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HPLC of plasmalogen‐containing phosphatidylcholine under reverse‐phase or argentation conditions
Author(s) -
Marmer William N.,
Foglia Thomas A.,
Vail Peter D.
Publication year - 1984
Publication title -
lipids
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.601
H-Index - 120
eISSN - 1558-9307
pISSN - 0024-4201
DOI - 10.1007/bf02534787
Subject(s) - plasmalogen , chemistry , chromatography , degree of unsaturation , phosphatidylcholine , high performance liquid chromatography , elution , yield (engineering) , ion exchange , organic chemistry , ion , phospholipid , biochemistry , materials science , membrane , metallurgy
Abstract Two approaches to the high pressure liquid chromatography (HPLC) isolation of intact plasmalogens were investigated. The first used reversed‐phase HPLC and sought to take advantage of subtle differences in the hydrophobicity of the alk‐1‐enyl chain from the acyl counterpart. On a C‐18 column, bovine heart phosphatidylcholine (PC), which was 47% plasmalogen, was separated into a number of fractions that differed in their molecular species composition. One combination of fractions amounted to a 26% yield of PC enriched to 82% plasmalogen. The second approach sought to take advantage of the uniquely electron‐rich functionality of the plasmalogens, the alk‐1‐enyl ether double bond, and its potential to coordinate with heavy metal ions. Specificially, bovine heart PC was applied to a cation‐exchange type HPLC column in the silver ion mode. Although complete exchange of all the active sites of the column with silver ion led to complete retention of PC, partial activation with silver ion resulted in the separation of the PC into fractions, according to the degree of unsaturation. Plasmalogen‐rich fractions eluted last and remained intact during the process. One combination of these fractions amounted to a 49% yield of PC enriched to 72% plasmalogen. Use of a cation‐exchange system in the mercuric ion mode led to on‐column hydrolysis of the plasmalogen; with palladium ion, the metallic species was stripped from the column by the eluting lipid.