To determine the antimicrobial and antioxidant activities of different extracts from Phyllanthus urinaria, the distribution of active components in different polarities solvents were clarified preliminarily. The secondary metabolites were extracted by methanol, by petroleum ether, and by ethyl acetate and n-butanol ordinally. Their antibacterial activities of the different extracts from P. urinaria L. were determined by TLC-MTT-bioautography assay. The inhibitory activities against Colletotrichum gloeosporioides were determined by mycelium growth rate method. The antioxidant activities were determined by a micro-plate spectrophotometric method based on the reduction of DPPH. Furthermore, HPLC was used to analyze and detect the distribution of secondary metabolites in different extracts from P. urinaria. The extracts of ethyl acetate and n-butanol showed antibacterial activities. Its inhibitory activity against Ralstonia solanacearum was significantly stronger than that of the positive control. In addition, the extract of ethyl acetate demonstrated the strongest antifungal and antioxidant activities, with the EC50 value of (1.02±0.15) mg/mL and IC50 value of (20.14±0.58) g/mL. It was found that the compounds of ethyl acetate extract from P. urinaria concentrated within the retention time of 9-12 min mainly according to the HPLC-UV analysis. In a word, the ethyl acetate extracts of P. urinaria displayed obvious antibacterial, antifungal and antioxidant activities. Ethyl acetate extract could be further used as a candidate resource for the separation and identification of active components.