食品科学 ›› 2021, Vol. 42 ›› Issue (18): 298-305.doi: 10.7506/spkx1002-6630-20200915-192

• 安全检测 • 上一篇    下一篇

基于D152树脂吸附蛋白质结合SERS测定鱼肉中的鸟嘌呤含量

黄栋玮,谷贵章,胡科娜,高兴杰,张进杰,杨文鸽,徐大伦   

  1. (1.宁波大学食品与药学学院,浙江 宁波 315000;2.湖州市食品药品检验研究院,浙江 湖州 313000)
  • 发布日期:2021-09-29
  • 基金资助:
    浙江省公益性行业项目(LGN19C200010;LGN20C200013);宁波市公益类科技计划项目(2019C10032;2019C10028)

Detection of Guanine in Fish Flesh by Protein Adsorption onto Macroporous Resin Combined with Surface-Enhanced Raman Spectroscopy

HUANG Dongwei, GU Guizhang, HU Kena, GAO Xingjie, ZHANG Jinjie, YANG Wenge, XU Dalun   

  1. (1. College of Food and Pharmaceutical Sciences, Ningbo University, Ningbo 315000, China; 2. Huzhou Institute for Food and Drug Control, Huzhou 313000, China)
  • Published:2021-09-29

摘要: 基于大孔树脂除杂与表面增强拉曼光谱的灵敏实时性,建立一种鱼肉中鸟嘌呤含量的快速检测方法。以D152树脂为填料,柱内直径1 cm,柱高20 cm,蛋白吸附率在83%~92%之间,鸟嘌呤滤除率大于95%。经单因素试验筛选和正交试验优化得出,上样流速为3 mL/min,洗脱液pH值为4.0以及洗脱流速为0.5 mL/min。基于银包金纳米颗粒(silver-coated gold nanoparticles,Au@Ag NPs)为表面增强基底,鸟嘌呤质量浓度在0.001~100 mg/L范围,鸟嘌呤质量浓度与拉曼强度呈线性关系,R2为0.969 9,检出限为0.1 mg/L。整个检测过程只需10 min,且无需复杂的样品处理。结果表明,D152树脂作为填料的层析柱能有效去除鱼肉中蛋白质等杂质的干扰,以Au@Ag NPs为基底的表面增强拉曼光谱技术的方法能够灵敏、快速的检测鱼肉中痕量嘌呤,检出限低于现有的高效液相色谱法,为开发设计水产品中嘌呤的快速检测方法提供研究基础。

关键词: 大孔树脂;鸟嘌呤;表面增强拉曼光谱;快速检测

Abstract: A rapid, sensitive real-time method for the detection of guanine in fish flesh was developed by impurity removal using microporous resin adsorption combined with surface-enhanced Raman spectroscopy (SERS). The percentage adsorption of protein onto a column packed with D152 resin (20?cm × 1?cm, i.d.) was 83%–92% and the recovery of guanine was higher than 95%. Using one-factor-at-a-time and orthogonal array design methods, the optimum adsorption and desorption conditions were established as follows: sample loading flow rate 3 mL/min, eluent pH 4.0, and elution flow rate 0.5?mL/min. Silver-coated gold nanoparticles (Au@Ag NPs) were used as a substrate for SERS. Guanine concentration in the range from 0.001 to 100?mg/L had a linear relationship with its Raman intensity, with a correlation coefficient (R2) of 0.969 9 and the detection limit of the method was 0.1?mg/L. The entire detection process took only 10 minutes without the requirement for any complicated sample preparation. These results show that the chromatographic column filled with D152 resin could effectively eliminate the interference of impurities such as proteins in fish flesh, and the SERS method could rapidly and sensitively detect trace purines in fish flesh with a detection limit lower than that of the existing HPLC method.

Key words: macroporous resin; guanine; surface-enhanced Raman spectroscopy; rapid detection

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