2.2. Analysis of Malondialdehyde (MDA) Concentration in Brain Cortex Homogenates

Experimentally, 0.2 g of the obtained brain cortex samples were homogenized in the volume of 5 mL of ice KCl (154 mM) by Teflon pestles homogenizers, and the supernatants were immediately harvested. Experimentally, 200 µL of the supernatant was mixed with 3 mL of the H₃PO₄ and 800 µL of the KCl solution followed by vortex well. The standard solution of 1, 1, 3, 3-tetra ethoxy propane was used to react with the thiobarbituric acid (TBA) substance and was boiled for 1 h. Finally, 4 mL of the butanol solution was added into the solution and vortex for 5 min followed collected the supernatant for MDA analysis. In general, the detective principle of this present assay relied on the measurement of the pink color that is generated through the reaction of MDA with TBA. The concentrations of brain tissue MDA were determined by spectrophotometer at 532 nm (U-1900, Hitachi, Japan).