Malondialdehyde Content Measurement

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 (E_x/E_m = 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.

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