基于适配体吸附金纳米颗粒比色传感检测组胺  

Colorimetric Sensor for Sensitive Detection of Histamine Based on Aptamer-Adsorbed Gold Nanoparticles

作　　者：罗倩 鲁迨 黄晨涛 石星波 LUO Qian;LU Dai;HUANG Chentao;SHI Xingbo(Laboratory of Micro&Nano Biosensing Technology in Food Safety,Hunan Provincial Key Laboratory of Food Science and Biotechnology,College of Food Science and Technology,Hunan Agricultural University,Changsha 410128,China)

机构地区：[1]湖南农业大学食品科学技术学院,食品科学与生物技术湖南省重点实验室,食品安全微纳生物传感技术课题组,湖南长沙410128

出　　处：《食品科学》2022年第8期303-309,共7页Food Science

基　　金：国家自然科学基金面上项目(31972155);湖南省自然科学基金项目(2019JJ40115;2020RC3050);中国博士后基金面上项目(2018M642980);长沙市杰出创新青年基金项目(kq1905017;kq2009058;kq2106048);湖南省重点研发项目(2021NK2021)。

摘　　要：建立一种基于核酸适配体吸附金纳米颗粒(gold nanoparticles,AuNPs)比色传感方法特异性检测组胺的方法。利用组胺的双重作用,1)能与其适配体特异性识别,暴露AuNPs表面;2)多余组胺中的咪唑环结构能取代AuNPs表面的柠檬酸根离子,破坏AuNPs间的相互静电作用,导致聚集现象,进而引起从红到蓝的颜色变化,从而实现组胺的定量检测。利用紫外-可见分光度计考察AuNPs的吸光度变化,得出比色方法的检测限为8.89 nmol/L,线性范围为50 nmol/L~1.2μmol/L(R^(2)=0.999)。同时,利用手机成像样品,并利用Image J软件分析各样品的红(R)、绿(G)、蓝(B)各通道的值,选用G/R作为检测信号,得出RGB方法的检测限为24.91 nmol/L,线性范围为150 nmol/L~1.0μmol/L(R^(2)=0.997)。此外,该方法对组胺具有很好的选择性,两种信号输出方式在水样中的加标回收率分别为91.82%~102.27%、98.96%~102.89%;在鱼肉样品中的加标回收率分别为89.77%~108.92%、88.96%~109.82%。本方法简单、快速、便携,尤其RGB方法无需借助精密仪器,即能实现现场即时分析样品的需求,以期该方法能推广于高灵敏监测动物源性食品的新鲜度。A new aptamer-adsorbed gold nanoparticles-based colorimetric method was developed for sensitive detection of histamine,by taking advantage of the facts:1)histamine can specifically recognize histamine aptamer,leading to exposure of the surface of gold nanoparticles(AuNPs);2)the imidazole structure of excess histamine can replace the citrate ions on the surface of AuNPs,and destroy the electrostatic interaction between the AuNPs,thereby leading to an aggregation phenomenon that causes a color change from red to blue.Based on the changes in the absorbance of AuNPs solutions,the limit of detection(LOD)was calculated to be 8.89 nmol/L,and the linear range was 50 nmol/L–1.2μmol/L(R^(2)=0.999).Meanwhile,a smart phone was used to take photos of the sample solutions,and the values of red(R),green(G)and blue(B)channels were obtained by the Image J software.Using G/R ratio as the detection signal,the LOD of the RGB method was 24.91 nmol/L,and good linearity was observed in the range from 150 nmol/L to 1.0μmol/L(R^(2)=0.997).In addition,this method has good selectivity to histamine.The recoveries of the spectrophotometric and RGB methods were 91.82%–102.27%and 98.96%–102.89%for spiked water samples and 89.77%–108.92%and 88.96%–109.82%for spiked fish samples,respectively.Both methods are simple,rapid and sensitive.Particularly,the RGB method,which can meet the requirements of on-site real-time detection without the need for any sophisticated instrumentation,is expected to be applied for sensitive monitoring of the freshness of animal-derived foods.

关 键 词：适配体 金纳米颗粒 聚集 组胺 比色传感器 RGB 

分 类 号：TS207.3[轻工技术与工程—食品科学]