Xu Yi, Peng Weilin, Liu Xinjian, Li Genxi
Department of Biochemistry and National Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing 210093, PR China.
Biosens Bioelectron. 2004 Oct 15;20(3):533-7. doi: 10.1016/j.bios.2004.02.017.
A novel and stable film made from polyethylene glycol (PEG) on pyrolytic graphite (PG) electrode was presented in this paper for incorporating horseradish peroxidase (HRP) to study the direct electrochemistry of the enzyme. In PEG film, HRP showed a thin-layer electrochemistry behavior. The apparent standard potential (E degrees ') was -0.379 V versus SCE at pH 7.2. Moreover, the PEG-HRP modified electrode exhibited excellent electrocatalytical response to the reduction of H2O2 with a calibration range between 2.0 x 10(-6) and 6.0 x 10(-4) M and a good linear relation from 2.0 x 10(-6) to 1.0 x 10(-4) M, on which an unmediated H2O2 biosensor was based. The detection limit of 6.7 x 10(-7) M was estimated when the signal-to-noise ratio was 3. The relative standard deviation (R.S.D.) was 4.7% for six successive determinations at a concentration of 4.0 x 10(-5) M. The apparent Michaelis-Menten constant (Km app) of the sensor was found to be 1.38 mM. Epinephrine, dopamine, and ascorbic acid did not interfere with the sensitive determination of H2O2.
本文介绍了一种在热解石墨(PG)电极上由聚乙二醇(PEG)制成的新型稳定薄膜,用于固定辣根过氧化物酶(HRP)以研究该酶的直接电化学。在PEG薄膜中,HRP呈现出薄层电化学行为。在pH 7.2时,相对于饱和甘汞电极(SCE)的表观标准电位(E°')为-0.379 V。此外,PEG-HRP修饰电极对H2O2的还原表现出优异的电催化响应,校准范围在2.0×10^(-6)至6.0×10^(-4) M之间,在2.0×10^(-6)至1.0×10^(-4) M之间具有良好的线性关系,基于此构建了一种无媒介的H2O2生物传感器。当信噪比为3时,估计检测限为6.7×10^(-7) M。在4.0×10^(-5) M浓度下连续六次测定的相对标准偏差(R.S.D.)为4.7%。该传感器的表观米氏常数(Km app)为1.38 mM。肾上腺素、多巴胺和抗坏血酸不干扰H2O2的灵敏测定。