Isolation and Partial Characterization of the Most Bioactive Metabolite from The Hexane Extract of the Aerial Part of Hydrocotyle Verticillata (WHORLED MARSHPENNYWORTH)
Several of thousands of bioactive compounds have been mined from the natural product resources both from the marine and terrestrial environment all over the world. Natural product has proved potentiality of many plants for diverse clinical conditions. Many plants were found to contain compounds, the bioactive components which are used in natural medicine. In this study, the plant of interest is Hydrocotyle verticillata, mistakenly identified to be “Gotu ko la” (Centella asiatica) by local folks of Iligan City and Butuan City, Philippines. The natural components of the aerial part of H. verticillata were extracted using solvent extraction. The crude ethanolic extract of the aerial part of Hydrocotyle verticillata was further subjected to solvent partitioning according to increasing polarity: n-hexane, ethyl acetate (EtOAc), dichloromethane (DCM) and water. Bioassays like Brine Shrimp Lethality Test was then conducted to test which of the extracts consist of bioactive components. Among the four partitioned solvent extracts, result showed that n – hexane extract showed to be the most bioactive since it has the lowest LC50 . Fractionation and purification of the hexane extract was done through series of Gravity Column Chromatography (GCC). Four isolates (HF5B1, HF5B2, HF5B3 and HF5B4) were obtained. Analysis using High Performance Liquid Chromatography (normal phase HPLC) showed three of the isolates (HF5B2, HF5B3 and HF5B4) to have similar profile, two peaks of closely similar retention time. HF5B2 was further purified in HPLC and yielded a subfraction HF5B2a with a single sharp peak on the HPLC chromatogram and with a retention time of 8.135 min. This subfraction was partially characterized using Perkin Elmer FTIR Spectrometer. The spectrum showed peaks for the following functional groups: NH, C=O, C-C(O)-C, and CN stretch (aliphatic amine). Considering the presence of these functional groups particularly the amine, the isolated metabolite HF5B2A from hexane ext
