The essential oils of Centaurea pulcherrima Willd. var. pulcherrima (Asteraceae) were isolated by hydrodistillation (HD) and a microwave distillation (MD), than characterised by GC-FID and GC-MS. A total of 58 and 57 compounds were identified, constituting over 93.7%, and 91.6% of volatile oil composition of C. pulcherrima var. pulcherrima, respectively. Sesquiterpene hydrocarbons were shown to be the main group of constituents (HD: 42.4% versus MD: 51.5%). The major component of the oils of C. pulcherrima var. pulcherrima was germacrene D (HD, 17.8% versus MD, 23.2%). The antimicrobial activity of the isolated essential oils of the plant was also investigated, and they showed good antibacterial activity against to tested Gram-positive bacteria, especially to M. smegmatis and a yeast-like fungus C. albicans.