Chromatographic lipid profiling of stress-exposed cells

Lipidomics is an emerging field of science not only due to its integral part of cell biology and biophysics but also due to the key role of lipids in the modulation of membrane physical properties, signaling, and cell death regulation. The aim of this study was to characterize changes in N-palmitoyl ceramide concentration and in the global lipid profile in macrophages challenged by oxidized low-density lipoprotein and nutrient deprived hepatocytes. For this purpose, a quantitative targeted method based on gas chromatography-mass spectrometry for the determination of total N-palmitoyl ceramide concentrations in the cellular membranes of cells under stress was used. Ultrahigh-performance liquid chromatography-quadrupole-time of flight mass spectrometry was applied for the comprehensive profiling of lipids. In essence, we found that both models of cellular stress caused an increase in N-palmitoyl ceramide levels. In addition, increased levels of other ceramides were observed as well as up- and down-regulation of several other lipid species.