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Quantitative characterization of phospholipids in milk fat via 31P NMR using a monophasic solvent mixture

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Lipids

Abstract

The phospholipids (PL) occurring in both ewe and cow milk fat globule membrane were identified and quantitatively determined using 31P NMR spectroscopy with inverse gated decoupled sequences, which allowed a rigorous quantitative analysis. A strict relation between amount and distribution of PL and type of feeding was found. The method was calibrated over a mixture of PL standards. A recently introduced solvent constituted by a monophasic dimethylformamide/triethylamine/guanidinium hydrochloride solvent mixture was used. Compared to the traditional chloroform/methanol/water-EDTA solvent, the new solvent mixture shows very similar accuracy and precision from a quantitative point of view. The monophasic solvent overcomes the partition problems related to a biphasic system, and slightly enlarges the range of 31P NMR chemical shifts, thus improving the resolution. In addition, the new solvent apparently displays a lower chemical shift dependence on the various PL concentrations. The limit of the method is mainly determined by the formation of adducts between triethylamine and some PL, namely, PE, monomethylphosphati-dylethanolamine, phosphatidylethanolamine plasmalogens, and some lyso-PL. However, the new 31P NMR signals arising from these adducts could be easily quantified in the determination of PE.

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Abbreviations

at:

acquisition time

C.S.:

chemical shift

1-D:

one-dimensional

2-D:

two-dimensional

D1:

delay time

DMF:

dimethylformamide

DMPE:

dimethylphosphatidylethanolamine

EPLAS:

phosphatidylethanolamine plasmalogens

Et3N:

triethylamine

GH+ :

guanidinium hydrochloride

LPC:

lysophosphatidylcholine

LPE:

lysophosphatidylethanolamine

MFGM:

milk fat globule membrane

MMPE:

monomethylphosphatidyl-ethanolamine

nOe:

nuclear Overhauser effect

PG:

phosphatidylglycerol

PL:

phospholipids

SM:

sphingomyelin

T1 :

spin lattice relaxation time

References

  1. Gunstone, F.D., and Padley, F.B. (1997) Lipid Technologies and Application, pp. 61–68, Marcel Dekker, New York.

    Google Scholar 

  2. Zeisel, S.H., and Canty, D.J. (1993) Choline Phospholipids: Molecular Mechanisms for Human Disease: A Meeting Report, J. Nutr. Biochem. 4, 258–263.

    Article  CAS  Google Scholar 

  3. Canty, D.J., and Zeisel, S.H. (1994) Lecithin and Choline in Human Health and Disease, Nutr. Rev. 52, 327–339.

    Article  PubMed  CAS  Google Scholar 

  4. Walstra, P., and Jenness, R. (1984) Dairy Chemistry and Physics, pp. 58–76, John Wiley & Sons, New York.

    Google Scholar 

  5. Gober, K.H., Günther, B.R., Lünebach, E.M., Repplinger, G., and Wiedemann, M. (1993) Isolation and Analysis of Phospholipids and Phospholipids Mixtures, in Phospholipids Handbook (Gregor, C., ed.), pp. 39–64, Marcel Dekker, New York.

    Google Scholar 

  6. Ibrahim, A.S., and Ghannoum, M.A. (1996) Chromatographic Analysis of Lipids, in Manual on Membrane Lipids (Prasad, R., ed.), pp. 52–79, Springer-Verlag, Berlin.

    Google Scholar 

  7. Bell, M.V. (1997) Separations of Molecular Species of Phospholipids by High-Performance Liquid Chromatography, in Advances in Lipid Methodology—Four (Christie, W.W., ed.), pp. 46–82, The Oily Press, Dundee, Scotland.

    Google Scholar 

  8. Meneses, P., and Glonek, T. (1988) High Resolution 31P NMR of Extracted Phospholipids, J. Lipid Res. 29, 679–689.

    PubMed  CAS  Google Scholar 

  9. Glonek, T., and Merchant, T.E. (1996) 31P Nuclear Magnetic Resonance Profiling of Phospholipids, in Advances in Lipid Methodology—Three (Christie, W.W., ed.), pp. 37–75, The Oily Press, Dundee, Scotland.

    Google Scholar 

  10. Larijani, B., Poccia, D.L., and Dickinson, L.C. (2000) Phospholipid Identification and Quantification of Membrane Vesicle Subfractions by 31P-1H Two-Dimensional Nuclear Magnetic Resonance, Lipids 35, 1289–1297.

    Article  PubMed  CAS  Google Scholar 

  11. Branca, M., Culeddu, N., Fruianu, M., and Serra, M.V. (1995) 31P Nuclear Magnetic Resonance Analysis of Phospholipids in a Ternary Homogeneous System, Anal. Biochem. 232, 1–6.

    Article  PubMed  CAS  Google Scholar 

  12. Bosco, M., Culeddu, N., Toffanin, R., and Pollesello, P. (1997) Organic Solvent Systems for 31P Nuclear Magnetic Resonance Analysis of Lecithin Phospholipids: Applications to Two-Dimensional Gradient-Enhanced 1H-Detected Heteronuclear Multiple Quantum Coherence Experiments, Anal. Biochem. 245, 38–47.

    Article  PubMed  CAS  Google Scholar 

  13. Culeddu, N., Bosco, M., Toffanin, R., and Pollesello, P. (1998) 31P NMR Analysis of Phospholipids in Crude Extracts from Different Sources: Improved Efficiency of the Solvent System, Magn. Reson. Chem. 36, 907–912.

    Article  CAS  Google Scholar 

  14. Folch, J., Lees, M., and Sloane Stanley, G.H. (1957) A Simple Method for the Isolation and Purification of Total Lipides from Animal Tissues, J. Biol. Chem. 226, 497–509.

    PubMed  CAS  Google Scholar 

  15. Fisher, E. (1920) Wanderung von Acyl bei den Glyceriden, Ber. Dtsch. Chem. Ges. 53, 1621–1633.

    Google Scholar 

  16. Patton, S., and Keenan, T.W. (1975) The Milk Fat Globule Membrane, Biochim. Biophys. Acta 415, 273–309.

    PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Dipartimento di Scienze Chimiche, Università di Cagliari, Cittadella di Monserrato, S.S. 554 Bio Sestu, 09042, Monserrato-Cagliari, Italy

    Sergio Murgia, Stefania Mele & Maura Monduzzi

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  1. Sergio Murgia
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  2. Stefania Mele
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  3. Maura Monduzzi
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Correspondence to Sergio Murgia.

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Murgia, S., Mele, S. & Monduzzi, M. Quantitative characterization of phospholipids in milk fat via 31P NMR using a monophasic solvent mixture. Lipids 38, 585–591 (2003). https://doi.org/10.1007/s11745-003-1500-3

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  • DOI: https://doi.org/10.1007/s11745-003-1500-3

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