Key Lab of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, P. R. China.
J Sep Sci. 2011 Jan;34(1):53-8. doi: 10.1002/jssc.201000625. Epub 2010 Dec 3.
An efficient method for purification of alkaloids from Corydalis yanhusuo W. T. Wang using HPLC was developed, featuring a polar-copolymerized stationary phase named C18HCE. As ionizable solutes, the crude alkaloid sample often suffered from serious peak tailing problem on conventional RP-LC columns, and the separation would rapidly became destroyed with the increasing of load amount. However, on the new stationary phase, good peak shapes (asymmetry factor <1.5) as well as good loadability were easily obtained in a commonly used acidic mobile phase condition. The loading amount could reach 10 mg per injection on an analytical C18HCE column for laboratory-scale purification. About 6.8 mg of palmatine (HPLC purity >98%) and 44.4 mg of dehydrocorydaline (HPLC purity >98%) were rapidly derived from 200 mg of the crude alkaloid sample, and the recoveries of these two compounds were 76.5 and 81.7%, respectively. The purified alkaloids were characterized by comparing retention times with standard compounds as well as (1)H-NMR data. The new method is simple and high yielding, and it may provide a promising tool for purification of alkaloids as well as other alkaline compounds.
建立了一种从延胡索（Corydalis yanhusuo W. T. Wang）中高效纯化生物碱的 HPLC 方法，所用的固定相是一种名为 C18HCE 的极性共聚固定相。作为可离解的溶质，粗生物碱样品在常规 RP-LC 柱上经常会出现严重的峰拖尾问题，并且随着上样量的增加，分离效果会迅速下降。然而，在新的固定相上，在常用的酸性流动相条件下，很容易获得良好的峰形（不对称因子<1.5）和良好的上样量。在分析型 C18HCE 柱上，每次进样的上样量可达到 10 mg，可用于实验室规模的纯化。从 200 mg 粗生物碱样品中快速得到 6.8 mg 延胡索乙素（HPLC 纯度>98%）和 44.4 mg 脱氢紫堇碱（HPLC 纯度>98%），这两种化合物的回收率分别为 76.5%和 81.7%。通过与标准化合物和（1）H-NMR 数据比较保留时间对纯化的生物碱进行了表征。该新方法简单、产率高，可能为生物碱以及其他碱性化合物的纯化提供一种有前途的工具。
