Abstract
A simple and reliable method for the identification and classification based on cultivation ages of Centella asiatica (CA) was developed. The developed method used a combination of a simultaneous quantification of four triterpenes (madecassoside, asiaticoside, madecassic acid, and asiatic acid) and a fingerprint analysis of methanol extract of CA using a high-performance liquid chromatography (HPLC). The four triterpenes were well separated under the optimized elution conditions. Fingerprint chromatogram of CA showed marked differences from those of closely related plants, namely Hydrocotyle sibthorpiodes and Hydrocotyle verticillata, so that it can be used for discrimination of CA from those plants. Also the concentration of four triterpenes was different among CA samples from different location and at 3, 4 and 5 months post-planting. Validity of the method was demonstrated by evaluating the system suitability, linearity, precision, accuracy, limit of detection, and limit of quantification, as well as by determining the stability of components in CA extract. All of the parameters met criteria for a reliable and valid method according to the Association of Official Analytical Chemists (AOAC) guidelines. In addition, the HPLC fingerprint with chemometrics analysis could be used for classification of CA according to their cultivation ages.
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