)~IC_(80)) for the detection of clenbuterol( CL) from 0.31 μg /L to 4.52 μg /L,with the median inhibitory concentration( IC_(50)) of 1.18 μg /L,the limit of detection( LOD,IC_(10)) was 0.14 μg /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. In 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%. The comparison experiment with ELISA and HPLC / MS tests showed that the test strip could be used in the multiresidue determination of beta- agonists in swine urine.
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