食品科学 ›› 2012, Vol. 33 ›› Issue (15): 252-256.

• 生物工程 • 上一篇    下一篇

苏丹红Ⅰ免疫亲和柱的研制与鉴定

朱立鑫1,2,裘雪梅1,陈兴龙1,2,刘仁荣1,*,徐 玲1,徐富勇1,2   

  1. 1.江西科技师范学院生命科学学院 2.南昌大学生命科学与食品工程学院
  • 收稿日期:2011-06-03 修回日期:2012-06-27 出版日期:2012-08-15 发布日期:2012-09-07
  • 通讯作者: 刘仁荣 E-mail:lilirenrong@hotmail.com
  • 基金资助:

    江西省教育厅课题

Preparation and Identification of Sudan I Immunoaffinity Column

  • Received:2011-06-03 Revised:2012-06-27 Online:2012-08-15 Published:2012-09-07

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。

关键词: 苏丹红, 免疫亲和柱, 高效液相色谱

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 water-acetonitrile-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.

Key words: Sudan I, immunoaffinity column, high performance liquid chromatography (HPLC)

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