Simplified Column Fractionation with DPPH Assay Guidance: Isolated a Potent Hepatoprotective Fraction from Cassia auriculata

Background: Hepatoprotection is a critical area for research due to limitations of synthetic drugs and their side effects in the treatment of liver diseases. Natural compounds, particularly those derived from herbs with hepatoprotective therapeutical potential are serving as a best alternative for the same indication. Cassia auriculata, used traditionally in Indian folk medicine, has having a promising pharmacological activities, including hepatoprotective effects. Objectives: Present study aimed to evaluate using antioxidant activity to find out hepatoprotective compounds present in C. auriculata leaf extract and its solvent fractions through in vitro, in vivo experiments and LCMS analysis. Materials and Methods: The DPPH assay was employed to assess in vitro antioxidant activity, while cytotoxicity and hepatoprotective effects against H₂O2-induced hepatotoxicity in HepG2 cell lines were investigated. Furthermore, the hepatoprotective activity of the most active Column fraction was evaluated against CCl₄-induced hepatotoxicity in albino Wistar rats. LCMS analysis was utilized to identify bioactive compounds in the most active Column fraction and correlations between antioxidant activity and phenolic, tannin and flavonoid contents were also explored. Results: The findings of the present study suggest that most active Column fraction from C. auriculata possess significant antioxidant and hepatoprotective properties, making them promising candidates for further investigation as potential therapeutic agents for liver diseases. Conclusion: This study aims to bridge the gap in understanding the correlation between antioxidant and hepatoprotective potential of C. auriculata extract and its fractions through rigorous experimental investigations. Overall, the present study contributes to the growing body of evidence supporting the therapeutic potential of natural compounds in liver disease management, paving the way for further research and development in this field. Keywords: Cassia auriculata, Hepatoprotective activity, LCMS, HepG2 cell lines, Column chromatography.