Eucalyptus leaves are a secondary product of wood production. The pressurized liquid extraction (PLE) technique is an attractive process due to its advantages. The objective of this work was to obtain extracts of Eucalyptus saligna leaves using the PLE technique, and to characterize different extraction time fractions in relation to bioactive compounds and antioxidant and antimicrobial activities. The extraction of E. saligna leaves was carried out in a jacketed extractor at 40 °C, pressure of 10.4 MPa, with a solvent flow of 2.5 mL.min-1 , using 95% ethanol. The system remained pressurized for 30 minutes in static extraction, followed by 180 minutes in dynamic extraction. The levels of total phenols, flavonoids, antioxidant activity and antimicrobial potential against Staphylococcus aureus and Escherichia coli were evaluated. The extraction showed an overall yield of 12.3% after 180 minutes. The total concentrations of phenols and flavonoids were 618.57 mgGAE.g-1 and 160.28 mgQE.g-1 respectively. The extracts showed good antioxidant and antimicrobial potential, with IC50 between 0.016 and 0.043 mg.mL-1 , and MIC against S. aureus and E. coli between 0.313 and 1.25 mg.mL-1. PLE demonstrated efficiency in obtaining extracts of E. saligna leaves with bioactive properties.