Taras Shevchenko National University of Kyiv, Volodymyrska Str., 64, 01003 Kyiv, Ukraine.
Taras Shevchenko National University of Kyiv, Volodymyrska Str., 64, 01003 Kyiv, Ukraine; Institute of Molecular Biology and Genetics NAS of Ukraine, Zabolotnogo Str., 150, 03680 Kyiv, Ukraine.
Bioelectrochemistry. 2018 Dec;124:40-46. doi: 10.1016/j.bioelechem.2018.07.002. Epub 2018 Jul 5.
A new conductometric biosensor based on coimmobilized urease and arginase has been developed for arginine determination in pharmaceutics. First, the main parameters of the selected method of immobilization (concentrations of arginase, urease, and glutaraldehyde, time of incubation) were optimized. An influence of the solution parameters (buffer ionic strength, capacity, pH, Mn concentration) on the biosensor operation was studied, working conditions were optimized. After biosensor optimization, the main analytical characteristics were as follows. The limit of detection - 2.5 μM, the linear range - 2.5-500 μM, the sensitivity to arginine 13.4 ± 2.4 μS/mM, the response time - 20 s. The signals repeatability and operational stability in continuous exploitation were studied over one working day and during one week. Additionally, the selectivity of the developed biosensor towards arginine was essayed relative to other amino acids. The developed biosensor has been used to measure arginine concentrations in some drugs. The results obtained were in high correlation with the characteristics declared by producers.
一种基于共固定化脲酶和精氨酸酶的新型电导生物传感器已被开发用于药物中精氨酸的测定。首先,优化了选择的固定化方法(精氨酸酶、脲酶和戊二醛的浓度、孵育时间)的主要参数。研究了溶液参数(缓冲离子强度、容量、pH 值、Mn 浓度)对生物传感器操作的影响,优化了工作条件。对生物传感器进行优化后,主要分析特性如下。检测限-2.5μM,线性范围-2.5-500μM,对精氨酸的灵敏度为 13.4±2.4μS/mM,响应时间-20s。在一个工作日和一周内研究了连续开发过程中信号的重复性和操作稳定性。此外,还相对于其他氨基酸测定了所开发生物传感器对精氨酸的选择性。已将所开发的生物传感器用于测量某些药物中的精氨酸浓度。所获得的结果与生产者所宣称的特征高度相关。