A fast liquid chromatography-mass Spectrometry methodology for membrane lipid profiling through hydrophilic interaction liquid chromatography

• A new HILIC method for separation of several membrane lipids is proposed.
• Lipids here investigated belong to different classes (PL, glyco- and betaine lipids).
• Lipids are obtained from different biological extracts.
• Analysis of lipids deriving from eukaryotic animals and plants is reported.
• ESI-MS carried out on TQ is used as detector of lipids separation.

In this paper, we report the development of a new method based on HILIC-ESI-MS for the separation of several different membrane lipid classes and their detection on a triple quadrupole mass spectrometer using Precursor Ion (PIS) and Neutral Loss (NL) scanning in positive ion mode. Four different columns were tested for their ability to separate, under different conditions, a mixture of 14 lipid standards containing 7 glycerophospholipids (GPL), 2 glycosphyngolipids (GSL), 3 glycolipids (GL) and 2 betaine lipids (BL).The best separation was obtained using a Lichrosphere DIOL column as stationary phase and water (10 mm ammonium acetate)/acetonitrile gradient elution as mobile phase which allows the separation of the 14 lipid classes within 35 min runtime. Our method was successfully tested for the separation and analysis of crude lipid extracts obtained from a green alga (Jaoa bullata), a dinoflagellate (Peridinium cinctum) and a plant (Vitis vinifera cv. Corvina)