The genus Santolina is represented by more than 10 species widely distributed in the Mediterranean area. Species of this genus are used in folk medicine as anthelmintic, antispasmodic and antinfectious. In the current study of the essential oil of Santolina rosmarinifolia L., 31 compounds were determined in the GC-MS, representing 93.40 % of the essential oil with the prevalence of germacrene-D (30.20%), -myrcene (12.00%), tricosane (10.60%), - pinene (10.10%), sabinene (7.00%) and pentacosane (6.70%). The essential oil exhibited a good antibacterial activity against almost food-borne pathogens specially Escherichia coli ATCC 25922, Escherichia coli, and Enterobacter aerogenes with 25 mm, 20 mm, 20 mm, inhibition zone diameters, respectively. Minimum inhibitory concentration (MIC) values for all the bacteria were ranged between 0.16 mg⁄ml and 0.64 mg⁄ml.