title: A Caco-2 cell-based quantitative antioxidant activity assay for antioxidants.
abstract: A Caco-2 cell-based antioxidant activity (CAA) assay for quantitative evaluation of antioxidants was developed by optimizing seeding density and culture time of Caco-2 cells, incubation time and concentration of fluorescent probe (2',7'-dichlorofluorescin diacetate, DCFH-DA), incubation way and incubation time of antioxidants (pure phytochemicals) and DCFH-DA with cells, and detection time of fluorescence. Results showed that the CAA assay was of good reproducibility and could be used to evaluate the antioxidant activity of antioxidants at the following conditions: seeding density of 5 × 10(4)/well, cell culture time of 24h, co-incubation of 60 μM DCFH-DA and pure phytochemicals with Caco-2 cells for 20 min and fluorescence recorded for 90 min. Additionally, a significant correlation was observed between CAA values and rat plasma ORAC values following the intake of antioxidants for selected pure phytochemicals (R(2) = 0.815, p < 0.01), demonstrating the good biological relevance of CAA assay.
authors.affiliation: Shenzhen Key Laboratory of Fermentation, Purification and Analysis, Shenzhen Polytechnic, Shenzhen 518055, People's Republic of China.
authors.forename: Hongxia
authors.initials: H
authors.lastname: Wan
authors.affiliation: Shenzhen Key Laboratory of Fermentation, Purification and Analysis, Shenzhen Polytechnic, Shenzhen 518055, People's Republic of China. Electronic address: liudongsz@szpt.edu.cn.
authors.forename: Dong
authors.initials: D
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Chinese Medicine and Molecular Pharmacology, Shenzhen Research Institute of The Hong Kong Polytechnic University, Shenzhen 518057, People's Republic of China.
authors.forename: Xiangying
authors.initials: X
authors.lastname: Yu
authors.affiliation: Shenzhen Key Laboratory of Fermentation, Purification and Analysis, Shenzhen Polytechnic, Shenzhen 518055, People's Republic of China. Electronic address: susan@szpt.edu.cn.
authors.forename: Haiyan
authors.initials: H
authors.lastname: Sun
authors.affiliation: Shenzhen Key Laboratory of Fermentation, Purification and Analysis, Shenzhen Polytechnic, Shenzhen 518055, People's Republic of China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000737:chemistry* / Q000494:pharmacology*; D018938:Caco-2 Cells; D018929:Cell Culture Techniques; D004353:Drug Evaluation, Preclinical / Q000379:methods; D006801:Humans; D008297:Male; D010084:Oxidation-Reduction; D010936:Plant Extracts; D011897:Random Allocation; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D000975:Antioxidants; D010936:Plant Extracts
keywords: Antioxidants; Caco-2 cell-based antioxidant activity (CAA); Caco-2 cells; Oxygen radical absorbance capacity (ORAC); Quantitative assay