Isolation and purification of arctigenin from Fructus Arctii by enzymatic hydrolysis combined with high‐speed counter‐current chromatography
2014
Liu, Feng | Xi, Xingjun | Wang, Mei | Fan, Li | Geng, Yanling | Wang, Xiao
Enzymatic hydrolysis pretreatment combined with high‐speed counter‐current chromatography for the transformation and isolation of arctigenin from Fructus Arctii was successfully developed. In the first step, the extract solution of Fructus Arctii was enzymatic hydrolyzed by β‐glucosidase. The optimal hydrolysis conditions were 40°C, pH 5.0, 24 h of hydrolysis time, and 1.25 mg/mL β‐glucosidase concentration. Under these conditions, the content of arctigenin was transformed from 2.60 to 12.59 mg/g. In the second step, arctigenin in the hydrolysis products was separated and purified by high‐speed counter‐current chromatography with a two‐phase solvent system composed of petroleum ether/ethyl acetate/methanol/water (10:25:15:20, v/v), and the fraction was analyzed by HPLC, ESI‐MS, and ¹H NMR spectroscopy. Finally, 102 mg of arctigenin with a purity of 98.9% was obtained in a one‐step separation from 200 mg of hydrolyzed sample.
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