高速逆流色谱原理及应用

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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
二维逆流色谱分离中药应用
实例6 二维CCC分离冬凌草
HSCCC条件: 溶剂系统: 一维:正己烷-乙酸乙酯-甲醇-水 1:5:1:5 二维:正己烷-乙酸乙酯-甲醇-水 3:5:3:5 流 速 :2ml min−1 转 速 :800 rpm 样 品 量 :250 mg 粗提物 机 型: 2D-CCC 保 留 值:一维: 40% 二维: 63% 1: oridonin (甲素) 95% 2: ponicidin (乙素) 95%
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,
Introduction
Hypericum perforatum L., a typical traditional Chinese medicine, is used all over the world for the treatment of depression. Additionally, it has been reported that the extracts of H. perforatum may have an inhibitory effect, on human bladder
1.5 ml/min
2.5 ml/min
Journal of Chromatography A, 1092 (2005) 216–221
HSCCC-HPLC-DAD联用分离中药应用
贯叶连翘中金丝桃苷
逆流色谱分离中药应用(三相溶剂系统)
三相CCC分离15种化合物
HEX—MeOAc—ACN--Water
Structural identification
Acknowledgements References
高速逆流色谱的延伸应用
pH区带逆流色谱分离中药单体中应用
实例1 pH区带CCC分离同分异构体
HSCCC条件:
溶剂系统:正丁醇-水(1:1)
固 定 相:上相pH=0.5(HCl) 流 动 相:下相pH=10.7(NH3) 流 转 速:2.0 ml min-1 速:715 rpm
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)
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)
弱极性(正己烷)
Preparative Isolation of Six Major Saponins from Platycodi
Radix by High-speed Counter-current Chromatography
Young Wan Ha and Yeong Shik Kim ABSTRACT:
Introduction – Platycosides, the primary constituents of Platycodi Radix, are known to have
Application of accelerated solvent extraction coupled with high-performance counter-current chromatography to extraction and online isolation of chemical constituents from Hypericum perforatum L.
Journal of Chromatography A, 1146 (2007) 125–130
HSCCC-ESI-MSn分离中药应用
实例7 分离白花前胡中香豆素
HSCCC条件:
溶剂系统: 石油醚-乙酸乙酯-甲醇-水 5:5:6:4
流 速 :2ml min−1 转 速 :900 rpm 样 品 量 :250 mg 粗提物 机 型 :TBE-300 保 留 值 :54.3% 1: 7.0 mg 99.3% 2: 7.4 mg 96.1% 3: 12.5mg 96.5% 4: 42.1mg 99.3% 5: 9.8 mg 99.7% 6: 65.2 mg 98.1% 7: 31.7mg, 92.8%
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
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
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)
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-enriched fraction. Their structures were characterized by
Journal of Chromatography B, 877 (2009) 2571–2578
扩展和想象
《中国猪业网》2010-12-27
湖州:活猪走进去 鲜肉运出来 全机械化操作
由于该定点屠宰迁建项目采用自动化同步流水线加工,活猪进去,出来便 是新鲜的猪肉,减少了手工的环节和过程,猪肉更卫生,肉品质量也会有 大幅提高。
electrospray ionisation mass spectrometry (ESI-MS), 1HNMR and 13C-NMR.
Introduction Experimental Apparatus. Reagents and materials.
Preparation of crude sample.
检测波长:206 nm 样 品 量:230 mg 粗提物

型:pH-zone-refining CCC
固定相保留: 60% 1: 3-sulfophthalic acid 99.0%
2: 4-sulfophthalic acid 99. 0%
Journal of Chromatography A, 966 (2002) 111–118
Journal of Chromatography A, 1061 (2004) 51–54
逆流色谱分离中药应用(V流动相)
实例3 CCC分离短瓣金莲花黄酮苷
HSCCC条件: 溶剂系统:乙酸乙酯-正丁醇-水(2:1:3) 流 速: 0 –190 min, 1.5ml min−1 190–480 min, 2.5ml min−1 转 速:800 rpm 检测波长:254 nm 样 品 量:500 mg 粗提物 机 型:CCC-1000 保 留 值:49% 1: quercetin-3-O-neohesperidoside 85.1% 9.3 mg 2: orientin(荭草素) 97% 95.8 mg 3: 未知化合物 97% 4: vitexin(牡荆素) 97% 11.6 mg
高速逆流色谱原理及应用
多级萃取技术
离 心 分 配 色 谱 平 面 图
Baidu Nhomakorabea
离心分配色谱立体图
同步行星式逆流色谱
高 速 逆 流 色 谱 运 行 平 面 图
分离区
混合区
分离区
HSCCC 系统连接工作图
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
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
cancer cells , and act as an antioxidant and anticonvulsant.
The extracts of H. perforatum contain many constituents with documented biological activity such as quinic acids, phloroglucinols and a broad range of flavonoids . Exposure of the extracts of H. perforatum to light may lead to the degradation of phloroglucinols, which are extremely sensitive to oxidation and unstable in solution on exposure to air, therefore phloroglucinols are difficult to separate and isolate by conventional method. In this case, amethod combining an extraction system with an isolation system online to avoid the exposure of the extracts to
逆流色谱分离中药应用(V流动相)
实例2 CCC分离蛇床子
HSCCC条件: 溶剂系统: 正己烷-乙酸乙酯-乙醇-水(5:5 :5:5) 流 速: 0 –180 min, 1ml min−1 180–480 min, 2ml min−1 转 速:1000 rpm 检测波长:254 nm 样 品 量:500 mg 粗提物 机 型:CCC-1000 相 保 留:58% A: bergapten 96.5% 45.8 mg B: imperatorin 98.2% 118.3 mg 92.1 、93.7%
Yuchi Zhang, Chunming Liu, Min Yu, Zhengkun Zhang, Yanjuan Qi, Jing Wang, Guimei Wu, Sainan Li, Jing Yu, Yang Hu
Journal of Chromatography A, 1218 (2011) 2827–2834
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