食品科学 ›› 2021, Vol. 42 ›› Issue (14): 270-275.doi: 10.7506/spkx1002-6630-20200625-342

• 安全检测 • 上一篇    下一篇

基于上转换纳米材料与磁分离的荧光免疫分析方法检测食品中酪胺

张彪,黄娜,张万利,王硕,生威   

  1. (1.天津科技大学食品科学与工程学院,食品营养与安全国家重点实验室,天津 300457;2.南开大学医学院,天津 300072)
  • 发布日期:2021-07-27
  • 基金资助:
    “十三五”国家重点研发计划重点专项(2016YFD0401204);天津市研究生科研创新项目(2019YJSB007)

Fluorescence Immunoassay Combined with Magnetic Separation Using Upconversion Nanoparticles as Label for the Detection of Tyramine in Food Samples

ZHANG Biao, HUANG Na, ZHANG Wanli, WANG Shuo, SHENG Wei   

  1. (1. State Key Laboratory of Food Nutrition and Safety, College of Food Science and Engineering, Tianjin University of Science and Technology, Tianjin 300457, China; 2. School of Medicine, Nankai University, Tianjin 300072, China)
  • Published:2021-07-27

摘要: 实验基于间接竞争反应原理,结合上转换纳米材料与磁分离技术,建立分析食品中酪胺荧光免疫的方法,并进一步应用于肉制品、水产品及发酵制品中酪胺的检测。最佳实验条件为12 μg抗体偶联NaYF4Yb,Tm上转换纳米材料制备信号探针,70 μg包被原偶联磁性聚苯乙烯微球制备捕获探针,信号探针添加量为70 μL,感应探针添加量为100 μL,孵育时间为30 min。方法线性范围为0.1~100.0 μg/L,检测限为0.05 μg/L,该方法可特异性识别酪胺,与其他生物胺及其结构类似物无交叉反应。样品添加回收率为86.44%~101.62%,变异系数小于10%,该方法适用于食品中酪胺的快速检测。

关键词: 酪胺;多克隆抗体;免疫分析;上转换纳米材料;磁分离

Abstract: Based on the principle of indirect competitive reaction combined with magnetic separation using up-conversion nanomaterials, a fluorescence immunoassay for the detection of tyramine in meat products, aquatic products and fermented products was established. The optimal working conditions were obtained as follows: 12 μg of the antibody was coupled with NaYF4Yb, Tm up-conversion nanomaterials to prepare signal probes, 70 μg of the coated antigen was coupled with magnetic polystyrene microspheres to prepare capture probes, the addition amounts of signal probes and sensing probes were 70 μL and 100 μL, respectively, and the competitive reaction time was 30 min. The linear range of the developed method was 0.1–100.0 μg/L, with a detection limit of 0.05 μg/L. The method could specifically recognize tyramine without cross-reaction with its structural analogs or other biological amines. The recoveries of tyramine from spiked samples ranged from 86.44% to 101.62%, with coefficient of variation less than 10%. The proposed fluorescence immunoassay can be applied for rapid detection of tyramine in foods.

Key words: tyramine; polyclonal antibody; immunoassay; upconversion nanoparticles; magnetic separation

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