Rapid and convenient detection of ascorbic acid using a fluorescent nitroxide switch
Rapid and convenient detection of ascorbic acid using a fluorescent nitroxide switch
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DOI：
10.1016/j.freeradbiomed.2012.09.032
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发表时间：
2012-12-01
影响因子：
7.4
通讯作者：
Yamada, Ken-ichi
中科院分区：
文献类型：
--
作者：
Matsuoka, Yuta;Yamato, Mayumi;Yamada, Ken-ichi
抗坏血酸是一种体内多功能的小分子还原剂。减少抗坏血酸的摄入会导致脯氨酸和赖氨酸缺乏羟基化，导致三螺旋结构更松散，从而导致坏疽。抗坏血酸也是一种抗氧化剂，可以防止氧化应激。由于抗坏血酸与疾病状态有关，快速、方便地检测抗坏血酸在诊断中应该是有用的。抗坏血酸将氮氧化物还原为相应的羟胺，一种灵敏而新颖的检测方法是利用氮氧化物与荧光团的共价偶联，通过电子交换作用在分子内猝灭荧光发射。在这里，我们开发了一种新的荧光团-氮氧化物探针。Naph-dify氮氧化物，用于抗坏血酸。NaH-dify氮氧化物与抗坏血酸反应迅速，并表现出荧光增强，但对其他还原剂或活性氧物种无反应。为了证实荧光团氮氧化物探针的实用价值，我们用Nahdify氮氧化物测定了缺抗坏血酸饲料喂养的成骨性疾病Shionogi大鼠血浆中抗坏血酸的含量。结果表明，这种新型荧光氮氧化物探针可以灵敏、简便地检测抗坏血酸，可作为疾病状态的诊断工具。(C)2012 Elsevier Inc.保留所有权利。
Ascorbic acid is a small-molecule reductant with multiple functions in vivo. Reducing ascorbic acid intake leads to a lack of hydroxylation of prolines and lysines, causing a looser triple helix and resulting in scurvy. Ascorbic acid also acts as an antioxidant to prevent oxidative stress. Because ascorbic acid is related to disease states, rapid and convenient detection of ascorbic acid should be useful in diagnosis. Nitroxide is reduced to the corresponding hydroxylamine by ascorbic acid and a sensitive and novel approach to its detection employs covalent coupling of nitroxide with a fluorophore, leading to intramolecular quenching of fluorescence emission by electron-exchange interactions. Here, we developed a new fluorophore-nitroxide probe. Naph-DiFy nitroxide, for ascorbic acid. Naph-DiFy nitroxide rapidly reacted with ascorbic acid and showed fluorescence enhancement, but not in response to other reductants or reactive oxygen species. To confirm the practical usefulness of the fluorophore-nitroxide probe, we demonstrated the use of Naph-DiFy nitroxide for the measurement of ascorbic acid in the plasma of osteogenic disorder Shionogi rats when fed an ascorbic acid-deficient diet. The results suggest that this novel fluorophore-nitroxide probe could sensitively and easily detect ascorbic acid and be useful as a tool for the diagnosis of disease states. (C) 2012 Elsevier Inc. All rights reserved.