Lipidomic analysis of porcine organs using hydrophilic interaction liquid chromatography and mass spectroscopy

Abstract

The identification and quantitation of all lipids in complex biological tissues is the first step towards the understanding how lipids function in a biological system and the elucidation of the mechanism of lipid-related diseases including obesity, atherosclerosis, cancer, cardiovascular diseases, etc. Our optimized nontargeted method using hydrophilic interaction liquid chromatography (HILIC) coupled to electrospray ionization mass spectrometry (ESI-MS) was used for the characterization of the lipidome, mainly glycerophospholipids and sphingolipids in porcine brain, heart, kidney, liver, lung, spinal cord, spleen and stomach. Individual lipid classes are quantified based on the peak integration of individual lipid classes separated in the HILIC mode multiplied by their response factors and correlated by sphingosylethanolamine (d17:1/12:0) as a single internal standard. Subsequently, relative abundances of deprotonated molecules [M-H]– in negativeion ESI mass spectra are used for determination of lipid species concentration inside individual classes. The approach using [M-H]– ions enables to obtain detailed information about phosphatidylethanolamines and their different forms of fatty acyl linkage on the glycerol skeleton (plasmalogens and ether analogs), phosphatidylinositols and hexosylceramides in studied organs. Our results provide important knowledge about lipid representations in vital porcine organs and can be applied for future metabolic studies on human to investigate serious lipid-related human disorders.