TBARS formation was measured following the method described by Adriana E Scoccia [3 (link)], malonaldihyde, a secondary metabolite and a marker of lipid peroxidation and oxidative stress, which reacts with Thiobarbituric Acid (TBA) to yield pink colored complex that can be measured spectophotometrically. Briefly, 50 μL plasma was diluted in 10 mL CuSO4 (5.0 mmol/L)solution and incubated at 37°C for 3 hours in ultrasonic waterbath with occasional exposure of ultrasonic wave (SonoSWISS SW6 H, Switzerland) to allow the oxidation process. Then 0.5 mL of 0.78% aqueous solution of thiobarbituric acid and 50 μL of acetic acid was added in 0.5 mL of the plasma-CuSO4 mixture. Control was prepared in the same way except CuSO4. The mixture was heated at 80°C for 40 min in a ultrasonic waterbath and absorbances were taken at 532 nm using a spectrophotometer (SHIMADZU UV 1650 pc, UV–vis. Spectrophotometer, Japan). Percentage of oxidation was calculated as follows-
Where Aps is the absorbance of plasma sample, Ac is absorbance of control.
Where Aps is the absorbance of plasma sample, Ac is absorbance of control.
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