*2.5. Lipid Peroxidation Measurement*

Lipid peroxidation was assessed using the levels of thiobarbituric acid reactive substances (TBARS). First, 0.5 mg/mL of the homogenate was resuspended in 1 mL of phosphate buffer (50 mM KH2PO4, pH 7.6) and incubated with 50 μM FeSO4 for 30 min at 4 ◦C to induce lipid peroxidation [27].

At the end of the incubation time, 2 mL of acid solution (trichloroacetic acid, thiobarbituric acid, and hydrochloric acid) were added to each sample, and they were incubated in boiling water for 30 min. Subsequently, the tubes were placed on ice for 5 min and centrifuged at 7500 rpm for 5 min. The absorbance of each sample was determined at 532 nm on a Shimadzu UV-2550 spectrophotometer. Data were expressed as TBARS/mg protein numbers.
