Introduction: Candidiasis therapy is a complicated concern because of the occurrence of resistance to antifungal agents. We studied the anti-fungal effects of Ferula macrecolea essential oil (FME) against Candida albicans resistant and sensitive strains, as well as its cytotoxic effects against normal and cancer cell lines. Methods: The minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of F. macrecolea essential oil against C. albicans ATCC 5027 and C. albicans ATCC 76616 were studied by broth-microdilution approach. The cytotoxicities of FME on HGF1-PI (normal gingival cell line) and HepG2 (liver cancer cell line) cells were also studied. Results: The main components of essential oil were terpinolene (71.25%), n-nonanal (6.32%), and linalool (3.95%), respectively. The MIC and MFC of FME on C. albicans sensitive to nystatin were 1.6 and 2.0 μg/mL, respectively. The MIC and MFC of FME on nystatin-resistant strains were 3.3 and 4 μg/mL, respectively. The MIC and MFC of terpinolene on C. albicans sensitive to nystatin were 0.8 and 1.0 μg/mL, respectively. The MIC and MFC of terpinolene on nystatin-resistant strains were 2 and 2.4 μg/mL, respectively. The essential oil and terpinolene had no significant cytotoxic effects against normal cells. Conclusion: We revealed the promising antifungal effect of F. macrecolea essential oil and its main component, terpinolene, against C. albicans sensitive and resistant to nystatin with no significant toxicity on normal cells.