【摘要】 以羧基化的琼脂糖凝胶4FF为载体,采用碳化二亚胺法偶联抗苏丹红Ⅰ(SudanⅠ)单克隆抗体,制备苏丹红Ⅰ免疫亲和柱(IAC)。将SudanⅠ样品过免疫亲和柱,乙腈洗脱后以高效液相色谱法(HPLC)检测,紫外检测波长为478nm,流动相为0.1%甲酸水溶液-乙腈-0.1%甲酸乙腈溶液-丙酮(体积比为21.25:3.75:60:15)。抗SudanⅠ单克隆抗体免疫亲和柱以0.3g带羧基的琼脂糖凝胶4FF与1mg SudanⅠ单抗偶联,柱容量为1.6μg。辣椒粉以0.25～3mg/kg水平添加SudanⅠ标准品,平均回收率为44.52%～77.40%,相对标准偏差为4.6%～8.3%。信噪比(RSN)为3:1时的最低检测限为15ng/mL。本实验成功制备出苏丹红免疫亲和柱。更多还原

【Abstract】 A Sudan I immunoaffinity chromatographic column(IAC) was prepared by carbodiimide method,in which a modified Sepharose 4FF containing carboxyl group was used as the carrier to couple with a monoclonal antibody against Sudan I.A reversed-phase HPLC method was established to determine Sudan I using a mobile phase composed of 0.1% formic wateracetonitrile-0.1% formic acetonitrile-acetone(21.25:3.75:60:15,V/V) at a flow rate of 1 mL/min.The detection wavelength was 478 nm.The retention time of Sudan I was found to be 7.97 min.The prepared IAC column as a conjugate between 0.3 g of Sepharose 4FF and 1 mg of purified anti-Sudan I monoclonal antibody showed a capacity of 1.6μg.The average recovery rates of Sudan I from chili powder spiked with Sudan I standard at levels varying from 0.25 to 3 mg/kg was 44.52%～77.40% with relative standard deviation of 4.6%～8.3%.The detection limit was 15 ng/mL based on a signal-to-noise ratio of 3:1.Therefore,a Sudan I IAC column has been prepared successfully.更多还原