*4.4. Total Sugars*

The total sugars were determined spectrophotometrically following the anthrone method with slight modifications [57]. The anthrone reagent (10.3 mM) was prepared by dissolving anthrone in ice-cold 95% H2SO4. The reagent was left to stand for 30–40 min before use. Then, 1 g of fresh sample was extracted in 3 mL of distilled water and centrifuged at 3000× *g* for 15 min at room temperature (RT). Next, 0.5 mL of extract was placed on top of 2.5 mL of anthrone reagent incubated in ice for 5 min. The reactions were heated to 95 ◦C for 10 min and left to cool in ice. The absorbance was read at 620 nm. A calibration curve was generated using a glucose solution (0 to 0.05 mg·mL−<sup>1</sup> ) (*R* <sup>2</sup> = 0.9995).
