Phytochemical Profiling of Plant Extracts

The fresh leaves were oven dried at 40°C then grind to fine powder. The ethanolic extract was prepared by adding ethanol (5 mL) to 0.5 g of powdered raw material, sonicated for 30 minutes, and then filtered using Whatman no. 1 filter paper. The supernatant is collected and used for HPTLC and HPLC analysis. The standard stock solution (1 mg/mL) of shaftoside, orientin, isovitexin, and vitexin was prepared in HPLC grade ethanol and stored at 4°C. Working solutions of lower concentration (shaftoside: 20, 40, 60, 80, 100, 150, 175, and 200 μg/mL, orientin, isovitexin, vitexin: 0.2, 0.4, 0.6, 0.8, 10.0, 20, 40, 60, 80, and 100 ug/mL) were prepared by appropriate dilution of the stock solutions in ethanol.