Lipids

, Volume 24, Issue 5, pp 396–407

Molecular species of glycerophospholipids and sphingomyelins of human erythrocytes: Improved method of analysis

  • J. J. Myher
  • A. Kuksis
  • S. Pind
Article

DOI: 10.1007/BF02535147

Cite this article as:
Myher, J.J., Kuksis, A. & Pind, S. Lipids (1989) 24: 396. doi:10.1007/BF02535147

Abstract

This study reports the application of modern methods of molecular species analysis in determination of the structure of both major and minor glycerophospholipids and sphingomyelins of human erythrocytes. Individual phospholipid classes were resolved from total lipid extracts by thin-layer chromatography. Diradylglycerols were released by phospholipase C and converted into trimethylsilyl ethers, which were resolved into the alkenylacyl, alkylacyl and diacylglycerol subclasses by normal phase high performance liquid chromatography. Molecular species of diradylglycerols and ceramides were quantitated according to carbon and double bond number by gas liquid chromatography using a fused silica capillary column wall-coated with bonded RTx-2330. The molecular species of ceramides were determined by GC/MS. The diradyl glycerophosphocholines contained 93.0% diacyl, 4.6% alkylacyl and 2.5% alkenylacyl, white the diradyl glycerophosphoethanolamines were made up of 48.8% diacyl, 47.8% alkenylacyl and 3.4% alkylacyl subclasses. Analysis of the molecular species showed that the long chain polyunsaturated acids were mainly combined with C16 in all diradyl GPC subclasses and in diacyl GPE, while in the alkylacyl and alkenylacyl GPE and in diacyl glycerophosphoinositol and diacyl glycerophosphoserine they were combined mainly with C18 saturated fatty chains. In addition to the C16 and C18 alkyl and alkenyl, the ether fractions also contained significant proportions of C20, C22 and C24 chains. The molecular species of the ceramide moieties of the SPH were made up largely of mono- and diunsaturated species. Over 200 molecular species were identified and quantitated in a representative sample of human red blood cells.

Abbreviations

CV

coefficients of variation

FAME

fatty acid methyl esters

FID

flame ionization detection

GC

gas chromatography

GC/MS

gas chromatography/mass spectroscopy

GLC

gas-liquid chromatography

GPC

glycerophosphocholines

GPE

glycerophosphoethanolamines

GPI

glycerophosphoinositol

GPL

glycerophospholipids

GPS

glycerophosphoserine

HPLC

high performance liquid chromatography

PI

phosphatidylinositol

PS

phosphatidylserine

SPH

sphingomyelins

TLC

thin-layer chromatography

TMS

trimethylsilyl

BDMS

tertiary-butyldimethylsilyl

Copyright information

© American Oil Chemists’ Society 1989

Authors and Affiliations

  • J. J. Myher
    • 1
  • A. Kuksis
    • 1
  • S. Pind
    • 1
  1. 1.Banting and Best Department of Medical Research and Department of BiochemistryUniversity of TorontoTorontoCanada

Personalised recommendations