Hypericum olympicum L. (Hypericaceae) thrives in Turkey, S. Balkans, Greece, and Serbia. While the decoction prepared is internally used for stomachache; used externally for inflamed wounds and cuts in traditional. This research aims to examine the in vitro antimicrobial of the essential oil (EO) obtained using H. olympicum aerial parts. The EOs from H. olympicum aerial parts and roots were distilled and characterized by GC-FID and GC/MS systems, respectively. Germacrene D (26.4%) and α-pinene (8.5%) were found as the main components of the aerial parts of H. olympicum EO. The EO of H. olympicum root were characterized with undecane (38.9%), α-pinene (14.2%), and 3-methyl nonane (9.2%). The antimicrobial activity of EO of H. olympicum aerial parts was observed S. pyogenes (219 µg/mL), E. coli (8.75 mg/mL), S. aureus (109 µg/mL) and C. albicans (55 µg/mL). Essential oil of H. olympicum aerial parts is differently reported in the literature while the chemical composition of EO of H. olympicum root is identified for the first time. According to our findings, the essential oil of the H. olympicum aerial parts showed a strong effect against skin wound pathogens.