2.13. Measurement of Inhibition of Lipid Peroxidation

Determination of the inhibition of lipid peroxidation (LPO) in linoleic acid system was measured by ferric thiocyanate assay (FTC) [37]. The tested solution, containing 0.6 mL of compound (10 mM in MeOH), 0.6 mL of linoleic acid emulsions (25 mg/mL in 99% ethanol), and 1.2 mL of 50 mM phosphate buffer (pH 7.4), was incubated in the dark at 40 °C. A 0.1 mL aliquot of it was then added to 3 mL of 70% (v/v) ethanol and 0.2 mL of 30% (w/v) ammonium thiocyanate. Precisely 3 min after the addition of 0.2 mL of 20 mM ferrous chloride in 3.5% (v/v) hydrochloric acid to the reaction mixture, the absorbance of the resulting red color was measured at 500 nm. Aliquots were assayed every 24 h for five days. BHT (10 mM in MeOH) was used as a positive control. MeOH was used as a blank.