IIn recent years, due to the possible side effects of synthetic drugs, people have turned to natural drugs in combating diseases. Plants are important natural resources with these properties. In our study, some biological activities of the aerial parts of Gentiana olivieri were determined. In this context, the plant was extracted with ethanol in a Soxhlet apparatus. The antioxidant activity of the plant was measured with Rel Assay TAS and TOS kits. Its antimicrobial activity was determined by the agar dilution method. Determination of total phenolic content was determined using the Folin-Ciocalteu reagent. Total flavonoid quantification was performed using aluminum chloride assay. As a result of the analyzes, the TAS value of the ethanol extract of the aerial parts of Gentiana olivieri was determined as 7.775±0.114 mmol/L, the TOS value as 12.252±0.094 μmol/L and the OSI value as 0.158±0.002. In addition, total phenolic content was measured as 104.92±1.40 mg/g, total flavonoid content as 73.83±1.29 mg/g. In addition, it was determined that the plant extract was effective against standard bacterial and fungal strains at extract concentrations in the range of 50-200 μg/mL. As a result, it was determined that Gentiana olivieri could be an important natural source in terms of antioxidant and antimicrobial activity.