色谱

• 特别策划 • 上一篇    下一篇

工业制备高效液相色谱法制备淫羊藿中的黄酮系列对照品

高明哲, 王莉, 彭杰, 肖红斌*   

  1. 中国科学院分离分析化学重点实验室, 中国科学院大连化学物理研究所, 辽宁 大连 116023
  • 收稿日期:2011-07-29 修回日期:2011-08-17 出版日期:2011-09-28 发布日期:2011-10-28
  • 通讯作者: 肖红斌,博士,研究员,主要从事中药药效物质的高效分离、分析检测. Tel: (0411)84379756
  • 基金资助:

    国家重大新药创制课题(2009ZX09502-023)、国家“十二五”计划项目(2011ZX11307)和中国科学院知识创新工程重要方向性项目(KSCX2-YW-R-209)

Preparation of flavonol glycoside reference standard series from Epimedium brevicornum Maxim using pilot-scale preparative high performance liquid chromatography

GAO Mingzhe, WANG Li, PENG Jie, XIAO Hongbin*   

  1. CAS Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China
  • Received:2011-07-29 Revised:2011-08-17 Online:2011-09-28 Published:2011-10-28

摘要: 淫羊藿甙和朝藿定A、B、C是淫羊藿中重要的活性成分,本研究应用工业制备高效液相色谱从淫羊藿粗提物中分离制备了这4个成分。淫羊藿粗提物经大孔吸附树脂粗分离获得相应的组分后,利用工业制备高效液相色谱完成精制纯化。采用自装填Chromatorex C18制备柱(220 mm×77 mm, 10 μm),乙腈-水(26:74或30:70, v/v)为流动相进行洗脱,在35 min内,实现了这4种成分的基线分离及规模制备。从300 g粗提物(总黄酮含量约20%)中获得淫羊藿甙33 g、朝藿定C 4.6 g、朝藿定B 3.7 g和朝藿定A 0.6 g,产品纯度均达到98%以上。此方法通过两步分离即可实现这4种成分的完全分离,具有快速高效、产品纯度高的特点,适于淫羊藿中淫羊藿甙、朝藿定A、B、C系列对照品的规模制备。

关键词: 朝藿定A、B、C, 对照品, 高效液相色谱, 工业制备, 黄酮类化合物, 淫羊藿, 淫羊藿甙

Abstract: Icariin and epimedins A, B, C are a series of active flavonol glycoside in Epimedium brevicornum Maxim. A pilot-scale preparative high performance liquid chromatographic (HPLC) method was developed to purify the four flavonol glycosides as reference standards from the crude extract of Epimedium brevicornum Maxim. After the crude extract containing ~20% flavonols was enriched using macropore resin, the obtained target fractions were subjected to pilot-scale preparative HPLC purification. With the aid of a self-packed pilot-scale preparative column (220 mm×77 mm, 10 μm), the 4 target compounds were separated well within 35 min in a single chromatographic run by the elution with acetonitrile-water (26:74 or 30:70, v/v). By repetitive injection of the enriched target fraction onto the preparative column, 33 g icariin as well as 4.6 g epimedin C, 3.7 g epimedin B, and 0.6 g epimedin A were obtained from 300 g crude extract. The purities of all products were greater than 98%. This pilot-scale preparative HPLC technique and the two step separation technology for 4 target compounds are quite useful for the production of the reference standard series with good purity like icariin, epimedins A, B, C standards due to its high performance, rapid separation and more amounts of products obtained.

Key words: epimedins A, B, C, flavonol glycosides, high performance liquid chromatography (HPLC), reference standard series, Epimedium brevicornum Maxim, icariin, pilot-scale preparation