4.7. Preparation of the Ethyl Acetate Extract

An amount (250 g) of P. subscutatum dried leaves were minced and then macerated in ethyl acetate for one hour at room temperature. The process was repeated three times and the resulting extracts were combined, filtered, and distilled under vacuum at 35 °C, to afford 4.19 g of an oily residue. Chlorophyll was then removed by SPE, passing the residue through a column packed with C₁₈ reversed phase (50 g). Elution with 80% aqueous methanol gave, after evaporation, 1.76 g of a chlorophyll-free residue.