Shenava Aashritha. Int. Res. J. Pharm.

2013, 4 (10)

INTERNATIONAL RESEARCH JOURNAL OF PHARMACY

ISSN 2230 8407

www.irjponline.com
Research Article

SYNTHESIS OF SILVER NANOPARTICLES BY CHEMICAL REDUCTION METHOD

AND THEIR ANTIFUNGAL ACTIVITY
Shenava Aashritha*
BDS, MDS (PhD), Department of Prosthodontics, AB Shetty Institute of Dental Sciences, Mangalore, India
*Corresponding Author Email: keepthefaith999@yahoo.com
Article Received on: 30/09/13 Revised on: 07/10/13 Approved for publication: 11/10/13
DOI: 10.7897/2230-8407.041024
IRJP is an official publication of Moksha Publishing House. Website: www.mokshaph.com
All rights reserved.
ABSTRACT
The aim of this study was to evaluate the antimicrobial activity of silvercolloidal nanoparticles which were synthesised by chemical reduction. Silver
nanoparticles were synthesized by reduction of silver nitrate with sodium citrate. The presence of silver nanoparticles was detected by atomic absorption
spectroscopy. Antifungal activity of silver nanoparticles was detected by the zone of inhibition. Silver nanoparticles exhibited a characteristic surface plasmon
resonance band that is measured by UV-Vis spectroscopy, showing a typical absorbance peak for nanoparticles centred at 430 nm. The antifungal activity of
silver nanoparticles was measured by the zones of inhibition by Kirby Bauer sensitivity testing which were measured after 24 h of incubation at 370C of
Candida albicans growth on sabouraud dextrose agar. This study, integrates nanotechnology leading to possible advances in the formulation of new types of
fungicide.
Keywords: Silver nanoparticles, surface Plasmon, UV-Vis absorption Spectrum, chemicals reduction.

INTRODUCTION
Silver ions are used in the formulation of dental resin
composites; in coatings of medical devices; as a bactericidal
coating in water filters; as an antimicrobial agent in air
sanitizer sprays, pillows, respirators, socks, wet wipes,
detergents, soaps, shampoos, toothpastes, washing machines,
and many other consumer products; as bone cement; and in
many wound dressings to name a few. Silver is generally
used in the nitrate form to induce antimicrobial effect, but
when silver nanoparticles are used, there is a huge increase in
the surface area available for the microbe to be exposed to.
There are many ways depicted in various literatures to
synthesize silver nanoparticles. These include physical,
chemical, and biological methods. The physical and chemical
methods are numerous in number, and many of these
methods are expensive1. The production of nanoparticles in
physical and chemical processes can be obtained by both the
so-called top-down and bottom-up methods. The top down method involves the mechanical grinding of bulk
metals and subsequent stabilization of the resulting nano
sized metal particles by the addition of colloidal protecting
agents2,3. The bottom-up methods, on the other hand, include
reduction of metals, electrochemical methods, and
sonodecomposition. The obtained nanoparticles with the size
range of 3 to 40 nm are characterized by UV-visible (UV
vis) absorption spectroscopy to evaluate their quality4. There
is the electrochemical method which involves the electro
reduction of AgNO3 (silver nitrate) in aqueous solution in the
presence of polyethylene glycol. The nanoparticles thus
produced are characterized by TEM, X-ray diffraction, and
UVvis absorption spectroscopy and are 10 nm in diameter5.
Sonodecomposition, to yield silver nanoparticles, involves
the usage of ultrasonic waves to induce cavitations, a
phenomenon whereby the passage of ultrasonic waves
through an aqueous solution yields microscopic bubbles that
expand and ultimately burst. The synthesis of silver
nanoparticles involves sonochemical reduction of an aqueous
silver nitrate solution in an atmosphere of argon-hydrogen.
The silver nanoparticles are then characterized by TEM, Xray diffraction, absorption spectroscopy, differential scanning

calorimetry, and spectroscopy and are found to be 20 nm in

diameter. The mechanism of the sonochemical reduction
occurs due to the generation of hydrogen radicals during
thesonication process6. There are also many more techniques
of synthesizing silver nanoparticles, such as thermal
decomposition in organic solvents7, chemical and photo
reduction in reverse micelles8,9, spark discharge10, and
cryochemical synthesis11. The problem with most of the
chemical and physical methods of nano silver production is
that they are extremely expensive and also involve the use of
toxic, hazardous chemicals, which may pose potential
environmental and biological risks. It is an unavoidable fact
that the silver nanoparticles synthesized have to be handled
by humans and must be available at cheaper rates for their
effective utilization; thus, there is a need for an
environmentally and economically feasible way to synthesize
these nanoparticles. This study is a quest for such a method
has led to the need for biomimetic production of silver
nanoparticles whereby biological methods are used to
synthesize the silver nanoparticles. The growing need to
develop environmentally friendly and economically feasible
technologies for material synthesis led to the search for bio
mimetic methods of synthesis12. Chemical reduction is the
most frequently applied method for the preparation of AgNPs
as stable, colloidal dispersions in water or organic solvents.
Commonly used reductants are borohydride, citrate,
ascorbate and elemental hydrogen. The reduction of silver
ions (Ag+) in aqueous solution generally yields colloidal
silver with particle diameters of several nano meters. A
multitude of chemical reduction methods have been applied
to synthesize stable and various shapes of silver nanoparticles
in water by the use of different reducing agents (ascorbic
acid13, hydrazine14, dry methane15, dimethyl formamide16 and
sodium borohydride17). The shape, size and the size
distribution strongly depended on the strong and weak
tendency of organic substrates to reduce the silver salts.

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Shenava Aashritha. Int. Res. J. Pharm. 2013, 4 (10)

MATERIAL AND METHOD
Synthesis of colloidal nanoparticles
Silver nanoparticles were synthesized by means of the
turkevich method through the reduction of silver nitrate
(AgNO3), Citrate of sodium (sigma aldrich). All chemicals
were used as received. Double-distilled deionised water was
used. The aqueous solution of silver nitrate (from 1, 0 mM to
6, 0 mM) and 8 % (w/w) and sodium citrate (1, 0 mM to 2, 0
mM) was kept at boiling temperature for a few of minutes
until the solution turned amber yellow, indicating the
formation of colloidal silver nanoparticles, confirmed by
UV/Visible spectroscopy (Spectrophotometer Shimadzu
MultSpec-1501, Shimadzu Corporation, Tokyo, Japan). The
dispersions of silver nanoparticles display intense colours due
to the plasmon resonance absorption. The surface of a metal
is like plasma, having free electrons in the conduction band
and positively charged nuclei. Surface Plasmon resonance is
a collective excitation of the electrons inthe conduction band;
near the surface of the nanoparticles. Electrons are limited to

specific vibrations modes by theparticles size and shape.

Therefore, metallic nanoparticles have characteristic optical
absorption spectrums in the UV-Vis region18.
Antimicrobial test
The antimicrobial efficacy of the silver nanoparticle colloidal
suspension and of Ag nano composites against Candida
albicans was evaluated by zone of growth inhibition and
number of C. albicans colonies in agar plates, respectively.
For contact biocidal property of silver nanoparticles, C.
albicans diluted in 0.9 % NaCl (1 0.2 10 CFU/mL) was
plated onto Sabouraud dextrose agar (Sabouraud Dextrose
Agar, Becton Dickinson France SAS, Le Pont de Claix,
France). Sterile paper disks were placed on the agar and
wetted with 10 L of silver colloidal nanoparticles. These
agar plates were incubated for 48 hours at 37C 2C. The
zones of inhibition were visualized using Kirby-Bauer
sensitivity test.

B
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0.12

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0.10
0.08
0.06
0.04
0.02
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300

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Figure 1: Chemical synthesis of silver nanoparticles

Figure 2: UVVis absorption spectrum of silver nanoparticles prepared

using chemical synthesis

Figure 3: (a) Sabouraud Agar Plate; (b) Control Group; (c) Zone of Inhibition against Candida

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Shenava Aashritha. Int. Res. J. Pharm. 2013, 4 (10)

RESULTS
Silver nanoparticles were synthesized according to the
Turkevich method described in the previous section, the
colloidal solution turned pale yellow indicating that the silver
nanoparticles were formed. (Figure 1) shows the photograph
of the chemical reduction procedure where uniform
temperature was obtained. Silver nanoparticles are known to
exhibit a characteristic surface plasmon resonance band that
can be measured by UV-Vis spectroscopy (Figure 2) shows
the plasmon band of the silver nanoparticle suspensions,
showing a typical absorbance peak for nanoparticles centred
at 430 nm. The symmetrical shape of the plasmon band can
indicate a relative sharp particle size distribution. UV-visible
spectroscopy is one of the most widely used techniques for
structural characterization of silver nanoparticles. The
particles range in size 14 nm with mean diameter 10 nm.
Zones of inhibition were measured after 24 h of incubation at
370C. The comparativestability of discs (a) Sabouraud
dextrose agar (b) Control group (c) Candidal suspension with
discs dipped in AgNPs (1 0.2 10 CFU/mL) shows the
presence of certain level inhibited bacterial growth by more
than 90 %, the zone of inhibition was 1 cm. The diameter of
inhibition zones (in millimeters) around the different silver
nanoparticles sols with against test strain are shown in
(Figure 3). Antimicrobial activities of the synthesized silver
colloidal sols were assessed using the standard dilution micro
method, determining the minimum inhibitory concentration
(MIC) leading to inhibition of bacterial growth (National
Committee for Clinical Laboratory Standards. Performance
standards for antimicrobial susceptibility testing; twelfth
informational supplement. NCCLS document M100-S12.
NCCLS, Wayne, Pennsylvania, 2002).Various regulatory
agencies and standards-writing organizations subsequently
published standardized reference procedures based on the
Kirby-Bauer method. Among the earliest and most widely
accepted of these standardized procedures were those
published by the U.S. Food and Drug Administration (FDA)
and the World Health Organization (WHO).
DISCUSSION
The size of metallic nanoparticles ensures that a significantly
large surface area of the particles is in contact with the
bacterial cells. Such a large contact surface is expected to
enhance the extent of candidal elimination. The synthesis and
characterisation of nano scaled materials in terms of novel
physico-chemical properties is of great interest in the
formulation of fungicidal materials. The extent of inhibition
depends on the concentration of the silver nanoparticles as
well as on the initial candidal population. This was supported
by Sondi and Salopek (2004) who, reported that the
interaction of these particles with intracellular substances
from lysed cells caused their coagulation and the particles
were thrown out of the liquid system. The mechanism of
inhibitory action of silver ions on microorganism shows that
upon Ag+ treatment, DNA loses its replication ability and
expression of ribosomal subunit proteins, as well as other
cellular proteins and enzymes essential to ATP production,
becomes inactivated (Yamanaka et al., 2005). It has also been
hypothesized that Ag+ primarily affects the function of
membrane bound enzymes, in the respiratory chain.
CONCLUSION
Silver has always been an excellent antimicrobial and has
been used for this purpose for ages. The unique physical and
chemical properties of silver nanoparticles only increase the

efficacy of silver. Finally, this study shows that silver

nanoparticles have excellent antibacterial activity against
Candida albicans. This work, integrates nanotechnology and
microbiology, leading to possible advances in the formulation
of new types of fungicides. However, future studies on the
biocidal influence of this nanomaterial on Candida albicans
are necessary in order to fully evaluate its possible use as a
new fungicidal material.
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Cite this article as:
Shenava Aashritha. Synthesis of silver nanoparticles by chemical reduction
method and their antifungal activity. Int. Res. J. Pharm. 2013; 4(10):111-113
http://dx.doi.org/10.7897/2230-8407.041024

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