Abstract: Objective To establish a rapid separation and purification method of asiatic acid from centella asiatical(L.) Urban and to develop a reversed-phase high-performance liquid chromatography(HPLC) method with UV-vis detection for the quantitative determination of asiatic acid. Methods Crude extract separated and purified by silica gel column chromatography with gradient elution of petroleum ether-acetone system was first extracted with ethanol from centella asiatical(L.) urban and then done by ethyl acetate. TLC was used as the monitor during the process. HPLC separation was achieved on a Symmetry C₁₈ column(4.6 mm×250 mm, 5 μm) with a solvent system of acetonitrile -10 mmol·L^-1 ammonium acetate(38∶62). The wavelength of detector was set at 210 nm, the flow rate was 1 mL·min^-1, the column temperature was set at 25 ℃, and the injection volume was 20 μL. Results A calibration curve ranging from 10 to 200 mg·L^-1 was shown to be linear(R²=0.999 5), and the lower limit of quantification(LLOQ) was 10 mg·L^-1. The inter- and intra-day RSD% which were determined by three different concentration(25, 100, 200 mg·L^-1) were lower than 5.4%. The content of asiatic acid in centella asiatical(L.) Urban was 0.99 g·kg^-1. Conclusion The method was successfully applied to the separation, purification and the quantitative determination of AA.

Key words: centella asiatical(L.) urban, asiatic acid, high-performance liquid chromatography, purification