Application of a thin-layer chromatography—flame-ionization detection system for the determination of complex lipid constituents

Abstract

A simple and rapid method for the determination of relative amounts of complex lipid constituents was developed using a thin-layer chromatography—flame ionization detection (TLC—FID) system. The methanolysis products from a sphingoglycolipid (ceramide monohexoside) were separated on Chromarod SII (silica gel) with doouble development to distances of 6 and 10 cm from the start using chloroform—methanol—15 N ammonia solution (60:10:1) and n-hexane—diethyl ether (50:2). The components separated on the rod were automatically scanned with a hydrogen flame-ionization detector (Iatroscan). The relative responses of methyl glucoside and methyl galactoside to sphingosine and a fatty acid methyl ester were in the same proportions as the molecular ratio. The methanolysis products of sphingomyelin were also separated by the same system. Methanolytes of glyceroglycolipids and glycerophospholipids were separated into individual components with triple development using ethyl acetate—formic acid (70:2) twice and n-hexane—diethyl ether (50:20) to distances of 5 and 10 cm, respectively, from the start.