Hot-pressurized fluid extraction of flavonoids and phenolic acids from Brazilian propolis and their cytotoxic assay in vitro

This work has examined the hot-pressurized fluid extraction of seven flavonoids, caffeic acid phenethyl ester and four phenolic acids from Brazilian propolis lumps generating, during the process, fat- and water-soluble extracts. The solid content of water-soluble extract obtained by hot-pressurized water in the presence of 29% natural surfactant was 35.2 mg/mL and was 44% greater than that obtained without natural surfactant. Furthermore the amount of the seven flavonoids and caffeic acid phenethyl ester in the fat-soluble extract exceeded those in the water-soluble sample while, on the other hand, the amount of the four phenolic acids in the water-soluble extract was more than those in the fat-soluble extract. Our findings show that the total solid content and the amount of these 12 active compounds produced by the emulsified hot-pressurized water are 36% and 7% higher, respectively, than those produced by emulsified water at atmospheric pressure. The EC₅₀ value of the free radical scavenging activity of 1,1-diphenyl-2-picrylhydrazyl of the emulsified hot-pressurized water extract was the lowest, and presented the strongest anti-oxidation ability among all the extracts. In vitro cytotoxicity indicated that the water-soluble extract strongly suppressed the growth of leukemia (HL-60, U937), lung cancer (A549, CH27) and liver cancer (Hep G2, Hep 3B) cells in a concentration-dependent behavior.