电化学生物传感器 2008

Facile and controllable preparation of glucose biosensor based on Prussian blue nanoparticles hybrid composites.

Bioelectrochemistry (Amsterdam, Netherlands) Li L, Sheng Q, Zheng J, Zhang H
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组成图示

Facile and controllable preparation o... 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);样品基质:人血清、PBS缓冲液

检测原理

固定化葡萄糖氧化酶(GOD)催化葡萄糖氧化,生成过氧化氢(H2O2)。PVP保护普鲁士蓝纳米粒子(PBNPs)作为人工过氧化物酶,在0.0 V低电位下电催化H2O2还原为水,电子经聚苯胺(PANI)/多壁碳纳米管(MWNTs)导电网络传递至玻碳电极(GCE),产生与葡萄糖浓度成正比的安培电流。PANI/MWNTs提高导电性和电子传递速率,PBNPs提供高催化活性与纳米高比表面,壳聚糖(CS)交联网络固定GOD并维持其活性。低电位工作有效抑制抗坏血酸、尿酸、L-半胱氨酸和对乙酰氨基酚等干扰。稳态电流随葡萄糖浓度升高而增大,实现快速灵敏检测。

检测灵敏度

LOD: 6×10−7 M (S/N=3);线性范围: 6.7×10−6–1.9×10−3 M;灵敏度: 6.28 μA mM−1;r = 0.9998

效应效果

传感器抗干扰能力良好:0.2 mM尿酸、L-半胱氨酸和对乙酰氨基酚对1 mM葡萄糖无可见干扰;按实际比例加入0.3 mM抗坏血酸与20 mM葡萄糖时,电流仅降低6.3%。重现性方面,5个传感器对0.5 mM葡萄糖的RSD为5.1%;连续5次测定20 mM葡萄糖相对偏差小于1.1%,RSD为3.2%;储存3周后仍保留约90%响应。人血清样品与血糖仪结果一致,加标回收率为95.0%–104.5%。表观Michaelis–Menten常数1.9 mM,低于溶液GOD的33 mM,显示高亲和性。作者认为该方法简便可控,适用于葡萄糖及其他氧化酶传感器。

传感器的构成

  • 基底/工作电极:玻碳电极(GCE),抛光后作为导电基底和换能器。
  • 纳米材料修饰层:多壁碳纳米管(MWNTs),超声分散涂覆,提高导电性、比表面积和电子传递。
  • 导电聚合物修饰层:聚苯胺(PANI),在MWNTs/GCE上电聚合,形成导电网络并负载PBNPs。
  • 纳米催化层:PVP保护普鲁士蓝纳米粒子(PBNPs),电沉积于PANI/MWNTs/GCE,作为H2O2电催化剂。
  • 识别/固定化层:葡萄糖氧化酶(GOD)包埋于壳聚糖(CS)网络,催化葡萄糖氧化并固定酶。
  • 交联剂:戊二醛(glutaraldehyde),交联CS网络,稳定GOD微环境。

中文摘要

本文报道了一种基于聚乙烯吡咯烷酮(PVP)保护普鲁士蓝纳米粒子(PBNPs)–聚苯胺(PANI)/多壁碳纳米管(MWNTs)杂化复合物的葡萄糖生物传感器,采用电化学方法简便可控地制备。利用PVP辅助的PBNPs电沉积新路线,扫描电镜表明电极表面普鲁士蓝呈纳米尺度分布。该传感器对葡萄糖具有快速电流响应(<6 s),在6.7×10−6–1.9×10−3 M范围内呈良好线性,灵敏度为6.28 μA mM−1,检出限为6×10−7 M(S/N=3)。酶催化反应表观活化能为23.9 kJ/mol,表观Michaelis–Menten常数为1.9 mM,表明固定化葡萄糖氧化酶与底物具有较高亲和性。该构建方法兼具杂化复合物的导电、催化与生物相容优势,可实现葡萄糖的快速灵敏检测。人血清样品测定回收率为95.0%–104.5%,显示其实际应用潜力。

英文摘要

A glucose biosensor based on polyvinylpyrrolidone (PVP) protected Prussian blue nanoparticles (PBNPs)-polyaniline/multi-walled carbon nanotubes hybrid composites was fabricated by electrochemical method. A novel route for PBNPs preparation was applied in the fabrication with the help of PVP, and from scanning electron microscope images, Prussian blue particles on the electrode were found nanoscaled. The biosensor exhibits fast current response (<6 s) and a linearity in the range from 6.7x10(-6) to 1.9x10(-3) M with a high sensitivity of 6.28 microA mM(-1) and a detection limit of 6x10(-7) M (S/N=3) for the detection of glucose. The apparent activation energy of enzyme-catalyzed reaction and the apparent Michaelis-Menten constant are 23.9 kJ mol(-1) and 1.9 mM respectively, which suggests a high affinity of the enzyme-substrate. This easy and controllable construction method of glucose biosensor combines the characteristics of the components of the hybrid composites, which favors the fast and sensitive detection of glucose with improved analytical capabilities. In addition, the biosensor was examined in human serum samples for glucose determination with a recovery between 95.0 and 104.5%.

关键词

普鲁士蓝纳米粒子葡萄糖生物传感器聚苯胺多壁碳纳米管葡萄糖氧化酶壳聚糖