Optimization of the extraction and purifcation of Corydalis yanhusuo W.T. Wang based on the Q-marker uniform design method
Optimization of the extraction and purifcation of Corydalis yanhusuo W.T. Wang based on the Q-marker uniform design method
复制标题
基于Q-marker均匀设计法优化延胡索W.T. Wang的提取纯化
作者：
Yongping Zhang;Zuhua Wang;Jian Xu;Fangfang Yang;Chuanyang Dai;Weijie Xie;Zhu Liang;Songbo Su
延胡索生物碱主要分为三大类：原小檗碱、原托品碱和阿朴吗啡。因此，我们在提取和纯化生药时考虑了这三种生物碱的含量，这对于控制延胡索及其衍生产品的质量是必不可少的。在此，我们考察了Q标记均匀设计法在延胡索提取纯化工艺优化中的可行性。本研究采用基于Q标记的综合评分法和均匀设计法对延胡索的提取纯化工艺进行了优化。考察因素包括溶剂浓度、pH值、液固比、提取时间和提取次数。以8个Q标记、总生物碱浸出量和浸出率为评价指标。结果表明，延胡索的最佳回流提取工艺为：加20倍量70%乙醇(稀氨溶液pH=10)，加热回流2次，每次回流提取60min。9种树脂筛选结果表明，NKA-9大孔吸附树脂对8种富集度较高的延胡索生物碱具有最好的分离纯化效果。在最佳的浓缩工艺和洗脱条件下，用5BV的蒸馏水以2BV/h的体积流率洗脱水溶性杂质，洗脱溶剂为70%乙醇，洗脱体积为1.5BV/h，洗脱剂量为12BV。产品中总生物碱含量在50%以上，其中(+)-延胡索乙素3.55%，延胡索乙素3.13%，黄连碱2.79%，盐酸巴马汀2.24%，脱氢延胡索乙素13.11%，(R)-(+)-滨豆碱2.37%，原阿片碱2.71%，白花碱14.03%。结果表明，优化的提取纯化工艺稳定可行，有望为延胡索的工业化生产提供实验依据和参考。
Corydalis yanhusuo W.T. Wang alkaloids are mainly divided into three categories: protoberberine, prototropine and aporphine alkaloids. Therefore, we have taken into account these three alkaloid contents when extracting and purifying crude drugs, which is essential for the quality control of C. yanhusuo and its derivative products. Herein, we investigated the feasibility of the Q-marker uniform design method in the optimization of the extraction and purifcation of C. yanhusuo. In this study, Q-marker-based comprehensive scoring (CS) and uniform design methods were used to optimize the extraction and purifcation of C. yanhusuo. The inspective factors included the solvent concentration, pH, liquid–solid ratio, extraction time and frequency. Then 8 Q-markers, the total alkaloid extraction and the extraction rate were considered as the evaluating indicators during the process. The results indicated that the optimal refux .extraction process of C. yanhusuo was as follows: a total amount of 20 times 70% ethanol (pH=10 of diluted ammonia), heating and refuxing twice, and extracting each time for 60 min. The results of nine-resin screening exhibited that NKA-9 macroporous adsorption resin had the best separation and purifcation efect on 8 kinds of C. yanhusuo alkaloids with stronger enrichment. During the optimal enrichment process and elution conditions, the water-soluble impurities were washed of with 5 BV distilled water at a volume fow rate of 2 BV/h, and the elution solvent was 70% ethanol with an elution volume fow rate of 1.5 BV/h and an elution dosage of 12 BV. Additionally, the total alkaloids of the obtained product were over 50%, of which eight quality markers were (+)-corydaline 3.55%, tetrahydropalmatine 3.13%, coptisine 2.79%, palmatine hydrochloride 2.24%, dehydrocorydaline 13.11%, (R)-(+)-corypalmine 2.37%, protopine 2.71% and glaucine 14.03%. Our data demonstrated that the optimal extraction and purifcation process was stable and feasible, which was expected to provide an experimental basis and reference for the industrial production of C. yanhusuo.