College of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou 310058, China.
Key Laboratory of on Site Processing Equipment for Agricultural Products, Ministry of Agriculture, Hangzhou 310058, China.
J Zhejiang Univ Sci B. 2018;19(9):726-734. doi: 10.1631/jzus.B1700510.
p-Nitrophenylphosphate (PNPP) is usually employed as the substrate for enzyme-linked immunosorbent assays. p-Nitrophenol (PNP), the product of PNPP, with the catalyst alkaline phosphatase (ALP), will passivate an electrode, which limits applications in electrochemical analysis. A novel anti-passivation ink used in the preparation of a graphene/ionic liquid/chitosan composited (rGO/IL/Chi) electrode is proposed to solve the problem. The anti-passivation electrode was fabricated by directly writing the graphene-ionic liquid-chitosan composite on a single-side conductive gold strip. A glassy carbon electrode, a screen-printed electrode, and a graphene-chitosan composite-modified screen-printed electrode were investigated for comparison. Scanning electron microscopy was used to characterize the surface structure of the four different electrodes and cyclic voltammetry was carried out to compare their performance. The results showed that the rGO/IL/Chi electrode had the best performance according to its low peak potential and large peak current. Amperometric responses of the different electrodes to PNP proved that only the rGO/IL/Chi electrode was capable of anti-passivation. The detection of cardiac troponin I was used as a test example for electrochemical immunoassay. Differential pulse voltammetry was performed to detect cardiac troponin I and obtain a calibration curve. The limit of detection was 0.05 ng/ml.
对硝基苯膦酸盐(PNPP)通常用作酶联免疫吸附测定的底物。PNPP 的产物对硝基苯酚(PNP)在碱性磷酸酶(ALP)的催化下会使电极钝化,从而限制了其在电化学分析中的应用。为了解决这个问题,提出了一种新型的抗钝化油墨,用于制备石墨烯/离子液体/壳聚糖复合材料(rGO/IL/Chi)电极。通过直接在单面带导电金条上书写石墨烯-离子液体-壳聚糖复合材料来制备抗钝化电极。比较了玻碳电极、丝网印刷电极和石墨烯-壳聚糖复合修饰的丝网印刷电极。使用扫描电子显微镜对四种不同电极的表面结构进行了表征,并进行了循环伏安法比较其性能。结果表明,根据其低峰电位和大峰电流,rGO/IL/Chi 电极的性能最佳。不同电极对 PNP 的安培响应证明只有 rGO/IL/Chi 电极具有抗钝化能力。以心肌肌钙蛋白 I 的电化学免疫分析检测为例。进行了差分脉冲伏安法检测心肌肌钙蛋白 I 并获得校准曲线。检测限为 0.05ng/ml。