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Malondialdehyde Content Measurement

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Malondialdehyde (MDA) is an organic compound with the nominal formula of CH2 (CHO)2. MDA can be generated by oxidizing agents that alter lipid structure, and cause lipid peroxidation of polyunsaturated fatty acids. MDA is one of the most widely measured non-enzymatically formed reactive electrophilic species (RES), and is usually used as a marker for oxidative stress.

Lifeasible, as a leading plant biotechnology company, provides advanced technologies for MDA content measurement. The most commonly used methods for the determination of MDA content as follows (Figure 1):

Figure 1. Simplified schematic descriptions of three different reactions that are utilized in MDA analysis (Tsikas, 2017).

  • Thiobarbituric acid (TBA) reactive substances assay. This assay is based on the principle that under acidic conditions (e.g., glacial acetic acid or sulfuric acid) and elevated temperatures (e.g., 95 °C) for extended reaction time (e.g., 60 min), the MDA with TBA can form a red-colored MDA-(TBA)2 adduct (Figure 1A). The MDA-(TBA)2 adduct can be easily quantified colorimetrically (OD = 532 nm) or fluorometrically (Ex/Em = 532/553 nm).
  • Dinitrophenyl hydrazine (DNPH) assay. This assay utilizes derivatization reagents such as 2, 4- dinitrophenyl hydrazine (DNPH) or 2, 4-diaminonaphthalene (DAN) to react with the carbonyl groups of MDA (Figure 1B). The resulted complex can be quantified by HPLC, GC-MS, or LC-MS/MS methods.
  • Pentafluorobenzyl (PFB) bromide assay. The PFB bromide can alkylate MDA at its central C atom, which causes both carbonylic groups to intact (Figure 1C). The MDA-(PFB)2 derivative can be quantified by GC-MS or GC-MS/MS analysis.

We deliver consistent, high-quality data with the flexibility to adapt protocols based on specific customer requirements. Welcome to contact us for technical consults, quotations, questions, and more.


  • Tsikas D. Assessment of lipid peroxidation by measuring malondialdehyde (MDA) and relatives in biological samples: Analytical and biological challenges. Anal Biochem. 2017, 524: 13-30.