Sensitive analysis of ethanolamine and serine containing phosphoglycerides by high performance liquid chromatography

Abstract

A highly sensitive method for the separation and quantitative measurement of phospholipids containing primary amino groups, such as phosphatidylethanolamine, phosphatidylserine and lysophosphatidylethanolamine, is described. The method involves a simple and quantitative derivative formation of the phospholipids containing amino groups to their u.v. absorbing biphenylcarbonyl derivatives. These have molar extinction coefficients of about 23000 at 268 nm. The phospholipid derivatives are then separated and non destructively determined by high performance liquid chromatography. The amino phospholipids containing vinyl ether bonds (plasmalogens) can be determined separately from the diacyl and alkylacyl amino phospholipids. The lower limit of detection by high performance liquid chromatographic analysis of the phospholipid derivatives is about 10 to 13 pmol or 0.3 to 0.4 ng of phospholipid P. The quantitative range of derivative formation and analysis by high performance liquid chromatography of the phospholipids containing amino groups was shown to be 10 to 500 nmol. The method was shown to be applicable to the analysis of phospholipids containing amino groups in tissue samples.