Background: Ferula tingitana L. (Apiaceae) has been considered to have abortive and menstruation-inducing properties. It used to treat sore throat, fever, indigestion, and pains. Objectives: The objective of this study is to establish the chemical composition of the essential oil of flower, leaves of F. tingitana, and to throw light on antimicrobial, cytotoxic activities of Libyan plant. Materials and Methods: The chemical composition of the essential oil of flower (0.06% w/v) and leaves (0.1% w/v) of F. tingitana was comparatively analyzed by gas chromatography/mass spectrometry using nonpolar column DB-5. Results: A total of 28–32 components were identified, 15 being common in both samples. The main constituents of both flower- and leave-derived oil samples were α -thujene (13.5%–2.3%), elemol (8.9%–8.3%), eudesmol (0.6%–9.7%) and cadinol (2.2%–13.8%), respectively. The principle difference was a considerably more pronounced sesquiterpenes presence in the leaves-oil, amounting to 74.0%, than in the flower counterpart (39.9%). Caryophyllene (5.6%) and elemol (8.9%) were the major sesquiterpenes detected in flower-oil while leaves-oil showed less amounts of sesquiterpenoid hydrocarbons (27.7%) and represented by eudesmadiene (9.0%). On the contrary, while remaining the dominant group in both oil samples, monoterpenoids are relatively more abundant in flower-derived oil constituting 57.7% versus 24.5% detected in leaves. Conclusion: Leaves-oil sample being mostly efficient as antibacterial against Bacillus subtilis and Neisseria gonorrhoeae with potency 48.3, 41.9% compared to tetracycline standard antibacterial drug. The essential oil samples revealed marked in vitro cytotoxicity against breast (MCF7), cervical (HELA) and liver(HEPG2) carcinoma cell lines with IC50% (6.9, 4.8), (8.6, 10.9), and (4.4, 4.2) for the flower-, leaves-derived oil sample, respectively.