电化学生物传感器 2009

Fabrication of an amperometric bienzyme biosensing system with neutral red functionalized carbon nanotubes.

The Analyst Jeykumari DR, Narayanan SS
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组成图示

Fabrication of an amperometric bienzy... 传感器构成示意图

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传感器类型

电化学生物传感器

检测对象

葡萄糖(glucose, D-glucose);样品基质:人血/人血清

检测原理

葡萄糖在GOx催化下与O2反应生成葡萄糖酸和H2O2;HRP以NR为介体催化H2O2还原,将NR还原态氧化为氧化态;氧化态NR在GCE表面接受电子还原为还原态,产生安培还原电流。NR共价连接于MWNTs,MWNTs提供电子传导通道,Nf促进电子转移并降低干扰。工作电位-0.35 V,低电位下NR氧化还原可逆,葡萄糖浓度升高使H2O2生成增加,NR氧化还原循环加快,稳态电流增大。无酶催化放大,但双酶级联和纳米导电介体提高灵敏度。

检测灵敏度

LOD: 4 nM (S/N = 3);线性范围: 12 nM–16 mM;线性回归方程: I(A) = 0.52[glucose] + 4 (A;相关系数: 0.9999

效应效果

传感器在-0.35 V下对0.1 mM葡萄糖的95%稳态响应时间为2 s。六个独立制备传感器对0.1 mM葡萄糖的RSD为5.1%,10次重复进样RSD为4.5%。4°C PBS或空气中保存60 d保留98%初始活性,之后下降3%;摘要称4°C下5个月储存稳定。对0.5 mM抗坏血酸、尿酸、尿素、甘氨酸、对乙酰氨基酚的干扰相对于1.0 mM葡萄糖可忽略。人血/血清样品与医院比色法结果一致:5个样品测定值分别为6.97±0.09、7.53±0.04、6.43±0.08、5.50±0.09、7.31±0.06 mM,对应医院值6.98、7.51、6.47、5.49、7.32 mM。作者认为其适合快速、敏感、稳定、抗干扰的葡萄糖分析。

传感器的构成

  • 基底/工作电极:玻璃碳电极(GCE),经砂纸和氧化铝抛光清洗,作为电子换能器。
  • 纳米材料修饰层:多壁碳纳米管(MWNTs),经硝酸氧化引入羧基,提供导电通道和比表面积。
  • 介体功能化层:中性红(NR),通过EDC/sulfo-NHS与MWNTs羧基共价连接,作为电子转移介体并促进HRP再生。
  • 识别/催化元件:葡萄糖氧化酶(GOx)和辣根过氧化物酶(HRP),分别催化葡萄糖氧化生成H2O2和催化H2O2还原。
  • 分散/固定基质:Nafion(Nf,5 wt%),分散NR/MWNTs并固定GOx/HRP,形成纳米生物复合膜,同时提供抗干扰选择性。

中文摘要

新型纳米材料在生物传感器应用中是纳米技术快速发展的领域。本文探索了功能化多壁碳纳米管(MWNTs)作为安培法测定葡萄糖的平台,采用葡萄糖氧化酶(GOx)和辣根过氧化物酶(HRP),并用中性红(NR)功能化MWNTs固定。描述了用于低浓度葡萄糖测定的葡萄糖生物传感器的制备与分析表征。由于碳纳米管的电催化效应,可在低电位下以可接受的灵敏度测量酶反应产生的法拉第响应。优化了影响传感器灵敏度的实验参数,如施加电位、pH、温度等,并考察了潜在干扰。葡萄糖生物传感器响应时间很快(2秒内),在4°C下具有5个月的储存稳定性。所提出的生物传感器具有响应时间短、灵敏度高、操作简便和传感器组装简单等特点。该生物传感器成功应用于人血样品中葡萄糖的测定,并获得可接受的结果。

英文摘要

Novel nanomaterials for biosensor applications represent a rapidly progressing field of nanotechnology. An exploration of functionalized multi-walled carbon nanotubes (MWNTs) as a platform for amperometric determination of glucose employing the enzymes glucose oxidase (GOx) and horseradish peroxidase (HRP), which were immobilized with neutral red (NR) functionalized MWNTs is presented. The fabrication and analytical characterization of the glucose biosensor for the measurement of low concentration of glucose is described. Owing to the electrocatalytic effect of carbon nanotubes, the measurement of faradic responses resulting from enzymatic reactions has been realized at low potential with acceptable sensitivity. Experimental parameters affecting the sensitivity of biosensors, e.g., applied potential, pH, temperature etc., were optimized and potential interferences were examined. The response time for the glucose biosensor was very fast (within 2 seconds) and it showed good storage stability at 4 degrees C over a 5-month period. The proposed biosensor exhibited short response time, high sensitivity, easy operation, and simple sensor assembly. The biosensor was successfully applied to the determination of glucose in human blood samples and acceptable results were obtained.

关键词

电化学生物传感器葡萄糖多壁碳纳米管中性红双酶安培法