Background and Aim: The present study investigated the antimicrobial, and cytotoxic activities of the methanol extract of Nectaroscordum tripedale leaves.Materials and Methods: Methanolic extract of Nectaroscordum tripedale was investigated for its phytochemical components, antimicrobial activity and cytotoxicity. The antibacterial potentialities of methanol extract of Nectaroscordum tripedale leaves were investigated by the disc diffusion and broth dilution method against five bacterial isolates including three food-borne pathogens (Staphylococcus aureus, Listeria monocytogenes and Escherichia coli) and two healthcare-associated pathogens (Methicillin resistance Staphylococcus and Pseudomonas aeruginosa) and cytotoxicity activity were evaluated on acute myeloid leukemia cell line (KG-1a) and normal lymphocyte cells and  the effect of Nectaroscordum tripedale extract and methotrexate in 0.01, 0.1, 1, and 2 mg/ml concentrations on these cells were compared.Results: The extract was found to contain 27 chemical compounds. The maximum zone of inhibition was observed in Methicillin Resistant Staphylococcus aureus (MRSA). The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of Nectaroscordum tripedale for E. coli and MSRA were similar (166 and 322 µg/ml, respectively). While the high level of MIC and MBC is related to Pseudomonas aeruginosa. The percentage of lysis for extract and methotrexate on the KG-1a was seen after 24 hours.Conclusions: In comparison with methotrexate, Nectaroscordum tripedale had minimal effect on normal cells. The present study revealed that the methanol extract of Nectaroscordum tripedale leaves has significant antibacterial activities along with moderate cytotoxicity's on the cancer cells that may lead to new drug development.