IDENTIFICATION OF POTENTIAL ANTIOXIDANT AND HEPATOPROTECTIVE CONSTITUENTS OF VITEX DONIANA BY UHPLC/+ESI-QQTOF-MS/MS ANALYSIS

Objective: This study evaluated the antioxidant and hepatoprotective activities of the dichloromethane (DCM-F) and n-butanol (BUT) fractions of Vitex doniana and identified the bioactive constituents using liquid chromatography-mass spectrometry (LC-MS) analysis. Methods: The methanol extract of the leaves was subjected to liquid-liquid fractionation and the BUT soluble separated by vacuum LC (VLC) using binary mixtures of DCM in methanol (1 L) sequentially in the ratios of 9:1 BF1–1:9 BF5. The antioxidant activity was investigated in vitro using 1, 1-diphenyl-2-picrylhydrazyl (DPPH) scavenging, phosphomolybdenum total antioxidant capacity, and ferric ion reducing antioxidant power assay methods. The in vivo antioxidant potential was performed by measuring the concentrations of both enzymatic and non-enzymatic parameters in carbon tetrachloride (CCl4)-induced hepatic injury. Results: The extract, DCM-F, and the VLC sub-fractions showed good antioxidant activity which is comparable with ascorbic acid (53.6%). DCM-F showed the highest antioxidant activity (71.6%), while the least activity was shown by BF5 (15.0%). Peak dereplication of LC-MS chromatograms identified several putative compounds in the BUT soluble. DCM-F caused a significant increase (p<0.05) in superoxide dismutase, catalase, glutathione peroxidase, and Vitamins C and E in CCl4-induced rats compared to standard. The levels of serum alkaline phosphatase, alanine aminotransferase and aspartate aminotransferase, and malondialdehyde that were elevated in CCl4-induced rats were also significantly reduced (p<0.05) by the DCM-F compared to standard. Conclusion: The ability of V. doniana leaves to mop up free radicals and reverse the CCl4-induced hepatic injury in rats suggests antioxidant potentials. The bioactive compounds identified in BUT fraction could support its ethnomedicinal uses in the treatment of illnesses associated with radical scavenging activity