高速逆流色谱原理及应用

References
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高速逆流色谱的延伸应用
高速逆流色谱原理及应用
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多级萃取技术
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离 心 分 配 色 谱 平 面 图
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离心分配色谱立体图
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同步行星式逆流色谱
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高 速 逆 流 色 谱 运 行 平 面 图
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分离区
混合区
分离区
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HSCCC 系统连接工作图
Young Wan Ha and Yeong Shik Kim
ABSTRACT:
Introduction – Platycosides, the primary constituents of Platycodi Radix, are known to have numerous and varied biological activities, exerting anti-inflammation, anti-allergy, anti-tumour, antiobesity and anti-hyperlipidemia effects. However, effective methods for isolating and purifying platycosides from Platycodi Radix are not currently available. Objective – To develop an efficient method for the preparative separation of six platycosides from Platycodi Radix by high speed counter-current chromatography (HSCCC) coupled with an evaporative light scattering detection (ELSD) system. Methodology – Preparative separation was performed by water extraction using reversed-phase C18 column chromatography on an HSCCC-ELSD system. A two-phase solvent system comprised hexane–n-butanol–water (1:40:20, v/v) and (1:10:5, v/v) was employed. Two other key parameters, revolution speed of the separation column and flow-rate of the mobile phase, were also investigated for optimum HSCCC performance. Each peak fraction obtained from separation of the platycosides was collected according to the ELSD elution profile and determined by HPLC. Results – Using the described method, six platycosides, all with purities of over 94%, could be isolated from 300mg of the platycoside-enriche精d选fra课c件tion. Their structures were characterized by15 electrospray ionisation mass spectrometry (ESI-MS), 1HNMR and 13C-NMR.
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HSCCC方法的建立过程:
1、HPLC方法的建立
尽可能使各峰得到良好的分离
2、溶剂系统的选择
根据样品的溶解特性选择溶剂体系 测定分配系数，依据分配系数确定溶剂系统 K2/K1:0.5-2.5
3、HSCCC分离条件优化
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色谱柱转速；固定相的保留；流动相流速；分离温度。
4、HPLC测定收集组分的纯度
Introduction
Experimental
Apparatus. Reagents and materials. Preparation of crude sample. Measurement of partition coefficients Preparation of the two-phase solvent system and sample solution. HSCCC separation HPLC analysis and identification of HSCCC peak fractions Results and Discussion Selection of the two-phase solvent system and HSCCC conditions Structural identification Acknowledgements
正己烷-乙腈 正己烷-甲醇-水 正己烷-乙酸乙酯-甲醇-水 (石油醚-乙酸乙酯-甲醇-水)
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Preparative Isolation of Six Major Saponins from Platycodi
Radix by High-speed Counter-current Chromatography
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HSCCC常用溶剂系统（依据组分的极性和溶解性）
水溶性和强极性
丁醇-醋酸-水丁醇-甲醇-水(缓冲液) 丁醇-乙酸乙酯-水(缓冲液) 乙酸乙酯-水(缓冲液)
中等极性(氯仿)
氯仿-甲醇-水(毒) 甲基异丁基酮-丙酮-水 正己烷-乙酸乙酯-甲醇-水 (石油醚-乙酸乙酯-甲醇-水)
弱极性(正己烷)