In the course of screening for antioxidant compounds by measuring the radical scavenging effect on DPPH (1,1-diphenyl- 2-picrylhydrazyl), a total extract of the twigs of Vitex rotundifolia (Verbenaceae) was found to show potent antioxidant activity. Subsequent activity-guided fractionation of the methanolic extract led to the isolation of three iridoid compounds, 10-O-vanilloylaucubin (1), 10-O-p-hydroxybenzoylaucubin (2) and aucubin (3), two C-glycoside flavones, vitexin (4) and orientin (5), and a quinic acid derivative, 3,4-di-O-caffeoylquinic acid (6). Their structures were elucidated by spectroscopic studies. Among them, compounds 5 and 6 showed the significant antioxidative effects on DPPH free radical scavenging test. In riboflavin-NBT-light and xanthine-NBT-xanthine oxidase systems, compounds 5 and 6 exhibited the formation of the blue formazan in a dose-dependent manner. Compounds 5 and 6 showed better superoxide quenching activities than vitamin C.