电化学生物传感器 2010

Synergistic contributions of fullerene, ferrocene, chitosan and ionic liquid towards improved performance for a glucose sensor.

Biosensors & bioelectronics Zhilei W, Zaijun L, Xiulan S, Yinjun F, Junkang L
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组成图示

Synergistic contributions of fulleren... 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

葡萄糖在GOx催化下氧化为葡萄糖酸内酯,GOx辅基由氧化态GOx(ox)还原为GOx(red)。固定化GOx(red)通过C60/Fc复合介体将电子传递至电极:Fc+/Fc可逆氧化还原对与C60电子受体协同加速电子中继,CS-IL网络维持酶活性并提供导电微环境。在+100 mV低电位下,Fc在电极表面再氧化产生安培催化电流,电流随葡萄糖浓度增加而增大。低电位避免抗坏血酸、尿酸等电活性物质氧化,提高选择性。C60/Fc介体与CS-IL网络构成信号放大与稳定机制,使响应快速且灵敏。

检测灵敏度

LOD: 3 × 10−9 M (S/N = 3);线性范围: 1 × 10−8 M–1 × 10−5 M;灵敏度: 234.67 nA nM−1 cm−2;R^2 = 0.9998

效应效果

传感器在+100 mV低电位下对100倍抗坏血酸和尿酸无显著干扰,选择性良好。对2.0 μM葡萄糖重复10次测定RSD为1.3%,精密度高。室温空气保存30周后响应保持初始值的95%,长期稳定性优异。表观KM为0.03 mM,低于游离GOx(27 mM)及多种报道传感器,表明酶亲和力和活性高。用于5份人血清样品检测,结果与医院己糖激酶法一致,F和t值均低于95%置信限(F=6.39,t=2.78),无显著差异。作者认为该传感器制备简单、成本低、响应快,适用于血清等实际样品中葡萄糖检测。

传感器的构成

  • 基底电极:玻璃碳电极(GCE),经氧化铝抛光、酸/乙醇超声和循环伏安活化,提供导电基底
  • 富勒烯修饰层:C60(99.5%,二氯甲烷溶液涂覆),作为电子受体/介体,促进电子转移
  • 二茂铁-离子液体层:Fc(乙醇饱和溶液)与IL([DBIM][TF2N])混合涂覆,Fc作为可逆氧化还原介体,IL增强导电与成膜
  • 壳聚糖-离子液体网络层:CS(1%乙酸溶液)与IL超声分散混合涂覆,形成生物相容网络,固定酶并提供微环境
  • 识别元件:葡萄糖氧化酶GOx(黑曲霉来源,136300 U/g,PBS溶液滴加),催化葡萄糖氧化
  • 信号读出:三电极体系(Ag/AgCl参比电极、铂丝对电极、电化学工作站),在+100 mV进行计时电流/伏安读出

中文摘要

本文报道了一种利用富勒烯C60、二茂铁Fc、壳聚糖CS和离子液体IL对葡萄糖氧化酶GOx的协同有利特性构建的葡萄糖传感器GOx/C60-Fc-CS-IL。采用循环伏安法、阻抗谱和计时电流法评价性能。由于GOx与电极间实现高效电子转移,传感器表现出高灵敏度234.67 nA nM−1 cm−2、低检出限3×10−9 M(S/N=3)、快速响应时间0.752 s、宽校准范围1×10−8 M至1×10−5 M以及优异长期稳定性(30周)。复合介质中GOx表观米氏常数KM为0.03 mM,表明固定化酶对葡萄糖氧化具有高生物电催化活性。由于工作电位低(100 mV),传感器对人血中常见电活性干扰物如抗坏血酸和尿酸相对不敏感。优异的电化学可逆性、高灵敏度与稳定性、制备简单且可在较短时间内完成是该类葡萄糖生物传感器的主要优势。

英文摘要

The paper describes an ingenious approach for the fabrication of a promising glucose sensor, GOx/C(60)-Fc-CS-IL, that exploits the synergistic beneficial characteristics of fullerene (C(60)), ferrocene (Fc), chitosan (CS) and ionic liquid (IL) for glucose oxidase (GOx). Cyclic voltammetry, impedance spectroscopy and chronoamperometry were used to evaluate performance of the biosensor, respectively. Since efficient electron transfer between GOx and the electrode was achieved, the biosensor exhibits a high sensitivity (234.67 nA nM(-1) cm(-2)), low detection limit (S/N=3) (3x10(-9) M), fast response time (0.752 s), wide calibration range (from 1x10(-8) M to 1x10(-5) M) and excellent long-term stability (30 weeks). The apparent Michaelis-Menten constant (K(M)) of GOx on the composite medium, 0.03 mM, shows high bioelectrocatalytic activity of immobilized enzyme toward glucose oxidation. Due to low operating potential (100 V), the biosensor is relatively insensitive to electroactive interfering species in human blood such as ascorbic acid, and uric acid, which are commonly found in blood samples. Excellent electrochemical reversibility, high sensitivity and stability, technically simple and possibility of preparation at short period of time are of great advantages of these glucose biosensors.

关键词

葡萄糖生物传感器电化学生物传感器富勒烯二茂铁壳聚糖离子液体