虎杖酶水解及模拟移动床色谱法纯化白藜芦醇

Optimization of Resveratrol from Polygonum cuspidatum by Enzyme-assisted Extraction and Purification of Resveratrol by Simulated Moving Bed Chromatography

目的确定复合酶水解法得到虎杖中白藜芦醇的最优工艺条件,以及模拟移动床色谱法纯化分离白藜芦醇的工艺参数。方法以白藜芦醇的提取率为指标,考察不同因素对酶水解率的影响规律,并在此基础上运用单因素分析法选出优化的条件,并用三带模拟移动床色谱法纯化白藜芦醇。结果最佳提取工艺为:提取温度45℃,酶解p H值3.0,酶解时间8.0h,植物水解酶的用量为20%(w/w)。采用三带模拟移动床色谱法纯化虎杖白藜芦醇,模拟移动床色谱纯化白藜芦醇的工艺参数为:流动相为V(甲醇)∶V(水)=1∶1;4根IDL=10mm×200mm ODS制备柱;柱连接方式1-2-1;洗脱带流量1.0m L/min,精制带流量1.5m L/min,吸附带流量1.6m L/min,进样流量0.1m L/min,切换时间19min。结论采用酶处理后,可显著提高虎杖中白藜芦醇的得率;采用模拟移动床色谱法分离白藜芦醇,萃取液中白藜芦醇的含量大于98.1%(高效液相色谱法(HPLC)面积归一化法)。

Objective To optimize the extraction techonology of resveratrol from Polygonum cuspidatum by enzyme - assisted extraction and confirm the optimum parameters of purification of resveratrot by simulated moving bed chroma- tography. Methods After the preliminary test, the optimum condition of enzyme - assisted extraction was obtained with an univariate analysis design; purification of resveratrol from Polygonum cuspidatum was accomplished with three zones simulated moving bed chromatography. Results The best experimental conditions were as follows：the extraction temper- ature was 45℃ , the enzymatic pH value was 3.0, the enzymatic time was 8.0h,and the dosage of the enzyme was 20% （w/w）. The parameters of purification for resveratrol with simulated moving bed chromatography were as follows ： mobile phase of methanol -water（ 1： 1, v/v ） ,4 columns （IDL = 10mm× 200mm, ODS stationary phase）, columns connection of 1 -2 - 1, the refine zone flow of 1.5mL/min, the extraction zone flow of 1.0mL/rain, the sorption zone flow of 1.6mL/min. the injection sample flow of 0. lmL/min and the switch time of 19 min. Conclusion Enzyme - assisted extraction can elevate the yield of resveratrol from Polygonum cuspidatum. The content of the resveratrol in the extract liquid （E） obtained by simulated moving bed chromatography was more than 98.1% （HPLC area normalization meth- od）.