Quantification and comparison of some natural sphingomyelins by on-line high-performance liquid chromatography/discharge-assisted thermospray mass spectrometry.

Sphingomyelins obtained from bovine brain, chicken egg yolk and bovine milk fat were analysed. Separation was performed by reversed-phase high-performance liquid chromatography utilizing a binary solvent gradient consisting of n-butanol-water-1-propanol-isooctane. Detection was accomplished by light scattering and on-line discharge-assisted thermospray (plasmaspray) ionization mass spectrometry. The positive ion mass spectra exhibit prominent ions related to the amine base structure and fragments which can be utilized for identification of molecular species. The abundance of the fragments containing the fatty acid reflects relatively well the fatty acid composition of natural sphingomyelin mixtures. Bovine brain sphingomyelin has, for example, two distinctive major molecular species composed of C24:1 and C18:0 acids as indicated by the m/z 630.6 and m/z 548.5 ions, respectively. While egg yolk exhibits ions of m/z 520.6 which indicate the presence of C16:0, bovine milk sphingomyelin has several prominent ions of m/z 632.5, 618.6 and 604.7, reflecting the proportions of C24:0, C23:0 and C22:0, respectively.