tThe influence of different parameters on the Supercritical Carbon Dioxide (SCO2) extraction of ItalianPropolis were studied with attention to extraction yield and chemical composition of obtained fractions.Operating parameters of SCO2were optimized using central composite design. Analysis by multipleregression indicated that pressure and time have a major linear effect on the extraction yield and extractcomposition.Propolis SCO2extracts present different chemical content compared to the ethanolic extract obtainedwith ultrasound assisted extraction (UAE). SCO2extract can be used to perform further UAE ethanolextraction obtaining a flavonoid containing extract. These findings indicate two possible applications ofsupercritical carbon dioxide for Propolis extraction, one to obtain lipophilic fractions enriched in specificconstituents, the other as pre-treatment of the crude material to facilitate the further extraction withethanol.