4.10. Lipid Peroxidation (MDA) Assay

Reactive oxygen species can generate the lipid peroxidation process in an organism. Malondialdehyde (MDA) is one of the final products of polyunsaturated fatty acids peroxidation in the cells. An increase in free radicals causes overproduction of MDA. In the lipid peroxidation assay protocol, the MDA in the sample reacts with thiobarbituric acid (TBA) to generate an MDA–TBA adduct. Cells (1 × 10⁶) were homogenized on ice in 300 µL of MDA lysis buffer (with 3 μL BHT (100×), then centrifuged (13,000× g, 10 min) to remove insoluble material. An aliquot of 200 μL of the supernatant was analyzed for lipid peroxidation with a TBARS Assay Kit (Cayman Chemical). Lipid peroxidation was determined through the reaction of malondialdehyde (MDA) and thiobarbituric acid (TBA), forming a colorimetric (532 nm) MDA–TBA adduct.