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Quantitative31P nuclear magnetic resonance analysis of the phospholipids of erythrocyte membranes using detergent
Authors:M Hossein Nouri-Sorkhabi  Lesley C Wright  David R Sullivan  Philip W Kuchel
Affiliation:(1) Department of Biochemistry, University of Sydney, 2006 Sydney, N.S.W.;(2) Department of Infectious Diseases, Westmead Hospital, 2145, N.S.W.;(3) Department of Clinical Biochemistry, Royal Prince Alfred Hospital, Camperdown, 2050, N.S.W., Australia
Abstract:31P Nuclear magnetic resonance (NMR) spectra of human erythrocyte lysates dissolved in sodium cholate were acquired. The narrow resonances of phospholipids were mostly well resolved, allowing identification and accurate quantitative analysis of phospholipid classes of the erythrocyte membranes. The ether-linked phosphatidylethanolamine components of the erythrocyte membranes were identified, based on the removal of plasmalogens by acidolysis and of diacyl phospholipid species by degradation using phospholipase A1. It was also shown that the introduction of double bonds on the acyl chains of phosphatidylcholine shifted the31P NMR resonances to lower frequencies. Quantitative analyses of phospholipids from the spectra were based on their apparent molar concentrations. The recoveries of phospholipids from erythrocytes were significantly higher than those using conventional extraction procedures.
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