4.6. Lipid Peroxidation

The level of lipid peroxidation in the brain was determined by the TBARS assay [52]. A supernatant of homogenized brain tissue (200 μL) was mixed with 1% phosphoric acid and 0.67% TBA solution. This mixture was boiled for 45 min, and then cooled on ice. n-Butanol was added to this mixture, and it was centrifuged at 3000 rpm for 10 min. The absorbance of the supernatant was detected at 540 nm using microplate reader (Thermo Fisher Scientific, Vantaa, Finland). The level of lipid peroxidation was calculated in terms of MDA contents using the standard curve of MDA.