Effects of Column Length and Particle Diameter on Phospholipid Analysis by Nanoflow Liquid Chromatography-Electrospray Ionization- Mass Spectrometry

The effects of column length and particle size on the efficiency of separation and characterization of phospholipids(PLs) are investigated using nanoflow liquid chromatography-electrospray ionization-tandem mass spectrometry (nLC-ESI-MS-MS).Since PLs are associated with cell proliferation, apoptosis, and signal transduction, it is of increasing interests in lipidomics toestablish reliable analytical methods for the qualitative and quantitative profiling of PLs related to biomarker development inadult diseases. Due to the complexity of PLs, the preliminary separation of PLs is necessary prior to MS analysis. In this study,length of capillary column and the particle size of reversed phase (C18) packing materials are varied to find a reliable conditionfor the high speed and high resolution separation using 8 PL standard mixtures. From experiments, it was found that a capillarycolumn of nLC-ESI-MS-MS analysis for PL mixtures can be minimized to a 5 cm long pulled tip column packed with 3 μm C18particles without losing resolution.