Lipid analysis by matrix-assisted laser desorption and ionization mass spectrometry: A methodological approach.

Whereas matrix-assisted laser desorption and ionization time-of-flight mass spectrometry (MALDI-TOF-MS) has gained high importance in the field of protein analysis, surprisingly few studies were published about the use of MALDI for lipid analysis. Lipids, however, are well-suited for MALDI since all experiments can be performed in a sole organic phase and, thus, extremely homogeneous matrix/analyte mixtures are formed. We report here for the first time the application of MALDI-TOF-MS for the analysis of diacylglycerols, phosphatidylcholines, and (poly)phosphoinositides. It is shown that in a matrix of 2,5-dihydroxybenzoic acid the molecular ions (M + 1) of phosphatidylcholines as well as the corresponding adducts of different phosphoinositides are easily detected even in complex mixtures, and thus, detailed data on the fatty acid composition are provided. In contrast, diacylglycerols are mainly detected as the corresponding sodium or potassium adducts, but not as the protonated forms. Fragmentation reactions of fatty acids on the double bonds and on the polar lipid head group are observed to a minor extent in the spectra of all investigated lipids. Generally, choline derivatives are most sensitive toward further fragmentation reactions. Due to its very high sensitivity (up to picomolar concentrations) MALDI-TOF-MS can be used for the direct investigation of biologically relevant lipid mixtures occurring, e.g. , in cell membranes. The analysis of the lipid composition of neutrophilic granulocytes is given as a representative example for future applications.