食品科学 ›› 2021, Vol. 42 ›› Issue (20): 246-251.doi: 10.7506/spkx1002-6630-20200729-378

• 安全检测 • 上一篇    下一篇

基于铜纳米簇模拟酶活性建立比色法测定鱼肉中的次黄嘌呤

赵子轩,杨雪,魏洁,陈晓梅   

  1. (集美大学海洋食品与生物工程学院,福建 厦门 361021)
  • 出版日期:2021-10-25 发布日期:2021-11-12
  • 基金资助:
    国家自然科学基金面上项目(21675062);国家自然科学基金青年科学基金项目(21305050); 福建省自然科学基金项目(2017J05024);福建省特支“双百计划”人才项目

Colorimetric Determination of Hypoxanthine in Fish Based on the Enzyme Mimetic Activity of Copper Nanoclusters

ZHAO Zixuan, YANG Xue, WEI Jie, CHEN Xiaomei   

  1. (College of Ocean Food and Biological Engineering, Jimei University, Xiamen 361021, China)
  • Online:2021-10-25 Published:2021-11-12

摘要: 基于铜纳米簇(copper nanoclusters,CuNCs)的过氧化氢(H2O2)模拟酶活性,建立鱼肉中次黄嘌呤(hypoxanthine,Hx)含量快速检测的比色分析方法。通过室温搅拌法得到半胱氨酸(cysteine,Cys)-CuNCs,利用透射电镜、光电子能谱、傅里叶变换红外光谱和荧光光谱,分别表征Cys-CuNCs的形貌、元素组成、表面官能团和发光特征。紫外-可见光谱实验表明,Cys-CuNCs能灵敏催化H2O2氧化3,3’,5,5’-四甲基联苯胺显色,实现对H2O2含量的比色测定。在此基础上,利用Hx在黄嘌呤氧化酶的作用下生成H2O2,建立了一种检测Hx的比色分析方法。当孵育温度30 ℃、孵育时间1 h时,Hx浓度在4~100 μmol/L范围内与吸光度呈良好的线性关系,检出限为1.2 μmol/L。将该方法应用于鱼肉中Hx含量的测定,并通过与总挥发性盐基氮建立相关性,实现对鱼肉新鲜度的判断。

关键词: 次黄嘌呤;铜纳米簇;模拟酶;过氧化氢;鲜度

Abstract: A colorimetric method for the rapid detection of hypoxanthine (Hx) content in fish was established using copper nanoclusters (CuNCs) with high intrinsic peroxidase-like activity. Cysteine-stabilized CuNCs were prepared by a simple stirring method at room temperature. The morphology, elemental composition, surface functional groups and luminescence property of Cys-CuNCs were characterized by transmission electron microscopy (TEM), photoelectron spectroscopy, Fourier transform infrared spectrometer (FTIR) spectroscopy and fluorescence spectroscopy, respectively. Ultraviolet-visible spectroscopic experiments showed that Cys-CuNCs sensitively catalyzed the oxidation of 3,3’,5,5’-tetramethylbenzidine by H2O2, allowing for the colorimetric sensing of H2O2 content. On this basis, a colorimetric assay for Hx determination was established by generating H2O2 under the catalysis of xanthine oxidase. Under the optimal conditions as follows: incubation temperature 30 ℃ and incubation time 1 h, good linearity between Hx concentration in the range of 4–100 μmol/L and absorbance was observed and the detection of limit was 1.2 μmol/L. Finally, the proposed method was applied to the determination of Hx content in fish, and the freshness of fish was judged by establishing its correlation with total volatile basic nitrogen (TVB-N) content.

Key words: hypoxanthine; copper nanoclusters; enzyme mimetic activity; hydrogen peroxide; freshness

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