2.2.2. Extraction of Curcumin

Curcuma longa rhizome dried powder (750 g) was extracted with acetone in a Soxhlet (4 h), filtered, and evaporated (Rotavap ® Hei-VAP, Heidolph, Germany). The remained extract (65 g) was defatted with petroleum ether (3 × 1 L) and run over silica gel CC (2.5 × 60 cm, 125 g) using methylene chloride and methanol mixture as solvent system at different ratios in increasing gradient of polarity. Similar fractions were collected after comparing the TLC silica gel 60 F254 plates. Fractions were dissolved in ethanol (150 ml) and heated then methylene chloride was added (30 ml) and kept at 5 °C overnight. Yellow powder of Curcumin that precipitated were filtered, and purified on Sephadex LH-20 (110 g, 4 × 77 cm) using pure methanol (Analytical grade) and tested by spotting against standard Curcumin (500 mg, Sigma Aldrich C7727) (Revathy et al., 2011, Nurjanah and Saepudin, 2019). The extracted Curcumin is dried and weighed about 17.5 g.