Numnuam Apon, Thavarungkul Panote, Kanatharana Proespichaya
Trace Analysis and Biosensor Research Center, Prince of Songkla University, Hat Yai, Songkhla, 90112, Thailand,
Anal Bioanal Chem. 2014 Jun;406(15):3763-72. doi: 10.1007/s00216-014-7770-3. Epub 2014 Apr 10.
A novel amperometric uric acid biosensor was fabricated by immobilizing uricase on an electrospun nanocomposite of chitosan-carbon nanotubes nanofiber (Chi-CNTsNF) covering an electrodeposited layer of silver nanoparticles (AgNPs) on a gold electrode (uricase/Chi-CNTsNF/AgNPs/Au). The uric acid response was determined at an optimum applied potential of -0.35 V vs Ag/AgCl in a flow-injection system based on the change of the reduction current for dissolved oxygen during oxidation of uric acid by the immobilized uricase. The response was directly proportional to the uric acid concentration. Under the optimum conditions, the fabricated uric acid biosensor had a very wide linear range, 1.0-400 μmol L(-1), with a very low limit of detection of 1.0 μmol L(-1) (s/n = 3). The operational stability of the uricase/Chi-CNTsNF/AgNPs/Au biosensor (up to 205 injections) was excellent and the storage life was more than six weeks. A low Michaelis-Menten constant of 0.21 mmol L(-1) indicated that the immobilized uricase had high affinity for uric acid. The presence of potential common interfering substances, for example ascorbic acid, glucose, and lactic acid, had negligible effects on the performance of the biosensor. When used for analysis of uric acid in serum samples, the results agreed well with those obtained by use of the standard enzymatic colorimetric method (P > 0.05).
通过将尿酸酶固定在覆盖有金电极上电沉积银纳米颗粒(AgNPs)层的壳聚糖-碳纳米管纳米纤维(Chi-CNTsNF)的电纺纳米复合材料上,制备了一种新型安培型尿酸生物传感器(尿酸酶/Chi-CNTsNF/AgNPs/Au)。在流动注射系统中,基于固定化尿酸酶氧化尿酸过程中溶解氧还原电流的变化,在相对于Ag/AgCl为-0.35 V的最佳施加电位下测定尿酸响应。该响应与尿酸浓度成正比。在最佳条件下,所制备的尿酸生物传感器具有非常宽的线性范围,即1.0 - 400 μmol L⁻¹,检测限极低,为1.0 μmol L⁻¹(信噪比 = 3)。尿酸酶/Chi-CNTsNF/AgNPs/Au生物传感器的操作稳定性极佳(高达205次进样),储存寿命超过六周。低米氏常数为0.21 mmol L⁻¹表明固定化尿酸酶对尿酸具有高亲和力。潜在的常见干扰物质,例如抗坏血酸、葡萄糖和乳酸的存在,对生物传感器的性能影响可忽略不计。当用于分析血清样品中的尿酸时,结果与使用标准酶比色法获得的结果非常吻合(P > 0.05)。