This paper focuses on the chemical composition, antioxidant, cytotoxicity (brine shrimp lethality and hemolytic assay) and antimicrobial activities of essential oil of mace (Myristica fragrans) oil. We have found 33 compounds as evidenced by GC-MS analysis. The most prominent of these compounds are safrole, β-phellandrene, 3-p-menthene and L-terpinen-4-ol. Antioxidant activity of the mace essential oil was examined using the free radical scavenging assay (21.95 μg/mL), β-carotene in linoleic acid and percent inhibition in linoleic acid (67.9 %) system. Four pathogenic fungi, 4 gram positive and 2 gram negative bacteria were used for antimicrobial activity of oil. Aspergillus niger and Streptococcus mutans proved to be most sensitive with MIC values i.e. 1.01 and 1.72 mg/mL, respectively. The mace essential oil exhibited toxicity with LC50 value 31.05 μg/mL cytotoxicity against nauplii of Artemia salina and minimum hemolytic activity was recorded as 5.21% at 0.5 mg/mL concentration.