*2.7. Measurement of the Lipid Peroxidation–MDA Content*

The MDA content was determined using Heath and Packer's technique [57]. Roughly 0.5 g fresh leaf samples were pulverized in 5 mL of trichloroacetic acid (10%) and 2-thiobarbituric acid (0.65%). The mixture was then heated for 1 h at 100 ◦C, cooled to ambient temperature, and centrifuged at 10,000 rpm for 10 min, then the absorbances of the samples were measured in triplicate at 532, 600, and 450 nm. The MDA content of the reaction solution was determined using the following equation:

$$\text{MDA} \left(\mu\text{mol g} - 1\text{ FW}\right) = 6.45 \left(\text{A532} - \text{A600}\right) - \left(0.56 \text{ A450}\right) \tag{1}$$

The MDA concentration was calculated using an extinction coefficient of 155 mmol L−<sup>1</sup> cm−<sup>1</sup> and expressed as nmol of MDA g−<sup>1</sup> FW.
