Sphingomyelin is more sensitively detectable as a negative ion than phosphatidylcholine: a matrix-assisted laser desorption/ionization time-of-flight mass spectrometric study using 9-aminoacridine (9-AA) as matrix

Phospholipids (PLs) are increasingly analyzed by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) and imaging MS. Different classes of PLs are preferentially detectable either as positive or negative ions depending on the charges of their headgroups. Sphingomyelin (SM) and phosphatidylcholine (PC) occur in virtually all biological samples and both are assumed to be detectable with the same sensitivity (in the positive ion mode) because their headgroups are identical. We will show here that the detectabilities of PC and SM depend on the matrix used. In the presence of 2,5-dihydroxybenzoic acid (DHB) SM is more sensitively detectable in positive ion mode than PC while the use of 9-aminoacridine (9-AA) as matrix inverts the detectabilities. Our explanation is that the preferred generation of negative ions from SM if 9-AA is used as matrix results in a reduced yield of positive ions. It will also be shown that this is not only valid if a simplified model system is investigated, but also if, for instance, extracts from human erythrocytes are investigated. It will also be outlined that this finding is particularly important in the context of imaging studies where no previous separation of the lipids of interest can be performed. Copyright (C) 2011 John Wiley & Sons, Ltd.