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Acidic hydrolysis of plasmalogens followed by high‐performance liquid chromatography
Author(s) -
Murphy Eric J.,
Stephens Ralph,
JurkowitzAlexander Marianne,
Horrocks Lloyd A.
Publication year - 1993
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/bf02536090
Subject(s) - glycerophospholipid , plasmalogen , glycerophospholipids , chemistry , hydrolysis , ethanolamine , chromatography , phospholipid , fatty acid , acid hydrolysis , organic chemistry , biochemistry , membrane
A simple, quantitative method for determining the plasmalogen content of small samples is reported here. The method uses the different susceptibility to acid‐catalyzed hydrolysis of the alkyl, alkenyl and acyl linkages to separate the plasmalogen subclass from the other two non‐labile subclasses. Hydrolysis of plasmenylethanolamine and plasmenylcholine was complete after 4 and 1 min of acid treatment, respectively. The acid‐catalyzed hydrolysis did not alter the phospholipid fatty acid composition, making this method useful for fatty acid compositional analysis of the plasmalogen subclass. High‐performance liquid chromatography was used for separations, and phospholipids were quantitated by assay of lipid phosphorus or by direct quantitation of peak area. Using this method, small amounts (10 nmol) of ethanolamine glycerophospholipid and choline glycerophospholipid are subjected to acid‐catalyzed hydrolysis and subsequent separation of the resulting lysocompounds obtained from plasmalogens from the more acid‐stable alkylacyl and diacyl glycerophospholipid fractions. Our values for plasmalogens from commercial preparations of choline and ethanolamine glycerophospholipids agree with literature values. The usefulness of the method is demonstrated for small glycerophospholipid samples that are equivalent to samples from cultured neural cells.