Analysis of Lipids by High Performance Liquid Chromatography

  • L. A. Smith
  • G. A. ThompsonJr.
Part of the Modern Methods of Plant Analysis book series (MOLMETHPLANT, volume 5)


High performance liquid chromatography (HPLC) of natural lipids has been remarkably slow to develop as an analytical tool. The absence of easily detectable functional groups on most lipids makes them difficult to measure quantitatively using the more common types of detectors. As a result, many of the published methods for underivatized lipid HPLC are semiquantitative at best and are primarily designed to separate rather than quantify the individual components. The structures of some lipids are amenable to derivatization, yielding light-absorbing products that may be quantified on a molar basis. Applications of this type have been quite successful, although extreme care must be taken to avoid degradation or selective losses of some lipid species during the derivatization steps.


High Performance Liquid Chromatography Molecular Species Lipid Class High Performance Liquid Chromatography Analysis Reverse Phase High Performance Liquid Chromatography 
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.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Alcock NJ, Eckers C, Games DE, Games MPL, Lant MS, McDowall MA, Rossiter M, Smith RW, Westwood SA, Wong HY (1982) High-performance liquid chromatography-mass spectrometry with transport interfaces. J Chromatogr 251: 165–174CrossRefGoogle Scholar
  2. Arpino PJ, Guiochon G (1982) Optimization of the instrumental parameters of a combined liquid chromatograph-mass spectrometer, coupled by an interface for direct liquid introduction. J Chromatogr 251: 153–164CrossRefGoogle Scholar
  3. Batley M, Packer NH, Redmond JW (1980) High-performance liquid chromatography of diglyceride o-nitrobenzoates; an approach to molecular analysis of phospholipids. J Chromatogr 198: 520–525PubMedCrossRefGoogle Scholar
  4. Batley M, Packer NH, Redmond JW (1982) Molecular analysis of the phospholipids of Escherichia coli. Biochim Biophys Acta 710: 400–405PubMedCrossRefGoogle Scholar
  5. Billheimer JT, Avart S, Milani B (1983) Separation of steryl esters by reversed-phase liquid chromatography. J Lipid Res 24: 1646–1650PubMedGoogle Scholar
  6. Bligh EG, Dyer WJ (1959) A rapid method of total lipid extraction and purification. Can J Biochem Physiol 37: 911–917PubMedCrossRefGoogle Scholar
  7. Booth RFG, Quinn PJ (1983) Principles and application of high performance liquid chromatography. In: Wrigglesworth JM (ed) Biochemical research techniques. Wiley, New York, pp 119–146Google Scholar
  8. Charlesworth JM (1978) Evaporative analyzer as a mass detector for liquid chromatography. Anal Chem 50: 1414–1420CrossRefGoogle Scholar
  9. Christie WW (1985) Rapid separation and quantification of lipid classes by high-performance liquid chromatography and mass (light scattering) detection. J Lipid Res 26: 507–512PubMedGoogle Scholar
  10. DiBussolo JM, Nes WR (1982) Structural elucidation of sterols by reversed phase liquid chromatography. I. Assignment of retention coefficients to various groups. J Chromatogr Sci 20: 193–202Google Scholar
  11. Dong MW, DiCesare JL (1983) Improved separation of natural oil triglycerides by liquid chromatography using columns packed with 3 µm particles. J Am Oil Chem Soc 60: 788–791CrossRefGoogle Scholar
  12. DuPont (1978) Get better performance from your HPLC system. In: Liquid chromatography report. Wilmington, DelawareGoogle Scholar
  13. Eichenberger W (1982) Distribution of diacylglyceryl-O-4’(N,N,N-trimethyl)homoserine in different algae. Plant Sci Lett 24: 91–95CrossRefGoogle Scholar
  14. Folch J, Lees M, Sloan-Stanley GH (1957) A simple method for the isolation and purification of total lipids from animal tissues. J Biol Chem 226: 497–509PubMedGoogle Scholar
  15. Halgunset J, Lund EW, Sunde A (1982) Improved separation of biologically relevant C14–C20 fatty acids by reverse-phase high-performance liquid chromatography. J Chromatogr 237: 496–499CrossRefGoogle Scholar
  16. Hamilton JG, Comai K (1984) Separation of neutral lipids and free fatty acids by high performance liquid chromatography using low wavelength ultraviolet detection. J Lipid Res 25: 1142–1148PubMedGoogle Scholar
  17. Jensen GW (1981) Improved separation of triglycerides at low temperatures by reversed-phase liquid chromatography. J Chromatogr 204: 207–411CrossRefGoogle Scholar
  18. Jungalwala FB, Evans JE, Kadowaki H, McCluer RH (1984 a) High performance liquid chromatography-chemical ionization mass spectrometry of sphingoid bases using moving-belt interface. J Lipid Res 25: 209–216Google Scholar
  19. Jungalwala FB, Evans JE, McCluer RH (1984b) Compositional and molecular species analysis of phospholipids by high performance liquid chromatography coupled with chemical ionization mass spectrometry. J Lipid Res 25: 738–749PubMedGoogle Scholar
  20. Kadowaki H, Evans JE, McCluer RH (1984) Separation of brain monoganglioside molec- ular species by high-performance liquid chromatography. J Lipid Res 25: 1132–1139PubMedGoogle Scholar
  21. Kaduce TL, Norton KC, Spector AA (1983) A rapid, isocratic method for phospholipid separation by high-performance liquid chromatography. J Lipid Res 24: 1398–1403PubMedGoogle Scholar
  22. Kates M (1972) Techniques of lipidology. In: Work TS, Work E (eds) Laboratory techniques in biochemistry and molecular biology, vol 3, II. Elsevier/North-Holland, Amsterdam, pp 265–610Google Scholar
  23. Kates M, Eberhardt FM (1957) Isolation and fractionation of leaf phosphatides. Can J Bot 35: 895–921CrossRefGoogle Scholar
  24. Katrangi N, Kaplan LA, Stein EA (1984) Separation and quantitation of serum ß-carotene and other carotenoids by high performance liquid chromatography. J Lipid Res 25: 400–406PubMedGoogle Scholar
  25. Krien P, Devant G, Hardy M (1982) Application of microbore columns to liquid chromatography-mass spectrometry. J Chromatogr 251: 129–139CrossRefGoogle Scholar
  26. Lynch DV, Thompson GA Jr (1982) Microsomal phospholipid molecular species alterations during low temperature acclimation in Dunaliella. Plant Physiol (Bethesda) 74: 193–197CrossRefGoogle Scholar
  27. Macrae R, Trugo LC, Dick J (1982) The mass detector: a new detection system for carbohydrate and lipid analysis. Chromatographia 15: 476–478CrossRefGoogle Scholar
  28. Morrison WR, Smith LM (1964) Preparation of fatty acid methyl esters and dimethylacetals from lipids with boron trifluoride-methanol. J Lipid Res 5: 600–608PubMedGoogle Scholar
  29. Nakagawa Y, Horrocks LA (1983) Separation of alkenylacyl, alkylacyl, and diacyl analogues and their molecular species by high performance liquid chromatography. J Lipid Res 24: 1268–1275PubMedGoogle Scholar
  30. Plattner RD (1981) High performance liquid chromatography of triglycerides: controlling selectivity with reverse phase columns. J Am Oil Chem Soc 58: 638–642CrossRefGoogle Scholar
  31. Quinn PJ, Williams WP (1978) Plant lipids and their role in membrane function. Prog Biophys Mol Biol 34: 109–173PubMedCrossRefGoogle Scholar
  32. Rouser G, Fleischer S, Yamamoto A (1970) Two dimensional thin layer chromatographic separation of polar lipids and determination of phospholipids by phosphorus analysis of spots. Lipids 5: 494–498PubMedCrossRefGoogle Scholar
  33. Sato N, Furuya M (1983) Distribution of diacylglyceryltrimethylhomoserine in green plants. Plant Cell Physiol 24: 1113–1116Google Scholar
  34. Smith LA, Norman HA, Cho SH, Thompson GA Jr (1985) Isolation and quantitative analysis of phosphatidylglycerol and glycolipid molecular species using reverse phase high performance liquid chromatography with flame ionization detection. J Chromatogr 346: 291–299PubMedCrossRefGoogle Scholar
  35. Snyder LR (1978) Classification of the solvent properties of common liquids. J Chromatogr Sci 16: 223–234Google Scholar
  36. Snyder LR, Kirkland JJ (eds) (1979) Introduction to modern chromatography, 2nd edn. Wiley, New York, p 290Google Scholar
  37. Stolyhwo A, Colin H, Guiochon G (1983) Use of light scattering as a detector in liquid chromatography. J Chromatogr 265: 1–18CrossRefGoogle Scholar
  38. Szakasits JJ, Robinson RE (1974) Disc conveyor flame ionization detector for liquid chromatography. Anal Chem 46: 1648–1652CrossRefGoogle Scholar
  39. Tsimidou M, Macrae R (1984) Influence of injection solvent on the reversed-phase chromatography of triglycerides. J Chromatogr 285: 178–181CrossRefGoogle Scholar
  40. Vestal ML (1983) Ion emissions from liquids (for mass spectroscopy). Mass Spectrom Rev 2: 447–452CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 1987

Authors and Affiliations

  • L. A. Smith
  • G. A. ThompsonJr.

There are no affiliations available

Personalised recommendations