© IJSM 2022.Essential oils have several biological activities such as antimicrobial, antioxidant, proliferative, and anti-inflammatory. This study aimed identification of bioactive compounds found in Elemi essential oil (EO) and to determine the anti-quorum sensing and cytotoxic activities of EO. In this study, bioactive compounds of EO were analyzed using GC-MS, and the antibacterial activity of elemi was screened against Staphylococcus aureus ATCC 25923, Methicillin-Resistant Staphylococcus aureus ATCC 43300, Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, Pseudomonas aeruginosa PAO1. Anti-biofilm activity and pyocyanin production on P. aeruginosa PAO1 were also investigated. The effect of EO on cell viability was also analyzed by thiazolyl blue tetrazolium bromide (MTT) and neutral red uptake (NR) assay in fibroblast cells. According to GC results, the major component of EO was determined as limonene (55%). A sub-MIC of elemi essential oil inhibited biofilm formation and pyocyanin production by 43% and 56%, respectively. On the other hand, EO also had an acute effect on the mitochondrial and lysosomal activities of fibroblast cell lines. Mitochondrial and lysosomal activities were significantly decreased when EO concentrations were applied for 24 and 48 hours (p<0.05). In conclusion, EO has inhibitory activity on biofilm formation and pyocyanin production, and also the lower doses of oil have no toxic effects on fibroblast cells. However, higher doses of EO have more cytotoxic effects on mitochondrial activity rather than the lysosomal activity of fibroblast cell lines. It is thought that EO exhibits these activities due to the amount of limonene in its content.