The free-radical-scavenging property, antibacterial activity and brine shrimp toxicity of petroleum ether (PE), dichloromethane (DCM) and methanol (MeOH) extracts of Centaurea persica, a Turkish medicinal plant, were assessed using the 2,2-diphenyl-1-picryl-hydrazyl (DPPH) assay, the resazurin microtiter plate based assay, and the brine shrimp lethality assay, respectively. Additionally, the disc diffusion assay was also used to assess antibacterial activity. Only the MeOH extract of C. persica exhibited a significant free-radical-scavenging property in the DPPH assay, with an RC50 value of 0.025 mg/mL. However, in the TLC-based qualitative assay, petroleum ether and DCM extracts showed an extremely low level of free-radical-scavenging property. Among the solid-phase extraction fractions of the MeOH extract, the fractions eluted with 60% and 80% MeOH in water exhibited the highest level of free-radical-scavenging activity (RC50 = 0.010 and 0.015 mg/mL, respectively). While DCM extract showed reasonable antibacterial activity against five out of the nine test strains both in the disc diffusion assay and in the resazurin assay, the MeOH extract was highly active against both Escherichia coli and ampicillin-resistant E. coli strains. Among the solid-phase extraction fractions of the MeOH extract, fractions eluted with 80% MeOH in water and 100% MeOH displayed significant antibacterial potencies against both E. coli species. None of the extracts showed any significant toxicity towards brine shrimps (LD50 = >1.00 mg/mL).