色谱

色谱 ›› 2017, Vol. 35 ›› Issue (1): 70-74.DOI: 10.3724/SP.J.1123.2016.08046

• 研究论文 • 上一篇    下一篇

固相萃取法纯化制备水飞蓟宾和异水飞蓟宾

赵晓舒1, 张娜珍2, 刘敏1, 邓付美1, 吴明火1   

  1. 1. 大连理工大学食品与环境学院, 辽宁 盘锦 124221;
    2. 大连理工大学文法学院, 辽宁 盘锦 124221
  • 收稿日期:2016-08-31 出版日期:2017-01-08 发布日期:2013-05-06
  • 通讯作者: 吴明火,Tel:(0427)2631788,E-mail:wumh@dlut.edu.cn.
  • 基金资助:

    国家自然科学基金项目（21205058）；中央高校基本科研业务费专项资金资助项目（DUT15RC（3）048）；大连理工大学大学科技园专项项目.

Purification and preparation of silybin and isosilybin by solid phase extraction

ZHAO Xiaoshu1, ZHANG Nazhen2, LIU Min1, DENG Fumei1, WU Minghuo1   

  1. 1. School of Food and Environmental, Dalian University of Technology, Panjin 124221, China;
    2. School of Humanities and Law, Dalian University of Technology, Panjin 124221, China
  • Received:2016-08-31 Online:2017-01-08 Published:2013-05-06
  • Supported by:

    National Natural Science Foundation of China (No. 21205058); Fundamental Research Funds for the Central Universities (No. DUT15RC(3)048); Special Project for University Science Park of Dalian University of Technology.

PDF

77

摘要：

水飞蓟宾和异水飞蓟宾是水飞蓟素中的主要有效成分，其纯化制备主要借助柱色谱法，制备量大，纯化效果好，但过程非常费时。该研究的主要目的是利用更为快速高效的固相萃取（SPE）法从水飞蓟粗提物中分离纯化水飞蓟宾和异水飞蓟宾。建立了用于分析水飞蓟宾和异水飞蓟宾的高相液相色谱法，通过优化洗脱梯度，实现了水飞蓟宾、异水飞蓟宾与其他组分的分离。试用了3种保留机理不同的SPE填料，包括亲水亲脂（HLB）填料、亲水色谱（HILIC）填料及反相C18硅胶填料。通过对比发现C18硅胶对目标化合物的选择性最佳。进一步控制SPE的淋洗及洗脱条件，收集相应的洗脱液，经氮吹干燥后得到纯化的样品。提纯后的水飞蓟宾和异水飞蓟宾混合物的纯度可达94%以上。水飞蓟宾和异水飞蓟宾的平均回收率分别为70.5%~81.7%和66.7%~81.8%，相对标准偏差分别为2.7%~9.4%和1.5%~6.1%。该方法简单、高效，免去传统分离纯化过程中长时间的柱色谱分离过程，适合制备纯度较高的水飞蓟宾和异水飞蓟宾的二元混合标准样品。

关键词: 分离纯化, 固相萃取, 水飞蓟宾, 液相色谱, 异水飞蓟宾

Abstract:

Silybin and isosilybin are mainly effective elements of silymarin. To improve the performance of treatment and decrease the side effects from the unknown components, purified silybin and isosilybin are in request. The main purpose of this research was to develop a solid phase extraction (SPE) method for the purification of silybin and isosilybin from silymarin extract. For the analysis of the samples, a liquid chromatography method with ultraviolet detector (LC-UV) was developed to separate the silybin and isosilybin from their impurities. The separation was achieved in 8 min and a total analytical time of 11 min with column washing and equilibration. In the SPE, three different SPE packings were tested, including hydrophilic-lipophilic balance (HLB), hydrophilic interaction chromatography (HILIC) and reverse phase C18 silica microspheres. Based on the selectivity against silybin and isosilybin, the C18 packings were chosen. After the optimization of the elution of SPE, the purity over 94% could be achieved for the silybin and isosilybin mixture with the recoveries of 70.5%-81.7% and 66.7%-81.8%, and the relative standard deviations (RSDs) of 2.7%-9.4% and 1.5%-6.1%, respectively. Though silybin and isosilybin could not be separated from each other by using SPE, this is a useful method for the purification of silybin and isosilybin mixture from the crude extract of their plant seeds due to its simplicity and efficiency.

Key words: isosilybin, liquid chromatography (LC), separation and purification, silybin, solid phase extraction (SPE)

中图分类号: