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Separation of Madecassoside and Madecassic Acid Isomers by High Performance Liquid Chromatography Using β -Cyclodextrin as Mobile Phase Additive

  • Kai, Guiqing (Engineering Research Center of Bio-process Ministry of Education, Hefei University of Technology) ;
  • Pan, Jian (Engineering Research Center of Bio-process Ministry of Education, Hefei University of Technology) ;
  • Yuan, Chuanxun (Engineering Research Center of Bio-process Ministry of Education, Hefei University of Technology) ;
  • Yuan, Yuan (Engineering Research Center of Bio-process Ministry of Education, Hefei University of Technology)
  • Published : 2008.03.20

Abstract

An improved HPLC method for the separation of madecassoside isomers (madecassoside and asiaticoside-B) has been developed. The isomers can be separated with high resolution from extracts of Centella asiatica by HPLC using $\beta$-cyclodextrin as a mobile phase added on a $C_{18}$ column. The result shows that the isomers can be separated with a mobile phase consisting of methanol:water (50:50, v/v) with 4 mmol/L $\beta$-CD. To elucidate the mechanism of the separation of madecassoside and asiaticoside-B, this paper studied the separation of their aglycon parts (madecassic acid and terminolic acid), another pair of isomers. The isomers can also be separated with high resolution with a mobile phase consisting of methanol:water (65:35, v/v) with 4 mmol/L $\beta$-CD and the pH of the mobile phase was adjusted to 4. The paper also studied the separation of the two isomers by HPLC using $\alpha$-CD and Glucosyl-$\beta$-CD as a mobile phase additive in order to elucidate the mechanism of the separation process.

Keywords

References

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