Quantitative Analysis of Glycerophospholipids in Mitochondria by Mass Spectrometry

Lipids draw increasing attention of cell biologists because of the wide variety of functions beyond their role as building blocks of cellular membranes. Mitochondrial membranes possess characteristic lipid compositions that are intimately associated with mitochondrial architecture and activities. Therefore, quantitative assessment of lipids in isolated mitochondria is of importance for mitochondrial research. Here, I describe our workflow for quantitative analysis of glycerophospholipids in mitochondria with a focus on purification of pure mitochondrial fractions from yeast and cultured mammalian cells as well as improved settings for the analysis of cardiolipin by nano-electrospray ionization mass spectrometry.