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Angelica acutiloba is one of the most intensively cultivated medicinal plants in Korea. The roots of this plant have been used as an important herbal drug, especially for the treatment of various female disorders, as the traditional therapy in Korea and other Asian countries. Consumption of its fresh leaves as a healthy vegetable has recently increased. In this study, essential oil fractions were extracted from the roots and leaves of this plant by steam distillation. Compositions of the two oils were compared by gas chromatography-mass spectrometry (GC-MS). The antibacterial activities of the essential oil were determined against three strains of Escherichia coli. DPPH radical scavenging and reducing power tests were performed to evaluateits antioxidant activities. The cytotoxic activities of the essential oil against a human breast and a uterine cancer cell line were estimated by MTT tests. Additionally, the morphological changes after treatment of the oil fraction were observed under a microscope. The essential oil fraction and its main components, Z-ligustilide and butylidene phthalide, inhibited the growth of three E. coli strains examined, with minimum inhibiting concentrations (MICs) ranging from 1.0 mg/ml to 8.0 mg/ml. Additionally, the essential oil fraction of A. acutiloba exhibited significant DPPH free radical scavenging activity and reducing power. Significant cytotoxic activities of the A. acutiloba essential oil were observed for human uterine (Hela) and breast (MCF-7) cancer cell lines.