ABTS solution was freshly prepared for each assay. 1.0 ml ethanol extract (1 mg/ml) was allowed to react with 1 ml of the ABTS solution and the absorbance was taken at 734 nm after 7 min using the spectrophotometer. The ABTS scavenging capacity of the extract was compared with that of ascorbic acid and calculated the percentage inhibition ABTS radical scavenging activity (%) = [(Abscontrol−Abssample)/Abscontrol] × 100 where Abscontrol is

the absorbance of ABTS radical + methanol; Abssample is the absorbance of ABTS radical + sample extract/standard. The standard test organisms for antibacterial activity included the Escherichia coli (ATCC 10586), Pseudomonas aeruginosa www.selleckchem.com/products/z-vad-fmk.html (ATCC 10662), Staphylococcus aureus (ATCC 18590), Proteus vulgaris (ATCC 12453) and Bacillus subtilis (ATCC 8590) were all pathogenic type and obtained commercially from Hi-media Pvt. Ltd and maintained at 4 °C in nutrient agar media. The subculture was done on regular interval of 2 months. The in-vitro testing for antibacterial property of the test samples (complexes

and ligands) was carried out by standard microbiological agar well method. A suspension of each bacterium with the cell density of approx. 1 × 107 colony forming units CFU/ml, prepared separately in nutrient broth media pre-sterilized INK-128 at 121 °C for 20 min was used as bacterial inoculums (BI). About 1.0 ml of BI from each test organisms was transferred to different conical flask containing 50 ml pre-sterilized nutrient agar medium (tempr ≤ 40 °C). After proper mixing, about 20 ml of the culture media in the conical flasks was distributed in two pre-sterilized Petri plates each and then allowed to settle for

solidification of the media. Wells measuring the diameter of 6.0 mm were bored at equidistant places in the nutrient agar media and STK38 each was impregnated with test compounds (100 μg/ml) dissolved in DMSO and incubated at 37 °C for 24 h. The antibacterial property was measured and expressed as diameter (mm) of the zone of inhibition (ZOI) caused by the extracts. All the observations were made in duplicate for each of the test samples. The average of two independent observations was recorded as data in the table. The minimum inhibitory concentration (MIC) of the ethanolic extract was determined by preparing solution of varying concentration (0.2, 0.4, 0.6, 0.8 and 1 mg/ml). The streptomycin (25 mcg/disc) sensitivity of the reference bacterial strains was assessed by the disc diffusion method. The phytochemical characters of all the samples are summarized in Table 1. Presence of alkaloids, tannins, saponin, terpenoid, flavonoid, phenol and cardiac glycoside and absence of anthraquinone and steroid were recorded in the sample. These phytochemicals are playing vital role for the treatment of different types of diseases and therefore they are still used in modern and traditional system of medicine.