Determination of Molecular Species of Phosphatidylcholines as 2-Anthrylurethanes by Reversed-phase HPLC with Fluorescence Detection and on-line Electrospray Ionization Mass Spectrometry

Abstract

A highly sensitive method for the determination of phosphatidylcholine (PC) molecular species was established. PCs from soybean, mackerel (Scomber japonicus) muscle, saury (Cololabis saira) muscle and salmon (Oncorhynchus keta) roes were hydrolyzed with phospholipase C and the diacylglycerols thus obtained were converted to 2-anthrylurethanes, which were then subjected to reversed-phase high-performance liquid chromatography (HPLC) with fluorescence detection and on-line electrospray ionization mass spectrometry (ESI/MS). PC anthrylurethanes were found to be clearly separated into molecular species at the femtomole level by fluorescence examination. Positive ESI/MS of the derivatives gave prominent pseudomolecular [M+Na] ⁺ ions, by which individual molecular species could be identified.