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

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.