Hydrogen Peroxide-Induced Deacetylation of Acetyl Resorufin as a Novel Indicator Reaction for Fluorometric Detection of Glucose Using Only Glucose Oxidase
| Autor: | Yuji Yamauchi, Hatsuo Maeda, Shinya Matsu-Ura, Mari Nishida, Hidenobu Ohmori, Toshihisa Senba |
|---|---|
| Rok vydání: | 2001 |
| Předmět: |
Ascorbic Acid
Catalysis Glucose Oxidase chemistry.chemical_compound Oxazines Drug Discovery Glucose oxidase Hydrogen peroxide chemistry.chemical_classification Chromatography biology Bilirubin Hydrogen Peroxide General Chemistry General Medicine Ascorbic acid Fluorescence Uric Acid Glucose Spectrometry Fluorescence Enzyme chemistry Biochemistry Dealkylation Acetylation biology.protein Uric acid Indicators and Reagents Anaerobic exercise |
| Zdroj: | Chemical and Pharmaceutical Bulletin. 49:294-298 |
| ISSN: | 1347-5223 0009-2363 |
| DOI: | 10.1248/cpb.49.294 |
| Popis: | Hydrogen peroxide (H2O2)-induced deacetylation of non-fluorescent acetyl resorufin (1) to fluorescent resorufin (2) as a novel indicator reaction for fluorometric detection of glucose using only glucose oxidase (GOD) is described. When a 1:1:1 mixture of 1 (in CH3CN), glucose, and GOD (each in pH 7.4 phosphate buffer) was incubated at 25 degrees C under aerobic conditions, the resulting solution turned yellow to fluorescent pink due to 2. The formation of 2 was markedly retarded on incubation under anaerobic conditions. When a mixture of 1 and H2O2 was incubated under aerobic conditions, the formation of 2 was noted as in the case of the enzymatic reaction of 1. These results demonstrated that the observed color change is brought about through deacetylation of 1 to 2 induced by H2O2 generated in GOD-catalyzed oxidation of glucose. With regard to the fluorometric traces of the enzymatic reaction with 1 (0.2 mM), GOD (0.5 mg/ml), and glucose at 25 degrees C, fluorescence intensity exhibited a linear relationship against glucose concentration between 0.2 and 2.0 mm, with a correlation coefficient of 0.997. Neither ascorbic acid, uric acid, nor bilirubin significantly interfered with the transformation of 1 to 2 through GOD-catalyzed oxidation of glucose. |
| Databáze: | OpenAIRE |
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