The fractionation of phosphatidylinositol into molecular species by thin-layer chromatography on silver nitrate-impregnated silica gel.

Abstract

1. Two methods for the fractionation of phosphatidylinositol into molecular species were developed. In addition to preserving the fatty acid moiety of the molecule, the first method preserves the phosphorus, and the second preserves both the phosphorus and inositol ring. 2. In the first method, phosphatidylinositol was oxidized with periodate and the products reacted with diazomethane. I.r. examination showed that the main product was identical with dimethylphosphatidic acid. Fractionation to molecular species was carried out on thin layers impregnated with silver nitrate. The fatty acid composition of the species was determined by gas-liquid chromatography, and their distribution in lamb liver phosphatidylinositol was studied by a method using [(3)H]methanol. 3. In the second method, phosphatidylinositol was acetylated under mild reaction conditions. The major product was the triacetylated derivative of this phospholipid. This was reacted with diazomethane and the methylated-triacetylated phosphatidylinositol was fractionated into molecular species on silver nitrate-impregnated thin layers. Solvent mixtures containing acetone and distilled chloroform were found most suitable for this purpose. The fatty acid composition of the molecular species was determined by g.l.c., and their distribution in lamb liver phosphatidylinositol was studied by a method using [1-(14)C]acetic anhydride during the acetylation reaction. 4. Results from both methods agree fairly well. The most predominant species of lamb liver phosphatidylinositol is the monoenoic (60%) followed by the tetraenoic (17%). The di- and tri-enoic species existed as minor components.