Protocol for rapid and accurate quantification of phospholipids in yeast and mammalian systems using LC-MS

We describe a protocol for measuring phospholipid class and fatty acid composition in the budding yeast Saccharomyces cerevisiae using a liquid chromatography-mass spectrometry (LC-MS)-based approach. We compile a mass spectral-retention time library verified for major phospholipids in the budding yeast and showcase the profiling process of phospholipid compositions in mutants with defective syntheses of phosphatidylethanolamine (PE) and phosphatidylcholine (PC). We further provide methods for extracting and quantifying phospholipids in mammalian systems. For complete details on the use and execution of this protocol, please refer to Fang et al. (2022).