抗氧化能力检测方法评估及微型化

 目的评估现有抗氧化能力检测方法,建立微型化抗氧化能力检测方法,加速抗氧化物质的筛选。方法在考察血浆铁还原能力(FRAP)、二苯基苦基苯肼(DPPH)方法学的基础上,建立微型FRAP、DPPH法,并通过微型FRAP、DPPH法对内源性抗氧化剂(维生素C、维生素E、谷胱苷肽、α-硫辛酸、β-胡萝卜素)和外源性抗氧化剂(白藜芦醇)进行抗氧化能力检测,以考察其灵敏度和专属性。结果微型FRAP、DPPH法具有良好的线性、精密度、回收率。与传统FRAP、DPPH方法相比,不仅简便、快速、样品用量大大减少,而且对水溶性维生素E类似物(Trolox)、白藜芦醇、茶多酚的检测结果非常相近,对维生素E、维生素C和白藜芦醇都有很好的灵敏度和专属性。结论①微型抗氧化能力检测方法可以加速评价物质的抗氧化能力;②微型抗氧化能力检测与传统紫外法相比,在评价质量上没有明显区别;③微型抗氧化能力检测方法对内源性和外源性抗氧化物质均显示很好的灵敏度和专属性。

Evaluation and Miniaturization for Antioxidant Capacity Measurement

OBJECTIVE To evaluate and miniaturize the assays for antioxidant capacity measurement and screen of herb-derived antioxidants.METHODS Based on the studies of ferric reduction ability plasma(FRAP) and 2,2′-diphenyl-1-picrylhydrazyl(DPPH) assays,miniaturized 96-well FRAP and DPPH assays were developed.The parameters such as sensitivity and specificity of miniaturized assays were evaluated via the measurement of antioxidant capacity of different substances,including endogenous antioxidants(vitamin C,vitamin E,GSH,α-lipoic acid,and β-carotene) and exogenous antioxidants(resveratrol and tea polyphenols).RESULTS 96-well FRAP assay and DPPH assay demonstrated considerable quality of linearity,accuracy,and recovery.Comparing with traditional FRAP assay and DPPH assay,the 96-well assays were markedly convenient,rapid,and small sample consumption,which were proved with our test of trolox,resveratrol and tea polyphenols.Furthermore,the 96-well assay showed excellent sensitivity and specificity towards the antioxidant capacity of vitamin E,vitamin C and resveratrol.CONCLUSION ①The evaluation of antioxidative capacity of chemical substances or biological samples can be significantly benefited by 96-well FRAP assay and DPPH assay.②Compared to traditional UV assays,96-well FRAP assay and DPPH assay demonstrate comparable quality.③96-well FRAP assay and DPPH assay show excellent sensitivity and specificity toward endogenous and exogenous antioxidants.