Liu Feng, Xi Xingjun, Wang Mei, Fan Li, Geng Yanling, Wang Xiao
Shandong Analysis and Test Center, Shandong Academy of Sciences, Jinan, Shandong, China.
J Sep Sci. 2014 Feb;37(4):376-81. doi: 10.1002/jssc.201301061. Epub 2014 Jan 6.
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 (1)H 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.
成功开发了酶解预处理结合高速逆流色谱法从牛蒡子中转化分离牛蒡子苷元的方法。第一步，牛蒡子提取液用β-葡萄糖苷酶进行酶解。最佳水解条件为40℃、pH 5.0、水解时间24小时、β-葡萄糖苷酶浓度1.25mg/mL。在此条件下，牛蒡子苷元含量从2.60mg/g转化为12.59mg/g。第二步，采用由石油醚/乙酸乙酯/甲醇/水（10:25:15:20，v/v）组成的两相溶剂体系，通过高速逆流色谱法对水解产物中的牛蒡子苷元进行分离纯化，并采用HPLC、ESI-MS和¹H NMR光谱对馏分进行分析。最后，从200mg水解样品中一步分离得到102mg纯度为98.9%的牛蒡子苷元。
