Abstract
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.
Similar content being viewed by others
Abbreviations
- ADP:
-
adenosine 5′-diphosphate
- AIN-76 diet:
-
American Institute of Nutrition-76 diet
- HPLC:
-
high-performance liquid chromatography
- MA:
-
malonaldehyde
- TBA:
-
2-thiobarbituric acid
- TBARS:
-
TBA reactive substances
References
Pryor, W.A., Stanley, J.P., and Blair, E. (1976)Lipids 11 370–379.
Frankel, E.N., and Neff, W.E. (1983)Biochim. Biophys. Acta 754, 264–270.
Yagi, K. (1982) inlipid Peroxides in Biology and Medicine (Yagi, K., ed.), pp. 223–242, Academic Press, New York.
Dillard, C.J., and Tappel, A.L. (1973)Lipids 8, 183–189.
Basu, A.K., and Marnett, L.J. (1983)Carcinogenesis 4, 331–333.
Shamberger, R.J., Andreone, T.L., and Willis, C.E. (1974)J. Natl. Cancer Inst. 53, 1771–1773.
Chio K.S., and Tappel, A.L. (1969)Biochemistry 8, 2827–2832.
Nair, V., Vietti D.E., and Cooper, C.S. (1981)J. Am. Chem. Soc. 103, 3030–3036.
Kwon, T.W., and Brown, W.D. (1965)Fed. Proc. 24, 592.
Landsberg, J.D., and Sinnhuber, R.O. (1965)J. Am. Oil Chem. Soc. 42, 821–824.
Shin, B.C., Huggins, J.W., and Carraway, K.L. (1972)Lipids 7, 229–233.
Nair, V., Cooper, C.S., Vietti, D.E., and Turner, G.A. (1986)Lipids 21, 6–10.
Brooks, B.R., and Klamerth, O.L. (1968)Eur. J. Biochem. 5, 178–182.
Buege, J.A., and Aust, S.D. (1978)Methods Enzymol. 52, 302–310.
Gutteridge, J.M.C. (1986)Free Rad. Res. Comms. 1, 173–184.
Sinnhuber, R.O., Yu, T.C., and Yu, Te Chang. (1958)Food Res. 23, 626–633.
Nair, V., and Turner, G.A. (1984)Lipids 19, 804–805.
Marcuse, R., and Johansson, L. (1973)J. Am. Oil Chem. Soc. 50, 387–391.
Ohkawa, H., Ohishi, N., and Yagi, K. (1979)Anal. Biochem. 95, 351–358.
Bigwood, T., and Read, G. (1989)Free Rad. Res. Comms. 6, 387–392.
Kosugi, H., Kojima, T., and Kikugawa, K. (1989)Lipids 24, 873–881.
Ichinose, T., Miller, M.G., and Shibamoto, T. (1989)Lipids 24, 895–898.
Bird, R.P., Hung, S.O.S., Hadley, M., and Draper, H.H. (1983)Anal. Biochem. 128, 240–244.
Hirayama, T., Yamada, N., Nohara, M., and Fukui, S. (1984)J. Sci. Food Agric. 35, 338–344.
Csallany, A.S., Guan, M.D., Manwaring, J.D., and Addis, P.B. (1984)Anal. Biochem. 142, 277–283.
Esterbauer, H., and Slater, T.F. (1981)IRCS Med. Sci. 9, 749–750.
Wade, C.R., Jackson, P.G., and Van Rij, A.M. (1985)Biochem. Med. 33, 291–296.
Bull, A.W. and Marnett, L.J. (1985)Anal. Biochem. 149, 284–290.
Therasse, J., and Lemonnier, F. (1987)J. Chromat. 413, 237–241.
Shamberger, R.J., Shamberger, B.A., and Willis, C.E. (1977)J. Nutr. 107, 1404–1409.
American Institute of Nutrition (1977)J. Nutr. 107, 1340–1348.
Hogeboom, G.H. (1955)Methods Enzymol. 1, 16–19.
Esterbauer, H., Lang, J., Zadravec, S., and Slater, T.F. (1984)Methods Enzymol. 105, 319–328.
Burk, R.F. (1983)Biochim. Biophys. Acta 757, 21–28.
Bradford, M.M. (1976)Anal. Biochem. 72, 248–254.
Thompson, J.N., and Hatina, G. (1979)J. Liquid Chromatog, 2, 327–344.
Behrens, W.A., and Madere, R. (1982)Nutr. Rep. Int. 25, 107–112.
Snedecor, G.W., and Cochran, W.G. (1980)Statistical Methods, 7th ed., Iowa State University Press, Ames.
Lee, H.S., and Csallany, A.S. (1987)Lipids 22, 104–107.
Draper, H.H., Polensek, L., Hadley, M., and McGirr, L.G. (1984)Lipids 19, 836–843.
Dhanakoti, C.N., and Draper, H.H. (1987)Lipids 22, 643–646.
Farris, M.W., Pascoe, G.A., and Reed, D.J. (1985)The Toxicologist 5, 149.
Author information
Authors and Affiliations
Additional information
Bureau of Nutritional Sciences Publication No. 348.
About this article
Cite this article
Behrens, W.A., Madère, R. Malonaldehyde determination in tissues and biological fluids by ion-pairing high-performance liquid chromatography. Lipids 26, 232–236 (1991). https://doi.org/10.1007/BF02543977
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02543977