电化学生物传感器 2012

Amperometric biosensor based on 3D ordered freestanding porous Pt nanowire array electrode.

Nanoscale Wang Y, Zhu Y, Chen J, Zeng Y
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组成图示

Amperometric biosensor based on 3D or... 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)、葡萄糖(glucose, Glc);实验基质为0.1 M PBS(pH 7.0,含0.1 M KCl)搅拌溶液,作者提出可应用于血液葡萄糖监测但未测试真实血清。

检测原理

该传感器以固定于PPNWAE表面的葡萄糖氧化酶(GOD)为识别元件。葡萄糖扩散至电极表面并被GOD催化氧化为葡萄糖酸,同时GOD由氧化态还原为还原态;还原态GOD被溶液中溶解氧重新氧化,按化学计量生成H2O2。生成的H2O2扩散进入三维有序多孔铂纳米线阵列,在+0.55 V下于Pt活性位点发生阳极氧化,产生法拉第电流。因多孔纳米线大幅提高有效电化学面积、缩短传质路径并促进电子转移,电流随H2O2及葡萄糖浓度升高而增大。直接检测H2O2时,H2O2同样在PPNWAE上电催化氧化,信号正比于其浓度。

检测灵敏度

H2O2:LOD: 3.6 mM;线性方程: I = 8.43324 + 364.2805CH2O2(CH2O2 = 4.5 mM–0.43 mM,R = 0.9998);I = 0.00152 + 0.25638CH2O2 − 0.0038C^2_H2O2(CH2O2 = 0.43–27.1 mM,R = 0.9985);灵敏度: 0.36 mA cm−2 mM−1(低浓段)、约0.26 mA cm−2 mM−1(高浓段);检测范围: 4.5 mM–27.1 mM。葡萄糖:LOD: 4.5 mM;校准方程: I = 0.61763 − 0.40723exp(−CGlc/204.97586) − 0.2055exp(−CGlc/11.73513),R = 0.9999;4.5–27.8 mM 灵敏度约8.74 mA cm−2 mM−1;总检测范围: 4.5 mM–189.5 mM;km: 14.6 mM。

效应效果

PPNWAE/GOD对葡萄糖响应快,稳态电流4–10 s达到;选择性方面,在5.6 mM葡萄糖存在下加入0.1 mM抗坏血酸(AA)和0.02 mM尿酸(UA),AA与UA的电流响应分别约为葡萄糖的24%和1%。与PNWAE/GOD相比,PPNWAE/GOD在高浓度葡萄糖下响应电流提高约100%,PPNWAE对H2O2响应提高约50%;同时具有更低检出限、更高灵敏度和更小米氏常数(14.6 mM对17.6 mM)。葡萄糖检测范围达4.5–189.5 mM,覆盖正常与糖尿病血糖水平,作者认为可用于医学和食品领域。未报告长期稳定性、RSD和真实样品回收率。

传感器的构成

  • 基底/换能器电极:铂圆盘电极(Pt disk electrode, PE),作为导电基底并与纳米线阵列直接生长集成,保证导电性与稳定性。
  • 模板/形貌控制层:多孔道蚀刻聚碳酸酯膜(PC membrane,孔径400 nm),限定Pt–Cu合金纳米线沉积,随后用CH2Cl2溶解去除。
  • 纳米线修饰层:三维有序自由多孔铂纳米线阵列(PPNWAE),由Pt–Cu合金电沉积并经25% HNO3脱除Cu形成,提供高活性面积和多孔粗糙表面。
  • 对照修饰层:铂纳米线阵列(PNWAE),仅电沉积Pt形成无孔纳米线,用于比较孔隙结构的贡献。
  • 识别元件:葡萄糖氧化酶(glucose oxidase, GOD),催化葡萄糖生成H2O2,是葡萄糖识别与转化的生物元件。
  • 交联固定剂:戊二醛(glutaraldehyde, GA,5 wt%),与GOD混合后在4 ℃交联12 h,将GOD固定于纳米线表面。
  • 电化学测量体系:Ag/AgCl参比电极、螺旋Pt丝对电极和0.1 M PBS(pH 7.0,含0.1 M KCl)支持电解质,用于三电极安培检测。

中文摘要

本文通过金属电沉积并将纳米线直接集成于铂圆盘电极,简便地制备了具有数纳米孔径的三维(3D)有序自由多孔铂(Pt)纳米线阵列电极(PPNWAE)以及无孔铂纳米线阵列电极(PNWAE)。PPNWAE具有高活性面积,对过氧化氢(H2O2)表现出优异灵敏度(0.36 mA cm−2 mM−1)和较宽检测范围(4.5 mM–27.1 mM)。进一步构建了基于葡萄糖氧化酶(GOD)的PPNWAE/GOD生物传感器,对葡萄糖具有显著较宽的检测范围(4.5 mM–189.5 mM)。与基于PNWAE的生物传感器相比,PPNWAE基生物传感器表现出更低的检出限、更高的灵敏度和更小的米氏–门滕常数km。特别是,由于颗粒状且更粗糙的多孔纳米线表面有利于提高GOD生物活性,PPNWAE/GOD对葡萄糖的响应电流比PNWAE/GOD高约100%,PPNWAE对H2O2的响应电流比PNWAE高约50%。还评估了PPNWAE/GOD葡萄糖生物传感器的选择性。

英文摘要

A three-dimensionally (3D) ordered freestanding porous platinum (Pt) nanowire array electrode (PPNWAE) with pores of several nanometers in size and a Pt nanowire array electrode (PNWAE) without pores were facilely fabricated by metal electrodeposition and direct integration with a Pt disk electrode. The unusual PPNWAE with high active area showed excellent sensitivity (0.36 mA cm(-2) mM(-1)) and a wide detection range (4.5 μM-27.1 mM) to hydrogen peroxide (H(2)O(2)). A glucose oxidase (GOD)-based biosensor (PPNWAE/GOD) with a considerably wide detection range (4.5 μM-189.5 mM) to glucose was demonstrated. Furthermore, a lower detection limit, higher sensitivity and smaller value of Michaelis-Menten constant k(m) were recorded for PPNWAE-based biosensors compared with PNWAE-based biosensors. Particularly, the response current to glucose of PPNWAE/GOD was ca. 100% higher than that of PNWAE/GOD and the response current to H(2)O(2) of PPNWAE was ca. 50% higher than that of PNWAE, owing to the granular and rougher porous nanowire surface enabling greater bioactivity for GOD. The selectivity of PPNWAE/GOD glucose biosensor was also estimated.