2.7. Measurement of Malondialdehyde (MDA) Contents

Lipid peroxidation was quantified by measuring malondialdehyde (MDA) levels using the TBARS method. Briefly, whole brain tissues were homogenized with 20 mM tris-HCl buffer (pH 7.4) and centrifuged at 12,000 × g for 15 min at 4 °C. Supernatants were precipitated using phosphoric acid and incubated with thiobarbituric acid (TBA) at 95 °C for 1 h. Organic complexes were extracted; n-butanol and absorbances were measured at 532 nm using a microplate reader. Lipid peroxidation was quantified using an MDA (Sigma Co., St. Louis, MO, USA) calibration curve.