The essential oils of the aerial parts and roots (APEO and REO) of Pulicaria vulgaris subsp. dentata (Sm.) Batt. were analyzed by GC and GC-MS. Twenty-one compounds (97.2% of the total oil composition) were identified in the REO, with Î³-irone (39.2%), 7-epi-silphiperfol-5-ene (19.3%), 2,5-dimethoxy-p-cymene (8.5%) and Î³-himachalene (8.0%) as main constituents. The APEO was rich in cis-Î²-guaiene (59.7%), together with eighteen other compounds, accounting for 93.2% of the total oil composition. Antiradical and enzymatic antioxidant properties of the isolated oils were also evaluated. The REO exhibited the highest antioxidant activity using DPPH, ABTS, catalase and paraoxonase tests. Antimicrobial, cytotoxic and tyrosinase inhibiting properties of these essential oils have been also studied. APEO exhibited significant activity towards Pseudomonas syringae and Agrobacterium tumefaciens. The REO showed the most significant cytotoxic effect against HeLa cell line and the highest tyrosinase inhibition activity.