A single run LC-MS/MS method for phospholipidomics.
Liquid chromatography coupled to tandem mass spectrometry has been compared to shotgun analysis with the objective of finding the best compromise for a single run analysis of whole cell phospholipids. Hydrophilic interaction liquid chromatography (HILIC), normal phase (NP), and reversed phase (RP) liquid chromatography were evaluated with reference phospholipids belonging to phosphatidic acid (PA), phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylglycerol (PG), phosphatidylinositol (PI), and phosphatidylserine (PS) classes. NP-HPLC- and RP-HPLC-ESI-MS/MS were applied to yeast phospholipidome analysis, using a wild-type strain and two strains defective for acyltransferases that are known to be involved in de novo phospholipid synthesis or phospholipid remodeling. The MRM mode was used for relative quantitation of individual compounds based on reference phospholipids bearing fatty acid chains with an odd number of carbon atoms. Combined LC-MS/MS was found superior to shotgun analysis, leading to a larger number of quantified species than shotgun analysis. Finally, RP-HPLC-MS/MS was the preferred method for its higher selectivity, robustness, and better repeatability.
Chimie Biologie des Membranes et Nanoobjets CBMN-UMR 5248 Centre de Génomique Fonctionnelle Université Bordeaux 2, Université de Bordeaux, 146, rue Léo Saignat, 33076 Bordeaux cedex, France. c.bure@cbmn.u-bordeaux.fr
Corinne
C
Buré
Sophie
S
Ayciriex
Eric
E
Testet
Jean-Marie
JM
Schmitter
D000217:Acyltransferases / Q000737:chemistry; D002244:Carbon / Q000737:chemistry; D002623:Chemistry Techniques, Analytical; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002853:Chromatography, Liquid / Q000379:methods*; D005658:Fungi / Q000378:metabolism; D008055:Lipids / Q000737:chemistry; D010712:Phosphatidic Acids / Q000737:chemistry; D010713:Phosphatidylcholines / Q000737:chemistry; D010714:Phosphatidylethanolamines / Q000737:chemistry; D010715:Phosphatidylglycerols / Q000737:chemistry; D010716:Phosphatidylinositols / Q000737:chemistry; D010718:Phosphatidylserines / Q000737:chemistry; D010743:Phospholipids / Q000737:chemistry*; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*
D008055:Lipids; D010712:Phosphatidic Acids; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D010715:Phosphatidylglycerols; D010716:Phosphatidylinositols; D010718:Phosphatidylserines; D010743:Phospholipids; C483858:phosphatidylethanolamine; D002244:Carbon; D000217:Acyltransferases