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Research Article| Volume 29, ISSUE 5, P679-689, May 1988

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High resolution 31P NMR of extracted phospholipids.

Open AccessPublished:May 01, 1988DOI:https://doi.org/10.1016/S0022-2275(20)38513-8
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      The common phospholipids from biological sources were quantitated using phosphorus-31 nuclear magnetic resonance (NMR) spectroscopy in conjunction with an analytical reagent composed of two parts: 1) 2 ml of reagent chloroform in which was dissolved 0.01-100 mg of crude tissue lipid extracted from tissue sources using chloroform-methanol 2:1, the extract having been washed with 0.2 vol. of 0.1 M KCl; 2) 1 ml of an aqueous methanol reagent composed of one part 0.2 M (ethylenedinitrilo)-tetraacetic acid in D2O titrated to pH 6 with CsOH and four parts of reagent methanol. In a magnetic field of 11.75 Tesla, the extracted phospholipids yield narrow signals (1.8-3.2 Hz at half-height), corresponding to each generic species, e.g., phosphatidylcholines, phosphatidylethanolamines, etc., permitting resolution among the various phospholipid families and their lyso and plasmalogen derivatives. The reagent permits assays of high precision and accuracy using a modest amount of NMR spectrometer time (ca. 15 min/assay). The procedures described, which are compared to high-performance liquid chromatography, are convenient for the routine analysis of phospholipids from biological sources.

      REFERENCES

        • Davis D.G.
        • Inesi G.
        Phosphorus and proton magnetic resonance studies in sarcoplasmic reticulum membranes and lipids.
        Biochim. Biophys. Acta. 1972; 282: 180-186
        • Yeagle P.L.
        Phospholipid-protein interactions and the structure of the human erythrocyte membrane: nuclear magnetic resonance studies.
        Erythrocyte Membr. 1984; 3: 153-175
        • Seelig J.
        3lP nuclear magnetic resonance and the head group structure of phosphonolipids in membranes.
        Biochim. Biophys. Acta. 1978; 515: 105-140
        • Dufourc E.J.
        • Smith I.C.P.
        • Dufourcq J.
        Molecular details of melittin-induced lysis of phospholipid membranes as revealed by deuterium and phosphorus NMR.
        Biochemistry. 1986; 25: 6448-6455
        • Michaelson D.M.
        • Horowitz A.F.
        • Klein M.P.
        Transbilayer asymmetry and surface homogeneity of mixed phospholipids in cosonicated vesicles.
        Biochemistry. 1973; 12: 2637-2645
        • Uhing M.C.
        A 31P NMR study of the thermal transition of dipalmitoyl lecithin vesicles.
        Chem. Phys. Lipids. 1975; 14: 303-308
        • Berden J.A.
        • Barker R.W.
        • Radda G.K.
        NMR studies on phospholipid bilayers: some factors affecting lipid distribution.
        Biochim. Biophys. Acta. 1975; 375: 186-208
        • Assmann G.
        • Brewer Jr., H.B.
        A molecular model of high density lipoproteins.
        Proc. Natl. Acad. Sci. USA. 1974; 71: 1534-1538
        • Glonek T
        • Henderson T.O.
        • Kruski A.W.
        • Scanu A.M.
        31P nuclear magnetic resonance: application to the study of human serum high density lipoproteins.
        Biochim. Biophys. Acta. 1974; 348: 155-161
        • Henderson T.O.
        • Glonek T.
        • Myers T.C.
        Phosphorus-31 nuclear magnetic resonance spectroscopy of phospholipids.
        Biochemistry. 1974; 13: 623-628
        • Brasure E.B.
        • Henderson T.O.
        • Glonek T.
        • Pattnaik N.M.
        • Scanu A.M.
        Action of α-phospholipase A2 on human serum high density lipoprotein-3: kinetic study of the reaction by 31P nuclear magnetic resonance spectroscopy.
        Biochemistry. 1978; 17: 3934-3938
        • London E.
        • Feigenson G.W.
        Phosphorus NMR analysis of phospholipids in detergents.
        J. Lipid Res. 1979; 20: 408-412
        • Bárány M.
        • Glonek T.
        Identification of diseased states by P-31 NMR.
        in: Gorenstein D. Phosphorus-31 NMR: Principles and Applications. Academic Press, New York1984: 511-545
        • Greiner J.V.
        • Kopp S.J.
        • Glonek T.
        Phosphorus nuclear magnetic resonance and ocular metabolism.
        Surv. Ophthalmol. 1985; 30: 189-202
        • Bárány M.
        • Glonek T.
        Phosphorus-31 nuclear magnetic resonance of contractile systems.
        Methods Enzymol. 1982; 85B: 624-676
        • Chen S.S-H.
        • Kow A.Y.
        Improved procedure for the separation of phospholipids by high-performance liquid chromatography.
        J. Chromatogr. 1982; 227: 25-31
        • Folch J.
        • Lees M.
        • Sloane Stanley G.H.
        A simple method for the isolation and purification of total lipids from animal tissues.
        J. Biol. Chem. 1957; 226: 497-509
        • Kirkpatrick D.S.
        • Bishop S.H.
        Simplified wet ash procedure for total phosphorus analysis of organophosphonates in biological samples.
        Anal. Chem. 1971; 43: 1707-1709
        • Mark V.
        • Dungan C.H.
        • Crutchfield M.M.
        • Van Wazer J.R.
        Grayson M. Griffith E.J. Compilation of P31 NMR data. In P31Nuclear Magnetic Resonance: Topics in Phosphorus Chemistry. 5. Interscience Publishers, New York1967: 227-458
        • Glonek T
        • Van Wazer J.R.
        Aqueous tetrahydroxyphosphonium perchlorate as a narrow-line 3IP NMR reference substance.
        J. Magn. Reson. 1974; 13: 390-391
        • Greiner J.V.
        • Kopp S.J.
        • Sanders D.R.
        • Glonek T.
        Organophosphates of the crystalline lens: a nuclear magnetic resonance spectroscopic study.
        Invest. Ophthalmol. & Visual Sci. 1981; 21: 700-713
        • Quin L.D.
        The natural occurrence of compounds with the carbon-phosphorus bond.
        in: Grayson M. Griffith E.J. Topics in Phosphorus Chemistry. 4. Interscience Publishers, New York1967: 23-48
        • Glonek T
        • Kopp S.J.
        Ex vivo P-31 NMR of lens, cornea, heart, and brain.
        Magn. Reson. Imaging. 1985; 3: 359-376
        • Van Wazer J.R.
        Phosphorus and Its Compounds. 1. Interscience Publishers, New York1958: 419-678
        • Costello A.J. R.
        • Glonek T
        • Van Wazer J.R.
        Phosphorus-31 chemical shift variations with countercation and ionic strength for the various ethyl phosphates.
        Inorg. Chem. 1976; 15: 972-974