Skip to main content

Measurement of malondialdehyde by high performance liquid chromatography with fluorescence detection

Abstract

A method has been developed to measure malondial-dehyde (MDA) in biological systems. MDA was reacted with 2-thiobarbituric acid (TBA) in the presence of butylated hydroxytoluene (BHT) to minimize formation of artifacts. Initial separation of the TBA-MDA adduct was accomplished by isobutanol extraction. Further elimination and separation of interfering substances was achieved by high performance liquid chromatography. The mobile phase consisted of a 1∶1 (v/v) mixture of methanol and water with 0.05% (w/v) tetrabutyl ammonium dihydrogen phosphate added as an ion pairing reagent. At a flow rate of 1 ml/min, the TBA-MDA adduct was eluted from a 15-cm, c-18, reversed phase column in approximately 4.9 min. The TBA-MDA adduct was quantitated with a fluorescence detector set at 515 nm excitation and 550 nm emission. Using this method, picomole quantities of MDA can be easily detected in plasma and liver samples.

This is a preview of subscription content, access via your institution.

Abbreviations

BHT:

butylated hydroxytoluene

HCI:

hydrochloric acid

HPLC:

high performance liquid chromatography

MDA:

malondialdehyde

TBA:

2-thiobarbituric acid

TMP:

1,1,3,3-tetramethoxypropane

References

  1. 1.

    Feher, J., G. Csomos, and A. Vereckei, (1987)Free Radical Reactions in Medicine, pp. 1–197, Springer-Verlag, Berlin.

    Google Scholar 

  2. 2.

    McBrien, D.C.H., and T.F. Slater, eds. (1989)Free Radicals, Lipid Peroxidation and Cancer, pp. 1–447, Academic Press, London.

    Google Scholar 

  3. 3.

    Chow, C.K., ed. (1988)Cellular Antioxidant Defense Mechanisms, Vols. 1–3, CRC Press, Boca Raton.

    Google Scholar 

  4. 4.

    Slater, T.F. (1984)Methods Enzymol. 105, 283–293.

    PubMed  CAS  Google Scholar 

  5. 5.

    Chow, C.K. (1979)Am. J. Clin. Nutr. 32, 1066–1081.

    PubMed  CAS  Google Scholar 

  6. 6.

    Smith, C., and R.E. Anderson, (1987)Free Rad. Biol. Med. 3, 341–344.

    PubMed  CAS  Google Scholar 

  7. 7.

    Aoshima, H. (1978)Anal. Biochem. 87, 49–55.

    PubMed  CAS  Article  Google Scholar 

  8. 8.

    Asakawa, T., and S. Matsushita, (1980)Lipids 15, 137–140.

    CAS  Article  Google Scholar 

  9. 9.

    Wong, S.H.Y., J.A. Knight, S.M. Hopfer, O. Zaharia, C.N. Leach, Jr., and F.W. Sunderman, Jr. (1987)Clin. Chem. 33, 214–220.

    PubMed  CAS  Google Scholar 

  10. 10.

    Wade, C.R., P.G. Jackson, and A.M. Van Rij, (1985)Biochem. Med 33, 291–296.

    PubMed  CAS  Article  Google Scholar 

  11. 11.

    Bird, R.P., S.O. Hung, M. Hadley, and H.H. Draper, (1983)Anal. Biochem. 128, 240–244.

    PubMed  CAS  Article  Google Scholar 

  12. 12.

    Lee, H.S., and A.S. Csallany, (1987)Lipids 22, 104–107.

    PubMed  CAS  Article  Google Scholar 

  13. 13.

    Esterbauer, H., J. Lnag, S. Zadravee, and T.F. Slater, (1984)Methods Enzymol. 105, 319–328.

    PubMed  CAS  Article  Google Scholar 

  14. 14.

    Bird, R.P., and H.H. Draper, (1984)Methods Enzymol. 105, 299–305.

    PubMed  CAS  Google Scholar 

  15. 15.

    Bull, A.W., and L.J. Marnett, (1985)Anal. Biochem. 149, 284–290.

    PubMed  CAS  Article  Google Scholar 

  16. 16.

    Kikugawa, K., T. Kato, and A. Iwata, (1988)Anal. Biochem. 174, 512–521.

    PubMed  CAS  Article  Google Scholar 

  17. 17.

    Esterbauer, H., and H. Zollner, (1989)Free Rad. Biol. Med. 7, 197–203.

    PubMed  CAS  Article  Google Scholar 

  18. 18.

    Yagi, K. (1976)Biochem. Med. 15, 212–216.

    PubMed  CAS  Article  Google Scholar 

  19. 19.

    Knight, J.A., S.E. Smith, V.E. Kinder, and Anstall, H.B. (1987)Clin. Chem. 33, 2289–2291.

    PubMed  CAS  Google Scholar 

  20. 20.

    Yamamoto, Y., M.H. Brodsky, J.C. Baker, and B.M. Ames, (1987)Anal. Biochem. 160, 7–13.

    PubMed  CAS  Article  Google Scholar 

  21. 21.

    Miyazawa, T., K. Yasuda, K. Fujimoto, and T. Kaneda, (1988)J. Biochem. 103, 744–746.

    PubMed  CAS  Google Scholar 

Download references

Author information

Affiliations

Authors

Corresponding author

Correspondence to Ching K. Chow.

About this article

Cite this article

Tatum, V.L., Changchit, C. & Chow, C.K. Measurement of malondialdehyde by high performance liquid chromatography with fluorescence detection. Lipids 25, 226–229 (1990). https://doi.org/10.1007/BF02535752

Download citation

Keywords

  • Linolenic Acid Oxidation
  • Part Distil Water
  • Soybean Lipoxidase
  • Tetrabutyl Ammonium Dihydrogen Phosphate