Abstract:

A high performance liquid chromatographic (HPLC) method was successfully used to separate asiaticoside and
madecassoside from the extract of Centella asiatica (L.) Urb. The raw material containing 20% asiaticoside and 45%
madecassoside was dissolved in methanol and used for the preparation of the pure compounds. The flow rate of the
mobile phase, injection volume and detection wavelength were optimized. The optimum operation parameters were
selected as follows: a C18 column (50 mm×200 mm, 5 μm) with methanol-water (60∶40, v/v) as the mobile phase at a
flow rate of 100 mL/min, the detection wavelength of 220 nm, and the injection volume of 1.5 mL at the concentration
of 40 mg/mL. Asiaticoside and madecassoside can be obtained in a 20 min runtime, and the purities of the products
were detected by melting point, thin layer chromatography (TLC) and HPLC method. The results showed that the
purities of products were above 98%. The effective and rapid method has been successfully applied to the preparation
of asiaticoside and madecassoside from Centella asiatica (L.) Urb.

Key words:
madecassoside, asiaticoside, Centella asiatica (L.) Urb , triterpenoid saponins, preparative high performance liquid chromatography (Pre-HPLC)