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Keywords:

  • 2;
  • 4-dinitrophenylhydrazine;
  • HPLC;
  • lipid peroxidation;
  • malondialdehyde;
  • polyunsaturated fatty acids

Abstract

Malondialdehyde (MDA) is a biomarker of lipid peroxidation and is present in foods and biological samples such as plasma. A high-performance liquid chromatography (HPLC) method was applied to determine MDA in fish liver samples after derivatization with 2,4-dinitrophenylhydrazine (DNPH) using a ODS2 column (10 cm × 4.6 mm, 3 μm) and a photodiode array detector. The mobile phase consisted of 0.2% acetic acid (v/v) in distilled water and acetonitrile (42:58, v/v). The present method was validated in terms of linearity, lower limit of quantification, lower limit of detection, precision, accuracy, recovery, and stability of MDA according to U.S. Food and Drug Administration (FDA) guidelines. The limit of quantification of MDA was 0.39 μmol/L, which is comparable to other methods. The recovery of the spiked MDA liver samples was in the range of 92.4% to 104.2%. This newly modified HPLC method is specific, sensitive, and accurate and allows the analysis of MDA within 4 min in fish liver but also in other tissues and plasma.

Practical Application

Malondialdehyde is an established biomarker of lipid peroxidation in foods. We developed a fast, simple, and sensitive HPLC method with photodiode array detection using DNPH for derivatization of malondialdehyde in fish liver.