强阳离子交换毛细管液相色谱-反相加压毛细管电色谱二维系统的构建及其在中药黄柏提取物分离中的应用

加压毛细管电色谱(pCEC)具有电泳和液相色谱的双重分离机理,其柱效高、选择性强、分辨率高和分离速度快并可进行梯度洗脱。我们在此基础上加入离子交换色谱模式,构建了强阳离子交换-反相加压毛细管液相色谱(micro strong cation exchange liquid chromatography/reversed phase pressurized capillary electrochromatography, μ-SCXLC/RP-pCEC)二维系统,并对中药黄柏的提取物进行了优化分离。第一维μ-SCXLC采用线性盐梯度分离,样品被切割成11个馏分洗脱收集后进入第二维,第二维脱盐后,采用RP-pCEC进行分离分析,梯度洗脱。以中药黄柏提取物为样品,此二维系统的分辨率和峰容量都较一维系统有很大提高,理论峰容量可达900左右,证明构建的二维体系非常适合复杂样品的分离分析。

Development of two-dimensional micro strong cation exchange liquid chromatography/reversed phase pressurized capillary electrochromatography and its application on Cortex Phellodendri extract

Pressurized capillary electrochromatography (pCEC) combines capillary electrophoresis (CE) with high performance liquid chromatography (HPLC),resulting in high separation efficiency,high selectivity,high resolution and fast speed.In addition,it provides a solvent gradient capability.Based on pCEC,a two-dimensional chromatography was constructed using micro strong cation exchange liquid chromatography and reversed phase pressurized capillary electrochromatography (μ-SCXLC/RP-pCEC) for the analysis of Cortex Phellodendri extract.A sample was separated by the first dimensional SCX column with a linear salt gradient elution into 11 plugs.Each plug was collected and injected into the second dimensional RP-pCEC column with a linear gradient for further separation.Compared with one-dimensional liquid chromatography,it has higher resolution and larger peak capacity.The two-dimensional chromatographic system is suitable for the separation and analysis of complex samples.