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highly sensitive determination of hydrogen peroxide and glucose by fluorescence correlation spectroscopy高度敏感的过氧化氢和葡萄糖的测定荧光相关光谱
Highly Sensitive Determination of Hydrogen Peroxide and Glucose by Fluorescence Correlation Spectroscopy 1 2 2 2 3 2 Satoshi Watabe , Yuki Sakamoto , Mika Morikawa , Ryuichi Okada , Toshiaki Miura , Etsuro Ito * 1 BL Co. Ltd., Numazu, Shizuoka, Japan, 2 Kagawa School of Pharmaceutical Sciences, Tokushima Bunri University, Sanuki, Kagawa, Japan, 3 Graduate School of Pharmaceutical Sciences, Hokkaido University, Kita-ku, Sapporo, Hokkaido, Japan Abstract Background: Because H O is generated by various oxidase-catalyzed reactions, a highly sensitive determination method of 2 2 H2O2 is applicable to measurements of low levels of various oxidases and their substrates such as glucose, lactate, glutamate, urate, xanthine, choline, cholesterol and NADPH. We propose herein a new, highly sensitive method for the measurement of H2O2 and glucose using fluorescence correlation spectroscopy (FCS). Methodology/Principal Findings: FCS has the advantage of allowing us to determine the number of fluorescent molecules. FCS measures the fluctuations in fluorescence intensity caused by fluorescent probe movement in a small light cavity with a defined volume generated by confocal illumination. We thus developed a highly sensitive determination system of H2O2 by FCS, where horseradish peroxidase (HRP) catalyzes the formation of a covalent bond between fluorescent molecules and proteins in the presence of H O . Our developed system gave a linear calibration curve for H O in the range of 28 to 2 2 2 2
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