电化学生物传感器 2011

Preparation and characterization of bismuth oxide nanoparticles-multiwalled carbon nanotube composite for the development of horseradish peroxidase based H₂O₂ biosensor.

Talanta Periasamy AP, Yang S, Chen SM
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组成图示

Preparation and characterization of b... 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

过氧化氢(Hydrogen peroxide, H2O2);样品基质:0.05 M PBS(pH 7)缓冲溶液

检测原理

该传感器基于 HRP 的直接电子转移与酶催化还原 H2O2 的电化学机制。Bi2O3 纳米颗粒具有多孔结构和大比表面积,MWCNT 提供导电网络,二者复合后修饰于 GCE,使 HRP 血红素 Fe(III/II) 中心与电极之间建立快速电子通道,在 −0.326 V 附近出现准可逆氧化还原峰。当 H2O2 扩散至电极表面时,被 HRP 催化还原,电子经 Bi2O3–MWCNT 传递至 GCE,在 −0.3 V 下产生阴极电流。H2O2 浓度升高时,酶催化还原速率增加,安培电流随之线性增大。Nafion 外层固定 HRP 并阻挡干扰物,使传感器在低过电位下获得高灵敏响应。

检测灵敏度

线性范围: 8.34–28.88 mM;R^2 = 0.9743;灵敏度: 26.54 μA mM−1 cm−2;CV线性范围: 19.2–222.1 μM;R^2 = 0.9845;CV灵敏度: 1.62 μA mM−1 cm−2

效应效果

该传感器在 −0.3 V 下对 H2O2 响应时间为 5 s,安培线性范围为 8.34–28.88 mM,灵敏度 26.54 μA mM−1 cm−2,R2=0.9743;CV 法线性范围为 19.2–222.1 μM,灵敏度 1.62 μA mM−1 cm−2,R2=0.9845。与裸 GCE 相比,H2O2 还原过电位降低 370 mV;与 MWCNT 和 Bi2O3–MWCNT 相比分别降低 189 和 176 mV。加入 100 μM 抗坏血酸、尿酸和多巴胺均无明显干扰。连续 5000 s 安培响应稳定;4 ℃ PBS 中储存一周后仍保留 94.4% 响应电流。重复性和重现性 RSD 分别为 2.8% 和 2.82%。作者认为该平台酶负载高、电子转移快、稳定性好,适合低成本 H2O2 检测。

传感器的构成

  • 基底/换能器电极:玻碳电极(GCE),提供电子传导与安培信号换能。
  • 纳米材料修饰层:氧化铋纳米颗粒–多壁碳纳米管(Bi2O3–MWCNT)复合膜,滴涂于 GCE,提供大比表面积、多孔结构和电子传导通道,促进 HRP 直接电子转移。
  • 识别元件:辣根过氧化物酶(HRP),固定于 Bi2O3–MWCNT 上,催化 H2O2 还原。
  • 封闭/固定层:1% Nafion(NF)薄层,作为结合剂与保护层,通过静电作用锚定 HRP 并降低干扰物影响。

中文摘要

本文报道了氧化铋(Bi2O3)纳米颗粒与多壁碳纳米管(MWCNTs)纳米复合材料的制备与表征。XRD 表明 Bi2O3 为结晶态 β 相,具单斜对称;FESEM 显示粒径约 50 nm;EDX 证实 Bi 与 C 共存,SEM 和 AFM 表征了复合膜形貌。在 Bi2O3–MWCNT 修饰的玻碳电极(GCE)上,辣根过氧化物酶(HRP)可实现直接电子转移,并用 1% Nafion(NF)薄层作为结合剂固定 HRP。NF/HRP/Bi2O3–MWCNT/GCE 在 0.05 M PBS(pH 7)中呈现准可逆氧化还原峰,形式电位为 −0.326 V(vs. Ag/AgCl)。该膜显著降低 H2O2 还原过电位,对 H2O2 的安培响应时间为 5 s,线性范围 8.34–28.88 mM,灵敏度 26.54 μA mM−1 cm−2,且具有良好的操作稳定性和高选择性。

英文摘要

In this work, preparation and characterization of a novel nanocomposite containing bismuth oxide (Bi(2)O(3)) nanoparticles and multiwalled carbon nanotubes (MWCNTs) was presented. Powder X-ray diffraction (XRD) studies revealed that as-synthesized Bi(2)O(3) nanoparticles are crystalline and belong to α-phase with monoclinic symmetry. Field emission scanning electron microscopy (FESEM) study results showed that the size of Bi(2)O(3) nanoparticles is 50 nm. Energy-dispersive X-ray (EDX) spectra of as-prepared Bi(2)O(3)-MWCNT nanocomposite displayed characteristic Bi and C peaks which confirmed the incorporation of Bi(2)O(3) with MWCNT. The prepared Bi(2)O(3)-MWCNT was also characterized by scanning electron microscopy (SEM) and atomic force microscopy (AFM) studies. The direct electron transfer of horseradish peroxidase (HRP) has been revealed at Bi(2)O(3)-MWCNT modified glassy carbon electrode (GCE). In order to firmly anchor the HRP molecules onto Bi(2)O(3)-MWCNT matrix, a thin layer of 1% nafion (NF) solution was coated as a binder. The fabricated NF/HRP/Bi(2)O(3)-MWCNT/GCE exhibits well defined quasi-reversible redox peaks at a formal potential (E(0)') of -0.326V vs. Ag/AgCl reference electrode in 0.05M phosphate buffer solution (PBS), pH 7. NF/HRP/Bi(2)O(3)-MWCNT film remarkably lowers the over potential for H(2)O(2) reduction than MWCNT, Bi(2)O(3)-MWCNT and unmodified GCEs. The proposed composite film exhibits quick amperometric i-t response (5s) towards H(2)O(2) in the linear range of 8.34-28.88mM with a sensitivity of 26.54 μA μM(-1)cm(-2). The developed NF/HRP/Bi(2)O(3)-MWCNT biosensor has a good operational stability and high selectivity towards H(2)O(2).