Bioassay-Guided Isolation of Active Constituents of Tinospora cordifolia and Validation of Anticancer Efficacy by In Vitro Assays

Abstract

Abstract: The primary objective of this research is to collect Tinospora cordifolia specimens and systematically extract their active constituents using various organic solvents and water. The multi-step extraction process is crucial for optimising the yield of bioactive compounds, which can vary based on the polarity of the solvent used. The selected solvents, including methanol, ethanol, chloroform, and water, are expected to target the bioactive phytochemicals, such as alkaloids, flavonoids, and terpenoids. Preliminary phytochemical screening was conducted post-extraction to confirm the presence of these secondary metabolites. This is create the foundation for future bioactivity investigations. To identify the active compounds demonstrating anticancer activity, bioassay-guided purification methods was employed following the preliminary cytotoxicity assessment. This approach employs silica gel column chromatography and liquid-liquid partitioning for the physical and chemical isolation of components. Further bioactivity investigations was performed on the obtained fractions to ascertain those having the greatest potential for anticancer efficacy. The safety profile of the bioactive fractions must be assessed concurrently. The anticancer properties of the bioactive fractions was validated by a series of in vitro assays, including apoptosis measurement, cell cycle analysis, and reactive oxygen species formation. These research will expand our understanding of how bioactive substances promote programmed cell death and regulate cellular responses to oxidative stress. Conversely, cell cycle analysis can reveal substances that disrupt the advancement of the cell cycle.