Optimization of methods for determining major triterpenoid compounds in Centella asiatica (L.) Urban

Abstract [Objective] In order to provide a basis for quality control of Centella asiatica (L.) Urban, a high-performance liquid chromatography method was established using cyclodextrin as a mobile phase additive for the determination of asiaticoside, madecassoside, and asiaticoside B. [Methods] Molecular design was used to optimize the type of the mobile phase additive-cyclodextrin. The response surface method was used to analyze the effects of cyclodextrin concentration, the proportion of acetonitrile in the mobile phase, and pH of the mobile phase on separation, as well as to optimize the chromatographic condition. The optimized chromatographic condition was used to determine the content of asiaticoside, madecassoside, and asiaticoside B in C. asiatica. [Results] In the optimization of determination method, γ-cyclodextrin was used as a mobile phase additive. The separation of asiaticoside, asiaticoside B, and madecassoside was carried out on CAPCELL PAK C18 (150 mm×4.6 mm, 5 μ m) with acetonitrile- 3 mmol/L γ-cyclodextrin solution (contained 0.10% phosphoric acid) (20∶80, V/V ). The detecting wavelength was 205 nm. The flow rate was set at 1.0 mL/min and the column temperature was 25 ℃. The resolution of madecassoside and asiaticoside B was 4.32. Linearity was good in the range of 200-2 000 ng for asiaticoside, 120-1 200 ng for asiaticoside B, and 200-1 200 ng for madecassoside. The precision, reproducibility, stability, and recovery rate of method were good. [Conclusion] The new method is simple, accurate, reliable, and environmentally friendly, and can be used as a quality control of major triterpenoid compounds in C. asiatica (L.) Urban.