The extraction yield was 26% of the dry weight. The results of phytochemical screening of the methanolic extract revealed the presence of saponins, flavonoids, steroids, cardiac glycosides, tannins and phenol. The test for alkaloid was negative. Total learn more phenol and flavonol content of the methanolic extract was 34 mg/g and 28.1 mg/g of dry sample. The zones of inhibition of H. japonicum methanolic extract against fourteen bacterial cultures are tabulated in Table 1. The extract had a broad spectrum antibacterial activity. Both Gram positive and Gram negative bacteria were inhibited by the extract except P. aeruginosa. The MIC of the extract was 1 mg/ml against all the test

cultures used except E. aerogenes and P. aeruginosa. Total antioxidant activity of the methanolic extract of H. japonicum was 37.28 ± 0.54 μg/mg of the extract as estimated by Molybdenum reduction assay. The antiradical power of the extract was determined by using DPPH stable free radicals. Dose dependent DPPH radical quenching by the extract and BHA were compared in Fig. 1. The IC50 values of the extract and BHA were 77.7 ± 5.6 μg/ml and 55.85 ± 6.89 μg/ml respectively. The extract and quercetin both inhibited β-carotene bleaching up to 25 h at three tested

concentrations (1000 μg/ml, 500 μg/ml and 100 μg/ml). Complete bleaching of β-carotene was observed after 17 h in absence of extract or standard. The antioxidant activity of the extract and quercetin after 25 h of incubation find more was 83.18% and 63.01% respectively at the concentration of 100 μg per assay. Dose dependent activity during of the extract is shown in Fig. 2A. The β-carotene bleaching with lapse of time in presence and absence of extract and quercetin was compared in Fig. 2B. The activity of the extract was significantly higher than control and quercetin (at P ≤ 0.001). The activity of

H. japonicum methanolic extract was 31% better than quercetin. The extract and quercetin inhibited the lipid peroxidation by 95.38% and 94.16% respectively at the concentration of 15 μg per assay. A dose dependent DNA protection activity was observed in H. japonicum extract ( Fig. 3). Smeared DNA band in control (without extract or quercetin) represents the hydroxyl radical mediated DNA damage. The band smearing was decreased with increase in the concentration of extract and quercetin from 100 μg/ml to 500 μg/ml. DNA bands were similar to that of native calf thymus DNA at the concentration of 500 μg/ml. The HPLC fingerprint of the methanolic extract is given in Fig. 4. Six phenolic acids and two flavonoids were identified based on retention time compared with that of reference standards. Percentage composition of each of the phenolic acids in the extract is given in Table 2. H. japonicum is a well known medicinal plant in China.