Shanghai Key Laboratory of Green Chemistry and Chemical Processes, School of Chemistry and Molecular Engineering, East China Normal University , Dongchuan Road 500, Shanghai 200241, People's Republic of China.
Anal Chem. 2017 Jun 20;89(12):6656-6662. doi: 10.1021/acs.analchem.7b00881. Epub 2017 Jun 9.
Glucose and pH are two important indicators of diabetes mellitus. However, their dynamic changes at the same time in brain are still not clear, mainly due to a lack of a single biosensor capable of simultaneous quantification of two species in a live rat brain. In this work, a selective and sensitive ratiometric electrochemical biosensor was developed for simultaneously quantifying glucose and pH using both current and potential outputs in a rat brain of diabetic model. Here, glucose oxidase was first employed as a specific recognition element for both glucose and pH because the active center (FAD) could undergo a 2H/2e process. Moreover, an insensitive molecule toward pH and glucose was used as an inner-reference element to provide a built-in correction to improve the accuracy. The ratio between the oxidation peak current density of glucose and that of ABTS gradually increased with increasing concentration of glucose, and showed a good linearity in the range of 0.3-8.2 mM. Meanwhile, the midpotential difference between glucose oxidase and 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) positively shifted with pH decreasing, leading to accurate determination of pH in the linear range of 5.67-7.65. Thus, combined with the unique properties of carbon fiber microelectrode, including easy to insert and good biocompatibility, the developed single biosensor was successfully applied to detect pH and glucose at the same time in hippocampus, striatum, and cortex in a live rat brain of diabetic model.
葡萄糖和 pH 值是糖尿病的两个重要指标。然而,它们在大脑中的动态变化尚不清楚,主要是因为缺乏一种能够同时定量检测活鼠脑内两种物质的单一生物传感器。在这项工作中,我们开发了一种选择性和灵敏的比率型电化学生物传感器,可通过电流和电位输出同时定量检测糖尿病模型大鼠脑内的葡萄糖和 pH 值。在此,我们首次使用葡萄糖氧化酶作为葡萄糖和 pH 值的特异性识别元件,因为其活性中心 (FAD) 可以经历 2H/2e 过程。此外,我们还使用了一种对 pH 值和葡萄糖不敏感的分子作为内参元件,提供内置校正以提高准确性。葡萄糖氧化酶与 ABTS 的氧化峰电流密度之比随葡萄糖浓度的增加而逐渐增加,并在 0.3-8.2 mM 的范围内表现出良好的线性关系。同时,葡萄糖氧化酶与 2,2'-联氮-双(3-乙基苯并噻唑啉-6-磺酸)(ABTS)之间的中电位差随着 pH 值的降低而正向移动,从而能够准确测定 5.67-7.65 范围内的 pH 值。因此,结合碳纤维微电极的独特特性,包括易于插入和良好的生物相容性,该开发的单生物传感器成功应用于检测糖尿病模型活鼠脑内海马体、纹状体和皮层中同时存在的 pH 值和葡萄糖。