RP-HPLC Method Development for Simultaneous Determination of Madecassoside and Asiaticoside in <i>Centella asiatica</i>

Centella asiatica contains therapeutic triterpenoid glycosides, especially madecassoside and asiaticoside, which need reliable analytical methods for quality control. The present study was undertaken to develop and validate a robust RP-HPLC method for simultaneous quantification and to assess its stability under different conditions. The extracts of Centella asiatica were prepared, filtered, and then analyzed by using optimized RP-HPLC chromatographic conditions. The developed method was validated for linearity, precision, accuracy, LOD, and LOQ. Stability studies were performed by exposing samples to different environmental conditions and monitoring analyte degradation as a function of time. The optimized RP-HPLC method completely separated madecassoside and asiaticoside in less than 25 minutes of runtime. The method showed excellent linearity with R2 > 0.999, acceptable LOD and LOQ values, high precision, and accuracy for the tested concentrations. The stability testing showed a negligible breakdown of the compounds, proving the method's robustness. The strong validation parameters of this method prove its suitability for quantitative evaluation of major triterpenoid glycosides in C. asiatica. The negligible variation observed during stability studies proves its reliability under routine laboratory and storage conditions, hence useful during quality assurance in herbal formulations. A rapid, accurate, and robust RP-HPLC method of quantification for the major triterpenoids madecassoside and asiaticoside was developed and accordingly validated in Centella asiatica. Its performance characteristics and stability make it well suited for routine quality control and standardization of Centella asiatica–based products.