Preferencje help
Widoczny [Schowaj] Abstrakt
Liczba wyników
2014 | 23 | 2 |

Tytuł artykułu

Improved derivatization of malondialdehyde with 2-thiobarbituric acid for evaluation of oxidative stress in selected tissues of chickens

Warianty tytułu

Języki publikacji



A selective and sensitive method based on derivatization with 2-thiobarbituric acid and ultra-fast liquid chromatographic separation is described for the determination of malondialdehyde (MDA) in chicken liver, muscles and adipose tissue, and in lard and fish oil. Preparation of samples involves acid hydrolysis and derivatization. Separation is achieved using an Accucore C18-column (2.6 μm, Hydro-RP, 150×3.0 mm), an acetonitrile gradient in water, and detection at 530 nm. The results indicate that external calibration based on standard solutions of MDA may be used for measuring the MDA concentration in adipose tissue, lard and fish oil due to the absence of matrix effects. The MDA concentration in protein-rich biological samples should be calculated as the difference between the MDA concentration measured in MDA-spiked and unspiked samples of the same specimen and mass and the known concentration of the MDA spike. For liver or muscle samples, we suggest using external calibration based on standard solutions of MDA added to the same mass of liver or muscles. The proposed method is suitable for rapid and sensitive analysis of MDA in samples of animal origin or in plant oils. The method can also be suitable for routine evaluation of oxidative stress in animal tissues and oxidative stability of biological materials and animal products

Słowa kluczowe








Opis fizyczny



  • The Kielanowski Institute of Animal Physiology and Nutrition, Polish Academy of Sciences, 05-110 Jablonna, Poland
  • The Kielanowski Institute of Animal Physiology and Nutrition, Polish Academy of Sciences, 05-110 Jablonna, Poland
  • The Kielanowski Institute of Animal Physiology and Nutrition, Polish Academy of Sciences, 05-110 Jablonna, Poland
  • The Kielanowski Institute of Animal Physiology and Nutrition, Polish Academy of Sciences, 05-110 Jablonna, Poland
  • The Kielanowski Institute of Animal Physiology and Nutrition, Polish Academy of Sciences, 05-110 Jablonna, Poland


  • Baldeiras I., Santana I., Proenca M.T., Garrucho M.H., Pascoal R., Rodrigues A., Duro D., Oliveira C.R., 2010. Oxidative damage and progression to Alzheimer’s disease in patients with mild cognitive impairment. J. Alzheimers Dis. 21, 1165–1177
  • Czauderna M., Kowalczyk J., 2001. Separation of some mono-, diand tri-unsaturated fatty acids containing 18 carbon atoms by high-performance liquid chromatography and photodiode array detection. J. Chromatogr. B 760, 165–178
  • Czauderna M., Kowalczyk J., Marounek M., 2011. The simple and sensitive measurement of malondialdehyde in selected specimens of biological origin and some feed by reversed phase high performance liquid chromatography. J. Chromatogr. B, 879, 2251–2258
  • Dizdaroglu M., Jaruga P., Birincioglu M., Rodriguez H., 2002. Free radical-induced damage to DNA: mechanisms and measurement. Free Radical Biol. Med. 32, 1102–1115
  • Esterbauer H., Schaur R.J., H. Zollner H., 1991. Chemistry and biochemistry of 4- hydroxyl-nonenal, malonaldehyde and related aldehydes. Free Radical Biol. Med. 11, 81–128
  • Grotto D., Santa Maria L.D, Valentini J., Paniz C., Schmitt G., Garcia S.C., Pomblum V.J., Rocha J.B.T., Farina M., 2009. Importance of the lipid peroxidation biomarkers and methodological aspects for malondialdehyde quantification. Quim. Nova 32,169–174
  • Hong Y., Yeh S., Chang C., Hu M., 2000. Total plasma malondialdehyde levels in 16 Taiwanese college students determined by various thiobarbituric acid tests and an improved high-performance liquid chromatography-based method. Clin. Biochem. 33, 619–625
  • Janero D.R., 1990. Malondialdehyde and thiobarbituric acid-reactivity as diagnostic indices of lipid peroxidation and peroxidative tissue injury. Free Radical Biol. Med. 9, 515–540
  • Karatas F., Karatepe M., Baysar A., 2002. Determination of free malondialdehyde in human serum by high-performance liquid chromatography. Anal. Biochem. 311, 76–79
  • Kong Q., Lin C.L., 2010. Oxidative damage to RNA: mechanisms, consequences, and diseases. Cell. Mol. Life Sci. 67, 1817–1829
  • Korchazhkina O., Exley C., Spencer S.A., 2003. Measurement by reversed-phase high-perfor-mance liquid chromatography of malondialdehyde in normal human urine following derivatisation with 2,4-dinitrophenylhydrazine. J. Chromatogr. B 794, 353–362
  • Lawlor J.B., Sheehy P.J.A., Kerry J.P., Buckley D.J., Morrissey P.A., 2000. Measuring oxidative stability of beef muscles obtained from animals supplemented with vitamin E using conventional and derivative spectrophotometry. J. Food Sci. 65, 1138–1141
  • Levine R.L., Williams J.A., Stadtman E.R., Shacter E., 1994. Carbonyl assays for determination of oxidatively modified proteins. Methods Enzymol. 233, 346–357
  • Lykkesfeldt J., 2001. Determination of malondialdehyde as dithiobarbituric acid adduct in biological samples by HPLC with fluorescence detection: comparison with ultraviolet-visible spectrophotometry. Clin. Chem. 47, 1725–1727
  • Mateos R., Bravo L., 2007. Chromatographic and electrophoretic methods for the analysis of biomarkers of oxidative damage to macromolecules (DNA, lipids, and proteins). J. Sep. Sci. 30, 175–191
  • Nielsen F., Mikkelsen B.B., Nielsen J.B., Andersen H.R., Grandjean P., 1997. Plasma malondialdehyde as biomarker for oxidative stress: reference interval and effects of life-style factors. Clin. Chem. 43, 1209–1214
  • Ohkawa H., Ohishi N., 1979. Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal. Biochem. 95, 351–358
  • Oter S., Jin S., Cucullo L., Dorman H.J.D., 2012. Oxidants and antioxidants: friends or foes? Oxid. Antioxid. Med. Sci. 1, 1–4
  • Pilz J., Meineke I., Gleiter H.C., 2000. Measurement of free and bound malondialdehyde in plasma by high-performance liquid chromatography as the 2,4-dinitrophenylhydrazine derivative. J. Chromatogr. B 742, 315–325
  • Tarladgis B.G., Watts B.M., 1960. Malonaldehyde production during the controlled oxidation of pure unsaturated fatty acids. J. Amer. Oil Chem. Soc. 37, 403–407
  • Wong S.H.Y., Knight J.A., Hopfer S.M., Zaharia O., Leach C.N., Sunderman F.W., 1987. Lipoperoxides in plasma as measured by liquid-chromatographic separation of malondialdehydethiobarbituric acid adduct. Clin. Chem. 33, 214–220

Typ dokumentu



Identyfikator YADDA

JavaScript jest wyłączony w Twojej przeglądarce internetowej. Włącz go, a następnie odśwież stronę, aby móc w pełni z niej korzystać.