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Many biological activities have been reported for the Ilex genus. However, few studies in the literature have reported on guayusa. To address this gap in our knowledge, chemical analysis of guayusa leaves was made. Extracts were obtained by applying Soxhlet, maceration, supercritical CO 2 and pressurised liquid extraction techniques, using water and ethanol as solvent/cosolvent.Extracts were evaluated for their phenolic content and antioxidant capacity. The chemical profile was obtained from HPLC. In raw guayusa leaves were identified caffeine (2.27±0.05 %), protein (15.31±0.07 %) and lipids (11.81±0.14 %). Extracts presented the highest phenolic content (156.56±1.32 mg GAE g -1 ) and the best antioxidant activity (EC 50 = 61.85±0.21 µg mL -1 ) when water was used as solvent/cosolvent. Through HPLC, three main substances were determined and quantified in the extracts: caffeine, theobromine and 5-caffeoylquinic acid. Based on these results, guayusa may be considered as a natural source of compounds with potential application in the food and pharmaceutical industries.