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724 S.N. Kharat, V.D. Mendhulkar / Materials Science and Engineering C 62 (2016) 719–724 5.6. DPPH assay for anti-oxidant activity The antioxidant activity of synthesized AgNPs and aqueous extract of E. scaber was evaluated by DPPH radical scavenging assay, using 5 mg/ml AgNPs synthesized particles and 5 mg/ml leaf extracts (the aliquots were prepared as 50, 100, 150, 200, 250 μg/ml for AgNPs and plant leaf extract), respectively. Ascorbic acid was used as a positive control (in the range of 20 to 140 μg/ml). The results obtained are sum- marized in Table 1. Our finding reveals that the synthesized AgNPs pos- sessed free radical scavenging activity like aqueous plant extract [25]. We observed significant shift in the DPPH radical scavenging ability for studied samples. The scavenging ability increased in a dose depen- dent manner. The recorded scavenging ability for the lowest concentra- tion of the synthesized AgNP (50 μg/ml) was 15.23 ± 0.04 and this scavenging ability was increased to 85.90 ± 0.08 (Table 1), when con- centration was increased to 250 μg/ml (average IC − 126.6 ± 0.06). 50 However, the scavenging ability was recorded for aqueous leaf extract at lowest concentration 13.03 ± 0.08 (50 μg/ml) and when concentra- tion was increased the scavenging ability was 72.80 ± 0.07 (250 μg/ ml) (Table 1) with average IC50 value, 113.06 ± 0.08. There are a num- ber of studies carried by researchers in nanotechnology field on various medicinal plants, but no report is available on nanoparticle synthesis in E. scaber. Hence, the attempt was aimed to study the potentiality of E. scaber mediated nanoparticles for its antioxidant activity. Silver nano- particle synthesis, characterization and there antimicrobial, antioxidant and cytotoxicity activities, were reported in Lantana camara (Ajitha et al., [1]), Momordica cymbalaria (Swamy et al., [2]), Plukenetia volubilis L. (Kumar et al., [23]), Ficus benghalensis (Saxena et al., [24]) and Iresine herbstii (Dipankar, and Murugan, [25]). The antioxidant DPPH radical scavenging activity of pet ether extract of E. scaber was reported by Subramanian et al., [26] with IC50 = 158.207. The anti-oxidant activity is generally due to the presence of phenolic compounds. We reported earlier (Mendhulkar and Kharat, [13]) that the plant E. scaber is a good source of phenolic compounds and flavonoids. 6. Conclusion On the basis of the results obtained it can be concluded that the approach to phytosynthesis of AgNPs from leaf extract of E. scaber is simple, cost effective, precise and eco-friendly. It is fast and convenient method. As E. scaber leaf extract has great medicinal importance and on other hand, AgNPs are potentially biocompatible, the results described here will be useful in biomedical applications as well as in the area of further research activities in nanobiotechnology. A color change from light to yellowish to reddish to colloidal during the formation of silver nanoparticles is confirmed by UV–Vis spectroscopy and further characterization was done using NTA, XRD, FTIR and TEM analysis. The antioxidant activity of both biologically synthesized silver nano- particles and plant leaf extract revealed increased shift in percent scav- enging ability for free radical. Silver nanoparticles might be useful for the development of newer and more potent antioxidants. Therefore, this green method is more economic and useful to produce promising and compatible metal nanoparticles using biological method at commer- cial level and to explore their potentials in drug delivery system using sur- face capping activities of medicinally useful plant metabolites. Acknowledgments The authors are thankful to the Sophisticated Analytical Instrument Facility (SAIF), Indian Institute of Technology Powai, Mumbai and the Head, Department of Botany for providing equipment facilities. We also thank Dr. Rajesh Raut and Dr. Ranjana Singh, for providing kind help for sample analysis. References [1] B. Ajitha, Y.A.K. Reddy, P.S. Reddy, Green synthesis and characterization of silver nanoparticles using Lantana camara leaf extract, Mater. Sci. Eng. C 49 (2015) 373–381. [2] M.K. Swamy, Mohd.S. Akhtar, S.K. Mohanty, U.R. Sinniah, Synthesis and characteri- zation of silver nanoparticles using fruit extract of Momordica cymbalaria and as- sessment of their in vitro antimicrobial, antioxidant and cytotoxicity activities, Spectrochim. Acta A Mol. Biomol. Spectrosc. 151 (2015) 939–944. [3] A.Sironmani,K.Daniel,Silvernanoparticles—universalmultifunctionalnanoparticles for bio-sensing, imaging for diagnostics and targeted drug delivery for therapeutic ap- plications, in: Dr. Izet Kapetanovic (Ed.), Drug Discovery and Development — Present and Future, InTech. 2011, pp. 477–478. [4] K.C. Bhainsa, S.F. D'Souza, Colloids Surf., B 47 (2006) 160. [5] K. Singh, M. Panghal, S. Kadyan, J.P. Yadav, Evaluation of antimicrobial activity of synthesized silver nanoparticles using Phyllanthus amarus and Tinospora cordifolia medicinal plants, J. Nanomed. Nanotechnol. 5 (6) (2014) 1–5. [6] P. Banerjee, M. Satapathy, A. Mukhopahayay, P. Das, Leaf extract mediated green synthesis of silver nanoparticles from widely available Indian plants: synthesis, characterization, antimicrobial property and toxicity analysis, Bioresour. Bioprocessing 1 (3) (2014) 1–10. [7] K. Sahayaraj, Novel biosilver nanoparticles and their biological utility and overview, Int. J. Pharm. 4 (2014) 26–39. [8] T.N.V.K.V.Prasad,E.K.Elumalai,S.Khateeja,Evaluationoftheantimicrobialefficacy of phytogenic silver nanoparticles, Asian Pac. J. Trop. Biomed. (2011) 82–85. [9] B.S. Geetha, S.Mangalam Nair, P.G. Latha, P. Remani, Sesquiterpene lactones isolated from Elephantopus scaber L. Inhibits human lymphocyte proliferation and the growth of tumour cell lines and induces apoptosis, In Vitro: J. Biomed. Biotechnol. 721285 (2012) 1–8. [10] K.R. Kirtikar, B.D. Basu, Indian Medicinal Plants, IIBishen Sing Mahendra Pal Singh Publisher, Dehradun, 1984. [11] M.G. Rajesh, M.S. Latha, Hepatoprotection by Elephantopus scaber L. CCL4. Induced liver injury, Indian J. Physiol. Pharmacol. 45 (2001) 481–486. [12] S.D.J. Singh, V. Krishna, K.L. Mankani, B.K. Manjunatha, S.M. Vidya, Y.M. Manohara, Wound healing activity of the leaf extracts and deoxyelephantopin isolated from Elephantopus scaber Linn, Indian J. Pharm. 37 (2005) 238–242. [13] V.D. Mendhulkar, S.N. Kharat, HPTLC assay for quercetin and rutin flavonoids in Elephantopus scaber [Linn.] grown under induced heat stress condition, Int. J. Pharm. Bio. Sci. 6 (B) (April 2015) 36–52. [14] M. Locatelli, R. Gindro, F. Travaglia, J.D. Coisson, M. Rinaldi, M. Arlorio, Study of the DPPH-scavenging activity: development of free software for the correct interpreta- tion of data, Food Chem. 114 (2009) 889–897. [15] S.F.S. Reihani, M.E. Azhar, Antioxidant activity and total phenolic content in aqueous extracts of selected traditional Malay salads (Ulam), Int. Food Res. J. 19 (2012) 1439–1444. 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