Ahmad N, et.al. I nt. J . Pharmacol. Pharm. Sci. (2014) 1;1:28-31
International Journal of Pharmacology and Pharmaceutical Sciences 2014; Vol: 1, Issue: 1, 28-31
Short Communication Novel rapid biological approach for synthesis of silver nanoparticles and its characterization Nabeel Ahmad 1 , Kavya Shree 1 , Monisha Srivastava 1 , Rajiv Dutta 2*
1. School of Biotechnology, IFTM University, Moradabad-244102 (UP), India 2. Institute of Bioscience and Bioengineering, Shri Ramswroop Memorial University, Barabanki (UP), India
INTRODUCTION Nanotechnology is multidisciplinary branch, being regarded as a new and emerging field of 21 st century which deals with biosynthesis at nano scale. Currently many ongoing developments at nano scale level imply that nanotechnology will undoubtedly play a very interesting role in several major technologies or conception, rearrangement and manipulation of the structure at the range of 1 nm to 100 nm [1, 2]. The tremendous growth and insight into the organization of nano scale structures into predefined superstructures ensures the critical role of nanotechnology in coming years.[3].There are many important applications of metal nanoparticles in medicine and pharmaceutical industry [4]. Many ongoing developments at nano scale level imply that nanotechnology will undoubtedly play a very interesting role in several major technologies. The metal nanoparticles exhibit many features due to which they are of great interest to the scientists and researchers such as biological, electrical, optical, magnetic and catalytic properties. The nanoparticles are of huge importance due to their minute size and large surface to volume ratio [5, 6]. Owing to this fact, the production and design of materials can be achieved by controlling the shape and size at nano scale. The most common metal nanoparticles used for the biomedical application are gold, silver and zinc nanoparticles. The gold nanoparticles are having numerous applications, especially in the detection of cancer cells [7]. On the other hand silver nanoparticles are of interest to the researchers because of their antimicrobial and antifungal activities. [8] There are several approaches available for the synthesis of silver nanoparticles, most of which require harsh and toxic chemicals which pose greater problems. Therefore scientists and researchers are trying to find out some novel eco-friendly methods for their synthesis via going green, i.e. either by employing plants or microorganisms. These methods are considered effective and valuable than those chemical methods [9]. Biosynthetic methods are the answer for the said problems and can be used for the synthesis of metal nanoparticles and are also very cost effective and appropriate for large scale commercial production of the same. The main aim of the work was the development and characterization of silver nanoparticles through biotransformation by Catharanthus roseus. Its an easily available plant known to possess antimicrobial, antifungal, antioxidant, antibiotic and cancer remedies properties.
Article Information Abstract Article History Received: 27/02/2014 Received in revised Form: 04/03/2014 Accepted: 06/03/2014 Published Online 07/03/2014 Keywords: Nanotechnology Biosynthesis Silver nanoparticles Catharanthus roseus C. roseus Biological methods are far superior to physical and chemical methods which are eco-friendly, manpower and time as well. Several research reports and processes were already known for the synthesis of metal nanoparticles. In this process we had developed a novel and rapid method for the generation of silver nanoparticles below the size of 30 nm through biological approach where C. roesus extract was used as reducing agent. Formation of Silver nanoparticles was confirmed by UV Vis spectra peak at ~425 nm. TEM images provide evidence for the accurate size of the synthesized nanoparticles. Home Page: http://ijppsjournal.org International Journal of Pharmacology and Pharmaceutical Sciences 29 Ahmad N, et.al. I nt. J . Pharmacol. Pharm. Sci. (2014) 1;1:28-31 MATERIAL AND METHODS Material: Fresh and healthy leaves of Catharanthus roseus (C. roseus) were harvested from School of Biotechnology, IFTM University Moradabad, Uttar Pradesh, INDIA. Chemicals: All chemicals used in this experiment were of highest purity and obtained from Sigma Aldrich India namely Silver nitrate. Distilled water was also used for the preparation of reagents. BIOGENSIS OF SILVER NANOPARTICLES Preparation of plant extract
Firstly, for the preparation of plant leaf extract, 10 g of the Catharanthus roseus (C. roseus ) leaves were taken and washed by dipping into tap water and further into distilled water for 30 minutes. After that the leaves were finely cut and kept overnight for drying. Next day, dried leaves powder were poured in 100 ml distilled water in 500 ml of Erlenmeyer flask and boiled for 20 min, then the extract was separated with the help of filter paper. The extract was stored at 4
C for further experiment.
Synthesis of Silver Nanoparticles by C. roseus The aqueous solution of 1mM silver nitrate (AgNO3) was prepared and used for the synthesis of silver nanoparticles. 10 ml of C. roseus extract was added into 90 ml of aqueous solution of 1 mM silver nitrate (AgNO3) for reduction into Ag + ions and its kept for incubation period of 15 min. under vigorous stirring by magnetic stirrer at 65C temperature. A rapid change of color was observed shows that the generation of silver Nanoparticles (Qualitative Analysis). Now the Silver Nanoparticles solution mixture was kept at 24 hours under mild stirring. Obtained mixture was then purified by centrifugation at 10,000 rpm for 15 min followed by dispersion of the pellet in deionized water. Characterization of Silver Nanoparticles
The characterization of silver nanoparticles was examined by UV spectrophotometer and Transmission Electron Microscopy (TEM). UV Vis Spectrophotometric Analysis
UV-Vis spectroscopy measurements of synthesized Silver Nanoparticles were characterized by Perkin-Elmer UV-VIS spectrophotometer. The scanning range for the samples was 200-800 nm range operated at a resolution of 1 nm. Transmission Electron Microscope Analysis The morphology and particle size of synthesized Silver Nanoparticles were determined by transmission electron microscopy (TEM) operating on Hitachi Model H-7500 using an accelerating voltage of 200 kV. RESULT AND DISCUSSION Visual Observation of Silver Nanoparticles through Biogenesis process Silver Nanoparticles unveil dark reddish brown color in aqueous solution which is due to excitation of Surface Plasmon Resonance. When the mixture of the extract was added with aqueous solution of silver nitrate complex under vigorous stirring by magnetic stirrer at 65C temperature, the color changes from colorless to brownish yellow within 5 min and after 15 min it turns in to dark reddish brown.. After 24 hours, no color change was found. The results are shown in Table 1. It was only due to the reduction of Ag+ which confirms qualitative analysis for the formation of Ag-nanoparticles as shown in Figure 1 (a and b). Characterization of Silver Nanoparticles UV-Vis spectrophotometric Analysis Biosynthesis of Silver nanoparticle may be easily characterized by UV-Vis absorption spectroscopy. From the qualitative analysis it is clear that the C. roseus is a medium for the reduction of Ag+ that generates silver nanoparticles. The UV-Vis absorption spectra of the Silver Nanoparticles were shown in Figure 2. Absorption spectra of Silver nanoparticles formed in the reaction media has absorbance maxima at ~425 nm. Transmission Electron Microscope Analysis Transmission Electron Microscopy (TEM) technique was used to visualize the size and shape of synthesized silver nanoparticles through C. roseus. It is evident from the figure 3 (a and b) that most of the synthesized particles were mono-dispersed, irregular in 30 Ahmad N, et.al. I nt. J . Pharmacol. Pharm. Sci. (2014) 1;1:28-31 shape and the average size estimated was ~11 nm. The maximum size and minimum size of synthesized nanoparticle were found to be 29.1 nm and 5.49 nm respectively.
Table 1: Rapid color change within 5 to 15 min S.No C. roseus Plant Extract color Aqueous AgNO3 Color Color change in 5 min Color change in 15 min Color change after 24 hours 1. Green White Transparent Brownish Yellow Dark Reddish Brown No change still Dark Reddish Brown
Figure 1 (a) Brownish yellow color within 5 min Figure 1 (b) Dark reddish brown color after 15 min
Figure 2: UV-Vis spectra of synthesized silver nanoparticles through C. roseus plant extract
Figure 3 (a): TEM micrograph of synthesized silver nanoparticles Figure 3 (b): TEM micrograph shows the size of synthesized silver nanoparticles CONCLUSION 31 Ahmad N, et.al. I nt. J . Pharmacol. Pharm. Sci. (2014) 1;1:28-31
We have developed a novel and rapid green biogenesis production of sliver nanoparticles by using extract of C. roseus. The developed process was fast and better from previously known processes and the size of the synthesized nanoparticles were much smaller than other previously known methods of synthesizing nanoparticles. The primary confirmation was done by qualitative analysis i.e change of brownish color within 5 min and after 15 min it turns in to dark reddish brown. The reduction of silver nanoparticles was also confirmed by UV-Vis spectrophotometer- as the maximum absorbance of UV-Vis spectra were found at around ~425 nm. The average size of the synthesized nanoparticles was approximately 11 nm. The study also proves as an evidence for the large scale commercial production of products in biotechnology industries. These nanoparticles can be used as a drug for various diseases like antimicrobial, antifungal and cancer treatment. Due to the smaller size these nanoparticles have great tendency to penetrate the tumors especially for cancer cell. CONFLICT OF INTEREST We declare that we have no conflict of interest. ACKNOLEDGMENT Mr. Nabeel Ahmad is grateful to Prof. R. M. Dubey, Vice Chancellor of IFTM University Moradabad Uttar Pradesh INDIA for providing necessary facilities and permission to carry out this research work. We would also like to extend our gratitude to the SAIF Chandigarh India for characterization of samples. REFERENCES 1. Bar H, Bhui DK, Sahoo GP, Sarkar P, De SP, Misra A. Green synthesis of silver nanoparticles using latex of Jatropha curcas. colloids surface Physiochem Eng Asp 2009; 339: 134-139. 2. Nabeel A, Kumar MS. Nanoparticles: Study of Preparation & its toxicity effects on living systems. Bionano frontier 2011; 4 (1) 1-5. 3. Prasad SB, Aeri V, Yashwant. Current Understanding of Synthesis and Pharmacological Aspects of Silver Nanoparticles. American Journal of Phytomedicine and Clinical Therapeutics 2013; 1 (7): 536-547. 4. Rassaei L, Sillanpaa, French RW, Compton RG, Marken F. Asenite determination in the presence of phosphate at electro aggregated gold nanoparticles deposits. Electroanalysis 2008; 20: 1286-1292. 5. Daniel MC, and Astruc D. Gold Nanoparticles: Assembly, Supramolecular Chemistry, Quantum-Size-Related Properties, and Applications toward Biology, Catalysis, and Nanotechnology Chem. Rev 2004; 104: 293-346. 6. Zharov V P J, Kim W, Curiel DT, Everts M. Self-assembling nanoclusters in living systems: application for integrated photothermal nanodiagnostics and nanotherapy Nanomed: nanotechnol Boi Med 2005; 1: 326-345. 7. Cai W, Gao TH, Hong JS. Applications of gold nanoparticles in cancer nanotechnology. Nanotechnol Sci Appl 2008; 1: 17-32. 8. Bhattacharya D, Gupta RK. Nanotechnology and Potential of M.Os. Crit Rev Biotechnol 2205; 25: 199-204. 9. Mohanpuria P, Rana NK, Yadav SK. Biosynthesis of nanoparticles technological concepts and future applications. Nanopart Res 2007; 10; 507-517
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