Background: The essential oil (EO) extracted from Citrus limon has gained attention for its potential bioactive properties due to its promising chemical composition. This study aimed to comprehensively evaluate the antioxidant and antimitotic activities of Citrus limon EO. Methods: Volatile compounds were thoroughly analyzed using headspace and gas chromatography techniques. Molecular docking studies identified D-limonene and α-bergamotene as having the highest antioxidant potential. Assessment of antioxidant capacity employed 2,2-diphenyl-1-picrylhydrazyl (DPPH) and β-Carotene bleaching assays, alongside the Total Antioxidant Capacity (TAC) assay. Absorption, distribution, metabolism, excretion and toxicity (ADMET) tests were conducted to confirm the EOs suitability for pharmaceutical applications. The antimitotic activity was evaluated using the Lepidium sativum test. Results: Results indicated a 1.99% essential oil yield, with D-limonene identified as the predominant constituent. In the DPPH assay, the half-maximal inhibitory concentration (IC 50 ) value was 1.27 ± 0.03 mg/mL, and the β-carotene bleaching assay exhibited an IC 50 value of 6.74 ± 0.12 mg/mL. The TAC assay determined the EOs total antioxidant capacity as 212.2 ± 1.35 μg Ascorbic acid (AA) eq/mg EO. The essential oil demonstrated significant antimitotic activity, reaching a maximum of 83.33% at a concentration of 250 μg/mL. Conclusions: These findings underscore the noteworthy antioxidant and antimitotic capabilities of Citrus limon essential oil, primarily attributed to D-limonene and other bioactive components.
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