This study was carried out to characterize antimicrobial, antioxidant and cytotoxic activities of Punica granatum rind extract as well as its chemical composition. The main objective of the present study is to reveal the medicinal values of P. granatum rind. Antimicrobial property of P. granatum rind extract was revealed by using two fold broth micro dilution method against Aeromonas hydrophila, Escherichia coli, Edwardsiella tarda, Flavobacterium spp., Klebsiella pneumonia,
Salmonella typhi, Vibrio alginolyticus, V. parahaemolyticus, V. cholerae and Pseudomonas
aeruginosa whereas antioxidant activity of the plant extract was determined with α, α-diphenyl-βpicrylhydrazyl (DPPH) radical scavenging method. Cytotoxic activity of the plant extract was evaluated with Colorimetric MTT (tetrazolium) assay against human breast cell line (MCF-7). Chemical compounds of the plant extract were screened and identified by using gas chromatography-mass spectrometry (GC-MS). The results of the present study showed that the MIC values of P. granatum rind extract ranged from 3.91 to 15.63 mg/l against the tested bacterial isolates. The value of IC50 of the plant extract against DPPH and MCF-7 cell line was 0.144 ± 0.087 ppt and 0.54 ± 0.03 µg/ml, respectively. A total of 14 chemical compounds were successfully identified from P. granatum rind extract and the major compound was Hexanamide, N-phenyl (68.52%.). The findings of the present study indicate promising medicinal values of the plant extract.