Few data are available about the effects of complexation of polyphenols with polysaccharide on their bioavailability. The complex of tea polyphenols (TP) with oat β-glucan was characterized by ultraviolet-visible spectrometry, Fourier transform infrared spectrometry, differential scanning calorimetry, atomic force microscopy, and solid-state (13)C NMR spectroscopy. The results indicated that the bonds which governed the interaction between TP and oat β-glucan were strong hydrogen bonds. The in vitro antioxidant activity of TP, β-glucan, their complex, and physical mixture was assessed using four systems, namely, DPPH(•), OH(•), and O(2)(•-) scavenging activities and reducing power. The complexation and blending of TP and β-glucan exhibited different impacts on the index of in vitro and in vivo antioxidant capacities. In the concentration range of 0.5-2.5 mg mL(-1), the complex had highest O(2)(•-) scavenging activity, whereas the highest OH(•) scavenging activity was found with the physical mixture. For antioxidant testing in vivo, there was no significant difference between the complex and the physical mixture in terms of glutathione peroxidase activity and levels of malondialdehyde and total antioxidant capacity in serums. However, the complex exhibited much higher activities of superoxide dismutase and glutathione peroxidase in livers than the physical mixture. The present study provided a deeper understanding of the influence of molecular interaction between TP and oat β-glucan on their antioxidant activities.