Journal home page: http://www.journalijiar.com INTERNATIONAL JOURNAL OF INNOVATIVE AND APPLIED RESEARCH RESEARCH ARTICLE GREEN SYNTHESIS OF SILVER NANOPARTICLES FROM DIPLOCYCLOS PALMATUS. (L.) C. Jeffrey *R. RETHINAM and R. JEYACHANDRAN Department of Botany, St. Joseph s College, (Autonomous) Tiruchirappalli 620 002, India. *Corresponding Author: rethinam87@yahoo.com Abstract: The reduction of silver ions to silver nanoparticles mediated through the plant leaves extract. The morphology and size of the silver nanoparticles were carried out by the Field Emission Scanning Electron Microscope. To identify the elemental composition of materials using Energy Dispersive X-Ray. X-ray diffraction analysis characterization of crystalline materials and the determination of the size is 35.66 nm. The qualitative UV-VIS NIR spectrum of the silver nanoparticles showed at wavelength 440.000 and peaks values 1.957 and proper baseline. The FT-IR spectrum was used to identify the functional group of the active components based on the peak values in the region of infrared radiation against around the silver nanoparticles. Key words: Diplocyclos palmatus, FE-SEM, EDAX, XRD, UV and FT-IR. 1. Introduction Nanotechnology is a rapidly growing science of producing and utilizing nano-sized particles that measure in nanometers. In nanotechnology, a particle is defined as a small object that behaves as a whole unit of properties. In terms of diameter, fine particles cover a range less than 100 nm (nm; 1 nm = 10 9 metre) (Mahendra Rai et al., 2009). Nanoparticles research is currently an area of intense scientific research due to a wide variety of potential applications in biomedical, optical and electronic fields. It is inferred that size, three dimensional shape, hydrophobicity and electronic configuration make nanoparticles an appealing subject in medicinal chemistry. The unique structure of nanoparticles coupled with immense shape for derivatization forms a base for exciting developments in therapeutics. (Nagender Reddy Panyala et al., 2008). In the field of electronics, energy, medicine, and life sciences, nanotechnology offers an expanding research, such as reproductive science and technology, conversion of agricultural and food wastes to energy and other useful byproducts through enzymatic nanobioprocessing, chemical sensors, cleaning of water, disease prevention, and treatment in plants using various nanocides (Carmen et al., 2003; Nair et al., 2010). There are different types of nanoparticles like those of metals, fibers, etc., among these silver nanoparticles have found many applications. Plant extracts are rich sources of secondary metabolites. Most of the extracts contain several metabolites that can easily reduce silver nitrate to silver nanoparticles (Jayapriya and Lalitha, 2013; Leela and Vivekanandan, 2008). 2. Materials and Methods 2.1 Preparation of plant powder The healthy plant leaves free from insect damaged and fungus-infected, dried at room temperature for 5-8 days or until they broke easily by hand. Once completely dry, plants were ground to a fine powder using an electronic blender. The powder was stored in a closed coloured glass bottle at room temperature until required. 2.2 Preparation of the aqueous extract The powder was weighing 1gram then it s transferred into a conical flask with 100ml of distilled water and boiled for 10minutes. It was then filtered through Whatman No.1 filter paper (pore size 25 μm). 2.3 Preparation of 1mM AgNO 3 solution 169.89 mg weighted for Ag NO 3 dissolved with 1000 ml of distilled water then stored in glass container at dark condition. 14
2.4 Synthesis of Silver Nanoparticles 1mM aqueous solution of Silver nitrate (AgNO 3 ) was prepared and used for the synthesis of silver nanoparticles. 10 ml of Diplocyclos palmatus Leaf extract was added into 90 ml of aqueous solution of 1mM Silver nitrate for reduction into Ag + ions and kept at room temperature for 24 hours. 2.5 CHARACTERIZATION 2.5.1 Field Emission Scanning Electron Microscope (FE-SEM) In standard electron microscopes, electrons are mostly generated by heating a tungsten filament. They are also produced by a crystal of LaB6. The use of LaB6 results in a higher electron density in the beam and a better resolution than that with the conventional device. In a field emission (FE) electron microscope, on the other hand, no heating but a so-called "cold" source is employed. Field emission is the emission of electrons from the surface of a conductor caused by a strong electric field. An extremely thin and sharp tungsten needle (tip diameter 10 100 nm) works as a cathode. The FE source reasonably combines with scanning electron microscopes (SEMs) whose development has been supported by advances in secondary electron detector technology. The acceleration voltage between cathode and anode is commonly in the order of magnitude of 0.5 to 30 kv, and the apparatus requires an extreme vacuum (~10 6 Pa) in the column of the microscope. Because the electron beam produced by the FE source is about 1000 times smaller than that in a standard microscope with a thermal electron gun, the image quality will be markedly improved; for example, resolution is on the order of ~2 nm at 1 kev and ~1 nm at 15 kev. Therefore, the FE scanning electron microscope (FE-SEM) is a very useful tool for high-resolution surface imaging in the fields of nonmaterial science. 2.5.2 Energy Dispersive X-Ray Spectroscopy (EDAX) Energy Dispersive X-Ray Analysis technique used to identify the elemental composition of materials. Energy Dispersive X-ray analysis is a technique to analyze near surface elements and estimate their proportion at different position, thus giving an overall mapping of the sample. This technique is used in conjugation with SEM. An electron beam strikes the surface of a conducting sample. The energy of the beam is typically in the range 10-20 kev. This causes X-rays to be emitted from the material. The energy of the X-rays emitted depends on the material under examination. By moving the electron beam across the material an image of each element in the sample can be obtained. Due to the low X-ray intensity, images usually take a number of hours to acquire. 2.5.3 X-Ray Diffraction (XRD) X-ray diffraction has been in use in two main areas, for the Fingerprint characterization of crystalline materials and the determination of the size, shape, and their structure unit cell for any compounds. X-ray diffraction analysis the dry powders of the silver nanoparticles were used for XRD analysis. Diffraction effects are observed when electromagnetic radiation impinges on periodic structures with geometrical variations on the length scale of the wavelength of the radiation. The interatomic distances in crystals and molecules amount to 0.15 0.4 nm which correspond in the electromagnetic spectrum with the wavelength of x-rays having photon energies between 3 and 8 kev. Accordingly, phenomena like constructive and destructive interference should become observable when crystalline and molecular structures are exposed to x-rays. X-ray diffract meters consist of three basic elements: an X-ray tube, a sample holder, and an X-ray detector. X Rays are generated in a cathode ray tube by heating a filament to produce electrons, accelerating the electrons toward a target by applying a voltage, and bombarding the target material with electrons. When electrons have sufficient energy to dislodge inner shell electrons of the target material, characteristic X-ray spectra are produced. These spectra consist of several components, the most common being Kα and Kβ. Kα consists, in part, of Kα1 and Kα2. Kα1 has a slightly shorter wavelength and twice the intensity as Kα2. AgNPs was calculated by employing Debye Scherrer formula. D K λ β cosθ Where D is the average crystalline diameter size (A ) K is constant (0.94) λ is the X-ray wavelength used (λ = 1.54A ), β is the angular line width at the half maximum of diffraction (radians) and θ is the braggs angle (degrees) 2.5.4 UV-VIS-NIR Spectrophotometer The qualitative UV-VIS NIR spectrum was used to identification of a compound, quantity present and structure based on the technique selected and the wavelength of Electromagnetic spectrum. Absorption of the ultra- 15
violet radiations results in the excitation of the electrons from the ground state to higher energy state. In UV-Vis spectroscopy, light is passed through a sample at a specific wavelength in the UV or visible spectrum. If the sample absorbs some of the light, not all of the light will be passing through, or be transmitted. Transmission is the ratio of the intensity of the transmitted light to the incident light, and is correlated to absorbance. The absorbance can be used in a quantitative manner, to obtain the concentration of a sample. It can also be used in a qualitative manner, to identify a compound by matching the measured absorbance over a range of wavelengths. 2.5.5 Fourier Transform Infrared Spectroscopy (FT IR) The FT-IR spectrum was used to identify the functional group of the active components based on the peak values in the region of infrared radiation. Place a small drop of the compound on one of the KBr plates. Place the second plate on top and make a quarter turn to obtain a nice even film. Place the plates into the sample holder and run a spectrum. If the sample is too concentrated, separate the plates and wipe one side clean before putting them back together. The KBr plates must be thoroughly cleaned after this procedure to prevent contamination of future samples. Wipe the windows with a tissue, and then wash several times with methylene chloride then ethanol. Use the polishing kit in the lab to polish the window surface. 3. Results and Discussion 3.1 FE-SEM The morphology and size of the silver nanoparticles were carried out by the Field Emission Scanning Electron Microscope was carried out Central Electro Chemical Research Institution, Karaikudi, India. During the process of vaporization, owing to the gas convection, the vaporized smog will rapidly loss its energy and condenses into nanoparticles due to its collision with the atoms of inert-gas. Along with the nanoparticles will then rush rapidly towards the surface of the very low-temperature collector, which then condense and form into nanoparticles. The collection process, liquid nitrogen is continuously injected into the collector to maintain a processing temperature at 196 C, as well as working pressure at 10 Torr. Besides, the working inert-gas is enabling the collection surface of the collector to maintain at a low temperature stably. Upon FE-SEM measurement, the result of the acquired nanoparticles is a spherical shaped structure and nanoparticles were analyzed in different magnifications 1µ, 2µ, 200 nm and 300 nm. (Ho Chang and Ming-Hsun Tsai, 2008) (Fig. 1). 3.2 EDAX Energy Dispersive X-Ray Analysis technique used to identify the elemental composition of materials. Energy Dispersive X-Ray Analysis was carried out Central Electro Chemical Research Institution, Karaikudi, India. In the X-ray range the energy of a single photon is just sufficient to produce a measurable voltage pulse X-ray, the output of an ultra low noise preamplifier connected to the low noise are a statistical measure of the corresponding quantum energy. Metallic silver nanocrystals generally show typical optical absorption peak approximately at 3 kev due to 0 15 (Venkateswarlu et al., 2010) (Fig. 2 and Table.1). 3.3 XRD X-ray diffraction has been in use in two main areas, for the Fingerprint characterization of crystalline materials and the determination of the size, shape, and their structure unit cell for any compounds. X-ray diffraction analysis the dry powders of the silver nanoparticles were used for XRD analysis. The synthesized Ag nanoparticles using aqueous leaf extract of synthesized silver nitrate solution was further confirmed by the characteristic peaks observed in XRD image. The XRD pattern showed different intensity peaks in the whole spectrum of 2θ values ranging from 0 to 80. The study was carried out Central Electro Chemical Research Institution, Karaikudi, India. Using a Shimadzu XRD-6000/6100 model with, Measurement Temperature [ C] 25.00, Start Position [ 2Th.] 10.0114, End Position [ 2Th.] 89.9794, Generator setting 30mA, 40kv with Anode material Cu, kα (Aº) 1.54060. X- ray diffraction analysis characterization of crystalline materials and the determination of the average grain crystalline size of Ag NPs were estimated to be approximately 35 nm. (Allafchian and Jalali, 2015) (Fig. 3 and Table.2). 3.4 UV-VIS spectrum The qualitative UV-VIS spectrum was used to identification of a compound, quantity present and structure based on the technique selected and the wavelength of Electromagnetic spectrum. The silver ions reduction confirmation was carried out Central Electro Chemical Research Institution, Karaikudi, India. Using a Cary 500 scan double beam mode, UV-VIS-NIR Spectrophotometer (Make: Varian), spectrum of the reaction solution after cooling at room temperature. The synthesized silver nitrate solution was selected at wavelength from 200 to 800 nm due to sharpness of the wavelength 440.000 and peaks values 1.957 and proper baseline. (Huang et al., 2007) (Fig. 4 and Table.3). 16
3.5 FT-IR The FT-IR spectrum was used to identify the functional group of the active components based on the peak values in the region of infrared radiation. FT-IR analysis was carried out Central Electro Chemical Research Institution, Karaikudi, India. Using a BRUKER OPTIK GMBH (Model: TENSOR 27) The samples scan time 64 scans, Inter fragram size 14220 points, Data from: 4000-1 to 400-1, Detector: RT DLaTGS, Spectral range: 370 to 7500 cm-1, The residues was dried and mixed with Potassium Bromide (KBr) the pellet was used for FTIR analysis in the range of 500-3500 cm-1(niraimathi et al., 2013) (Fig. 5 and Table.4). Figure-1 FE-SEM images Synthesized Silver Nanoparticles from Diplocyclos palmatus. 17
9 cps/ev Figure-2 EDAX - Spectrum Synthesized Silver Nanoparticles from Diplocyclos palmatus. 8 7 6 5 Cl K Ag O C Ca Al Mg Na Si Cl K Ag Ca 4 3 2 1 0 2 4 6 8 10 12 14 kev Spectrum: Acquisition 4940 Figure-3 XRD- Spectrum Synthesized Silver Nanoparticles from Diplocyclos palmatus. Counts outside students_58 100 50 0 20 30 40 50 60 70 80 Position [ 2Theta] (Copper (Cu)) 18
3486.89 Transmittance [%] 92 93 94 95 96 97 98 99 100 1646.24 2351.91 837.18 757.51 3856.55 3796.57 3731.99 2996.29 2889.18 2821.68 1498.94 2401.64 1771.12 1381.07 1235.52 1143.82 502.09 92 93 94 95 96 97 98 99 100 ISSN 2348 0319 International Journal of Innovative and Applied Research (2016), Volume 4, Issue (10): 14-22 Figure-4 UV-VIS-NIR Spectrum Synthesized Silver Nanoparticles from Diplocyclos palmatus. Figure-5 FT-IR Spectrum Synthesized Silver Nanoparticles from Diplocyclos palmatus. 3500 3000 2500 2000 1500 1000 500 3500 3000 2500 2000 Wavenumber cm-1 1500 1000 500 Sample Name: D5 03-03-16 19
Table-1 EDAX Spectrum Synthesized Silver nanoparticles using leaf extract of Diplocyclos palmatus. El AN Series unn. C norm. C Atom. C Error (1 Sigma) K fact. Z corr. A corr. F corr. [wt.%] [wt.%] [at.%] [wt.%] ------------------------------------------------------------------------------------------------ O 8 K-series 36.05 42.63 67.00 4.58 0.472 0.903 1.000 1.000 Ag 47 L-series 26.97 31.89 7.43 0.88 0.252 1.252 1.000 1.012 K 19 K-series 5.04 5.96 3.83 0.19 0.031 1.923 1.000 1.015 Cl 17 K-series 4.78 5.65 4.01 0.19 0.030 1.807 1.000 1.054 C 6 K-series 3.77 4.46 9.33 0.70 0.070 0.637 1.000 1.000 Si 14 K-series 2.42 2.87 2.57 0.13 0.018 1.570 1.000 1.015 Ca 20 K-series 1.96 2.32 1.45 0.10 0.013 1.785 1.000 1.014 Na 11 K-series 1.59 1.88 2.05 0.14 0.016 1.143 1.000 1.003 Mg 12 K-series 1.12 1.33 1.38 0.09 0.010 1.316 1.000 1.006 Al 13 K-series 0.87 1.03 0.96 0.07 0.007 1.411 1.000 1.010 -------------------------------------------------------------------------------------------------- Total: 84.57 100.00 100.00 Table -2 XRD Peak list of synthesized Silver Nanoparticles from Diplocyclos palmatus. S. No Pos. [ 2Th.] Height [cts] FWHM [ 2Th.] d-spacing [Å] Rel. Int. [%] Size (nm) 1 23.7134 26.88 0.5497 3.75217 21.99 20.0 2 27.8135 65.39 0.5353 3.20766 53.49 67.0 3 32.1920 122.25 0.4015 2.78068 100.00 22.0 4 38.1107 106.55 0.5353 2.36135 87.16 15.0 5 46.1646 51.33 0.8160 1.96478 41.99 22.0 6 54.5344 8.35 0.0010 1.68275 6.83 27.0 7 57.4924 19.12 0.0010 1.60301 15.64 93.0 8 64.4794 14.35 0.0010 1.44516 11.74 12.0 9 77.1614 20.38 0.3010 1.23624 16.67 43.0 Average particles size is 35.66 nm Table -3 UV-VIS-NIR Spectrum Synthesized Silver Nanoparticles from Diplocyclos palmatus. S. No Wave length Peak values 1 206.000 4.870 2 440.000 1.957 20
S. No Table -4 FT-IR Peak list of Synthesized Silver Nanoparticles from Diplocyclos palmatus. Wave Number Wave Number Ranges (Cm- 1 ) Molecular Motion Functional Group 502.09 C Br stretch alkyl halides 1 500-1000 757.51 C Cl stretch alkyl halides 837.18 C Cl stretch alkyl halides 1143.82 C N stretch aliphatic amines 2 1000-1500 1235.52 C H wag ( CH2X) alkyl halides 1381.07 C H bend alkanes 1498.94 N O asymmetric stretch nitro compounds 3 1500-2000 1646.24 N H bend 1 amines 1771.12 C C stretch alkynes 4 2000-2500 2351.91 H C=O: C H stretch aldehydes 2401.64 H C=O: C H stretch aldehydes 2821.68 H C=O: C H stretch aldehydes 5 2500-3000 2889.18 C H stretch alkanes 2996.29 C H stretch alkanes 3486.89 O H stretch, H bonded alcohols, phenols 6 3000-3500 3731.99 O H stretch, free hydroxyl alcohols, phenols 3796.57 O H stretch, free hydroxyl alcohols, phenols 3856.55 O H stretch, free hydroxyl alcohols, phenols 21
Conclusion Studies concerning, Green synthesis of silver nanoparticles from Diplocyclos palmatus, (L) Jeffry was carried out using the synthesized silver nanoparticles. The particles size was spherical shaped structure based on FE- SEM. To identify the elemental composition of materials using EDAX analysis and XRD analysis, size of AgNPs were estimated to be approximately 35 nm. The preliminary knowledge from the UV and FT-IR testing, and information regarding common sources of the analysts were able to set up parameters for additional testing. Acknowledgment The authors are thankful to the management of St. Joseph s College, (Autonomous) Tiruchirappalli-620 002 for providing necessary infrastructure facilities to carry out this research work. References Allafchian,A.R, Jalali, S.A.H (2015). Synthesis, characterization and antibacterial effect of poly (acrylonitrile/maleic acid) silver nanocomposite. J.Taiwan Inst. Chem.Eng. 57:154-159. Carmen IU, Chithra P, Huang Q, Takhistov P, Liu S, Kokini JL (2003). Nanotechnology: a new frontier in food science. Food Technol. 57:24-29. Jayapriya, E and Lalitha, P (2013). Synthesis of Silver Nanoparticles using Leaf Aqueous Extract of Ocimum basilicum (L.) International Journal of Chetek Research, 5(6):2985-2992. Ho Chang and Ming-Hsun Tsai, (2008). Synthesis and characterization of ZnO nanoparticles having prism shape by a novel gas condensation process. Rev. Adv. Mater. Sci. 18: 734-743. Huang J, Li Q, Sun D, Lu Y, Su Y, Yang X, Wang H, Wang Y, Shao W, He N, Hong J and Chen C (2007). Biosynthesis of silver and gold nanoparticles by novel sundried Cinnamomum campora leaf. Nanotechnology, 18:104-105. Leela Arangasamy and Vivekanandan Munusamy, (2008). African Journal of Biotechnology, 7(17):3162-3165. Mahendra Rai, Alka Yadav, Aniket Gade, (2009). Biotechnology Advances, 27(1):76-83. Nagender Reddy Panyala, Eladia María Peña-Méndez, Josef Havel (2008). Silver or silver nanoparticles: a hazardous threat to the environment and human health. J. Appl. Biomed, 6: 117-129. Nair R, Varghese SH, Nair BG, Maekawa T, Yoshida Y, Kumar DS (2010). Nanoparticulate material delivery to plants. Plant Sci, 179:154-163. Niramathi, K.L., V. Sudha, R. Lavanya, and Brindha,P. (2013). Biosysnthesis of silver nanoparticles using Alternanthera sessilis (Linn.) extract and their antimicrobial, antioxidant activities, Colloids surface B: Biointer. 102: 288-291. Venkateswarlu P, Ankanna S, Prasad TNVKV, Elumalai EK, Nagajyothi PC, Savithramma, (2010). Green synthesis of silver nanoparticles using Shorea tumbuggaia stem bark, Int J Drug Dev Res, 2:720-723. 22