Mădăraş M B, Buck R P
Department of Chemistry, University of North Carolina, Chapel Hill 27599-3290, USA.
Anal Chem. 1996 Nov 1;68(21):3832-9. doi: 10.1021/ac960239r.
Miniaturized, disposable amperometric biosensors for determination of creatinine in human serum are described. The base electrodes are fabricated using micro-electronics techniques, to build a multilayer film structure on a polyimide foil. By using a thin electropolymerized film of poly(1,3-diaminobenzene), the electrochemical interferences from ascorbate, urate, acetaminophen, and other oxidizable species are greatly diminished. The multienzyme system (creatininase, creatinase, sarcosine oxidase) is immobilized on top of the permselective layer using cross-linking of the proteins with glutaraldehyde. The electropolymerization conditions for obtaining almost ideal permselectivity of the inner layer are defined, as well as the optimal enzyme layer preparation. A composite polymeric outer membrane [Nafion + poly-(2-hydroxy-ethyl methacrylate) is used for diffusion control and to protect the enzyme layer from fouling. The reagentless planar sensors for creatinine and creatine have fast response time (t95 = 1 min), linear response up to 1.2 mM in batch-type and 2.0 mM in flow injection analysis and a detection limit of 10-20 muM. They are applied in a differential setup for creatinine assay in control and hospital human serum samples and are suitable for incorporation in a portable analyzer.
本文描述了用于测定人血清中肌酐的小型化一次性安培生物传感器。基底电极采用微电子技术制造，在聚酰亚胺箔上构建多层膜结构。通过使用聚（1,3 - 二氨基苯）的薄电聚合膜，可大大减少来自抗坏血酸盐、尿酸盐、对乙酰氨基酚和其他可氧化物质的电化学干扰。利用蛋白质与戊二醛交联的方法，将多酶系统（肌酐酶、肌酸酶、肌氨酸氧化酶）固定在选择性渗透层的顶部。确定了获得内层几乎理想选择性的电聚合条件以及最佳酶层制备方法。复合聚合物外膜[Nafion + 聚（甲基丙烯酸2 - 羟乙酯）]用于扩散控制并保护酶层免受污染。用于肌酐和肌酸的无试剂平面传感器具有快速响应时间（t95 = 1分钟），在分批式分析中线性响应高达1.2 mM，在流动注射分析中高达2.0 mM，检测限为10 - 20 μM。它们应用于差分设置中，用于检测对照和医院人血清样本中的肌酐，适用于集成到便携式分析仪中。
