Supercritical fluid extraction (SFE) was used for fractionation of grape seed to be applied in natural products of the food, cosmetic, or pharmaceutical industry. A two-step process was developed, consisting of conventional alcoholic extraction followed by SFE with carbon dioxide. The effects of pressure and concentration of co-solvent (ethanol) at constant temperature and solvent to feed-ratio were evaluated. The antioxidant activity of extracts and raffinates was measured with the DPPH* free radical scavenging assay. It was observed that supercritical carbon dioxide can be used in the described process for fractionation of an alcoholic grape seed extract to obtain a product with noticeably high antioxidant activity and a second one with no antioxidant activity or pro-oxidant activity