Department of Chemistry, North Carolina State University, Raleigh, North Carolina 27695, USA.
Anal Chem. 2010 Jun 15;82(12):5205-10. doi: 10.1021/ac100536s.
Hydrogen peroxide is a reactive oxygen species that is implicated in a number of neurological disease states and that serves a critical role in normal cell function. It is commonly exploited as a reporter molecule enabling the electrochemical detection of nonelectroactive molecules at electrodes modified with substrate-specific oxidative enzymes. We present the first voltammetric characterization of rapid hydrogen peroxide fluctuations at an uncoated carbon fiber microelectrode, demonstrating unprecedented chemical and spatial resolution. The carbon surface was electrochemically conditioned on the anodic scan and the irreversible oxidation of peroxide was detected on the cathodic scan. The oxidation potential was dependent on scan rate, occurring at +1.2 V versus Ag/AgCl at a scan rate of 400 V.s(-1). The relationship between peak oxidation current and concentration was linear across the physiological range tested, with deviation from linearity above 2 mM and a detection limit of 2 muM. Peroxide was distinguished from multiple interferents, both in vitro and in brain slices. The enzymatic degradation of peroxide was monitored, as was peroxide evolution in response to glucose at a glucose oxidase modified carbon fiber electrode. This novel approach provides the requisite sensitivity, selectivity, spatial and temporal resolution to study dynamic peroxide fluctuations in discrete biological locations.
过氧化氢是一种活性氧物质,与许多神经疾病状态有关,并在正常细胞功能中起着关键作用。它通常被用作报告分子,使在经过基底特异性氧化酶修饰的电极上,能够电化学检测非电活性分子。我们首次在未涂层碳纤维微电极上对快速过氧化氢波动进行了伏安特性表征,展示了前所未有的化学和空间分辨率。碳表面在阳极扫描时进行电化学处理,在阴极扫描时检测到过氧化物的不可逆氧化。氧化电位取决于扫描速率,在 400 V/s 的扫描速率下,相对于 Ag/AgCl 为 +1.2 V。在测试的生理范围内,峰氧化电流与浓度之间呈线性关系,在 2 mM 以上偏离线性,检测限为 2 μM。过氧化氢与多种干扰物(包括体外和脑切片)区分开来。监测了过氧化物的酶促降解,以及在葡萄糖氧化酶修饰的碳纤维电极上响应葡萄糖的过氧化物演化。这种新方法提供了必需的灵敏度、选择性、空间和时间分辨率,以研究离散生物位置的动态过氧化物波动。