摘要
目的:构建了一种核酸适配体传感器,用于食品中组胺的快速检测。方法:通过G-四链体和适配体5’端修饰的巯基和纳米金共价结合制备成DNA-金纳米颗粒(Gold nanoparticles,AuNPs)复合物。将组胺静电吸附在电极的表面,孵育纳米复合物,复合物中的适配体与组胺进行特异性结合。纳米复合物上的G-四链体序列与底物中的hemin结合形成G-四链体结构,能够催化底物中的氢醌转变为苯醌,在一定的电压下,苯醌被重新还原,产生可以被检测的电化学信号,实现组胺的定量检测。结果:对新构建的组胺传感器性能研究发现,在组胺浓度为1×10^(–6)~1×10^(–4)μg/mL时,线性相关系数为0.9922,检测限为0.5μg/mL。对该传感器特异性分析发现,干扰物(酪胺、L-半胱氨酸及组氨酸等)的响应信号远低于组胺的响应信号。稳定性分析显示,传感器在储存30 d后,依旧保持其不低于初始响应信号的95.1%左右。实际样品检测中,样品回收率为91.26%~107.6%,与高效液相色谱检测相比,结果无差异性。结论:新构建的核酸适配体传感器是一种无酶电化学传感器,可以选择性地捕获和识别组胺,具有良好的特异性和稳定性,可以有效地降低实验成本和操作难度。
Objective:To construct a nucleic acid aptamer sensor for detection rapid of histamine in food.Methods:DNA-gold nanoparticles(AuNPs)nanocomposites were prepared by covalent binding of thiol groups and gold nanoparticles modified at the 5'end of G-quadruplex and adapter.Electrostatically adsorb histamine onto the surface of the electrode,incubate the nanocomposite,and the adapter in the composite binds specifically to histamine.The G-quadruplex sequence on nanocomposites combined with hemin in the substrate to form a G-quadruplex structure,which could catalyze the conversion of hydroquinone in the substrate to benzoquinone,under a certain voltage,benzoquinone was then reduced to produce detectable electrochemical signals,achieving quantitative detection of histamine.Results:Research on the performance of the newly constructed histamine sensor has found that at a histamine concentration of 1×10^(–6)–1×10^(–4)μg/mL,the linear correlation coefficient of 0.9922 and the detection limit of 0.5μg/mL.Specific analysis of the sensor revealed that the response signals of interferents(such as tyramine,L-cysteine,and histidine)were much lower than those of histamine;Stability analysis showed that after 30 days of storage,the sensor still maintained a response signal of no less than 95.1%of the initial response signal.In actual sample detection,the sample recovery rate ranged from 91.26%to 107.60%,and compared with high performance liquid chromatography detection,showing no difference in the results.Conclusion:The newly constructed nucleic acid aptamer sensor is an enzyme electrochemical sensor that can selectively capture and recognize histamine,has good specificity and stability,and can effectively reduce experimental costs and operational difficulties.
作者
张永芳
郭楠楠
ZHANG Yongfang;GUO Nannan(Zhengzhou Key Laboratory for Fast Food Safety Testing,College of Food Science and Engineering,Zhengzhou University of Science and Technology,Zhengzhou 450000,China)
出处
《食品科技》
CAS
北大核心
2024年第4期345-352,共8页
Food Science and Technology
关键词
核酸适配体
纳米复合物
电化学传感器
无酶
组胺检测
nucleic acid aptamers
nanocomposites
electrochemical sensors
enzyme-free
histamine detection
