Screening and evaluation of antioxidant, antimicrobial, cytotoxic, thrombolytic and membrane stabilizing properties of the methanolic extract and solvent-solvent partitioning effect of Vitex negundo Bark

Objective: To investigate different biological activities of the methanolic extract and solventsolvent partitioning of Vitex negundo (V. negundo) bark.\udMethods: In-vitro anti-oxidant study was determined using total phenolic compound analysis\udand DPPH radical scavenging assay. In vitro antimicrobial study was measured by observing\udzone of inhibition. The cytotoxic activity was studied using brine shrimp lethality bioassay and\udthrombolytic activity by clot disruption method.\udResults: In the evaluation of anti-oxidant activity, the amount of total phenolic content differed\udin different extractives as from 12.94 mg of GAE/g of extractives to 54.19 mg of GAE/g of extractives\udof V. negundo. Among all extractives, the highest phenolic content was found in n-hexane\udfraction (N-HXN) (54.19 mg of GAE/g of extractives). Significant amount of phenolic compounds\udwere present in methanol extract (ME) (35.19 mg of GAE/g of extractives) and aqueous soluble\udfraction (AQSF) (12.94 mg of GAE/g of extractives). The free radical scavenging activity of ME and\udits different partitions were assessed using DPPH free radicals assay. The highest free radical\udscavenging activity was found in AQSF (IC50=7.78 µg/mL). The antimicrobial screening of the bark\udof V. negundo exhibited mild to moderate activity in test microorganisms. The chloroform soluble\udfraction demonstrated the highest inhibition against microbial growth having zone of inhibition\udas 8-12 mm. In the brine shrimp lethality bioassay, The LC50 values of n-hexane and chloroform\udsoluble fractions were found 3.713 6 µg/mL, and 0.910 µg/mL respectively while the LC50 values of\udstandard Vincristine sulphate was 0.408 µg/mL. The methanol extract and their different organic\udsoluble of V. negundo bark inhibited hypotonic solution-induced haemolysis of erythrocytes in\udin-vitro membrane stabilizing activity test. The AQSF and N-HXN showed inhibition of haemolysis\udas 55.05% and 29.89% that were lower than 71.9% of aspirin (0.10 mg/mL). Methanol extract of V.\udnegundo and all of its different partitions exhibited moderate thrombolytic activity of 36.95%-\ud22.98%.\udConclusions: These experiments were able to show the several biological activity of methanolic\udextract and its soluble fractions of V. negundo at a time