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720 S.N. Kharat, V.D. Mendhulkar / Materials Science and Engineering C 62 (2016) 719–724 Previous phytochemical investigations on E. scaber have also resulted in the isolation of flavonoids [14]. Nanobiotechnology has become one of the important sectors with focused attention on plant metabolite mediat- ed synthesis of nanoscale particles. The biological constituents of plant origin play pioneer role in reduc- ing salt in the plant extract to synthesis nanoparticles and contributing for their stability. We report an effortless, low-cost, convenient, green synthesis approach to obtain large quantities of silver nanoparticles by reduction of silver ions using E. scaber leaf extract. The importance of the present piece of work is viewed specially with respect to the novel aspect of surface capping of AgNPs with plant secondary metabolites of medicinal interest which may generate suggestive scope for re- search in the field of nanoparticles assisted plant metabolomics. This study is also important to understand the phytosynthesized AgNPs (synthesis of nanoscale silver particles using plant material extract) in scavenging the free radicals. The silver nanoparticle technology is awarded with newer applications in commercial and industrial front especially in the field of drug industry due to the useful potentials of nanoscale size. 2. Materials and methods 2.1. Plant material and chemicals The fresh and healthy leaves of the experimental plant E. scaber L. were collected from the Botanical garden of The Institute of Science, Mumbai. Leaves were rinsed thoroughly with tap water followed by distilled water to remove dust, pathogens and adhered matter. Silver nitrate (AgNO3) was procured from the Merck, and 1,1-Diphenyl-2- picrylhydrazyl (DPPH) from Sigma Aldrich, L-Ascorbic acid (Himedia), Methanol (SD-Fine), HPLC grade water was used in the present analysis. 2.2. Preparation of the E. scaber leaf extract Leaves weighing 10 g were thoroughly washed in distilled water for 5 min, dried, cut into fine pieces and were boiled in a conical flask with 100 ml of double distilled water up to 15 min and filtered through Whatmann No. 1 filter paper. The extract was filtered twice to obtained clear solution. 2.3. Synthesis of silver nanoparticles For the synthesis of AgNPs, 50 ml of aqueous leaf extract of E. scaber and 50 ml of 1 mM silver nitrate (AgNO3) solution were mixed with 400 ml of distilled water in the conical flask. Immediately the flask was kept in the sunlight. A change in the color was observed indicating the beginning of the mechanism of the formation of silver nanoparticles Fig. 1. The color change of the mixture from light to yellowish to reddish to colloidal was monitored periodically. After the synthesis and comple- tion of reaction the solution was centrifuged at 10,000 rpm for 20 min. The transparent solution was discarded and the pellets of silver nano- particles were collected. The pellets were dried in the oven at 45 to 50 °C to obtain constant weight (Fig. 2). 2.4. UV–Vis spectra analysis The bioreduction of Ag+ ions in each sample was observed periodi- cally followed by the dilution with 2 ml of double distilled water. This AgNP synthesized mixture was scan, in the range of 200 to 800 nm wavelengths under UV–visible spectrophotometer (Model-Shimadzu UV 1800, Germany). The spectra were recorded at the intervals of 1 min to 10 min and the distilled water was used as a baseline. Fig. 1. Change in the color indicating synthesis of Nanoparticles (A) 1 mM AgNO3 without plant extracts, (B) Plant aqueous extract and (A + B) 1 mM AgNO3 with E. scaber leaf extract after 5 min. 3. Characterization of silver nanoparticles (AgNPs) 3.1. Nanoparticle tracking analysis (NTA) NTA analysis was carried out using Nanosight UK-LM20 instrument. The NTA technique is a light-scattering technique and allows nanoparti- cles to be sized in suspension on a particle-by-particle basis. NTA use a laser light source to illuminate metal particles. Enhanced by a near- perfect black background, the particles appear individually as point- scatterers moving under Brownian motion. The NTA image analysis software automatically tracks and records the size of many particles. The viewing unit is configured with a temperature controller and is equipped with a 640 nm diode laser. The tracking of the Brownian motion of the particles was captured using the monochrome Marlin CCD camera (Allied Vision Technologies, Germany). The samples were injected by sterile syringe. 3.2. Transmission electron microscopy (TEM) Transmission electron microscopy (TEM) technique was used to visualize the morphology of the AgNPs. A TEM sample of the phyto- synthesized AgNPs using E. scaber aqueous extract was prepared by sonicating the sample in Sonicator (Vibronics VS 80) for 10 min. A drop of the SNPs colloidal solution was placed on carbon coated copper grids, Fig. 2. Pellete of synthesized AgNPs in E. scaber Linn.PDF Image | Antioxidant activity of Silver Nanoparticles using leaf extract
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