1.Murad Research Institute for Modernized Chinese Medicine,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;2. E-Institute of Shanghai Universities,Division of Nitric Oxide and Inflammatory Medicine,Shanghai 201203,China;3.Department of Integrative Biology and Pharmacology,Institute of Molecular Medicine,University of Texas Medical School,TX 77030,USA
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.
GO Yue-hong;ZHENG Jin-pu;ZHU Chun-yun;LI Y-jun;KE Yn;IN K;.
Evaluation and Miniaturization for Antioxidant Capacity Measurement [J]. Chinese Pharmaceutical Journal, 2008, 43(24): 1863-1866
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