Behaviour of plasmalogens during high-performance liquid chromatography on a silica column with a mobile phase containing phosphoric acid

Silica high-performance liquid chromatographic separation of phospho- and sphingolipids of biological origin using a mobile phase containing phosphoric acid leads to gradual hydrolysis of plasmalogens during their passage through the column. The resulting 2-acyl lyso analogues give rise to peaks that tail in the direction of the parent intact plasmalogen. Tailing can be prevented by previous complete acid hydrolysis of plasmalogens. Direct high-performance liquid chromatographic profiling of phospholipids, their plasmalogens (as 2-acyl lyso analogues) and sphingolipids is probably the method of choice for the diagnosis of patients with deficient plasmalogen biosynthesis caused by peroxisomal abnormalities.