Quantitative analysis of aminophospholipids by high-performance liquid chromatography using succinimidyl 2-naphthoxyacetate as a fluorescent label.

A simple and rapid high-performance liquid chromatographic procedure for the quantitative analysis of ethanolamine- and serine-containing phospholipids in tissue is described. The technique involves reaction of lipid extracts with succinimidyl 2-naphthoxyacetate to give fluorescent derivatives of aminophospholipids. Reaction products are separated by a silica gel column with gradient elution. The eluate is monitored by fluorescence detection at 228 nm (excitation) and 342 nm (emission). Ethanolamine and serine plasmalogens can be measured indirectly by converting their derivatives into lysophosphatidylethanolamine and lysophosphatidylserine derivatives with exposure to hydrogen chloride fumes. The method is highly sensitive and selective.