Background: The biological activities of Tradescantia pallida grown in Korea have not been well determined, thus the aim of this study was to investigate the possibility of using it as a medicinal plant. Methods and Results: To investigate the antioxidant activity, α-glucosidase inhibitory effect and antimicrobial activity of T. pallida, we performed the 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activity and reducing power assay. This assay for T. pallida leaf extract showed the highest antioxidant activity for the ethyl acetate fraction (RC50= 14.55 ± 0.16㎍/㎖ and Abs = 0.613 at 300㎍). Further, the ethyl acetate fraction exhibited higher α-glucosidase inhibitory effect with an IC50 value of 14.1 ± 0.1㎍/㎖ and showed antimicrobial activity against gram positive bacteria (minimum inhibitory concentration = 1,000㎍/㎖). Conclusions: The ethyl acetate fraction of the crude methanol extract of T. pallida showed remarkable antioxidant activity, α-glucosidase inhibitory effects and antimicrobial activity. These activities might be related to the flavonoid content in the T. pallida leaf extract.