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Acidic hydrolysis of plasmalogens followed by high-performance liquid chromatography

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Lipids

Abstract

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.

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Abbreviations

ChoGpl:

choline glycerophospholipid

EtnGpl:

ethanolamine glycerophospholipid

FAME:

fatty acid methyl esters

GLC:

gas-liquid chromatography

HPLC:

high-performance liquid chromatography

HUVE:

human umbilical vein endothelial

PBS:

phosphate buffered saline

PlsCho:

choline plasmalogen

PlsEtn:

ethanolamine plasmalogen

TLC:

thin-layer chromatography

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Authors and Affiliations

  1. Department of Medical Biochemistry and Neuroscience Program, The Ohio State University, 1645 Neil Avenue, Rm. 471, 43210, Columbus, Ohio

    Eric J. Murphy, Marianne Jurkowitz-Alexander & Lloyd A. Horrocks

  2. Department of Pathology, The Ohio State University, 43210, Columbus, Ohio

    Ralph Stephens

Authors
  1. Eric J. Murphy
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  2. Ralph Stephens
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  3. Marianne Jurkowitz-Alexander
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  4. Lloyd A. Horrocks
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Murphy, E.J., Stephens, R., Jurkowitz-Alexander, M. et al. Acidic hydrolysis of plasmalogens followed by high-performance liquid chromatography. Lipids 28, 565–568 (1993). https://doi.org/10.1007/BF02536090

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  • DOI: https://doi.org/10.1007/BF02536090

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