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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.
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International Journal of Pharmacology and
Pharmaceutical Sciences
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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
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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
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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.
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