Soybean phospholipids-phosphatidylethanolamine (PE), phosphatidylinositol (PI),, and phosphatidylcholine (PC)-were separated by normal-phase high performance liquid chromatography. The system was operated in a gradient mode. The experimental variables were gradient time and mobile phase composition. The experimental results showed that PE, PI and PC were resolved by three step-change gradient modes which employed ternary systems of hexane/1-propanol/water (58/40/2 and 56/40/4 by vol.%) and methanol/1-propanol/water (80/18/2, by vol.%) for the gradient times of 10, 30 and 76 min, respectively.