Simultaneous quantitation of three major triterpenoid glycosides in Centella asiatica extracts by high performance liquid chromatography with evaporative light scattering detection

Abstract A high‐performance liquid chromatography method with evaporative light scattering detection was established for simultaneous determination of three major triterpenoid glycosides, i.e. asiaticoside, madecassoside and asiaticoside‐B, in Centella asiatica extracts. The optimal chromatographic conditions were achieved on a COSMOSIL 5C 18 ‐MS‐II column by constant elution with water (0.01% trifluoroacetic acid, v/v) and acetonitrile (1.0% methyl tert‐butyl ether, 0.01% trifluoroacetic acid, v/v) (78:22) as mobile phase at a flow rate of 1.0 mL/min; the column temperature was 30°C. The evaporative light scattering detector was set at an evaporating temperature of 40°C and nitrogen gas pressure of 3.5 bar. The validation of the method included tests of linearity, sensitivity, precision, repeatability, stability and accuracy. All calibration curves showed good linear regression ( r 2 > 0.9993) within test ranges. The established method showed good precision and accuracy with overall intra‐day and inter‐day variations of 1.73–3.06 and 3.89%–4.92%, respectively, and overall recoveries of 97.63–99.39% for the three compounds analyzed. The method developed was successfully applied to quantify the main triterpenoid glycosides in Centella asiatica extracts from different companies. Copyright © 2007 John Wiley & Sons, Ltd.