Resolution of diacylglycerol moieties of natural glycerophospholipids by gas–liquid chromatography on polar capillary columns

Abstract

A rapid and practical method was developed for the GLC determination of the sn-1,2-diacylglycerol moieties of natural glycerophospholipids using polar wall-coated open tubular columns. The method gives complete resolution and quantitative estimates for all species according to MW and degree of unsaturation, including stearoyl docosahexaenoylglycerol and related polyunsaturates. For this purpose the sn-1,2-diacylglycerols are obtained from the glycerophospholipids by hydrolysis with phospholipase C and are converted into the trimethylsilyl or tert-butyldimethylsilyl ethers. The silyl ethers are separated by GLC on the capillary glass columns coated with a polar cyanopropylsiloxane polymer, in the temperature range 175-250.degree. C, using hydrogen as the carrier gas. Practical applications of the method are illustrated by analyses of the sn-1,2-diacylglycerol moieties of the phosphatidylcholines of soybean phosphatides, egg yolk, and rat liver. The method of analysis is applicable to other classes of glycerophospholipids and the total time requirements for the analysis of any one phospholipid class are comparable to those for a fatty acid analysis.