Purifiication of Artepillin C (ARC) with Anticancer Activities from Supercritical CO2-Ethanol Extracts of Brazilian Green Propolis by Using Adsorption Column Chromatography

Abstract

Phenolic compounds with varying phenolic structures are present in both terrestrial and marine organisms. Artepillin C (ARC) is a prenylated phenolic compound produced specifically in Brazilian green propolis. Earlier studies have demonstrated the potential of ARC as a dietary supplement for cancer prevention due to its ability to suppress tumor growth and metastasis. Here we report the development of an effective adsorption chromatography combined with supercritical CO2 extraction for the massive purification of ARC. A mixture of supercritical CO2 and ethanol extract of Brazilian green propolis was passed through a stainless chromatographic column packed with protruding metals at 4000 psi and 50 degrees C to remove wax from the column's base. ARC mixed with flavonoids collected from the top of the column was applied to another stainless column filled with silica gel under similar working conditions. After flavonoids were removed, strongly adsorbed ARC at the top portion of the column was collected by ethanol elution. The purity of ARC isolated using this method was between 90% and 95%, as indicated by HPLC chromatograms. Purified ARC strongly inhibited cancer cell division based on its cytotoxic effects on HCT116 human colon cancer cells, resulting in a signif-icantly increased distribution of cells in the S phase. ARC also blocks the motility of HCT116 cells, indicating its ability to suppress tumor metastasis. The developed chromatographic purification protocol can be used to isolate prenylated phenolic compounds from marine organisms.