Liu Yanjuan, Chen Xiaofen, Liu JunXi, Di Duolong
Key Laboratory of Chemistry of Northwestern Plant Resources and Key Laboratory for Natural Medicine of Gansu Province, Chinese Academy of Sciences, Lanzhou Institute of Chemical Physics, Lanzhou, China.
Graduate University of the Chinese Academy of Sciences, Beijing, China.
J Sep Sci. 2015 Jun;38(12):2038-45. doi: 10.1002/jssc.201401466. Epub 2015 Jun 1.
A three-phase solvent system was efficiently applied for high-speed counter-current chromatography to separate secondary metabolites with a wide range of hydrophobicity in Dicranostigma leptopodum. The three-phase solvent system of n-hexane/methyl tert-butyl ether/acetonitrile/0.5% triethylamine (2:2:3:2, v/v/v/v) was selected for high-speed counter-current chromatography separation. The separation was initiated by filling the column with a mixture of intermediate phase and lower phase as a stationary phase followed by elution with upper phase to separate the hydrophobic compounds. Then the mobile phase was switched to the intermediate phase to elute the moderately hydrophobic compounds, and finally the polar compounds still retained in the column were fractionated by eluting the column with the lower phase. In this research, 12 peaks were eluted out in one-step operation within 110 min, among them, eight compounds with acceptable purity were obtained and identified. The purities of β-sitosterol, protopine, allocryptopine, isocorydione, isocorydine, coptisine, berberrubine, and berberine were 94.7, 96.5, 97.9, 86.6, 98.9, 97.6, 95.7, and 92.8%, respectively.
一种三相溶剂系统被有效地应用于高速逆流色谱法，以分离秃疮花中具有广泛疏水性的次生代谢产物。选择正己烷/甲基叔丁基醚/乙腈/0.5%三乙胺（2:2:3:2，v/v/v/v）的三相溶剂系统用于高速逆流色谱分离。分离开始时，用中间相和下相的混合物作为固定相填充柱，然后用上相洗脱以分离疏水性化合物。然后将流动相切换到中间相以洗脱中等疏水性化合物，最后通过用下相洗脱柱来分离仍保留在柱中的极性化合物。在本研究中，在110分钟内一步操作洗脱出色谱峰12个，其中获得并鉴定出8种纯度合格的化合物。β-谷甾醇、原阿片碱、别隐品碱、异紫堇二酮、异紫堇定、黄连碱、小檗红碱和小檗碱的纯度分别为94.7%、96.5%、97.9%、86.6%、98.9%、97.6%、95.7%和92.8%。
