Determination of phospholipids in milk using a new phosphodiester stationary phase by liquid chromatography-matrix assisted desorption ionization mass spectrometry

•New stationary bonded phase was applied in analysis of phospholipids from milk.•Phospholipids were separated for classes on the base of the polarity.•Omega-3 and omega-6 fatty acids were identified using MALDI-TOF MS.•The 46 combination of fatty acids in phospholipids were indentified in milk samples. A methodology employing high performance liquid chromatography coupled with matrix-assisted laser desorption and ionization time-of-flight mass spectrometry has been utilized to determine the quality of phospholipid classes. Home-made phosphoester chemically bonded stationary phase containing diol, phosphate and octadecyl groups (Diol-P-C18) has been employed in the separation of polar lipids from milk. Each phospholipid fraction was collected manually and identified by MALDI-TOF MS.