Effects of Picrorhiza kurroa (Katuki rhizome) on Antioxidant Activity: A clinical study
The present study was carried out to investigate the efficacy and safety of the Effects of Picrorhiza kurroa (Katuki rhizome) on Antioxidant Activity
Extraction, fractionation, and isolation:
The fresh plant material was dried at 40±5° and crushed properly. One kilogram of powdered material was extracted with ethanol: water (95:5, v/v) (1×7 l, 1×3.5 l, 6×1 l). The ethanol solutions were combined and dried in a rota evaporator at 40±5° (130 g). The crude ethanol extract (100 g) was suspended in water and successively extracted with hexane (3×250 ml), chloroform (3×250 ml), ethyl acetate (3×250 ml), and n-butanol (3×250 ml) and evaporation of the solvents at reduced pressure gave 6.6 g of n-hexane, 7.0 g of chloroform, 1.5 g of ethyl acetate and 23.2 g of n-butanol extract. These extracts were lyophilized and kept in the dark at +4° until tested. Isolation of compounds through column chromatography was started by using 20.0 g of extract from the mixed obtained fractions of n-butanol. A slurry of the extract was prepared by dissolving it in a minimum volume of methanol followed by adsorbing the extract over silica gel (mesh size 230-400, 20.0 g). The slurry of the extract was uniformly packed over a dry silica gel column (mesh size 230-400, 455.0 g, 35×7.0 cm) for column chromatography. Elution of components was started through column chromatography using an isocratic solvent system (ethyl acetate:chloroform:methanol: water; 15:8:2:0.5). Fractions of 100 ml each were collected in a conical flask. TLC (silica gel F254) of all individual fractions were developed using a solvent system (ethyl acetate:chloroform: methanol: water; 15:8:4:1; 12:8:8:2) and then viewed under UV chamber at both the wavelengths (254 and 366 nm) followed by spraying with iodine in iodine chamber and finally sprayed with vanillin sulfuric acid as visualizing reagent. Based on the TLC profile of the fractions, similar fractions were pooled and then dried in rotavapor under reduced pressure at a temperature of about 40±5°. After drying all the fractions, pooled fractions were obtained. Fraction no. 70-80 were precipitated to obtain luteolin-5-O-glucoside (compound 1, fig. 1a).
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