Quantification of Ponceau 4R in Foods by Indirect Competitive Enzyme-Linked Immunosorbent Assay (icELISA).
Quantification of Ponceau 4R in Foods by Indirect Competitive Enzyme-Linked Immunosorbent Assay (icELISA).
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DOI：
10.1021/acs.jafc.5b02129
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发表时间：
2015-07
影响因子：
6.1
通讯作者：
Ya-qing Dong;Jie Zhang;Yue Xing;Zhaorui Song;Yufen Wang;Meng Meng-Meng;Chuan Deng;Z. Tong;Yongmei Yin;Rimo Xi
中科院分区：
文献类型：
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作者：
Ya-qing Dong;Jie Zhang;Yue Xing;Zhaorui Song;Yufen Wang;Meng Meng-Meng;Chuan Deng;Z. Tong;Yongmei Yin;Rimo Xi
丽春红4 R是一种很少被允许的偶氮染料，在一些国家可以添加到某些食品中。然而，由于其潜在的危害性，因此有必要开发一种可靠、快速的分析方法来对其进行监测。通过在丽春红4 R的结构中引入一个伯胺基团，首次设计合成了丽春红4 R的半抗原。在制备好的半抗原的基础上，用戊二醛将丽春红4 R连接到载体蛋白上，制备丽春红4 R免疫原。获得触发的多克隆抗体并通过ELISA测试以优化适当的稀释度。建立了丽春红4 R的间接ELISA方法，IC_（50）为36.82 ng/mL。方法的检出限为0.80 ng/mL，线性范围为1-10000 ng/mL。5个结构类似物与丽春红4 R多克隆抗体无交叉反应（<0.3）。在葡萄汁、碳酸饮料和RIO鸡尾酒3种食品样品中，该方法具有良好的稳定性和重现性，回收率为93.87- 103.77%，批内和批间变异系数均小于11.73%。结果表明，该方法灵敏、准确、特异、简便，可用于食品中胭脂红4 R的检测。
As one of the rarely allowable azo dyes, ponceau 4R can be added in some foods in some countries. However, it is necessary to develop a credible and rapid analytical method for its monitoring, because of its potentially harmful risk. The hapten of ponceau 4R was first designed and synthesized by introducing a primary amine group into the structure of ponceau 4R. Based on the well-prepared hapten, the immunogen of ponceau 4R was prepared using glutaraldehyde to link ponceau 4R to the carrier protein. The triggered polyclonal antibody was obtained and tested by ELISA to optimize the proper dilution. An icELISA was developed for ponceau 4R, and the IC50 of the method is 36.82 ng/mL. The limit of detection is 0.80 ng/mL, and the linear range is 1-10000 ng/mL. Five selected structural analogues have no cross-reactivity with the anti-ponceau 4R polyclonal antibody (<0.3). In three food samples (grape juice, carbonated beverage, and RIO cocktail), the assay exhibits good stability and reproducibility with a recovery range of 93.87-103.77%, and the intra- and interassay coefficients of variation were <11.73%. The results indicate that the proposed icELISA is sensitive, accurate, specific, and simple, which provides an alternative for the detection of ponceau 4R in foods.