Studies on isolated crude bioactive compound(s) from ficus deltoidea for smooth muscle contractility potential / Muhammad Nasrul Hadi Azhar

The isolation of the bioactive compound(s) from Ficus deltoidea was done by performing two different methods; traditional extraction technique using boiling step and conventional chemistry method. The former produced the crude water extract which will then partitioned with ethyl acetate producing water and ethyl acetate fractions. The latter involved sequential partioning of the crude methanol extract with solvent of different polarity (hexane, chloroform, ethyl acetate and distilled water). Angiotensin converting enzyme (ACE) and acetylcholinesterase (AChE) enzyme assays were used as bio-guided assays to search for the bioactive compound(s). Samples that have good activity were further separated and purified using size exclusion-, silica gel- and Celite® gel- column chromatography. Samples with good activity were also subjected to protein identification,using SDS - PAGE and LC/QToF MS approaches. From the traditional extraction method, water fraction (WF) and water fraction subfraction 2 (WFSF2) showed good inhibitory activity towards ACE (60.360±0.508% and 72.646±15.012% respectively). Both samples have very minimal inhibitory activity towards AChE (5.535±0.849% and 13.835±3.249% respectively). Water fractions showed the presence of rubber elongation factor protein, cdc2 kinase and non-specific serine/threonine protein kinase. Meanwhile, water fraction subfraction 2 only showed the presence of actin. The extract and fractions from the conventional solvent based method did not show good response towards the assays. Hexane soluble extract fraction 5 subfraction 1 (HSEF5SF1) and chloroform soluble extract fraction 1 subfraction l(CSEFlSFl) were sent for GCMS analysis.