To investigate the natural antioxidant and antimicrobial phytochemicals from black bamboo (Phyllostachys nigra MUNRO) leaves, the solvent fractions from crude methanol extract were made with n-hexane, ethyl acetate, and butanol, and their antioxidant and antimicrobial activities were determined. The antioxidant activities were examined by 1,1-diphenyl-1-picrylhydrazyl (DPPH) method and ferric ion reducing antioxidant power (FRAP) method, and the antimicrobial activities against Staphylococcus aureus were tested by paper disc agar diffusion method. Total phenolic contents and total flavonoid contents of the solvent fractions were also determined. The ethyl acetate fraction with the highest total phenolic contents among all fractions showed the strong antioxidant activities by DPPH method and FRAP method, and antimicrobial activities against S. aureus at all test concentrations. Caffeic acid, ferulic acid, quercetin, and kaempferol were analyzed by HPLC in the ethyl acetate fraction from black bamboo leaves by the comparison with the standard chemicals. It is supposed that the ethyl acetate fraction from black bamboo leaves could be used as natural preservatives in the food industry.