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Preliminary phytochemical screening of the various Ficus extract revealed the presence of triterpenoids, flavonoids, glycosides, steroids, alkaloids, saponins (Table 1).
Table 5 shows the reductive capabilities of methanolic extract of four different ficus species when compared to the standard, BHT. Like the anti oxidant activity, the reducing power increased significantly (P<0.01) with increasing amount of extracts. The FBBM showed the highest reducing power ability than the other extracts tested. However, the activity of the various extracts was less than the standard. The order of reducing ability of the extracts are FBBE>FGBE>FRBE>FMBE.
The content of total phenolics in the extracts of FBBE, FGBE, FRBE and FMBE were determined using Folin-Ciocalteu assay, calculated from regression equation of calibration curve (Fig.11) and expressed as µg pyrocatechol equivalents (PCE).The amount of total phenolic content in the extract correlated with the antioxidant activity. Among all the extract the significantly higher phenolic content amount was found in FBBE ().The phenolic content of other fractions were FGBE (), FRBE (), and FMBE respectively(Table 12).
5.4 Nitric oxide scavenging assay
Incubation of solutions of sodium nitroprusside in phosphate buffered saline at 25 0c for 150 min resulted in generation of nitric oxide. All the extracts effectively reduced the generation of nitric oxide radicals. The FGBE exhibited superior nitric oxide scavenging activity action with IC 50 103.7µg/ml which was lower than the standard ascorbic acid (46.8µg/ml).The IC 50 of FBBE, FMBE, FRBE were 105.1, 360.6 and 233.8 µg/ml and scavenging activity was lesser than that of standard (Table 8).
5.5. Super oxide anion radical scavenging assay
The extracts of four ficus species were found to be a scavenger of superoxide anion in NBT reduction system (Table 10). All the extracts showed significant (P<0.001) superoxide inhibiting activity at a concentration range of 25-400 µg/ml. The IC 50 of FBBE, FGBE, FMBE and FRBE were found to be 138.2, 183.4, 205.6 and 266.8 µg/ml respectively.
5.6. Hydroxyl radical scavenging assay
The degradation of deoxyribose by Fe 3+ -ascorbate - EDTA-H2O2 system was markedly decreased by the all extracts of ficus tested (Table 9) indicating the significant (P<0.01) hydroxyl radical scavenging activity. The 50% inhibitory concentration were higher for FGBE (24.23 µg/ml) than the other extracts employed in the study.
5.7. Total anti oxidant power assay (Phosphomolybdate method)
The phosphomolybdenum method is quantitative, and the total antioxidant capacity is expressed as α Tocopherol equivalents. Among the extracts tested, the FBBE contains x/ α Tocopherol equivalent /100 µg. The antioxidant activity increased in the order of FBBE, FGBE, FMBE and FRBE (Fig; Table 11).
5.8. Metal chelating capacity
Addition of the extracts of ficus species interfered with the ferrous-ferrozine complex and this was decreased with the increasing concentration of the drug. Among the extracted tested, the FBBM showed more pronounced ferrous chelating ability than those of other extracts(IC 50 166.8 µg/ml).The chelating action for FGBE, FMBE and FRBE were 183.6, 244.1 and 285.1 µg/ml respectively. EDTA expressed the highest ferrous ion chelating ability(IC 50 33.9 µg/ml) (Table 6).
5.9 Total flavonoid content
The total flavonoid content in the extracts of ficus was expressed as µg quercetin equivalent .The flavonoid content was higher for FBBE () and for other extracts it was found to be FGBE (), FRBE () , and FMBE() quercetin equivalent (fig ;table )
5.10. Hydrogen peroxide scavenging assay
The methanolic extracts four ficus species were scavenged hydrogen peroxide in a concentration dependent manner which could be seen by its graded increase in percentage inhibition (Table 11).
5.11. Total peroxy radical trapping potential
The peroxy radical scavenging activity was determined for four ficus species and results were compared using α Tocopherol synthetic analogue Trolox (Table 7).Addition of increasing concentration of fractions to the solution containing AAPH decreased the luminescence produced by DCF and absorbance decreased in a linear fashion .Only FBBM exhibited significant activity with IC 50 of 166.7 µg/ml
5.12. AAPH induced hemolysis
The peroxyl radical generated by AAPH on addition to erythrocyte suspension and its subsequent scavenging action produced by graded higher concentration of the extract are given in Table 14 .The results of the study were compared with the standard ascorbic acid and an increase in inhibition was noticed for all the extracts tested. FBBE exhibited significant IC 50 value (171.1 µg/ml) compared to other extracts.
5.13 Hydrogen peroxide induced hemolysis
Lipid oxidation of erythrocyte membrane mediated by H2O2 induces membrane damage and subsequently hemolysis. The antihemolytic activity of the various extracts on erythrocytes is presented in Table. All the sample extracts were found to have antihemolytic activity and the values are significantly different. Interestingly, FBBE registered the highest antihemolytic activity. The antihemolytic activity of various extracts and standards are in order of ASCORIC ACID>FBBE>FGBE>FMBE>FRBE.
5.14 Osmotic stress induced hemolytic activity
Hemolysis of human erythrocytes was induced by hypotonic stress (Figure 8). The antihemolytic activity of FBBE, FMBE, FRBE and FGBE on human erythrocytes are presented in the table. All the four extract have antihemolytic activity. The antihemolytic action for FBBE, FGBE, FMBE and FRBE were 164.1, 222.8, 242.3 and 234.1 µg/ml respectively. Ascorbic acid expressed the highest activity(IC 50 78.6 µg/ml) (Table 4).
5.15 Antimicrobial activity.
The results of antimicrobial assay of four ficus species under study by agar well diffusion assay as shown on the figure 12-24.the antimicrobial activity was measured by zone of inhibition (mm).The results of antimicrobial assay showed that methanolic extract of four ficus species have potential antimicrobial activity. FRBM have significant activity against S.aureus, P.aeruginosa and E. coli (21mm,25mm and 20mm at 4mg conc)compared to FBBE ,FMBE and FRBE. However The activity of FBBE,FMBE,FRBE and FGBE were less than the standard ciprofloxacin