Lipid peroxidation

The extent of lipid peroxidation was determined by the method of Ohkawa et al. 1979, the principle of this method being that malondialdehyde (MDA), an end product of lipid peroxidation, reacts with thiobarbituric acid (TBA) to form a pink chromogen. To this assay, 0.2 ml of 8.1% sodium dodecyl sulfate, 1.5ml of 20% acetic acid (pH 3.5), and 1.5ml of 0.81% thiobarbituric acid aqueous solution were added in succession in a reaction tube. To this reaction mixture, 0.2 ml of each homogenate will be added. The mixture will then be heated in boiling water for 60 min. After cooling to room temperature, 5ml of butanol:pyridine (15:1, v/v) solutions should be added. The mixture will then be centrifuged at 5000rpm for 15min. The upper organic layer will be separated, and the intensity of the resulting pink color can read at 532 nm. Tetramethoxypropane will be used as an external standard. The level of lipid peroxides can be expressed as nmoles of MDA formed/mg protein. Dr. Thanasekaran jayakumar (tjaya_2002@yahoo.co.in)