The use of the matrix 9-aminoacridine has been recently introduced in matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry analysis of both anionic and cationic phospholipids. In the present study, we take advantage of this technique to analyze the lipids of porcine olfactory mucosa and a membrane fraction enriched in cilia. Thin-layer chromatography (TLC) and (31)P-NMR analyses of the lipid extracts were also performed in parallel. MALDI-TOF-MS allowed the identification of lipid classes in the total lipid extract and individual lipids present in the main TLC bands. The comparison between the composition of the two lipid extracts showed that: (1) cardiolipin, present in small amount in the whole olfactory mucosa lipid extract, was absent in the extract of membranes enriched in olfactory cilia, (2) phosphatidylethanolamine species were less abundant in ciliary than in whole epithelial membranes, (3) sulfoglycosphingolipids were detected in the lipid extract of ciliary membranes, but not in that of epithelial membranes. Our results indicate that the lipid pattern of ciliary membranes is different from that of whole-tissue membranes and suggest that olfactory receptors require a specific lipid environment for their functioning.