色谱

色谱

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β-环糊精流动相添加剂法分离测定积雪草总苷元中的羟基积雪草酸

潘见,开桂青,袁传勋,周蓓蓓,金日生,袁媛   

  1. Engineering Research Center of Bio-process of Ministry of Education, Hefei University of Technology, Hefei 230069, China
  • 收稿日期:2007-01-11 修回日期:2007-02-11 出版日期:2007-05-30 发布日期:1985-09-25
  • 通讯作者: 潘见

Separation and Determination of Madecassic Acid in Triterpenic Genins of Centella asiatica by High Performance Liquid Chromatography Using β-Cyclodextrin as Mobile Phase Additive

PAN Jian, KAI Guiqing, YUAN Chuanxun, ZHOU Beibei, JIN Risheng, YUAN Yuan   

  1. Engineering Research Center of Bio-process of Ministry of Education, Hefei University of Technology, Hefei 230069, China
  • Received:2007-01-11 Revised:2007-02-11 Online:2007-05-30 Published:1985-09-25
  • Contact: PAN Jian

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摘要： 以β-环糊精为流动相添加剂,采用反相高效液相色谱法于C18反相柱上拆分了羟基积雪草酸及其同分异构体,建立了积雪草样品中羟基积雪草酸含量的测定方法,探讨了β-环糊精浓度、流动相pH对同分异构体分离度的影响。结果表明:当流动相为甲醇-水（体积比为65∶35）,pH为4时,同分异构体的分离度随着β-环糊精浓度的增加而增大;羟基积雪草酸在0.1~5.0 g/L范围内,峰面积与浓度呈线性关系（r2=0.9989）,说明该法适用于羟基积雪草酸的含量检测及有关药品的质量控制。

关键词: terminolic acid, β-环糊精, 反相高效液相色谱法, 积雪草 , 羟基积雪草酸, 同分异构体

Abstract:

Centella asiatica (L.) Urban is a tropical medicinal plant with a long history of therapeutic uses. Madecassic acid and terminolic acid, which are a pair of structural isomers, are two constituents of Centella asiatica. A method using reversed-phase high performance liquid chromatography in which β-cyclodextrin (β-CD) was the additive in mobile phase has been developed for separation of the structural isomers and determination of madecassic acid. The two compounds can be isolated with high resolution on a C18 reversed-phase column with the addition of β-CD in the mobile phase. The separation mechanism of the isomers was discussed. It was assumed that the separation of the isomers might have been resulted from different inclusion forces of complexes with β-CD. The effects of β-CD concentration and the pH of mobile phase on resolution were investigated. It was found that the resolution of the isomers increased with the increase of β-CD concentration when the mobile phase consisted of methanol-water (65∶35, v/v) at pH 4. The correlation coefficient (r2) of the linear calibration curve between peak area and concentration of madecassic acid was 0.9989 in the range of 0.1-5.0 g/L. This method was successfully used to determine the madecassic acid in triterpenic genins of Centella asiatica.

Key words: Centella asiatica , isomer, madecassic acid, terminolic acid, β-cyclodextrin, reversed-phase high performance liquid chromatography (RP-HPLC)