A normal high performance liquid chromatography with UV detection was developed for the determination of phosphatidyl cholines（PC），phosphatidyl ethanolamines（PE） and phosphatidyl inositol（PI） in soybean and soybean oil samples.Effects of extraction solvent，ultrasonic power，ultrasonic time，extracting temperature and purification method on the extraction efficiency were investigated.Effects of different stationary phases on the separation efficiency were studied.The optimal extraction conditions were as follows：extraction solvent:chloroform-methanol（2∶1，by volume）；ultrasonic extraction power：1 500 W；extraction time：30 min.The purification and separation of three phospholipid compounds were performed on an amine solid phase extraction column and a silica gel column with a mobile phase of hexane-isopropanol-1% HAc solution（8∶8 ∶1，by volume）.Under the optimized conditions，the linear ranges for the PC，PE and PI were in the range of 0.08-8.00，0.15-15.00,0.30-20.00 g?L-1，respectively.The limits of quantitation for the PC，PE and PI were 0.021，0.050,0.060 g?L-1，respectively,and the limits of detection were in the range of 8-23 mg?L-1.The recoveries of three phospholipids in fortified soybean and soybean oil sample ranged from 85% to 108%.The intra day RSDs and inter day RSDs of three phospholipid compounds were no more than 4.7% and 8.6%，respectively.