One ml of TBA (1%) and 1 ml of TCA (2.8%) were added to above mixture and incubated at 100 °C for 20 min. The development of pink color was measured at 532 nm and % inhibition was calculated. Lipid peroxidation inhibition was evaluated using

modified Halliwell and Gutteridge24 method. Freshly C59 wnt mouse excised goat liver was minced using glass Teflon homogenizer in cold phosphate buffered saline (pH 7.4). 10% homogenate was prepared and filtered to obtain a clear homogenate and this process was carried on ice. Varying concentrations (200–1000 μg/ml) of the extracts were added to the liver homogenate and lipid peroxidation was initiated by adding 100 μl ferrous sulfate (15 mM) to 3 ml of the tissue homogenate. After 30 min, 100 μl aliquot was taken in a tube containing 1.5 ml of 10% TCA. After 10 min, tubes were centrifuged and supernatant was mixed with 1.5 ml of 0.67% TBA in 50% acetic acid. The mixture was heated for 30 min in a boiling water bath. The intensity of the pink colored complex was measured at 535 nm. The degree of lipid peroxidation was assayed by estimating the TBARS

(TBA-reactive species) content and results were expressed as percentage inhibition. The ability of different extracts to protect DNA (pBR322, Merck, India) from damaging effects of hydroxyl radicals generated by Fenton’s reagent (FR) was assessed NVP-BGJ398 cell line by modified DNA nicking assay.25 The reaction mixture contained 2.5 μl of DNA (0.25 μg) and 10 μl FR (30 mM H2O2, 500 μM ascorbic acid and 800 μM FeCl3) followed by the addition of 5 μl of extracts and the final volume was made 20 μl with DW. The reaction mixture was then incubated for 45 min at 37 °C and followed by addition of 2.5 μl loading buffer (0.25% bromophenol blue, 50% glycerol). The results were analyzed on 0.8% agarose gel

electrophoresis using EtBr-staining. Oxidation of BSA (5 μg) in phosphate buffer was initiated by 25 mM AAPH26 and SB-3CT inhibited by different H. isora extracts (50 μg/ml). After incubation of 2 h at 37 °C, 0.02% BHT was added to prevent the formation of further peroxyl radical. The samples were then electrophoresed using 12% SDS-PAGE using the Protean® II System (Bio-Rad, USA) and the gel was stained with 0.25% CBB R-250. The results are presented as means of 3 replicates ± standard error (SE). Means were compared through Duncan’s Multiple Range Test (DMRT) at P ≤ 0.05, using MSTAT-C software. The graphs were plotted using Microcal Origin 6.0. Results depicted in Table 1 revealed that the plant is a rich source of phenols, flavonoids and ascorbic acid; and their quantities showed solvent-type-dependent variations. Several reports have shown a correlation between higher amounts of polyphenols in plants and correspondingly their higher antioxidant potential16, 25, 26 and 27 as they inhibit free radical formation and/or interrupt propagation of autoxidation.28 Our results supported these hypotheses. Phenolic contents were found in the range of 17.3–40.