Profiling of phospholipids in lipoproteins by multiplexed hollow fiber flow field-flow fractionation and nanoflow liquid chromatography-tandem mass spectrometry

This Study describes a Coupled analytical method to carry out the systematic profiling of phospholipids (PLs) in high-density lipoproteins (HDL) and low-density lipoproteins (LDL) from human blood plasma. HDL and LDL of healthy human plasma samples were separated by size and collected on a semi-preparative scale Using multiplexed hollow fiber now field-flow fractionation (MxHF5). Phospholipid mixtures contained in the resulting HDL and LDL fractions were analyzed by shotgun nanoflow liquid chromatography-tandem mass spectrometry (nLC-ESI-MS-MS). We utilized a dual scan method for the separation and Simultaneous characterization of complicated PL mixtures by nLC-ESI-MS-MS, such that phosphatidylcholine (PC) and phosphatidylethanolamine (PE) molecules were detected in positive ion mode in a first LC run. In a second LC run, phosphatidylinositol (PI), phosphatidlglyceron (PG), and phosphatidic acid (PA) were detected in negative ion mode. In this Study, a total of 56 PLs from HDL and 52 PLs from LDL particles were characterize(] by their molecular structures from data dependent collision-induced dissociation (CID) experiments, and their relative abundances were compared. (C) 2010 Elsevier B.V. All rights reserved.