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The supernatant was removed and incubated with 2 ml of thiobarbituric acid 0.2% (Merck) for 60 minutes at 90°C.
The MDA generated was measured by adding 1.6 ml of thiobarbituric acid solution and the reaction mixture heated at 95°C for 15 min.
The supernatant was incubated with 1 ml of PBS (pH: 7.2) and 1 ml of thiobarbituric acid 0.6% (Merck) for 20 minutes at 90°C.
After disappearance of yellow color derived from the released iodine, the 1.5 mL of thiobarbituric acid (0.1 M, pH 9.0) was added to a reaction mixture.
The reaction mixture was then incubated for 1 h at 37°C before the addition of 1 ml of thiobarbituric acid solution and 1 ml of trichloroacetic acid.
For TBARS analysis, 1 mL of saliva sample and known concentrations of malondialdehyde (MDA) standard were mixed with 2 mL of thiobarbituric acid (TBA) solution and digested for 60 min at 95 °C in a water bath.
Similar(47)
0.50 g of frozen soybean leaves was homogenized in 3 mL of 0.5% thiobarbituric acid (TBA) in 20% TCA and then centrifuged at 12,000×g for 20 min at 4 °C to select the extract.
The mixture contained 0.2 mL tissue sample and 2 mL of 0.6% thiobarbituric acid (TBA, w/v), after heating in boiling water for 15 min, cooling rapidly, then centrifuged at 3000 r/min for 10 min, and the supernatant was used for the determination of MDA level.
The reagent was prepared by mixing 375 mg of thiobarbituric acid, 20 g of Trichloroacetic acid, 10 mg of butyl-hydroxytoluene, 20 mL of HCl 1 N, in 50 mL of distilled water.
The homogenate was centrifuged at 4 000× g for 10 min at 25°C and 3 mL of 2-thiobarbituric acid in 20% trichloroacetic acid was added to a 2 mL aliquot of the supernatant.
The final sample concentration from 4 mg in 4 mL of ethanol from the same samples prepared for FTC assay was used. 2 mL of 20% trichloroacetic acid and 2 mL of 0.67% of thiobarbituric acid were added to 1 mL of sample solution from the FTC method.
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