Journal of the American Oil Chemists’ Society

, Volume 71, Issue 11, pp 1179–1187 | Cite as

Preparation of fatty acid methyl esters for gas-chromatographic analysis of lipids in biological materials

  • Ke-Shun Liu


Theoretically, preparation of fatty acid methyl esters (FAMEs) deals with reversible chemical reactions in a complex system. Methodologically, there are numerous ways, generally characterized by the type of catalysts used and steps involved. Although there are more than a half dozen common catalysts, the majority fall into either acidic (HCl, H2SO4 and BF3) or alkaline types (NaOCH3, KOH and NaOH), with each having its own catalytic capability and application limitations. In terms of steps, many conventional methods, including those officially recognized, consist of drying, digestion, extraction, purification, alkaline hydrolysis, transmethylation/methylation and postreaction work-up. Although these methods are capable of providing reliable estimates if some precautions are taken, they are cumbersome, time-consuming and cost-inefficient. A new approach has been to transmethylate lipidsin situ. Due to its simplicity, high sensitivity, comparable reliability and capability to determine total fatty acids, the method of direct transmethylation is finding a unique place in lipid determination. Regardless of which method is used, quantitative methylation requires chemists to take precautions at every step involved, particularly during FAME formation and subsequent recovery steps. Evidently, there is an urgent need for more systematic studies, guided by the chemical principle of reactions involved and physicochemical properties of regents and end products, into factors affecting these steps. Hopefully, this will lead to an improved method, which measures lipid composition in biological materials not only with high accuracy but also with high efficiency and minimum costs.

Key Words

Derivatization fatty acid methyl esters gas chromatography lipid analysis 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Anon., inGC Derivatization Guide, Alltech Associates, Inc., Applied Science Labs, Deerfield, 1993, Bulletin No. 126.Google Scholar
  2. 2.
    Craske, J.D.,J. Am. Oil Chem. Soc. 70:325 (1993).Google Scholar
  3. 3.
    Darbre, A., inHandbook of Derivatives for Chromatography, edited by K. Blau, and G.S. King. Heyden & Son, London, 1978, pp. 36–103.Google Scholar
  4. 4.
    Official Methods of Analysis, Association of Official Analytical Chemists, 10th edn., Washington, D.C., 1965, Section 26.052.Google Scholar
  5. 5.
    Ibid. Official Methods of Analysis, Association of Official Analytical Chemists, 15th edn., 1990, Sections 954.02 and 969.33.Google Scholar
  6. 6.
    Official Methods and Recommended Practices of the American Oil Chemists' Society, AOCS, Champaign, 1969, Method Ce. 2-66.Google Scholar
  7. 7.
    Ibid. Official Methods and Recommended Practices of the American Oil Chemists' Society, AOCS, Champaign, 1992, Method Ce 2-66.Google Scholar
  8. 8.
    International Organization for Standardization, International Standards Organization, Geneva, 1st edn., 1978, Ref. No. ISO-5509-1978 (E), Sec. 7.Google Scholar
  9. 9.
    International Union of Pure and Applied Chemistry, 6th edn., IUPAC, Pergamon Press, Oxford, 1979, Method 2.301, Section 3.Google Scholar
  10. 10.
    Abel, K., H. de Schmertzing and J.I. Peterson,J. Bact. 85:1039 (1963).Google Scholar
  11. 11.
    Dugan, L.R., G.W. McGinnins and D.V. Vahedra,Lipids 1:305 (1966).CrossRefGoogle Scholar
  12. 12.
    Barnes, P.C. and C.E. Holiday,J. Chromatogr. Sci. 10:181 (1972).Google Scholar
  13. 13.
    Welch, R.W.,J. Sci. Food Agric. 26:429 (1975).CrossRefGoogle Scholar
  14. 14.
    Kramer, J.K.G. and H.W. Hulan,J. Lipid Res. 17:674 (1976).Google Scholar
  15. 15.
    Outen, G.E., D.E. Beever and J.S. Fenlon,J. Sci. Fd. Agric. 27:419 (1976).CrossRefGoogle Scholar
  16. 16.
    Shimasaki, H., F.C. Phillips and O.S. Privett,J. Lipid Res. 18:540 (1977).Google Scholar
  17. 17.
    Lepage, G. and C.C. Roy,25:1391 (1984).Google Scholar
  18. 18.
    Browse, J., P.J. McCourt and C.R. Somerville,Anal. Biochem. 152:141 (1986).CrossRefGoogle Scholar
  19. 19.
    Long, A.R., S.J. Massie and W.J. Tyznik,J. Food Sci. 53:940 (1988).CrossRefGoogle Scholar
  20. 20.
    Sukhija, P.S. and D.L. Palmquist,J. Agric. Food Chem. 36:1202 (1988).CrossRefGoogle Scholar
  21. 21.
    Dahmer, M.L., P.D. Fleming, G.B. Collins and D.F. Hildebrand,J. Am. Oil Chem. Soc. 66:543 (1989).Google Scholar
  22. 22.
    Wlez, W., W. Sattler, H.-J. Leis and E. Malle,J. Chromatogr. 526:319 (1990).Google Scholar
  23. 23.
    Sattler, W., H. Puhl, M. Hayne, G.M. Kostner and H. Esterbauer,Anal. Biochem. 198:184 (1991).CrossRefGoogle Scholar
  24. 24.
    Ulberth, F. and M. Henninger,69:174 (1992).Google Scholar
  25. 25.
    Christie, W.W., inTopics in Lipid Chemistry, edited by F.D. Gunstone, and Paul Elek, (Scientific Books) Ltd., London, 1972, Vol. 3, pp. 171–197.Google Scholar
  26. 26.
    Christie, W.W., inLipid Analysis, Pergamon Press, Oxford, 1973, pp. 85–102.Google Scholar
  27. 27.
    Sheppard, A.J. and J.L. Iverson,J. Chromatogr. Sci. 13:448 (1975).Google Scholar
  28. 28.
    Jupille, T.,17:161 (1979).Google Scholar
  29. 29.
    Bannon, C.D., G.J. Breen, J.D. Craske, N.T. Hai, N.L. Harper and K.L. O'Rourke,247:71 (1982).CrossRefGoogle Scholar
  30. 30.
    Wood, R., inAnalyses of Fats, Oils and Derivatives, edited by E.G. Perkins, AOCS Press, Champaign, 1993, pp. 236–269.Google Scholar
  31. 31.
    Christie, W.W., inAdvances in Lipid Methodology—Two, edited by W.W. Christie, The Oily Press, Dundee, 1993, pp. 69–112.Google Scholar
  32. 32.
    Bannon, C.D., J.D. Craske, N.T. Hai, N.L. Hai, N.L. Happer and K.L. O'Rourke,J. Chromatogr. 247:63 (1982).CrossRefGoogle Scholar
  33. 33.
    Bannon, C.D., J.D. Craske and A.E. Hilliker,62:1501 (1985).Google Scholar
  34. 34.
    Craske, J.D. and C.D. Bannon,J. Am. Oil Chem. Soc. 64:1413 (1987).Google Scholar
  35. 35.
    Craske, J.D., C.D. Bannon and L.M. Norman,65:262 (1988).Google Scholar
  36. 36.
    Morrison, W.R., S.L. Tan and K.D. Tan,J. Sci. Food Agric. 31:329 (1980).CrossRefGoogle Scholar
  37. 37.
    Bligh, E.G. and W.J. Dyer,Can. J. Biochem. Physiol. 37:911 (1959).Google Scholar
  38. 38.
    Hougen, F.W. and V. Bodo,J. Am. Oil Chem. Soc. 50:230 (1973).Google Scholar
  39. 39.
    O'Keefe, S.F., V.A. Wiley and D.A. Knauft,70:489 (1993).Google Scholar
  40. 40.
    Singh, B.B., H.H. Hadley and F.I. Collins,Crop Sci. 8:171 (1968).CrossRefGoogle Scholar
  41. 41.
    Morrison, W.R. and L.M. Smith,J. Lipid Res. 5:600 (1964).Google Scholar
  42. 42.
    Stoffel, W., F. Chu and E.H. Ahrens, Jr.,Anal. Chem. 31:307 (1959).CrossRefGoogle Scholar
  43. 43.
    Kishimoto, Y. and N.S. Radin,J. Lipid Res. 1:72 (1959).Google Scholar
  44. 44.
    Kishimoto, Y. and N.S. Radin,6:435 (1965).Google Scholar
  45. 45.
    Jham, G.N., F.F.F. Teles and L.G. Campos,J. Am. Oil Chem. Soc. 59:132 (1982).Google Scholar
  46. 46.
    Mitchell, J. Jr., D.M. Smith and W.M.D. Bryant,J. Am. Chem. Soc. 62:4 (1940).CrossRefGoogle Scholar
  47. 47.
    Metcalfe, L.D. and A.A. Schmitz,Anal. Chem. 33:363 (1961).CrossRefGoogle Scholar
  48. 48.
    Metcalfe, L.D., A.A. Schmitz and J.R. Pelka,38:514 (1966).CrossRefGoogle Scholar
  49. 49.
    Dawidowowicz, E.A. and T.E. Thompson,J. Lipid Res. 12:636 (1971).Google Scholar
  50. 50.
    Koritala, S. and W.K. Rohwedder,Lipids 7:274 (1972).CrossRefGoogle Scholar
  51. 51.
    Medina, I., S. Augourg, J.M. Gallardo and R. Perez-Martin,Inter. J. Food Sci. Technol. 27:597 (1992).CrossRefGoogle Scholar
  52. 52.
    Christopherson, S.W. and R.L. Glass,J. Dairy Sci. 52:1289 (1969).CrossRefGoogle Scholar
  53. 53.
    Christie, W.W.,J. Lipid Res. 23:1072 (1982).Google Scholar
  54. 54.
    Ayorinde, F.O., J. Clifton, Jr., O.A. Afolabi and R.L. Sheppard,J. Am. Oil Chem. Soc. 65:942 (1988).Google Scholar
  55. 55.
    Schlenk, H. and J.L. Gellerman,Anal. Chem. 32:1412 (1960).CrossRefGoogle Scholar
  56. 56.
    Krisnangkura, K. and R. Simamaharnnop,J. Am. Oil Chem. Soc. 69:166 (1992).Google Scholar
  57. 57.
    Drucker, D.B.,Microbios, 5:109 (1972).Google Scholar
  58. 58.
    Lepage, G. and C.C. Roy,J. Lipid Res. 27:114 (1986).Google Scholar
  59. 59.
    Lepage, G. and C.C. Roy,29:227 (1988).Google Scholar
  60. 60.
    Ozgul, S. and S. Turkay,J. Am. Oil Chem. Soc. 70:145 (1993).Google Scholar

Copyright information

© AOCS Press 1994

Authors and Affiliations

  • Ke-Shun Liu
    • 1
  1. 1.Soyfood LaboratoryJacob Hartz Seed Co., Inc.Stuttgart

Personalised recommendations