Formation of 4-hydroxynonenal, a toxic aldehyde, in soybean oil at frying temperature


The formation of 4-hydroxy-2-trans-nonenal (HNE), a mutagenic and cytotoxic product of the peroxidation of linoleic acid, was monitored in soybean oil that was heated at 185°C for 2, 4, 6, 8, and 10 h. Unheated soybean oil contained no HNE and a relatively low concentration of polar lipophilic secondary oxidation products (aldehydes and related carbonyl compounds), measured as 2,4-dinitrophenylhydrazine derivatives by HPLC. An increase in the concentration of both HNE and total lipophilic polar oxidation products was observed with increased exposure to frying temperature. A considerable concentration of HNE had already formed at 2 h and the concentration continued to increase at 4 and 6 h of heating. After 6 h the concentration of HNE decreased, possibly due to degradation of the aldehyde with further exposure to high temperature. The loss of endogenous tocopherols was also monitored in the heated oil, and the tocopherol concentration decreased as the secondary lipid oxidation products increased.

This is a preview of subscription content, log in to check access.


  1. 1.

    Kaneda, T., and T. Miyazawa, Lipid Peroxides and Nutrition, World Rev. Nutr. Diet. 50:186–214 (1987).

    CAS  Google Scholar 

  2. 2.

    Crampton, E.W., R.H. Common, F.A. Farmer, A.G. Wells, and D. Crawford, Studies to Determine the Nature of the Damage to the Nutritive Value of Some Vegetable Oils from Heat Treatment, J. Nutr. 60:13–24 (1953).

    Google Scholar 

  3. 3.

    Chow, C.K., Biological and Toxicological Properties of Fatty Acids and Their Derivatives, in Fatty Acids in Industry, edited by R.W. Johnson and E. Fritz, Marcel Dekker, New York, 1989, pp. 617–658.

    Google Scholar 

  4. 4.

    Sanders, T.A.B., Nutritional Aspects of Randicity, in Rancidity in Foods, edited by J.C. Allen and R.J. Hamilton, Elsevier, London, 1989, pp. 125–139.

    Google Scholar 

  5. 5.

    Gurr, M.I., Lipids: Products of Industrial Hydrogenation, Oxidation and Heating, in Nutritional and Toxicological Aspects of Food Processing, edited by R. Walter and E. Quattrucci, Taylor and Francis, London, 1988, pp. 139–155.

    Google Scholar 

  6. 6.

    Giani, E., I. Masi and C. Galli, Heated Fat, Vitamin E, and Vascular Eicosanoids, Lipids 20:439–448 (1985).

    CAS  Article  Google Scholar 

  7. 7.

    Billek, G., Heated Fats in the Diet, in The Role of Fats in Human Nutrition, edited by F.G. Padley and J. Podmore, Ellis Horwood, Chichester, United Kingdom, 1985, pp. 163–172.

    Google Scholar 

  8. 8.

    Cortesi, R., and O.S. Privett, Toxicity of Fatty Ozonides and Peroxides, Lipids 27:715–721 (1992).

    Google Scholar 

  9. 9.

    Holman, R.T. and S.I. Greenberg, A Note on the Toxicities of Methyl Oleate Peroxide and Ethyl Linoleate Peroxide, J. Am. Oil Chem. Soc. 35:707 (1958).

    CAS  Google Scholar 

  10. 10.

    Olcott, H.S., and A. Dolev, Toxicity of Fatty Acid Ester Hydroperoxides, Proc. Soc. Exp. Biol. Med. 114:820 (1963).

    CAS  Google Scholar 

  11. 11.

    Billek, G., Health Aspects of Thermoxidized Oils and Fats, Eur. J. Lipid Sci. Technol. 102:587–593 (2000).

    CAS  Article  Google Scholar 

  12. 12.

    Esterbauer, H., H. Zollner and R.J. Schaur, Hydroxyalkenals: Cytotoxic Products of Lipid Peroxidation, ISI Atlas of Sci. Biochem. 1:311–317 (1988).

    CAS  Google Scholar 

  13. 13.

    Kim, S.-S., D.D. Gallaher, and A.S. Csallany, Lipophilic Aldehydes and Related Carbonyl Compounds in Rat and Human Urine, Lipids 34:489–496 (1999).

    Article  Google Scholar 

  14. 14.

    Grootveld, M., M.D. Atherton, A.N. Sheerin, J. Hawkes, D. Blake, T.E. Richens, C.J.L. Silwood, E. Lynch, and A.W.D. Claxson, In vivo Absorption, Metabolism, and Urinary Excretion of α,β-Unsaturated Aldehydes in Experimental Animals, J. Clin. Invest. 101:1210–1218 (1998).

    CAS  Google Scholar 

  15. 15.

    Kanazawa, K., E. Kanazawa, and M. Natake, Uptake of Secondary Autoxidation Products of Linoleic Acid by the Rat, Lipids 20:412–419 (1985).

    CAS  Article  Google Scholar 

  16. 16.

    Witz, G., Biological Interactions of α,β-Unsaturated Aldehydes, Free Rad. Biol. Med. 7:333–349 (1989).

    CAS  Article  Google Scholar 

  17. 17.

    Comporti, M., Lipid Peroxidation: Biopathological Significance, Free Rad. Biol. Med. 7:333–349 (1993).

    Google Scholar 

  18. 18.

    Kanazawa, E., and H. Ashida, Target Enzymes on Hepatic Dysfunction Caused by Dietary Products of Lipid Peroxidation, Arch. Biochem. Biophys. 288:71–78 (1991).

    CAS  Article  Google Scholar 

  19. 19.

    Kaneko, T., K. Kaji, and M. Matsuo, Cytotoxicities of a Linoleic Acid Hydroperoxide and Its Related Aliphatic Aldehydes Toward Cultured Human Umbilical Vein Endothelial Cells, Chem. Biol. Interact. 67:295–304 (1988).

    CAS  Article  Google Scholar 

  20. 20.

    Esterbauer, H., R.J. Schaur, and H. Zollner, Chemistry and Biochemistry of 4-Hydroxynonenal, Malonaldehyde and Related Aldehydes, Free Rad. Biol. Med. 11:81–128 (1991).

    CAS  Article  Google Scholar 

  21. 21.

    Esterbauer, H., Aldehydic Products of Lipid Peroxidation, in Free Radicals, Lipid Peroxidation and Cancer, edited by D.C.H. McBrien and T.F. Slater, Academic Press, New York, 1982, pp. 101–128.

    Google Scholar 

  22. 22.

    Esterbauer, H., Cytotoxicity and Genotoxicity of Lipid Oxidation Products, Am. J. Clin. Nutr. 57:779S-786S (1993).

    CAS  Google Scholar 

  23. 23.

    Lang, J., C. Celotto, and H. Esterbauer, Quantitative Determination of the Lipid Peroxidation Product 4-Hydroxynonenal by High-Performance Liquid Chromatography, Anal. Biochem. 150:369–378 (1985).

    CAS  Article  Google Scholar 

  24. 24.

    Liu, Y.-M., J.-R. Miao, and T. Toyo-oka, Determination of 4-Hydroxy-2-nonenal by Precolumn Derivatization and Liquid Chromatography with Laser Fluorescence Detection, J. Chromatogr. A 719:450–456 (1996).

    CAS  Article  Google Scholar 

  25. 25.

    Miyake, T., and T. Shibamoto, Simultaneous Determination of Acrolein, Malonaldehyde, and 4-Hydroxy-2-nonenal Produced from Lipids Oxidized with Fenton's Reagent, Food Chem. Toxicol. 34:1009–1011 (1996).

    CAS  Article  Google Scholar 

  26. 26.

    Seppanen, C.M., and A.S. Csallany, Simultaneous Determination of Lipophilic Aldehydes by High-Performance Liquid Chromatography in Vegetable Oil, J. Am. Oil Chem. Soc. 78:1253–1260 (2001).

    CAS  Google Scholar 

  27. 27.

    Thomas, M.J., T.W. Robinson, M. Samuel, and H.J. Forman, Detecting and Identifying Volatile Aldehydes as Dinitrophenyl-hydrazones Using Gas Chromatography-Mass Spectroscopy, Free Rad. Biol. Med. 18:553–557 (1995).

    CAS  Article  Google Scholar 

  28. 28.

    Csallany, A.S., S.-S. Kim, and D.D. Gallaher, Response of Urinary Lipophilic Aldehydes and Related Carbonyl Compounds to Factors That Stimulate Lipid Peroxidation in vivo, Lipids 35:855–862 (2000).

    CAS  Article  Google Scholar 

  29. 29.

    Esterbauer, H., and H. Zollner, Methods for Determination of Aldehydic Lipid Peroxidation Products, Free Rad. Biol. Med. 7:197–203 (1989).

    CAS  Article  Google Scholar 

  30. 30.

    Carpenter, A.P., Determination of Tocopherols in Vegetable Oils, J. Am. Oil Chem. Soc. 56:668–671 (1979).

    CAS  Google Scholar 

  31. 31.

    Buege, J.A., and S.D. Aust, Microsomal Lipid Peroxidation, Meth. Enzymol. 52:302–310 (1978).

    CAS  Article  Google Scholar 

  32. 32.

    Mehlenbacher, V.C., T.H. Hopper, E.M. Sallee, W.E. Link, R.O. Walker, R.C. Walker, and D. Firestone, Official Methods and Recommended Practices of the American Oil Chemists' Society, American Oil Chemists' Society, Champaign, 1990, Recommended Practice Cd 8B-90.

    Google Scholar 

  33. 33.

    Metcalf, L.D., and A.A. Schmitz, The Rapid Preparation of Fatty Acid Esters for Gas Chromatographic Analysis, Anal. Chem. 33:363–364 (1961).

    Article  Google Scholar 

  34. 34.

    Sonntag, N.O.V., Composition and Characteristics of Individual Fats and Oils, in Bailey's Industrial Oil and Fat Products, 4th edn., edited by D. Stern, John Wiley & Sons, New York, 1979, pp. 289–477.

    Google Scholar 

  35. 35.

    Stauffer, C.E., Fats and Oils, Eagan Press, St. Paul, 1996, 131 pp.

    Google Scholar 

  36. 36.

    U.S. Department of Agriculture, Agricultural Research Service, USDA Nutrient Database for Standard Reference, Release 12, Nutrient Data Laboratory Home Page, (1998).

  37. 37.

    Chow, C.K., and H.H. Drapei, Oxidative Stability and Antioxidant Activity of the Tocopherols in Corn and Soybean Oils, Int. J. Vitam. Nutr. Res. 44:396–403 (1974).

    CAS  Google Scholar 

  38. 38.

    Pokorný, J., Major Factors Affecting the Autoxidation of Lipids, in Autoxidation of Unsaturated Lipids, edited by H.W.-S. Chan, Academic Press, London, 1987, pp. 141–206.

    Google Scholar 

  39. 39.

    Lea, C.H., and R.J. Ward, Relative Antioxidant Activity of the Seven Tocopherols, J. Sci. Food Agric. 10:537–548 (1959).

    CAS  Article  Google Scholar 

  40. 40.

    Govind Rao, M.K., and K.T. Achaya, Role of Tocopherol as an Antioxidant in Safflower Oil, Fette Seifen Anstrichm. 69: 711–714 (1967).

    Article  Google Scholar 

  41. 41.

    Kovats, T.K., and E. Berdorfer-Krazner, On the Antioxidative Mechanism of α-, β-, γ-, and δ-Tocopherols in Lard, Nahrung 12:47–414 (1968).

    Google Scholar 

Download references

Author information



Corresponding author

Correspondence to A. Saari Csallany.

About this article

Cite this article

Seppanen, C.M., Saari Csallany, A. Formation of 4-hydroxynonenal, a toxic aldehyde, in soybean oil at frying temperature. J Amer Oil Chem Soc 79, 1033–1038 (2002).

Download citation

Key Words

  • Hydroxyaldehydes
  • 4-hydroxynonenal
  • lipid peroxidation
  • soybean oil
  • thermal oxidation
  • α,β-unsaturated aldehydes