Development and validation of an improved oxygen radical absorbance capacity assay using fluorescein as the fluorescent probe
Development and validation of an improved oxygen radical absorbance capacity assay using fluorescein as the fluorescent probe
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DOI：
10.1021/jf010586o
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发表时间：
2001-10-01
影响因子：
6.1
通讯作者：
Prior, RL
中科院分区：
文献类型：
--
作者：
Ou, BX;Hampsch-Woodill, M;Prior, RL
以荧光素（3 '，6'-dihydroxyspiro[isobenzofuran-1[3 H]，9 '[9 H]-xanthen]-3-one）为荧光探针，建立了一种改进的氧自由基吸收能力（ORAC）测定方法。我们的研究结果表明，荧光素（FL）是上级比B-藻红蛋白。通过LC/MS鉴定了过氧自由基诱导的FL氧化产物，并确定反应机理遵循经典的氢原子转移机理。此外，还讨论了ORAC（FL）与其他广泛使用的方法的方法学和机制比较。可以得出结论，与其他流行的方法，改进的ORAC（FL）测定提供了一个直接的措施，对过氧自由基的亲水链断裂抗氧化能力。
An improved method of oxygen radical absorbance capacity (ORAC) assay has been developed and validated using fluorescein (3',6'-dihydroxyspiro[isobenzofuran-1[3H],9'[9H]-xanthen]-3-one) as the fluorescent probe. Our results demonstrate that fluorescein (FL) is superior to B-phycoerythrin. The oxidized FL products induced by peroxyl radical were identified by LC/MS, and the reaction mechanism was determined to follow a classic hydrogen atom transfer mechanism. In addition, methodological and mechanistic comparison of ORAC(FL) with other widely used methods was discussed. It is concluded that, unlike other popular methods, the improved ORAC(FL) assay provides a direct measure of hydrophilic chain-breaking antioxidant capacity against peroxyl radical.