辣椒、花椒中罗丹明B免疫层析分析

doi:10.3969/j.issn.1000-1565.2021.06.008

摘要/Abstract

摘要： 为建立以胶体金标记抗体为基础的罗丹明B(RB)快速检测技术,利用戊二醛法将RB的衍生物罗丹明123(R123)与牛血清白蛋白(BSA)、卵清白蛋白(OVA)分别合成完全抗原(R123-BSA)和包被抗原(R123-OVA),对雌性大耳白兔皮下注射R123-BSA获得多克隆抗体.胶体金作为示踪标记物标记到硝酸纤维素膜(NC膜)上,涂覆R123-OVA作为检测线,用羊抗兔IgG作为质控线,通过优化实验条件,开发了渗滤式胶体金免疫快速检测试纸条,研制出RB标准品浓度比色卡.以比色卡作为判定依据,以辣椒、花椒为研究对象,消除基质影响,进行试纸条检测.结果表明:RB线性为0.01~20 ng/mL,其最低检出限为0.5 ng/mL,试纸条显色结果明显.该试纸条实验成功,无需特殊设备即可实现现场快速检测,适用于快速定性检测香料中RB.

关键词: 罗丹明B, 罗丹明123, 胶体金, 快速检测试纸

Abstract: In order to establish a rapid detection technique of rhodamine B(RB)based on colloidal gold labeled antibody, the complete antigen(R123-BSA)and coated antigen(R123-OVA)were synthesized from RB derivative R123(R123), bovine serum albumin(BSA)and ovalbumin(OVA)by glutaraldehyde method. Female white rabbits were subcutaneously injected with R123-BSA to obtain polyclonal antibodies. Colloidal gold was labeled on the nitrocellulose membrane as a tracer colloidal gold, coated with R123-OVA was used as the detection line and colloidal gold coated with sheep anti-rabbit IgG was used as the control line. By optimizing the experimental conditions, the percolation colloidal gold immunoassay strip was developed, and the RB standard concentration colorimetric card was also developed, based on the colorimetric card. Taking pepper and wild pepper as research objects, the influence of matrix was eliminated and the test strip was tested. The results showed that the linear range of RB was 0.01~20 ng/mL, and the lowest detection limit was 0.5 ng/mL. The color result of the test strip was obvious. The experiment of the test strip is successful, and the on-site rapid detection can be realized without special equipment, which is suitable for the rapid qualitative detection of rhodamine B.

Key words: rhodamine B, rhodamine 123, colloidal gold, quick test strip

中图分类号:

Q5-33

郑佳妍,周丽岩,韩晓利,李昳晴,王庭欣. 辣椒、花椒中罗丹明B免疫层析分析[J]. 河北大学学报(自然科学版), 2021, 41(6): 682-689.

ZHENG Jiayan, ZHOU Liyan, HAN Xiaoli, LI Yiqing, WANG Tingxin. Immunochromatographic analysis of rhodamine B in pepper and wild pepper[J]. Journal of Hebei University(Natural Science Edition), 2021, 41(6): 682-689.

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