Measurement of malondialdehyde (MDA) accumulation by thiobarbituric acid (TBA) reactive substances (TBARS) assay

The MDA content was determined according to Nareshkumar et al. (2020) with minor modifications. The samples were collected and homogenized in 2 ml of 0.1% (w/v) trichloroacetic acid (TCA). The homogenate was centrifuged at 10,000 rpm for 5 min. 0.5 ml of supernatant was added to the solution containing 4% TCA (w/v) and 0.5% TBA (w/v). The mixture was heated at 95°C for 15 min, then cool down to room temperature, and centrifuged at 10,000 rpm for 5 min. The clear solution’s absorbance was recorded at 532 nm and corrected for non-specific turbidity by subtracting the absorbance at 600 nm.