电化学生物传感器 2012

Development of a sandwich format, amperometric screen-printed uric acid biosensor for urine analysis.

Analytical biochemistry Kanyong P, Pemberton RM, Jackson SK, Hart JP
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组成图示

Development of a sandwich format, amp... 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

尿酸(uric acid, UA);样品基质:尿液(urine),校准使用磷酸盐缓冲液

检测原理

尿酸(UA)透过外层聚碳酸酯(PC)膜进入尿酸酶(UOX)层,被 UOX 催化氧化:UA + O2 + H2O → 尿囊素 + H2O2 + CO2。生成的 H2O2 透过内层醋酸纤维素(CA)膜到达掺钴酞菁(CoPC)的丝网印刷碳电极(CoPC–SPCE)。在 +0.5 V 下,H2O2 将 CoPC 的 Co2+ 还原为 Co+,Co+ 随即在电极表面再氧化为 Co2+ 并释放电子,形成与 UA 浓度相关的安培电流。CA 膜阻止 UA 直接氧化,PC 膜阻挡抗坏血酸、对乙酰氨基酚等干扰物,从而提高选择性。该设计以酶催化和 CoPC 电催化 H2O2 为核心,无需额外标记或信号放大。

检测灵敏度

LOD: 0.015 mM;线性范围: 0.015–0.25 mM;灵敏度: 2.10 μA/mM;标准加入法 R^2 = 0.9989

效应效果

该传感器对 2 mM 抗坏血酸(AA)和对乙酰氨基酚(PA)均无响应,CA 膜可消除 UA 直接氧化干扰。4 ℃干燥器保存 7 天后,0.2 mM UA 标准响应无下降。未加标尿液(n=6)CV 为 5.82%,加标 0.225 mM UA 后 CV 为 4.21%,平均回收率 97.3%;校准曲线 CV 为 4.3%。与部分安培尿酸传感器相比,其线性范围 0.015–0.25 mM 优于 0.0025–0.085 mM,检出限 0.015 mM 优于 0.07 mM。作者认为其可低成本丝网印刷、批量生产,适合尿液尿酸临床监测和家用检测。

传感器的构成

  • 换能器电极:CoPC–SPCE(掺钴酞菁 CoPC 的水性碳墨丝网印刷碳电极),电催化 H2O2 并输出安培信号
  • 内层选择性膜:CA(1.5% 醋酸纤维素,丙酮溶液滴涂),阻止 UA 直接接触电极,允许 H2O2 透过
  • 识别元件:UOX(尿酸酶,0.6 U 滴涂),催化 UA 生成 H2O2
  • 外层选择性膜:PC(0.45 μm 聚碳酸酯膜),覆盖并固定酶层,阻挡干扰物
  • 参比电极:Ag/AgCl(丝网印刷参比电极),提供稳定电位参考
  • 对电极:Pt 线(铂丝对电极),完成三电极电化学回路

中文摘要

本文报道了一种用于尿液分析的三明治式安培丝网印刷尿酸生物传感器。以掺入钴酞菁(CoPC)的水性碳墨丝网印刷碳电极(CoPC–SPCE)作为基础换能器,依次在其表面滴涂1.5%醋酸纤维素(CA)膜、尿酸酶(UOX)层,并覆盖聚碳酸酯(PC)膜,构成PC–UOX–CA–CoPC–SPCE。采用计时安培法优化尿液分析条件:温度35 ℃、pH 9.2缓冲液、离子强度50 mM、尿酸酶用量0.6 U、孵育时间180 s。该传感器用于健康人尿液检测,未加标尿液(n=6)精密度为5.82%;尿液加标0.225 mM尿酸后,精密度和回收率分别为4.21%和97.3%。传感器线性工作范围为0.015–0.25 mM,其中0.015 mM为检出限,灵敏度为2.10 μA/mM。

英文摘要

A screen-printed carbon electrode (SPCE) incorporating the electrocatalyst cobalt phthalocyanine (CoPC), fabricated using a water-based ink formulation, has been investigated as the base transducer for a uric acid biosensor. A sandwich biosensor was fabricated by first depositing cellulose acetate (CA) onto this transducer (CoPC-SPCE), followed by uricase (UOX) and finally a polycarbonate (PC) membrane; this device is designated PC-UOX-CA-CoPC-SPCE. This biosensor was used in conjunction with chronoamperometry to optimize the conditions for the analysis of urine: temperature, 35°C; buffer, pH 9.2; ionic strength, 50 mM; uricase, 0.6 U; incubation time, 180 s. The proposed biosensor was applied to urine from a healthy subject. The precision determined on unspiked urine (n=6) was 5.82%. Urine was fortified with 0.225 mM UA, and the resulting precision and recovery were 4.21 and 97.3%, respectively. The linear working range of the biosensor was found to be 0.015 to 0.25 mM (the former represents the detection limit), and the sensitivity was calculated to be 2.10 μA/mM.