YEAST LIPID ANALYSIS AND QUANTIFICATION BY MASS SPECTROMETRY

The systematic and quantitative analysis of the different lipid species within a cell or an organism has recently become possible and the general approach has been termed "lipidomics." Traditional methods of identification and quantification of lipid species were laborious processes and it was necessary to use a wide variety of techniques to analyse the different lipid species, especially concerning the assigning of particular acyl chain lengths, hydroxylations, and desaturations to the diverse lipid species. While it is still not possible to quantitatively analyze all lipid species in one fell swoop, great progress has been made with the intensive use of quantitative mass spectrometry approaches. It is now relatively simple to quantify most of the lipid species, including all of the major ones, in a yeast cell. Different degrees of sophistication of mass spectrometric analysis exist and the available techniques and instrumentation are evolving rapidly. Therefore, we have decided to present robust, simple methods to quantify the major yeast lipids by mass spectrometry that should be accessible to anyone who has access to a standard mass spectrometry equipment. The methods to identify and quantify yeast glycero-phospholipids and sphingolipids involve electrospray ionization mass spectrometry using fragmentation to characterize the lipid species. A simplified gas chromatographic method is used to quantify the major sterols that occur in wild-type yeast cells and ergosterol biosynthesis mutants.