表面增强拉曼光谱对鱼肉中组胺的快速定量分析  

Rapid and Quantitative Analysis of Histamine in Fish Using Surface Enhanced Raman Spectroscopy

作　　者：郭小莹 邱立 张进杰[1] 杨丹婷 唐春兰 徐大伦[1] 楼乔明[1] 杨文鸽[1] 胡奇杰 GUO Xiao-ying;QIU Li;ZHANG Jin-jie;YANG Dan-ting;TANG Chun-lan;XU Da-lun;LOU Qiao-ming;YANG Wen-ge;HU Qi-jie(College of Food and Pharmaceutical Sciences,Ningbo University,Ningbo 315000,China;School of Medicine,Ningbo University,Ningbo 315000,China;Huzhou Institute for Food and Drug Control,Huzhou 313000,China)

机构地区：[1]宁波大学食品与药学学院,浙江宁波315000 [2]宁波大学医学院,浙江宁波315000 [3]湖州市食品药品检验研究院,浙江湖州313000

出　　处：《光谱学与光谱分析》2019年第8期2561-2567,共7页Spectroscopy and Spectral Analysis

基　　金：国家自然科学基金项目(31201284,21405138);浙江省自然科学基金项目(LY17H260003);浙江省食品药品监管系统科技计划项目(SP2018016);浙江省湖州市公益性应用研究项目(2017GY11);宁波大学王宽诚幸福基金项目;浙江省公益性行业项目(LGN19C200010)资助

摘　　要：基于表面增强拉曼光谱(SERS)拟建立一种适用于水产鱼制品中组胺含量的快速检测方法。采用银纳米颗粒(Ag NPs)作为活性基底和氯化钠溶液作为聚集剂获取组胺的SERS特征峰,并结合线性回归算法对鱼肉中的组胺含量进行测定分析。首先对固体组胺、组胺水溶液以及鱼肉提取液中组胺的SERS特征峰及归属进行表征分析,然后对以Ag NPs的浓缩倍数与氯化钠溶液的浓度作为SERS基底的反应条件进行优化,最后在该优化条件下对鱼肉中组胺进行定量分析。结果表明, Ag NPs在400 nm处有最大吸光度,通过透射电子显微镜观察颗粒的形状主要为球形,均匀尺寸为30 nm左右。利用4-巯基苯甲酸作为探针分子对其进行拉曼测试,所得拉曼峰具有良好的重复性,且拉曼强度很高。因此该活性基底的合成方法不仅用时少、易操作,且合成的Ag NPs可作为可靠的增强基底应用于SERS试验中。此外通过紫外-可见分光光度计检测得出氯化钠溶液使Ag NPs在溶液中发生团聚,形成热点,可实现SERS信号增强。固体组胺的拉曼光谱图反映出1 167 cm^-1 处出现的特征峰主要是由N—H面内弯曲引起的;1 236 cm^-1 处的特征峰主要是咪唑中C—H平面内弯曲和环呼吸引起的;1 291 cm^-1 处主要与环伸展有关;1 474 cm^-1 处的特征峰主要是由咪唑N—H面内弯曲振动和环伸展引起的。优化反应条件在Ag NPs的浓缩倍数为15倍、氯化钠溶液浓度为1 mol·L^-1 时表现出最高的增强效应,并在该优化条件下检测了浓度为5~250 mg·L^-1 的组胺水溶液,得出在该优化条件下检测到组胺水溶液的最低浓度为5 mg·L^-1 。同时在该优化条件下采集了10~100 mg· L^-1 范围的鱼肉提取液中组胺的SERS光谱,并建立组胺溶液的特征拉曼位移峰强度与浓度之间的线性回归模型。得出在1 180, 1 258和1 425 cm^-1 处的特征峰与对应的拉曼峰强度值所建立的标准曲线有良�A rapid method for the detection of histamine in fish products was developed in this study based on Surface-enhanced Raman spectroscopy (SERS).Silver nanoparticles (Ag NPs) were used as the active substrate and sodium chloride solution as an aggregating agent to obtain histamine SERS peaks.The histamine content in fish extracts was determined by linear regression algorithm.In this research,the normal Raman spectra of histamine powder,the SERS characteristic peaks and vibrational assignment of histamine standard solutions as well as histamine in fish extracts were analyzed.The SERS reaction conditions for the concentration of silver nanoparticles and sodium chloride solution were optimized.Lastly,the optimization conditions were used to quantitatively analyze the histamine in fish extracts.The results showed that the silver nanoparticles had the maximum absorbance at 400 nm.The shape of the nanoparticles was mainly spherical and the average size was about 30 nm.Then using 4-MBA as a probe molecule to detect the stability and sensitivity of Ag NPs and obtained Raman peak has good repeatability and high Raman intensity.Therefore,not only is the synthesis method of the active substrate less time-consuming and easy to operate,but the synthesized Ag NPs can be used as a reliable reinforcing substrate in the SERS test.In addition,it was proved by UV-Vis spectrophotometer that the NaCl solution can arouse the agglomeration of Ag NPs to form hot spots,which can enhance SERS signal.The Raman spectrum of solid histamine reflects that the characteristic peak appeared at 1 167 cm^-1 due to the in-plane bending of N-H,the in-plane bending of imidazole in C-H plane and ring breathing (1 236 cm^-1 ),the ring stretching (1 291 cm^-1 ),the in-plane bending vibration of imidazole N-H and ring stretching (1 474 cm^-1 ).Then the optimized reaction conditions showed the highest enhancement effect when the concentration multiple of silver nanoparticle was 15× and the concentration of sodium chloride solution was 1 mol·L^-1 ,and the o

关 键 词：表面增强拉曼光谱 组胺 银纳米颗粒 鱼肉 

分 类 号：TS254.7[轻工技术与工程—水产品加工及贮藏工程]