Some factors in the salting-out extraction (SOE) method play a major role. The aim of this study was to investigate the interaction effects of the phase forming components and consequently select the best conditions to achieve a highly efficient recovery of phenolic compounds from walnut green husks (Juglans regia L.) using mixtures of ethanol and aqueous ammonium sulfate solutions. According to the results that were analyzed by response surface methodology, the optimal extraction conditions were obtained at ethanol: salt: water ratio of 34.8: 15.1: 54.4 (w/w) at a pH of 6-6.5 and 25 °C. At the optimal conditions, the overall phenolic and flavonoid content, and antioxidant activity were significantly higher than obtained by the conventional method. In addition, at a higher scale (i.e., 5 kg), similar results were obtained. Thus, it can be concluded that SOE has the potential to be scaled up for the simultaneous separation and purification of compounds from plant biomass. This paper is addressing extraction techniques, measurement, and characterization of new natural phenolic compounds from an agricultural by-product and valorization of waste.
Keywords: Agricultural by-product; extraction; phenolics; two phase separation; walnut green husk.