Hydrogen Peroxide-Induced Deacetylation of Acetyl Resorufin as a Novel Indicator Reaction for Fluorometric Detection of Glucose Using Only Glucose Oxidase.
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- MAEDA Hatsuo
- Graduate School of Pharmaceutical Sciences, Osaka University
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- MATSU-URA Shinya
- Graduate School of Pharmaceutical Sciences, Osaka University
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- NISHIDA Mari
- Graduate School of Pharmaceutical Sciences, Osaka University
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- SENBA Toshihisa
- Graduate School of Pharmaceutical Sciences, Osaka University
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- YAMAUCHI Yuji
- Graduate School of Pharmaceutical Sciences, Osaka University
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- OHMORI Hidenobu
- Graduate School of Pharmaceutical Sciences, Osaka University
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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 °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 °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.
収録刊行物
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- CHEMICAL & PHARMACEUTICAL BULLETIN
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CHEMICAL & PHARMACEUTICAL BULLETIN 49 (3), 294-298, 2001
公益社団法人 日本薬学会
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詳細情報 詳細情報について
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- CRID
- 1390282679137500928
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- NII論文ID
- 110003615890
- 30023110589
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- NII書誌ID
- AA00602100
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- COI
- 1:CAS:528:DC%2BD3MXhs1OhsLo%3D
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- ISSN
- 13475223
- 00092363
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- NDL書誌ID
- 5682974
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- PubMed
- 11253919
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- 本文言語コード
- en
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- データソース種別
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- JaLC
- NDL
- Crossref
- PubMed
- CiNii Articles
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- 抄録ライセンスフラグ
- 使用不可