An Approach for Developing a Simple and Quick Method for Separation of Asiatic Acid and Asiaticoside Rich Fraction From Centella Asiatica and Simultaneous Determination by Reversed-Phase High-Performance Liquid Chromatography

In Asian countries, Centella asiatica is exploited for abundant types of pharmacological activities due to the presence of opulent phytochemicals. Asiaticoside, madecassoside, and their sapogenin triterpene acids such as asiatic acid and madecassic acid are the most noticeable triterpenes present in C. asiatica. To date, numerous techniques/methods are used to extract and isolate the different kinds of phytoconstituents from C. asiatica. Still, most methods require some special requirements, and some procedures are monotonous and time-consuming. Meanwhile, previously reported methods used for the extraction and isolation were not validated for large-scale production, yield, and purity. The study's primary goal is to develop the methodology for extracting and isolating the Asiaticoside and asiatic acid from C. asiatica at the minimum time with the highest yield and purity. Asiaticoside and asiatic acid extraction and isolation involved the acid hydrolysis method and recovered in alcohol. The Reverse Phase-High Performace Liquid Chromatography (RP-HPLC) method was developed and validated as per ICH guidelines for quantifying both compounds. The obtained results indicate that the developed method produces asiaticoside and asiatic acid with good purity. As per the ICH guidelines, the RP-HPLC was developed and validated. The proposed method can be used to isolate the asiaticoside and asiatic acid from C. asiatica. Some modification in this method leads to the large-scale production of highly pure asiatic acid and asiaticoside for their versatile application in the area of cosmetics and phytopharmaceuticals.

Keywords:

Centella asiatica, Extraction, Isolation, asiatic acid, asiaticoside,

PDF

___

Idris, F. N., & Nadzir, M. M. (2021). Comparative Studies on Different Extraction Methods of Centella asiatica and Extracts Bioactive Compounds Effects on Antimicrobial Activities. Antibiotics, 10, 457.
Kaur, I., Suthar, N., Kaur, J., Bansal, Y., & Bansal, G. (2016). Accelerated Stability Studies on Dried Extracts of Centella asiatica Through Chemical, HPLC, HPTLC, and Biological Activity Analyses. Journal of Evidence-Based Complementary and Alternative Medicine, 21(4), NP127–NP137. doi: 10.1177/2156587216661468
Lu, C., Lin, T., Pan, T., Wang, P., Chiu, K., Lee, M., & Wang, S. (2021). Asiatic Acid Prevents Cognitive Deficits by Inhibiting Calpain Activation and Preserving Synaptic and Mitochondrial Function in Rats with Kainic Acid-Induced Seizure. Biomedicines, 9, 284.
Monton, C., Settharaksa, S., Luprasong, C., & Songsak, T. (2019). An optimization approach of dynamic maceration of Centella asiatica to obtain the highest content of four centelloids by response surface methodology. Revista Brasileira de Farmacognosia, 29(2), 254–261. doi: 10.1016/j.bjp.2019.01.001
Nouri, S., Nejad, S., Sonboli, A., & Hossein, M. (2021). Variability, association and path analysis of centellosides and agro- morphological characteristics in Iranian Centella asiatica ( L .) Urban ecotypes. South African Journal of Botany, 139, 254–266. doi: 10.1016/j.sajb.2021.03.006
Pingyod, C., Waranuch, N., Phrompittayarat, W., Boonnoun, P., & Ingkaninan, K. (2021). Extraction of Centella asiatica leaves using a mixture of subcritical dimethyl ether and ethanol : Optimization of conditions by response surface methodology Extraction of Centella asiatica leaves using a mixture of subcritical dimethyl ether and ethanol. Songklanakarin J. Sci. Technol., 43(3), 711–718. doi: 10.14456/sjst-psu.2021.94
Ribeiro, A. B., Ozelin, S. D., Lucas, H. D., Rinaldi, F., Karoline, N., Heloiza, S. F., Souza, L. D. R. De, Tavares, D. C., Furtado, R. A., & Cunha, W. R. (2021). Influence of Asiatic acid on cell proliferation and DNA damage in vitro and in vivo systems. J Biochem Mol Toxicol, 35(August 2020), 22712. doi: 10.1002/jbt.22712
Sabaragamuwa, R., Perera, C. O., & Fedrizzi, B. (2022). Ultrasound assisted extraction and quantification of targeted bioactive compounds of Centella asiatica ( Gotu Kola ) by UHPLC-MS / MS MRM tandem mass spectroscopy. Food Chemistry, 371(March 2021), 131187. doi: 10.1016/j.foodchem.2021.131187
Songvut, P., Chariyavilaskul, P., Khemawoot, P., & Tansawat, R. (2021). Pharmacokinetics and metabolomics investigation of an orally modified formula of standardized Centella asiatica extract in healthy volunteers. Scientific Reports, 1–13. doi: 10.1038/s41598-021-86267-2
Thong, W., Pathomwichaiwat, T., & Boonsith, S. (2021). Green extraction optimization of triterpenoid glycoside ‑ enriched extract from Centella asiatica ( L .) Urban using response surface methodology ( RSM ). Scientific Reports, 0123456789, 1–11. doi: 10.1038/s41598-021-01602-x