Novel fluorometric assay for hydroxyl radical scavenging capacity (HOSC) estimation
Novel fluorometric assay for hydroxyl radical scavenging capacity (HOSC) estimation
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DOI：
10.1021/jf052555p
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发表时间：
2006-02-08
影响因子：
6.1
通讯作者：
Yu, LL
中科院分区：
文献类型：
--
作者：
Moore, J;Yin, JJ;Yu, LL
开发并验证了一种新的荧光方法，用于使用荧光素作为探针估计羟基自由基清除能力（HOSC）。使用类芬顿 Fe3+/H2O2 反应在生理 pH 下产生恒定通量的纯羟基自由基。使用电子自旋共振和 DMPO 自旋捕获测量来评估和确认实验条件下纯羟基自由基的产生。通过测量存在或不存在抗氧化剂的荧光衰减曲线下的面积来量化所选抗氧化剂样品的羟基自由基清除能力，并表示为每单位抗氧化剂的 Trolox 当量。可以使用带有荧光检测器的读板器进行测定，以进行高通量测量。该测定的线性、精密度、准确性、重现性及其与使用选定的纯抗氧化剂化合物和植物提取物的流行的过氧自由基清除能力测定的相关性经过验证。这种方法可以为食品、营养和医学领域的研究人员提供一种易于使用的方案，以评估纯抗氧化剂和天然提取物在体外针对活性极高的羟基自由基的自由基清除能力，羟基自由基可能与许多退行性疾病和病症有关。
A novel fluorometric method was developed and validated for hydroxyl radical scavenging capacity (HOSC) estimation using fluorescein as the probe. A constant flux of pure hydroxyl radical is generated under physiological pH using a Fenton-like Fe3+/H2O2 reaction. The generation of pure hydroxyl radicals under the experimental conditions was evaluated and confirmed using electron spin resonance with DMPO spin-trapping measurements. The hydroxyl radical scavenging capacity of a selected antioxidant sample is quantified by measuring the area under the fluorescence decay curve with or without the presence of the antioxidant and expressed as Trolox equivalents per unit of the antioxidant. The assay may be performed using a plate reader with a fluorescence detector for high-throughput measurements. The assay was validated for linearity, precision, accuracy, reproducibility, and its correlation with a popular peroxyl radical scavenging capacity assay using selected pure antioxidant compounds and botanical extracts. This method may provide researchers in the food, nutrition, and medical fields an easy to use protocol to evaluate free radical scavenging capacity of pure antioxidants and natural extracts in vitro against the very reactive hydroxyl radical, which may be linked to numerous degenerative diseases and conditions.