MDA contents:

MDA in the leaves was investigated by the method of Carmak & Horst (1991). This reaction was based on Thiobarbituric acid (TBA). Fresh leaves (0.5g) were ground properly in 10 ml of 0.1% tri-chloroacetic acid solution and centrifuged for 10 minutes at 12,000rpm. 1ml of the supernatant was reacted with 1 ml of 20% TCA solution comprised of 0.5% thiobarbituric acid and then it was heated for 30 minutes at 100oC in a water bath and then cooled. The absorbance of the supernatant was read at 450, 532, and 600 nm. The absorption coefficient of 155,000 nmol mol-1 was used to calculate the MDA contents as;

MDA (nmol ml-1) = [(A532-A600)/155,000]106

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