A single run LC-MS/MS method for phospholipidomics

• Published in: Analytical and bioanalytical chemistry 2013, Vol.405 (1), p.203-213
• Main Authors: Buré, Corinne, Ayciriex, Sophie, Testet, Eric, Schmitter, Jean-Marie
• Format: Article
• Language: English
• Subjects: Acyltransferases - chemistry; Analytical Chemistry; Atomic properties; Biochemistry; Biochemistry, Molecular Biology; Carbon; Carbon - chemistry; Characterization and Evaluation of Materials; Chemical properties; Chemical Sciences; Chemistry; Chemistry and Materials Science; Chemistry Techniques, Analytical; Chromatography; Chromatography, High Pressure Liquid - methods; Chromatography, Liquid - methods; Fatty acids; Food Science; Fungi - metabolism; Genomics; Identification and classification; Laboratory Medicine; Life Sciences; Lipids; Lipids - chemistry; Liquid chromatography; Mass spectrometry; Methods; Microbiological chemistry; Monitoring/Environmental Analysis; Neurobiology; Neurons and Cognition; Original Paper; Phosphatidic Acids - chemistry; Phosphatidylcholines - chemistry; Phosphatidylethanolamines - chemistry; Phosphatidylglycerols - chemistry; Phosphatidylinositols - chemistry; Phosphatidylserines - chemistry; Phospholipids; Phospholipids - chemistry; Physiological aspects; Polyethylenes; Quantitative Methods; Reproducibility of Results; Scientific imaging; Shotguns; Strain; Structural Biology; Tandem Mass Spectrometry - methods; Yeast; Yeast fungi; Yeasts
• ISSN: 1618-2642; 1618-2650
• DOI: 10.1007/s00216-012-6466-9

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.