Excellent combination of counter-current chromatography and preparative high-performance liquid chromatography to separate galactolipids from pumpkin

Galactolipids in the fruits of Cucurbita moschata (pumpkin) could not be completely separated by high-performance liquid chromatography (HPLC). Preparative HPLC was not available for preparing major galactolipid monomers in pumpkin. In the present paper, a combination of high-speed counter-current chromatography (HSCCC) and preparative HPLC was used for preparing the galactolipids. A fraction containing galactolipids (Fr60) from the purification of the n-butanol extract of pumpkin by macro-porous resin column chromatography was first separated by HSCCC to result in three sub-fractions of each containing two galactolipid monomers. The three sub-fractions were further separated by preparative HPLC respectively to yield six galactolipid monomers with purity more than 96%. The method is a good one for preparing galactolipid monomers from plant materials for the studies of bioactivities.