International Centre of Biodynamics, 1B Intrarea Portocalelor, 060101, Bucharest, Romania.
Institute of Biology, 296 Splaiul Independentei, 060031, Bucharest, Romania.
Mikrochim Acta. 2020 Sep 5;187(10):550. doi: 10.1007/s00604-020-04477-3.
Carbon nanofibers (CNF) are efficient electrode modifiers in electrochemical biosensors that enhance the electrochemical active area, induce electrocatalytic effect toward the oxidation of the enzymatic cofactor nicotinamide adenine dinucleotide (reduced form, NADH), and enable the quantitative immobilization of enzymes. Combining CNF with efficient and stable mediators radically augments the speed of electron transfer between NADH and solid electrodes and leads to electrochemical sensors characterized by high sensitivity and stability. The main aim of this work was to investigate the performance of a novel mediator for NADH with advantageously low solubility in an electrochemical detector based on a screen-printed CNF electrode as well as its potential in biosensing. Using a mediator, prepared from Meldola Blue and Ni hexamine chloride, a stable and sensitive electrochemical NADH sensor is provided with a detection limit of 0.5 μmol L. Further on, covalent immobilization of a recently described aldehyde dehydrogenase from the Antarctic Flavobacterium PL002 strain on the surface of the mediator-modified electrode produced a stable biosensor for the detection of aldehydes. When integrated in a flow injection analysis (FIA) setup with amperometric detection at 0.1 V vs. Ag/AgCl, the measurement of benzaldehyde with a detection limit of 10 μmol L over a linear range of 30-300 μmol L is possible. Determination of trace benzaldehyde impurities in a pharmaceutical excipient was also demonstrated and results compared with a chromatographic method. Graphical abstract.
碳纳米纤维(CNF)是电化学生物传感器中有效的电极修饰剂,可增大电化学活性面积,诱导对酶辅因子烟酰胺腺嘌呤二核苷酸(还原形式,NADH)氧化的电催化作用,并实现酶的定量固定化。将 CNF 与高效且稳定的介体结合,可极大地增强 NADH 与固体电极之间的电子转移速度,从而产生具有高灵敏度和稳定性的电化学传感器。本工作的主要目的是研究一种新型 NADH 介体在基于丝网印刷 CNF 电极的电化学检测器中的性能,以及其在生物传感中的潜力。使用介体(由 Meldola 蓝和 Ni 六氨合氯化物制备),提供了一种稳定且灵敏的电化学 NADH 传感器,检测限为 0.5 μmol L。进一步地,将最近描述的来自南极黄杆菌 PL002 菌株的醛脱氢酶共价固定在介体修饰电极的表面上,产生了用于检测醛的稳定生物传感器。当与安培检测(0.1 V 对 Ag/AgCl)集成在流动注射分析(FIA)装置中时,可以在 30-300 μmol L 的线性范围内测量 10 μmol L 的苯甲醛,检测限为 10 μmol L。还证明了在药物赋形剂中痕量苯甲醛杂质的测定,并将结果与色谱法进行了比较。