Analysis of Phospholipids by High Performance Liquid Chromatography

  • Hafez A. Ahmed
Part of the Methods in Molecular Biology book series (MIMB, volume 19)


Phospholipids are amphipathic molecules of great importance and widespread in biological material. They play an essential role in the structure and function of biological membranes. They are classified according to the nature of the “backbone” residue (glycerol or sphingosine), the type of nitrogenous base (or hexahydric alcohol) attached to it, and the nature of the chemical bonds (ester or ester and ether) linking the hydrocarbon chains to the backbone molecule. Within the same class, an enormous number of subclasses is possible through variation in the number of carbon atoms in the hydrocarbon chains, their degree of unsaturation, and, in the case of the glycerophos-pholipids, the position (C1 or C2) of the particular fatty acid (or ether) residues on the glycerol backbone. High performance liquid chromatography (HPLC) has emerged as one of the most powerful and versatile forms of separation technique that can be applied to the efficient separation and determination of phospholipids.


High Performance Liquid Chromatography High Performance Liquid Chromatography Guard Column Choline Chloride Phospholipid Class 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


  1. 1.
    Willard, H. H., Merrit, L. L., Dean, J. A., and Sheille, F. A. (1988) High-performance liquid chromatography methods, in Instrumental Methods of Analysis (Willard, H. H., ed.), Wadsworth, Belmont, CA, pp. 580–613.Google Scholar
  2. 2.
    Patton, G. M., Fasulo, J. M., and Robins, S. J. (1982) Separation of phospholipids and individual molecular species of phospholipids by high performance liquid chromatography. J. Lipid Res. 23, 190–196.PubMedGoogle Scholar
  3. 3.
    Geurts Van Kessel, W. S. M., Hax, W. M. A., Demel, R. A., and De Gier, J. (1977) High performance liquid chromatographic separation and direct ultraviolet detection of phospholipids/ Biochim. Biophys. Acta 486, 524–530.CrossRefGoogle Scholar
  4. 4.
    Wilson, K. and Goulding, K. H. (1986) Chromatographic techniques, in A Biologist’s Guide to Principles and Techniques of Practical Biochemistry, 3rd ed., Edward Arnold, p. 235.Google Scholar
  5. 5.
    Jungalwala, F. B., Turel, R. J., Evans, J. E., and McCluer, R. H. (1975) Sensitive analysis of ethanolamine and serine containing phosphoglycerides by high performance liquid chromatography. Biochem. J. 145, 517–526.PubMedGoogle Scholar
  6. 6.
    Hsieh, J. Y. K., Welch, D. K., and Turcotte, J. G. (1981) High pressure liquid chromatographic separation of molecular species of phosphatidic acid dimethyl esters derived from phosphatidyl choline. Lipids 16, 761–763.CrossRefGoogle Scholar
  7. 7.
    Kaduce, T. L., Norton, K. C., and Spector, A. A. (1983) A rapid isocratic method for phospholipid separation by high performance liquid chromatography. J. Lipid Res. 24, 1398–1403.PubMedGoogle Scholar
  8. 8.
    Jungalwala, F. B., Evans, J. E., and McCluer, R. H. (1976) High performance liquid chromatography of phosphatidyl choline and sphingomyelin with detection in the region of 200 nm. Biochem. J. 155, 55–60.PubMedGoogle Scholar
  9. 9.
    Privett, O. D. and Erdahl, W. L. (1978) An improved flame ionization detector for high performance liquid chromatography. Anal. Biochem. 84,449–461.PubMedCrossRefGoogle Scholar
  10. 10.
    Phillips, F. C., Erdahl, W. L., and Privett, O. S. (1982) Quantitative analysis of lipid classes by liquid chromatography via a flame ionization detector. Lipids 17, 992–997.CrossRefGoogle Scholar
  11. 11.
    Snyder, L. R. (1968) Principles of Absorption Chromatography: the separation of nonionic compounds. Marcel Dekker, New York.Google Scholar
  12. 12.
    Jungalwala, F. B., Hayssen, V., Pasquini, J. M., and McCluer, R. H. (1979) Separation of molecular species of sphingomyelin by reversed-phase high performance liquid chromatography. J. Lipid Res. 20, 579–587.PubMedGoogle Scholar
  13. 13.
    Bartlett, G. R. (1959) Phosphorus assay in column chromatography. J. Biol. Chem. 234, 466–468.PubMedGoogle Scholar

Copyright information

© Humana Press Inc. 1993

Authors and Affiliations

  • Hafez A. Ahmed
    • 1
  1. 1.Department of Biochemical MedicineSt. George’s Hospital Medical SchoolLondonUK

Personalised recommendations