Separation and Determination of Asiaticoside, Asiaticoside-B and Madecassoside in Centella asiatica Total Triterpenoid Saponins by HPLC

Abstract

A high performance liquid chromatographic method was developed for the separation and determination of asiaticoside and madecassoside isomers (asiaticoside-B and madecassoside) in Centella asiatica products. The method involved separation by reversed phase chromatography on an Atlantis T3 C₁₈ column using water-acetonitrile-methyl tert-butyl ether (80:18:2:0.1, 0.1% acetic acid, by volume) as mobile phase. The isomers of asiaticoside-B and madecassoside were separated with high resolution by the developed method. The method's linearity, precision, and recovery were validated. The effects of stationary phase and mobile phase composition on the separation of madecassoside isomers were also investigated. Experimental results indicated that the Atlantis T3 C₁₈ column offered enhancing retention and resolution to the polar isomers, and the method could be referenced for the separation of other triterpenoid saponins isomers.

Keywords:

• Asiaticoside
• Centella asiatica
• HPLC
• Isomer
• Madecassoside
• Separation

ACKNOWLEDGMENTS

The authors are grateful for financial support from the Zhejiang Provincial Key Science and Technology Program (No. 2006C13009) and the Ministry of Science and Technology of the People's Republic of China (Nos. 2006BAD27B03 and 2007AA100404).