The extraction of phenolic compounds from eucalyptus leaves was optimized using heat and ultrasound-assisted
techniques, and the bioactive potential of the resulting extract was assessed. The independent variables,
including time (t), solvent concentration (S), and temperature (T) or power (P), were incorporated into a fivelevel
central composite design combined with Response Surface Methodology. Phenolic content was determined
by HPLC-DAD-ESI/MS and used as response criteria. The developed models were successfully fitted to the
experimental data to identify the optimal extraction conditions. Heat-assisted extraction proved to be the most
efficient method for phenolic recovery, yielding 27 ± 2 mg/g extract under optimal conditions (120 min,
76.5 ◦C, and 25 % ethanol, v/v). The extracts exhibited a high concentration of phenolic glycoside derivatives,
including gallotannin, quercetin, and isorhamnetin. These findings suggest that the extracts hold promise as
natural additives in food technology, owing to their moderate antimicrobial activity and strong antioxidant
properties.