CHARACTERIZATION OF NEWLY SYNTHESIZED SILVER NANOPARTICLE USING UV VISIBLE SPECTROSCOPIC TECHNIQUE RASHMI DWIVEDI

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1 International Journal of Bio-Technology and Research (IJBTR) ISSN(P): ; ISSN(E): X Vol. 5, Issue 6, Dec 2015, 9-16 TJPRC Pvt. Ltd. CHARACTERIZATION OF NEWLY SYNTHESIZED SILVER NANOPARTICLE USING UV VISIBLE SPECTROSCOPIC TECHNIQUE ABSTRACT RASHMI DWIVEDI Guru Ghasidas University, Bilaspur, Chhattisgarh, India Nanotechnology in present era gaining value due to capability of modulating metals in to their nanoparticles. There are few studies which are paying attention on the effects and mechanisms of nanomaterials on plants. The present research prominence on the biological synthesis of silver nanoparticle using herbal extract a natural biopolymer, acting as a reducing and stabilizing agent. In green synthesis we used natural reducing agents to synthesize nanoparticles (in this case silver nanoparticles). The leaves were found to be a good reducing as well as capping agent which can rapidly reduces silver ions (Ag + to Ag). The characteristic color changes from pale yellow to dark brown in the reaction due to their specific properties (Surface Plasmon Resonance). Characterization of newly synthesized silver nanoparticle was made using UV Visible spectroscopic technique. The UV-Vis spectrum of green synthesized silver nanoparticle from Jasminum grandiflorum and Cymbopogon citrullus have maximum absorbance peaks at and respectively. This study focus on a cost efficient ecofriendly and safe technique for the synthesis of silver nanoparticle using leaves of important medicinal plants. KEYWORDS: Silver Nanoparticles, Medicinal Plant, Bioreduction, Green Synthesis Received: Nov 06, 2015; Accepted: Nov 24, 2015; Published: Nov 26, 2015; Paper Id.: IJBTRDEC20152 INTRODUCTION Original Article The development of green processes for the production of nanoparticles is evolving into a significant branch of nanotechnology (Raveendran et al., 2006; Virender et al., 2009). Nanotechnology is expected to be the basis of many technological innovations in the 21 st century. The synthesis of metal nanoparticles is a promising research field due to the possible applications for the extension of novel technologies. Production of nanoparticles can be achieved mainly through three methods such as Chemical, Physical and Biological methods. Since noble metal nanoparticles such as gold, silver and platinum nanoparticles are expansively useful to human contacting areas, there is a growing call for to develop environmentally friendly route for nanoparticles synthesis that do not use toxic chemicals. Biological methods of nanoparticles synthesis using microorganisms, enzymes, and plant or plant extract have been suggested as possible ecofriendly alternatives to chemical and physical methods (Ahmad et al., 2004). The studies of bactericidal nanomaterials are gaining significance because of increase in new resistant strains of bacteria against most potent antibiotics. Metal nanoparticles are of importance due to their potential applications in catalysis, photonics, biomedicine, antimicrobial activity and optics (Wang et al., 2004; Biswas et al., 2004; Shipway et al., 2001; Nie et al., 1997; Govindraju et al., 2008; Govindraju et al., 2009). Silver nanoparticles are silver particles of between 1 nm and 100 nm in size and have captivated exhaustive research concern. Historically, silver has been known to have a disinfecting effect and has been found in applications ranging from traditional medicines to culinary items. It has been reported that silver nanoparticles editor@tjprc.org

2 10 Rashmi Dwivedi (SNPs) are non-toxic to humans and most efficient against bacteria, virus and other eukaryotic microorganisms at low concentrations without any side effects (Jonge et al., 2005). Moreover, several salts of silver and their derivatives are commercially manufactured as antimicrobial agents (Krutyakov et al., 2008). The most important application of silver and SNPs is in medical industry such as tropical ointments to prevent infection against burn and open wounds (Ip et al., 2006). Biological synthesis of nanoparticles by plant extracts is at present under exploitation as some researchers worked on it (Bhyan et al., 2007; Calvo et al., 2006) and testing for antimicrobial activities (Saxena et al., 2010; Khandelwal et al., 2010; Thirumurgan et al., 2010). It is a need of today to develop reliable, non-toxic, clean and eco-friendly experimental protocols for the synthesis of NPs, which is likely through ambient biological resources. Several microorganisms such as bacteria, fungi and yeasts have come up as nanofactories for synthesizing metal NPs of Ag. However, use of plants for the production of nanoparticles has drawn concentration for researchers because of its rapid, economical, eco-friendly protocol and also it provides a single step technique for the biosynthesis process (Huang et al., 2007). Hence, the present work deals with the simple, effective, low cost biological (green) synthesis of silver nanoparticles using different leaf extracts. Further, the bioreduction process was monitored by the UV-visible spectroscopy. MATERIALS AND METHODS Boiling /Collection of the Extracts Two medicinal plants, such as Jasminum grandiflorum and Cymbopogon citrullus included in this study were collected from the University campus. Primarily they were thoroughly washed with distilled water to remove dirt particles. Cleaned herbal parts (leaves) were dried with water absorbent paper (filter paper). 10g chopped leaves of every plant was dispensed in 100 ml of milli Q water and boiled for min. at 80ºC using water bath. Filtered the aqueous plant extract through Whatman filter paper no.1 and used further for the synthesis of silver nanoparticle. Synthesis of Silver Nanoparticles (Medicinal Plants Mediated) 1mM aqueous silver nitrate solution was prepared and stored in brown bottles. 10 ml of herbal extracts was taken in conical flask separately and to this 90 ml of 1mMAgNO 3 solution was added.the same protocol was followed for all the three herbal extracts. The conical flasks were incubated at room temperature. The color change from pale yellow to dark brown was checked periodically. The change in colour visually indicates the formation of Silver Nanoparticles (SNPs) which was used for further characterization study. UV-Vis Spectra Analysis The synthesized phytonanoparticles were characterized by UV-Vis spectroscopy, which is widely used technique for characterization of silver nanoparticles (Sun et al., 2001). The reduction of pure Ag + into Ag was monitored by measuring the UV-Vis spectrum by diluting a small aliquot of the sample into distilled water. UV-Vis spectral analysis was done by using UV-Vis spectrophotometer at the range of nm and observed the absorption peaks at nm regions indicating the presence of Ag nanoparticles. RESULTS AND DISCUSSIONS Synthesis of Silver Nano Particles Two medicinal plants were used to produce silver nanoparticles (Table. 1) and the reduction of silver ions into silver particles during exposure to the plant extract is followed by colour change from colourless to different color, Impact Factor (JCC): NAAS Rating: 2.75

3 Characterization of Newly Synthesized Silver Nanoparticle 11 Using UV Visible Spectroscopic Technique depending on the medicinal plant extract. As the plant extract was mixed in the aqueous solution of the silver ion complex, it started to change from watery to yellowish brown due to reduction of silver ion, which may be the indication of formation silver nanoparticles (Jain et al., 2009; Linga et al., 2011) (Figure 1). The aqueous silver ions when exposed to plant extracts were reduced and thus resulted in the formation of silver nanoparticle. The leaf extracts were initially pale yellow or yellow in colour. After the addition of aqueous silver nitrate solution, the colour changed to dark brown to black colour within 5-20 minutes due to excitation of surface plasmon vibrations in silver nanoparticles (Jha et al., 2010). The time duration taken to change in colour can be varies from species to species. Boswellia ovalifoliolata took 10 min. whereas Shorea tumbuggaia took 15 min to synthesize silver nanoparticles. The findings obtained are useful in terms of selection of medicinal plants for the synthesis of silver nanoparticles (Gilaki, 2010). Color changes appear after the completion of the reaction (Table 2), it is well known that silver nanoparticles exhibit yellowish brown based on their size (Li et al. 2009). Noble metal particles specially silver and gold exhibit a strong absorption band in the visible region and giving specific color to the solution (Ching et al., 2010). As the leaf extracts were mixed with the aqueous solution of the silver ion complex, it was changed into reddish brown color due to excitation of surface plasmon vibrations, which indicated the formation of Ag nanoparticles (Garry et al., 2006). UV Visible Spectroscopy for the Herbal Synthesized Silver NanoParticles In this research we have fruitfully synthesized silver nanoparticles using plant leaves extract. The UV visible spectroscopy of the synthesized nano particles were in the range of nm. All the two plants were showed to synthesize silver nanoparticles by the indication of suitable surface plasmon resonance (SPR) showing peaks under visible spectrum. The nanometer of various synthesized nano particles are Jasminum grandiflorum nm (Figure 2) and Cymbopogon citrullus nm (Figure 3). Broadening of peak indicated that the particles are poly dispersed and the weak absorption peak at shorter wavelengths owing to the presence of numerous organic compounds which are known to intermingle with silver ions. The mechanism of the reaction is the reduction of aqueous metal ion with plant leaves, and the formation of silver nanoparticles. The nanoparticles were primarily characterized by UV Visible spectroscopy, which was proved to be a very useful technique for the analysis of nanoparticles. Color changes appear after the completion of the reaction, it is well known that silver nanoparticles exhibit yellowish brown based on their size (Prabhu et al., 2010). UV-Vis spectrograph of the colloid of Ag nanoparticles has been recorded as a function of time by using a quartz cuvette with silver nitrate as the reference. The green leaves were selected for synthesis of SNPs because they are the site of photosynthesis and availability of more H + ions to reduce the silver nitrate into silver nanoparticles. The molecular basis for the biosynthesis of these silver crystals is speculated that the organic matrix contain silver binding proteins that provide amino acid moieties that serve as the nucleation sites (Sathyavathi et al., 2010). Green synthesis provides advancement over chemical and physical method as it is cost effective, environment friendly, easily scaled up for large scale synthesis and in this method there is no need to use high pressure, energy, temperature and toxic chemicals. CONCLUSIONS In conclusion, the medicinal plant could be used as an excellent and resourceful green material for the rapid and consistent synthesis of silver nanoparticles. The preliminary confirmation for the synthesis of silver nanoparticles (SNPs) was color changes and Uv-Vis absorption spectra of silver nanoparticles formed at specific peak at specific wavelength. editor@tjprc.org

4 12 Rashmi Dwivedi The present procedure is simple, safe, economic, non-toxic and eco-friendly as compare to toxic chemical process. Plant leaves could thus be used as an efficient choice to the cost intensive conventional methods. This study opens up a new opportunity of very conveniently synthesizing Ag nanoparticles using natural products which could be useful in various applications. Our findings could be targeted for the promising potential applications including drug formulation and biomedical applications in future. REFERENCES 1. Raveendran, P., Fu, J. and Wallen, S.L. (2006). A simple and green method for the synthesis of Au, Ag, and Au-Ag alloy nanoparticles. Green Chem, 8: Virender, K. S., Ria, A.Y. and Yekaterina, L. (2009). Silver nanoparticles: Green Synthesis and their antimicrobial activities. J. Mat. Sci, 9: Ahmad, A. and Sastry, M.A. Biological synthesis of triangular gold nanoprisms. Nat. Mater, 3, 2004, Wang, C., Flynn, N.T. and Langer, R. (2004). Controlled structure and properties of thermoresponsive nanoparticle-hydrogel composites. Journal of Advanced Materials, 16: Biswas, A., Aktas, O.C., Schumann, U., Saeed, U., Zaporjtchenko, V. and Faupel, F. (2004). Tunable multiple plasmon resonance wavelengths response from multicomponent polymer-metal nanocoposite systems. Applied Physical Letter, 84: Shipway, A.N. and Willner, I. (2001). Nanoparticles as structural and functional units in surface confined architectures. Chemical Communication, 20: Nie, S. and Emory, S.R. (1997). Probing single molecules and single nanoparticles by surface enhanced Raman Scattering. Science, 275: Govindraju, K., Kiruthiga, V. and Singaravelu, G. (2008). Evaluation of biosynthesized silver nanoparticles against fungal pathogens of mulberry Morus indica. Journal of Biopesticides, 1: Govindraju, K., Kiruthiga, V., Ganesh Kumar, V. and Singaravelu, G. (2009). Extracellular synthesis of silver nanoparticles by a marine alga, Sargassum wightii Grevilli and their antibacterial effects. Journal of Nanoscience and Nanotechnology, 9: Jonge, S.H., Yeo, S.Y. and Yi, S.C. (2005). The effect of filler particle size on the antibacterial properties of compounded polymer silver fibers. J. Mat. Sci, 40: Krutyakov, Y.A., Kudrynskiy, A., Olenin, A.Y. and Lisichkin, G.V. (2008). Extracellural biosynthesis and antimicrobial activity of silver nanoparticles. Russ Chem Rev, 77: Ip, M., Lui, S.L., Poon, V.K.M., Lung, I. and Burd, A. (2006). Antimicrobial activities of silver dressings: An in-vitro comparison. J. Medical Microbial, 55: Bhyan, S.B., Alam, M.M. and Ali, M.S. (2007). Effect of plant extracts on Okra mosaic virus incidence and yield related parameters of Okra. Asian J. Agri. Res, 1: Calvo, M.A., Angulo, E.C., Batllori, P., Shiva, C., Adelantado, C. and Vicente, A. (2006). Natural plant extracts and organic acids: synergism and implication on piglet s intestinal microbiota. Biotechnology, 5: Saxena, A., Tripathi, R.M. and Singh, R.P. (2010). Biological Synthesis of silver nanoparticles by using Onion (Allium cepa) extract and their antibacterial activity. Digest J Nanomater Biostruct, 5: Impact Factor (JCC): NAAS Rating: 2.75

5 Characterization of Newly Synthesized Silver Nanoparticle 13 Using UV Visible Spectroscopic Technique 16. Khandelwal, N., Singh, A., Jain, D., Upadhyay, M.K. and Verma, H.N. (2010). Green synthesis of silver nanoparticles using Argimone mexicana leaf extract and Evaluation of their antimicrobial activities. Digest J Nanomater Biostruct, 5: Thirumurgan, A., Tomy, N.A., Jai Ganesh, R. and Gobikrishnan, S. (2010). Biological reduction of silver nanoparticles using plant leaf extracts and its effect an increased antimicrobial activity against clinically isolated organism. De Phar Chem, 2: Huang, J., Li, Q., Sun, D., Lu, Y., Su, Y., Yang, X., Wang, H., Wang, Y., Shao, W., He, N. and Hong, J. (2007). Chen C, Biosynthesis of silver and gold nanoparticles by novel sundried Cinnamomum camphora leaf. Nanotechnology, 18: Sun, Y.P., Atorngitjawat, P. and Meziani, M.J. (2001). Preparation of Silver Nanoparticles via Rapid Expansion of Water in Carbon Dioxide Microemulsion into Reductant Solution. Langmuir, 17: Jain, D., Kumar Daima, S., Kachhwaha, S. and Kothari, S.L. (2009). Synthesis of plant mediated silver nanoparticles using Papaya Fruit Extract and Evaluation of their Antimicrobial Activities. Digest Journal of Nanomaterials and Biostructures, 4: Linga Rao, M. and Savithramma, N. (2011). Biological synthesis of silver nanoparticles using Svensonia hyderobadensis leaf extract and evaluation of their antimicrobial efficacy. J. Pharm. Sci. Res, 3: Jha, A.K. and Prasad, K. (2010). Green Synthesis of Silver Nanoparticles Using Cycas Leaf. Int. J. Green Nanotech: Phys and Chem, 1: Gilaki, M. (2010). Biosynthesis of silver nanoparticles using plant extracts. J. Biol. Sci, 10: Li, L.C., Liu, L., Liu, R.T. and Liu, S. (2009). Identification of phenyl ethanoid Glycosides in plant extract of Plantago asiatica by liquid chromatography-electrospray ionization mass spectrometry. Chinese J. Chem, 27: Ching, T., Hou, J. and Shaw, F. (2010). Biocatalysis and Biomolecules Engineering, John Willey & sons, Hoboken, New Jersey, pp Garry, R., Tim, L. and Kei, M. (2006). Electron transfer in nanomaterials, the electrochemical society, New Jersey, Prabhu, N., Divya, T.R. and Yamuna, G. (2010). Synthesis of silver phyto nanoparticles and their antibacterial efficacy. Digest J Nanomater Biostruct, 5: Sathyavathi, R., Balamurali Krishna, M., Venugopal Rao, S., Saritha, R. and Narayana Rao, D. (2010). Biosynthesis of Silver APPENDICES Nanoparticles Using Coriandrum Sativum Leaf Extract and Their Application in Nonlinear Optics. Adv Sci Lett, 3: Jasminum grandiflorum editor@tjprc.org

6 14 Rashmi Dwivedi Cymbopogon citratus Figure 1: Synthesis of Silver Nanoparticle Indicated by Change in Colour (a) AgNO 3, (b) Plant Extract & (c) AgNO 3 + Leaf Extract Figure 2: Jasminum grandiflorum No. P/V Wavelength (nm.) Abs Figure 3: Cymbopogon Citratus No. P/V Wavelength (nm.) Abs Impact Factor (JCC): NAAS Rating: 2.75

7 Characterization of Newly Synthesized Silver Nanoparticle 15 Using UV Visible Spectroscopic Technique Table 1: List of Medicinal Plants Collected for the Synthesis of Silver Nanoparticles S.no. Scientific Name Family Common Name Medicinal Uses Cymbopogon citratus Jasminum grandiflorum Poaceae/Gramineae Oleaceae lemongrass jasmine Promotes good digestion, and a preparation of lemon grass with pepper has been used for relief of menstrual troubles and nausea. It induces perspiration, to cool the body and reduce a fever. It is well known a mild insect repellent (citronella) and the essential oil is used in perfumery Plant pacifies vitiated vata, migraine, paralysis, wounds, ulcers, constipation, flatulence, skin diseases and stomatitis Table 2: Indication of Color Change in Synthesis of Silver Nano Particle (SNPs) Plant Leaf Color Change ph Change Color S. No. Extract +AgNO 3 Time Result Intensity Scientific Name Before After Before After Cymbopogon Light Dark min Positive citratus yellow yellow Jasminum Light 2. Brown min Positive grandiflorum yellow Color intensity: ++ = Dark color, +++ = very dark color editor@tjprc.org

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