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    The extracts were diluted in ethanol to a final volume of 50 mL. Twenty-five milliliters of this solution were added to 0. This mixture was refluxed for 30 min and filtered at room temperature. Twenty milliliters of this solution was diluted in distilled water 30 mL and extracted with ethyl acetate 15 mL. This procedure was repeated three times. The ethyl acetate fraction was concentrated in a rotary evaporator and resuspended in 20 mL methanol. HPLC analysis evaluated apigenin, catechin, epicatequin, kaempferol, luteolin, naringin, quercetin, rutin, and taxifolin content using a series HP equipped with a UV detector and quaternary pumping system as described previously [ 23 ].

    The following gradient conditions were used: Flavonoids were monitored by UV absorbance at nm. The quantities of the main flavonoids were estimated from the standard curves of standard compounds all from Sigma-Aldrich.

    Briefly, the ethanolic stock solutions of the extracts were diluted to different concentrations 0. Tubes were stored on dark for 20 min, and the absorbance was measured at nm. Catalase-like assay determined the hydrogen peroxide decomposition rate at nm [ 25 ]. All efforts were made to minimize animal suffering and the number of animals necessary to produce reliable scientific data. Ten male rats were euthanized by decapitation without anesthesia, and the brain and liver were rapidly excised on an iced Petri dish. The cerebellum, cerebral cortex, hippocampus, and liver were dissected.

    Tissues from all rats were mixed and homogenized in PBS pH 7. Lipid and protein oxidative damage, and activity of the antioxidant enzymes, superoxide dismutase EC1. Lipid peroxidation was monitored using the formation of thiobarbituric acid reactive species TBARS during an acid-heating reaction. This method has been widely adopted as a sensitive method for the measurement of lipid peroxidation.

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