biogenesis of silver nanoparticles using alternanthera ficoidea leaf

Int J Pharm Bio Sci 2013 July; 4(3): (P) 104 - 110
Research Article
Nanotechnology
International Journal of Pharma and Bio Sciences
ISSN
0975-6299
BIOGENESIS OF SILVER NANOPARTICLES USING ALTERNANTHERA
FICOIDEA LEAF EXTRACT AND ITS ANTIBACTERIAL POTENTIAL
MRINAL KUMAR BAISHYA1, DEBABRAT BAISHYA2*
AND MOHAN CHANDRA3 KALITA
1
Department of Biotechnology, Gauhati University, Guwahati- 781 014, Assam, India
Department of Bioengineering and Technology, Institute of Science and Technology,
Gauhati University, Guwahati- 781 014, Assam, India
3
Department of Biotechnology, Gauhati University, Guwahati- 781 014, Assam, India
2
ABSTRACT
The synthesis of nanomaterials is a cornerstone of nanotechnology. Most of the
synthesis methods encompassing chemical routes have led to environmental pollution,
especially the bi-products associated with metal production. Here we report an
environmentally benign and cost effective approach in synthesizing silver nanoparticles
using leaf extracts of Alternanthera ficoidea. The surface plasmon resonance of silver
nanoparticles produced a peak centered near 410nm was characterized by UV-Visible
spectrophotometer. Silver nanoparticles were circular in shape with maximum particles
in size range within 4–22 nm as observed under Transmission electron microscope
(TEM). As-synthesized silver nanoparticles were also found to inhibit the growth of two
gram negative clinical bacterial isolates Escherichia coli, and Serratia marcescens.
KEYWORDS: Alternanthera ficoidea, silver nanoparticles, transmission electron microscope,
antibacterial potential
DEBABRAT BAISHYA
Department of Bioengineering and Technology, Institute of Science and Technology,
Gauhati University, Guwahati- 781 014, Assam, India
*Corresponding author
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Int J Pharm Bio Sci 2013 July; 4(3): (P) 104 - 110
INTRODUCTION
The field of nanotechnology is one of the most
active research areas in modern material
science. There have been impressive
developments in the field of nanotechnology in
the recent past years, with numerous
methodologies developed to synthesize
nanoparticles of particular shape and size
depending
on
specific
requirements.
Moreover, new applications of nanomaterials
are emerging rapidly. Silver nanoparticles are
one of the promising products in the
nanotechnology industry. The development of
consistent processes for the synthesis of silver
nanomaterials is an important aspect of
current
nanotechnology
research.
The
methods currently being used encompass
chemical routes. A big impediment to
encouragement of these methods is that the
byproducts associated with metal production
have become a great concern with respect to
environmental pollution, additionally; some of
these processes are expensive. In addition,
the synthetic procedures involve conditions
such as high temperature, pressure and
environmental inertness, which are cost
intensive1. Ever increasing pressure to
develop environmentally benign technique for
nanoparticle synthesis has lead to a renewed
interest in biotransformation as a route to
growth of nano-scale structures. Biological
systems have a unique ability to control the
structure,
phase
and
nano-structural
topography of the inorganic crystals2. It is well
known that microbes such as bacteria3,4,
Yeast5, fungi6,7 and algae8,9 are able to adsorb
and accumulate metals and can be used in
the reduction of environmental pollution and
also for the recovery of metals from waste.
The biosynthetic method employing plant
extracts have drawn attention as a simple and
viable alternative to chemical procedures and
physical methods. Bioreduction of gold and
silver ions to yield metal nanoparticles using
living plants10,11, Geranium leaf broth12, Neem
leaf broth13 lemongrass extract14, Tamarind
leaf extract15 and Aloe vera plant extracts16
have been reported. In the present study A.
ficoidea leaf extract was used to synthesize
silver
nanoparticles.
As-synthesized
nanoparticles were
antibacterial activity.
then
tested
for
its
MATERIALS AND METHODS
(i) Collection of chemicals and plant
material:
Silver nitrate was purchased from HiMedia
Laboratories Pvt. Ltd., Mumbai. All other
chemicals were purchased from Merck,
Mumbai. The plant material i.e Alternanthera
ficoidea was collected from the Botanical
garden of Gauhati University. The bacterial
strains were obtained from Guwahati Medical
College Hospital, Guwahati, Assam.
(ii) Preparation of Alternanthera leaf extract
20 g fresh leaves of Alternanthera ficoidea
were collected and washed three times with
distilled water to remove dust particles.
Leaves were finely chopped and added to 100
mL of ethanol and filtered with Whatman
paper No. 1. Filtrate was collected and was
stored at 4 0C for further analysis.
(iii) Biosynthesis of Silver nanoparticles
550 µL of leaf extract was added to 2.45 mL of
3mM AgNO3 solution at room temperature.
The mixture was then kept in the dark and
change in the colour was noted.
(iv)
Characterization
of
prepared
nanoparticles
To observe the optical property of
biosynthesized silver nanoparticles, samples
were analyzed for UV–VIS spectroscopic
studies
(SHIMEDZU-1800)
at
room
temperature operated at a resolution of 1 nm
between 250 and 500 nm ranges..
Characterization of size and shape of
biosynthesized silver nanoparticles was done
by Transmission electron microscopy (TEM,
Philips CM 200). The sample was first
sonicated (Vibronics VS 80) for 15 min. A drop
of this solution was loaded on carbon-coated
copper grids, and solvent was allowed to
evaporate under Infrared light for 30 min. TEM
was operated at an accelerating voltage of
200 kV.
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Int J Pharm Bio Sci 2013 July; 4(3): (P) 104 - 110
(v) Antimicrobial Activity
The silver nanoparticles synthesized using
Alternanthera ficoidea leaf extract was tested
for antimicrobial activity by agar well diffusion
method
against
two
clinical
isolates
Escherichia coli, and Serratia marcescens.
The pure cultures of bacteria were subcultured on nutrient agar medium. Each
isolates was swabbed uniformly onto the
individual plates using sterile cotton swabs.
Wells of 5mm diameter were made on nutrient
agar plates using gel puncture. Using a
micropipette, 10 µL of nanoparticles solution
was poured onto each well on all plates. After
incubation at 37 0C for 24 hours, the different
levels of zone of inhibition of bacteria were
measured.
RESULTS AND DISCUSSION
The addition of Alternanthera ficoidea leaf
extract to silver nitrate solution resulted in
change in colour from dark greenish to dark
yellow (Fig 1). These color changes arise
because of the excitation of surface plasmon
vibrations with the silver nanoparticles17.
Figure 1
Synthesis of silver nanoparticles using A. ficoidea. a) A. ficoidea leaf extract;
b) Silver nanoparticles synthesis as seen by dark yellow colour
The surface plasmon resonance of silver nanoparticles produced a peak centered near 410nm,
which indicates the reduction of AgNO3 into silver nanoparticles (Fig 2). UV-VIS spectroscopy
could be used to examine size and shape controlled nanoparticles in aqueous suspensions18. The
UV-VIS spectrum was recorded after the completion of the reaction.
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Int J Pharm Bio Sci 2013 July; 4(3): (P) 104 - 110
0.2
S1
Abs.
410
0.1
0.0
350
400
450
500
wavelength (nm)
Figure 2
UV-VIS Spectroscopic analysis of Silver Nanoparticles synthesized
from leaf eaxtract of Alternanthera ficoidea
The TEM images of the prepared silver nanoparticles at 50, 20 and 5 nm scales are shown in the
Fig. 4 a, b, and c, respectively. It was observed that Ag nanoparticles were circular in shape with
maximum particles in size range within 4–22 nm It was also observed that silver nanoparticles
were evenly distributed in the sample.
a)
b)
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Int J Pharm Bio Sci 2013 July; 4(3): (P) 104 - 110
c)
Figure 4
TEM images of biosynthesized silver nanoparticles using Alternanthera
ficoidea leaf extracts at a) 50nm, b) 20nm and c) 5nm scales
Antibacterial activity was observed against E. coli and S. marsescens and the diameter of zone of
inhibition was recorded to be 5mm and 2mm respectively. However, the leaf extract of
Alternanthera ficoidea, taken as control, found to have no inhibitory effect on the tested bacterial
strains (Fig.5). Although the exact mechanism of inhibitory action of nanoparticles against bacteria
was not well known, it is thought that formations of free radicals from the surface of the silver
nanoparticles were responsible for the antibacterial function.19
a)
b)
c)
d)
Figure 5
Antimicrobial activity of biosynthesized silver
nanoparticles using Alternanthera ficoidea. a)
Alternanthera ficoidea leaf extract against E.
coli, b) Alternanthera ficoidea leaf extract
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Int J Pharm Bio Sci 2013 July; 4(3): (P) 104 - 110
against S. marsescens, c) Biosynthesized
silver nano particles against E. coli and d)
Biosynthesized silver nano particles against S.
marsescens
CONCLUSION
The present study of silver nanoparticles
employing the alcoholic extract of leaf of
Alternanthera ficoidea is advantageous in
many aspects. It is devoid of any such
synthetic reducing agents which are harsh and
toxic. The ethanolic extract was found to be
highly efficient in synthesizing the silver
nanoparticles under the set laboratory
conditions. The UV-VIS analysis of the surface
plasmon resonance property suggested the
formation of silver nanoparticles with an
absorption peak at 410nm. TEM analysis
shows the spherical shape of the silver
nanoparticles with size ranging between 4-22
nm. The biosynthesized silver nanoparticles
can be good candidate as antimicrobial agent
as evidence from its activity against E. coli
and S. marsescens. Thus, the silver
nanoparticles synthesized using the leaf
extract of Alternanthera ficoidea is believed to
have immense potential for application in the
field of pharmaceutical industries and in the
other fields as well.
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