Separation of cellular nonpolar neutral lipids by normal-phase chromatography and analysis by electrospray ionization mass spectrometry

• DOI: 10.1194/jlr.m700521-jlr200
• Publication Date: 2008-01-26T02:35:04Z
• Authors: Patrick M. Hutchins; Robert M. Barkley; Robert C. Murphy
• Keywords: Molecular mass

Neutral lipids are an important class of hydrophobic compounds found in all cells that play critical roles from energy storage to signal transduction. Several distinct structural families make up this class, and within each family there numbers individual molecular species. A solvent extraction protocol has been developed efficiently isolate neutral without complete more polar phospholipids. Normal-phase HPLC was used for the separation cholesteryl esters (CEs), monoalkylether diacylglycerols, triacylglycerols, diacylglycerols a single run extract. Furthermore, minor such as ubiquinone-9 could be detected RAW 264.7 cells. Molecular species lipid can analyzed both qualitatively quantitatively by on-line LC-MS LC-MS/MS strategies. The quantitation >20 CE revealed challenging with Toll-like receptor 4 agonist caused >20-fold increase content CEs cells, particularly those contained saturated (14:0, 16:0, 18:1) fatty acyl groups. Longer chain did not change response activation these