Background L. used to treat dysentery cuts body aches joint pain diarrhea venereal diseases (Kinoshuta and Fireman 1996; Parrotta 2001; Xiao and Wang 1991). Kong et al. (1985) reported the contraceptive properties of this herb. Yuehchukene an alkaloid isolated from this herb showed the Bosutinib significant anti-implantation effects in female mice when given orally or subcutaneously. Low dose of yuehchukene were also reported to have anti-tumor effects (Leung et al. 2000). A coumarin i.e. murrangatin derived from the leaves showed antithyroid house (Khare 2007). Keeping all in mind coupled with the fact that is a rich source of polyphenolic compounds and has also not been much explored for the antioxidant and antimutagenic activity the present study was conducted. Methods Extraction and phytochemical analysis New leaves (disease free) of were collected in the month of December 2011 from Mata Kaulan botanical garden of Guru Nanak Dev University or college campus Amritsar. Herb specimen (accession no. 7315) was recognized and deposited in herbarium of Department of Botanical and Environmental Sciences Guru Nanak Dev University or college Amritsar. Air dried leaves powder was extracted via using sequential extraction method to obtain the hexane chloroform ethyl acetate was measured in terms of hydrogen donating or radical scavenging ability via using DPPH radical scavenging assay (Blois 1958) and ABTS radical cation decolorization assay given by Re et al. (1999). The reductive capacity of iron (III) was decided using the method of Oyaizu (1986) Cu(II)-Nc by CUPRAC Bosutinib assay given by Apak et al. (2007) and the molybdate ion by the method explained by Prieto et al. (1999). Hydroxyl radical scavenging Rabbit Polyclonal to AIG1. ability of extract/fractions was determined by using DNA nicking assay which safeguard the supercoiled pBR 322 plasmid DNA from devastating effects of hydroxyl radicals generated by Fenton’s reagent as explained by Lee et al. (2002). Briefly 0.3 of plasmid DNA was mixed with 10?μl of Fenton’s reagent (30?mM H2O2 50 ascorbic acid and 80?μM FeCl3) followed by the addition of 10?μl of extract/fraction. The final volume of the combination was adjusted to 20.5?μl with distilled water. After incubate the reaction combination for 30?min at 37?°C the DNA was loaded on a 1?% agarose gel (prepared by dissolving 0.5?g of agarose in 50?ml of 1X TBE Buffer followed by ethidium bromide staining) and electrophoresis was accomplished. Rutin was taken as a positive control and the densitometric analysis was done by using AlphaEaseFC 4.0 software for determining the hydroxyl radical scavenging Bosutinib ability of extract/fraction. Antimutagenicity assay The Ames Salmonella histidine reversion assay explained by Maron and Ames (1983) with slight modifications given by Bala and Grover (1989) was used to elucidate the antimutagenic activity of different extract/fractions for TA98 (frame shift mutation) and TA100 (base pair substitution) tester strains of were procured from Prof. B.N. Ames University or college of California Berkeley. Different concentrations of extract/fractions (100 250 500 1000 and 2500?μg/0.1?ml of DMSO/plate) were tested against direct acting mutagens i.e. nitro-o-phenylenediamine (NPD 20 DMSO/plate) and sodium azide (NaN3 2.5 DMSO/plate) for TA98 and TA100 respectively by using two experimental procedures co-incubation and pre-incubation experiment. In the co-incubation experiment a mixture of 0.1?ml of bacterial culture 0.1 of mutagen 0.1 of different concentrations of extract/fractions was Bosutinib added to 2?ml of top agar (0.5?% NaCl 0.5 Agar and 0.5?mM Histidine/Biotine solution) which was then poured onto minimal agar plates. In the pre-incubation experiment the mixture of 0.1?ml of mutagen 0.1 of different concentrations of extract/portion and 0.5?ml of S9 mix (in case of indirect mutagen) was incubated for 30?min at 37?°C. After incubation the combination along with 0.1?ml of culture was added to 2?ml top agar and then poured onto minimal agar plates. Concurrently spontaneous (only 0.1?ml bacterial culture in 2?ml top agar) positive control (0.1?ml bacterial culture along with 0.1?ml mutagen in 2?ml top agar) and unfavorable control (0.1?ml bacterial culture along with 0.1?ml extract/portion concentration in 2?ml top agar) were also kept. Simultaneously the antimutagenic activity of extract/fractions was also.