色谱

色谱 ›› 2013, Vol. 31 ›› Issue (5): 447-450.DOI: 10.3724/SP.J.1123.2012.12054

• 研究论文 • 上一篇    下一篇

高效液相色谱手性流动相添加剂法分离3种黄酮类化合物对映体

孙亚男, 李彤, 马辰*   

  1. 中国医学科学院&北京协和医学院药物研究所, 北京 100050
  • 收稿日期:2012-12-29 修回日期:2013-01-21 出版日期:2013-05-28 发布日期:2013-05-16
  • 通讯作者: 马辰,Tel:(010)63165239,E-mail:mach@imm.ac.cn.
  • 基金资助:

    科技部创新方法工作专项项目(2011IM030200).

Separation of racemic flavonoids by high performance liquid chromatography using chiral mobile phase additive

SUN Yanan, LI Tong, MA Chen*   

  1. Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China
  • Received:2012-12-29 Revised:2013-01-21 Online:2013-05-28 Published:2013-05-16

PDF

276

被引次数

25

摘要：

采用反相高效液相色谱法(RP-HPLC),以磺丁基醚-β-环糊精(SBE-β-CD)作为手性流动相添加剂,建立了二氢黄豆苷原(dihydrodaidzein)、雌马酚(equol)和山姜素(alpinetin)3种黄酮类化合物的手性拆分方法。考察了环糊精的种类和浓度、有机相的种类和比例、缓冲盐的种类和浓度以及pH对3种化合物手性拆分效果的影响。结果表明:采用Kromasil 100-5C18(250 mm×4.6 mm, 5 μm)色谱柱,流动相为乙腈-10 mmol/L SBE-β-CD水溶液(含20 mmol/L KH2PO4, pH值到4.0)(体积比为20:80)的条件下,二氢黄豆苷原、雌马酚和山姜素的对映体都达到了基线分离,分离度分别为1.8, 1.9和1.4。该方法简便,分离效果好,对黄酮类化合物的拆分具有应用价值。

关键词: 反相高效液相色谱, 黄酮类化合物对映体, 磺丁基醚-β-环糊精, 手性分离

Abstract:

To perform chiral separation of three flavonoid racemates, including dihydrodaidzein, equol and alpinetin, a simple and effective method was developed by using sulfobutyl ether-β-cyclodextrin (SBE-β-CD) as a mobile phase additive in HPLC system. The effects of different factors on the chiral separation were investigated, including cyclodextrin (CD) types (α-CD, γ-CD, β-CD, methyl β-CD), CD concentration (5, 10, 20 mmol/L), pH value, nature of organic solvents (methanol, ethanol, acetonitrile) and phosphate buffer concentration. The analysis was carried out on a Kromasil 100-5C18 column (250 mm×4.6 mm, 5 μm). The mobile phase consisted of acetonitrile and 10 mmol/L SBE-β-CD aqueous solution(containing 20 mmol/L KH2PO4, at pH 4.0 adjusted with phosphoric acid)(20:80, v/v). The flow rate was set at 1 mL/min, the column temperature was 25 ℃, and the UV detections of dihydrodaidzein, equol and alpinetin were carried out at 276, 280 and 290 nm, respectively. The established method was able to separate the three racemates, and the resolutions of the enantiomers (dihydrodaidzein, equol and alpinetin) were 1.8, 1.9 and 1.4, respectively. This method might be applied to separate various types of flavanoid enantiomers in chemical structure.

Key words: chiral separation, flavonoid enantiomers, reversed-phase high performance liquid chromatography (RP-HPLC), sulfobutyl ether-β-cyclodextrin (SBE-β-CD)

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