, Volume 26, Issue 3, pp 232–236

Malonaldehyde determination in tissues and biological fluids by ion-pairing high-performance liquid chromatography

  • Willy A. Behrens
  • René Madère


A method for the analysis of malonaldehyde by ion pairing high-performance liquid chromatography is described. The method is direct; no thiobarbiturate chromogen formation is required, and sample preparation is simple. After deproteinization with 50% ethanol and removal of particulate by centrifugation samples were passed through a small silica amino column to remove contaminants. Diluted samples (20 μL) were injected onto an octadecylsilane column (25 cm×4.6 mm ID, 5 μm) which is eluted with 30 mM sodium phosphate buffer, pH 6.5 containing 30% ethanol and 1 mM tetradecyltrimethylammonium bromide. Detection was accomplished by monitoring absorbance at 267 nm. The lower limit for reliable quantification was 5 pmol per injection. The method has been successfully applied to the quantification of malonaldehyde present in plasma, urine and tissues of rats kept under different dietary conditions as well as afterin vivo treatment with CCl4 and iron-dextran. The method was also applied to the quantification of malonaldehyde during liver microsomal lipid peroxidation and was compared to the thiobarbituric acid test.



adenosine 5′-diphosphate

AIN-76 diet

American Institute of Nutrition-76 diet


high-performance liquid chromatography




2-thiobarbituric acid


TBA reactive substances


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Pryor, W.A., Stanley, J.P., and Blair, E. (1976)Lipids 11 370–379.PubMedCrossRefGoogle Scholar
  2. 2.
    Frankel, E.N., and Neff, W.E. (1983)Biochim. Biophys. Acta 754, 264–270.Google Scholar
  3. 3.
    Yagi, K. (1982) inlipid Peroxides in Biology and Medicine (Yagi, K., ed.), pp. 223–242, Academic Press, New York.Google Scholar
  4. 4.
    Dillard, C.J., and Tappel, A.L. (1973)Lipids 8, 183–189.PubMedGoogle Scholar
  5. 5.
    Basu, A.K., and Marnett, L.J. (1983)Carcinogenesis 4, 331–333.PubMedGoogle Scholar
  6. 6.
    Shamberger, R.J., Andreone, T.L., and Willis, C.E. (1974)J. Natl. Cancer Inst. 53, 1771–1773.PubMedGoogle Scholar
  7. 7.
    Chio K.S., and Tappel, A.L. (1969)Biochemistry 8, 2827–2832.PubMedCrossRefGoogle Scholar
  8. 8.
    Nair, V., Vietti D.E., and Cooper, C.S. (1981)J. Am. Chem. Soc. 103, 3030–3036.CrossRefGoogle Scholar
  9. 9.
    Kwon, T.W., and Brown, W.D. (1965)Fed. Proc. 24, 592.Google Scholar
  10. 10.
    Landsberg, J.D., and Sinnhuber, R.O. (1965)J. Am. Oil Chem. Soc. 42, 821–824.PubMedGoogle Scholar
  11. 11.
    Shin, B.C., Huggins, J.W., and Carraway, K.L. (1972)Lipids 7, 229–233.PubMedCrossRefGoogle Scholar
  12. 12.
    Nair, V., Cooper, C.S., Vietti, D.E., and Turner, G.A. (1986)Lipids 21, 6–10.PubMedCrossRefGoogle Scholar
  13. 13.
    Brooks, B.R., and Klamerth, O.L. (1968)Eur. J. Biochem. 5, 178–182.PubMedCrossRefGoogle Scholar
  14. 14.
    Buege, J.A., and Aust, S.D. (1978)Methods Enzymol. 52, 302–310.PubMedGoogle Scholar
  15. 15.
    Gutteridge, J.M.C. (1986)Free Rad. Res. Comms. 1, 173–184.Google Scholar
  16. 16.
    Sinnhuber, R.O., Yu, T.C., and Yu, Te Chang. (1958)Food Res. 23, 626–633.Google Scholar
  17. 17.
    Nair, V., and Turner, G.A. (1984)Lipids 19, 804–805.CrossRefGoogle Scholar
  18. 18.
    Marcuse, R., and Johansson, L. (1973)J. Am. Oil Chem. Soc. 50, 387–391.PubMedGoogle Scholar
  19. 19.
    Ohkawa, H., Ohishi, N., and Yagi, K. (1979)Anal. Biochem. 95, 351–358.PubMedCrossRefGoogle Scholar
  20. 20.
    Bigwood, T., and Read, G. (1989)Free Rad. Res. Comms. 6, 387–392.Google Scholar
  21. 21.
    Kosugi, H., Kojima, T., and Kikugawa, K. (1989)Lipids 24, 873–881.CrossRefGoogle Scholar
  22. 22.
    Ichinose, T., Miller, M.G., and Shibamoto, T. (1989)Lipids 24, 895–898.PubMedCrossRefGoogle Scholar
  23. 23.
    Bird, R.P., Hung, S.O.S., Hadley, M., and Draper, H.H. (1983)Anal. Biochem. 128, 240–244.PubMedCrossRefGoogle Scholar
  24. 24.
    Hirayama, T., Yamada, N., Nohara, M., and Fukui, S. (1984)J. Sci. Food Agric. 35, 338–344.CrossRefGoogle Scholar
  25. 25.
    Csallany, A.S., Guan, M.D., Manwaring, J.D., and Addis, P.B. (1984)Anal. Biochem. 142, 277–283.PubMedCrossRefGoogle Scholar
  26. 26.
    Esterbauer, H., and Slater, T.F. (1981)IRCS Med. Sci. 9, 749–750.Google Scholar
  27. 27.
    Wade, C.R., Jackson, P.G., and Van Rij, A.M. (1985)Biochem. Med. 33, 291–296.PubMedCrossRefGoogle Scholar
  28. 28.
    Bull, A.W. and Marnett, L.J. (1985)Anal. Biochem. 149, 284–290.PubMedCrossRefGoogle Scholar
  29. 29.
    Therasse, J., and Lemonnier, F. (1987)J. Chromat. 413, 237–241.Google Scholar
  30. 30.
    Shamberger, R.J., Shamberger, B.A., and Willis, C.E. (1977)J. Nutr. 107, 1404–1409.PubMedGoogle Scholar
  31. 31.
    American Institute of Nutrition (1977)J. Nutr. 107, 1340–1348.Google Scholar
  32. 32.
    Hogeboom, G.H. (1955)Methods Enzymol. 1, 16–19.Google Scholar
  33. 33.
    Esterbauer, H., Lang, J., Zadravec, S., and Slater, T.F. (1984)Methods Enzymol. 105, 319–328.PubMedCrossRefGoogle Scholar
  34. 34.
    Burk, R.F. (1983)Biochim. Biophys. Acta 757, 21–28.PubMedGoogle Scholar
  35. 35.
    Bradford, M.M. (1976)Anal. Biochem. 72, 248–254.PubMedCrossRefGoogle Scholar
  36. 36.
    Thompson, J.N., and Hatina, G. (1979)J. Liquid Chromatog, 2, 327–344.Google Scholar
  37. 37.
    Behrens, W.A., and Madere, R. (1982)Nutr. Rep. Int. 25, 107–112.Google Scholar
  38. 38.
    Snedecor, G.W., and Cochran, W.G. (1980)Statistical Methods, 7th ed., Iowa State University Press, Ames.Google Scholar
  39. 39.
    Lee, H.S., and Csallany, A.S. (1987)Lipids 22, 104–107.PubMedCrossRefGoogle Scholar
  40. 40.
    Draper, H.H., Polensek, L., Hadley, M., and McGirr, L.G. (1984)Lipids 19, 836–843.PubMedCrossRefGoogle Scholar
  41. 41.
    Dhanakoti, C.N., and Draper, H.H. (1987)Lipids 22, 643–646.PubMedGoogle Scholar
  42. 42.
    Farris, M.W., Pascoe, G.A., and Reed, D.J. (1985)The Toxicologist 5, 149.Google Scholar

Copyright information

© American Oil Chemists’ Society 1991

Authors and Affiliations

  • Willy A. Behrens
    • 1
  • René Madère
    • 1
  1. 1.Food Directorate, Bureau of Nutritional SciencesHealth Protection Branch, Health and Welfare CanadaOttawaCanada

Personalised recommendations