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Jagpreet Singh1, Gurleen Kaur1, Pawanpreet Kaur1, Rajat Bajaj1 and Mohit Rawat1*
1
Department of Nanotechnology, Sri Guru Granth Sahib World University, Fatehgarh
Sahib, Punjab.140406.
ABSTRACT
Article Received on
09 May 2016, Nanoparticles are the spearheads of the rapidly expanding field of
Revised on 29 May 2016,
Accepted on 19 June 2016
nanotechnology. An array of physical and chemical methods is used
DOI: 10.20959/wjpps20167-7227 for the synthesis of nanoparticles. The development of immaculate
protocols for the synthesis of highly monodisperse nanoparticles of
various sizes, geometries and chemical composition is one of the most
*Corresponding Author
Mohit Rawat challenging obstructions in the field of nanotechnology. The use of
Assistant Professor, toxic chemicals and non-polar solvents in synthesis leads to the
Department of inability to use nanoparticles in clinical fields. Therefore, development
Nanotechnology, Sri Guru
of clean, non-toxic, biocompatible and eco-friendly method for
Granth Sahib World
synthesis of nanoparticles deserves recognition. Silver nanoparticles
University, Fatehgarh
Sahib, Punjab.140406. are of interest because of the unique properties (e.g., size and shape
depending optical, electrical and magnetic properties) which can be
incorporated into antimicrobial applications, biosensor materials, composite fibers, cryogenic
superconducting materials, cosmetic products, and electronic components. Several physical
and chemical methods have been used for synthesizing and stabilizing silver nanoparticles.
The present review explores the immense plant diversity to be employed towards rapid and
single step protocol preparatory method with green principles over the conventional ones and
describes the antimicrobial activities of silver nanoparticles.
KEYWORDS: size and shape depending optical, electrical and magnetic properties, silver
nanoparticles, antimicrobial etc.
Graphical Abstract
INTRODUCTION
Nanotechnology can be defined as the understanding of phenomena and manipulation of
matter of matter at the atomic, molecular or macromolecular levels at dimensions between
approximately 1–100 nanometer. Due to its small size, it produces structures, devices, and
systems with atleast one novel/superior characteristic property. These properties may differ in
important ways from the properties of bulk materials and single atoms and molecules.
Currently, there are two main approaches for the synthesis of nanomaterials and fabrication
of nanostructure: “top down” and “bottom up” approaches.
Top-down approach is a physical method and also refers to microfabrication method where
tools are used to cut, mill and shape materials into the desired shape and order. Integrated
circuits is an example. It include preparation by lithographic techniques, etching, grinding in
a ball mill, sputtering, etc. However, the most acceptable and effective approach for
nanoparticle preparation is the „„bottom up‟‟ approach, where devices are „„grown‟‟ from
smaller building blocks such as atoms or molecules by self assembly. In this way, it is
possible to control the size and shape of the nanoparticle depending on the subsequent
application through variation in precursor concentrations, reaction conditions (temperature,
pH, etc.), functionalizing the nanoparticle surface, using templates, etc. Thus, it produces
products with high precision accuracy. Colloidal dispersion is an example.
Figure 1: Top-down and Bottom-up synthesis approaches. Reprinted from Sci. Tot.
Environ., Vol. 400 (1-3), Ju-Nam, Y. and Lead, J.R., Manufactured nanoparticles: An
overview of their chemistry, interactions and potential environmental implications,
pp396-414, Copyright 2008 with permission from Elsevier.
Antibacterial properties
Nanosilver kill the gram- positive and gram- negative bacteria very effectively, so it can
called as killing agent.[13,14,15], including antibiotic-resistant strains.[16] Gram-negative bacteria
are the bacteria which retain the colour of the stain even after washing with alcohol or
acetone and include genera such as Acinetobacter, Escherichia, Pseudomonas, Salmonella,
and Vibrio. Acinetobacter species are associated with nosocomial infections, i.e., infections
that are the result of treatment in a hospital or a healthcare service unit, but secondary to the
patient‟s original condition. Gram-positive bacteria are those which lose the colour of the
stain after wash with alcohol or acetone and include many well-known genera such as
Bacillus, Clostridium, Enterococcus, Listeria, Staphylococcus and Streptococcus. Antibiotic-
resistant bacteria are the bacteria that are not controlled or killed by antibiotics which include
strains such as methicillin-resistant and vancomycin-resistant Staphylococcus aureus and
Enterococcus faecium. To enhance the antibacterial activity of various antibiotics[17],
penicillin G, amoxicillin, erythromycin, clindamycin and vancomycin against Staphylococcus
aureus and Escherichia coli. silver nanoparticles (diameter 5-32 nm, average diameter 22.5
nm) play very prominent role. Size-dependent (diameter 1-450 nm) The antimicrobial activity
of silver nanoparticles depends on their size[18] and Gram-positive bacteria. Small
nanoparticles with a large surface area to volume ratio provide a more efficient means for
antibacterial activity even at very low concentration. Also the antimicrobial activity of silver
nanoparticles depend upon the concentration and shape.[19] The different shapes silver
nanoparticles of (spherical, rod-shaped, truncated triangular nanoplates) have been developed
by synthetic routes. Due to their large surface area to volume ratios, truncated triangular
silver nanoplates display the strongest antibacterial activity.
Medical Applications
Nanosilver is used for coating such as incorporated in wound dressings, diabetic socks,
scaffolds, sterilization materials in hospitals, medical textiles etc. One website claims that
“the number of people using colloidal silver as a dietary supplement on a daily basis is
measured in the millions”.
Industrial Applications
Catalysis
The silver nanomaterials and silver nanocomposites are used as catalyst in many such as CO
oxidation, benzene oxidation to phenol, photodegradation of gaseous acetaldehyde and the
reduction of the p-nitrophenol to p-aminophenol.[41] To catalyze the reduction of dyes by
sodium borohydride (NaBH4), silver nanoparticles immobilized on silica spheres[42] etc.
Electronics
Nanosilver has high electrical and thermal conductivity along with the enhanced optical
properties leads to various applications in electronics. In nanoelectronics, silver nanowires
are used as nanoconnectors and nanoelectrodes.[43] Other applications include),
optoelectronics, nanoelectronics (such as single-electron transistors and electrical
connectors), data storage devices, the preparation of active waveguides in optical devices
high density recording devices, intercalation materials for batteries, making micro-
interconnects in integrated circuits (IC) and integral capacitors etc.
Figure 4: Ascorbic acid reduction mechanism of gold and silver ions to obtain Ag0 and
Au0 nanoparticles.
Literature Review
Green Synthesis of Silver nanoparticles using plant extract is a very simple and cost-
effective way that satisfies the demand of research community and eliminates the possibility
of environment hazards simultaneously. Till date several groups had reported the synthesis of
silver nanoparticles using plant extract which are listed in Table 2. such as bioreduction of
silver ions to yield metal nanoparticles with the help of living plants, geranium, Neem leaf,
Aloevera plant extracts, Emblicaofficinalis (amla, Indian Gooseberry. The green synthesis of
silver nps using Capsicum annuum leaf extract has been reported. These biogenic silver
nanoparticles show high antimicrobial activity against gram-positive and gram-negative
bacteria. Table 3.
In a recent report, these nanoparticles have been synthesized on irradiation using an aqueous
mixture of Ficuscarica leaf extract.[104] The silver nanoparticles were formed after three hour
of incubation at 370C using aqueous solution of 5mM silver nitrate. Cymbopogan citratus
(DC) stapf (commonly known as lemon grass) a native aromatic herb from India and also
cultivated in other tropical and subtropical countries showed strong antibacterial effect
against P. aeruginosa, P. mirabilis, E. coli, Shigella flexaneri, S. Somenei and Klebsiella
pneumonia.[105] Many plants such as Pelargonium graveolens[106], Medicagosativa[107],
Azadirachta indica[108], Lemongrass[109], Aloevera[110], Cinnamomum Camphora[111], Emblica
officinalis[112], Capsicum annuum[113], Diospyros kaki[114], Caricapapaya[115], Coriandrum
sp.[116], Boswellia ovalifoliolata[117], Tridax procumbens, Jatropha curcas, Solanum
melongena, Datura metel, Citrus aurantium[118] and many weeds[129,120] have shown the
potential of reducing silver nitrate to give formation of AgNPs.
The bio-synthesis of silver nanoparticles from a silver nitrate solution using two medicinal
plant extract namely Ocimum tenuiflorum(Ag NP1) and Catharanthus roseus(Ag NP2)
has been reported by Dulen Saikia.[121] The plant leaf extracts were prepared by mixing 5 g
of dried leaves in 100 ml of deionized water in an Erlenmeyer flask, boiled for 30 minutes at
desired temperature and then filtered through a Whatman 42 no. filter paper. In a typical
reaction, 9 mL of the leaf extracts was added to the desired amount of aqueous AgNO 3
solution (1×10 -3 mol dm-3, 2×10 -3 mol dm-3, 3×10-3 mol dm-3, 4×10-3 mol dm-3 and 5×10 -3 mol
dm-3 AgNO3) and the reaction mixture was left at room temperature for the reduction process
to take place. When the leaf extract was mixed in the aqueous solution of the silver ion
complex, it started to change colour (within 30 minutes) from brown to reddish brown.
Figure 6: Colour change of AgNO3 solution with the change in concentration of AgNO3
solution (1mM to 5mM) containing Ocimum tenuiflorum leaf extract; (b) Colour
change of AgNO3 solution with the change of concentration of AgNO3 solution (1mM to
5mM) containing Catharanthus roseus leaf extracts; (c) UV-Vis absorption spectra of
AgNP1 using different concentrations of aqueous AgNO3 solution; (1) 1×10-3 mol dm-3,
(2) 2×10-3 mol dm-3, (3) 3×10-3 mol dm-3, (4) 4×10-3 mol dm-3, (5) 5×10-3 mol dm-3; (d) UV-
Vis absorption spectra of AgNP2 using different concentrations of aqueous AgNO3
solution; (1) 1×10-3 mol dm-3, (2) 2×10-3 mol dm-3, (3) 3×10-3 mol dm-3, (4) 4×10-3 mol dm-
3
, (5) 5×10-3 mol dm-3
RESULTS
The reduction of the silver ions through leaf extracts leading to the formation of AgNPs of
fairly well-defined dimensions is being demonstrated in UV-Vis, XRD analysis. The XRD
pattern reveals the face-centred crystal structure of AgNPs. The average grain size obtained
for AgNP1 is 29 nm and for AgNP2 is 19 nm.
Figure 9: Optical image. (A) Gradual colour change for the formation of AgNPs by 7%
of O. sanctum leaf extract at different time intervals. (B) AgNPs formation with O.
sanctum leaf extract at different concentrations (10%, 7%, 5% and 3%) measured at 60
min.
Figure 10: (a) UV-visible spectra for different concentrations of O. sanctum Linn. leaf
extract (PLE) with 10−3M AgNO3 measured at 60 min. (b) UV-visible spectra of 7%
aqueous leaf extract (PLE) of three different plants (Curve A) Ocimum sanctum Linn.
(Curve B) Citrus limon L (Curve C) Justicia adhatoda L with 10−3 M AgNO3
measured at 60 min.
Figure 11: (a) TEM image of biosynthesized silver nanoparticles using O. sanctum leaf
extract at 100 nm scale (b) Effect of the treatment of actively growing cells. With AgNPs
(dispersed in the aqueous leaf extract) formed by 7% leaf extracts of O. Sanctum on
growth pattern of Staphylococcus aureus [(●—●) for untreated and (○—○) for treated]
and Pseudomonas aeruginosa [(▲—▲) for untreated and (Δ—Δ) for treated]. All
values are means of three sets of experimental data.
In one more study, the Green Synthesis of Silver nanoparticles by Mulberry Leaves Extract
has been reported by Akl M. Awwad1, Nidà M. Salem[123] by utilizing the reduced property
of mulberry leaves extract and silver nitrate to synthesize a silver nanoparticles (Ag NPs) at
room temperature and characterized by using UV-visible absorption spectroscopy, scanning
electron microscopy (SEM) and X-ray diffraction (XRD). Further, silver nanoparticles
showed effective antibacterial activity toward Staphylococcusaureus and Shigella sp.
Figure 12: (a) Effect of contact time on AgNPs synthesised by mulberry leaves extract
(b) FT-IR spectra of Mulberry leaves powder.
Figure 13: (a) XRD pattern of AgNPS synthesized by mulberry leaves extract (b) SEM
image of silver nanoparticles.
Figure14: Activity of silver nanoparticles against: (a) Staphylococcus aureus and (b)
Shigella sp.
DISCUSSION
It was found that the average size of silver nanoparticles was 20 ± 2.8 nm that was calculated
by using Debye-Scherer equation. The presence of structural peaks in XRD patterns and
average crystalline size around 20nm clearly illustrates the crystalline nature of AgNPs.
The SEM image of silver nanoparticles, showed cubical and relatively uniform shape of
nanoparticle formation with diameter range 20-40 nm. The larger silver particles may be due
to the aggregation of the smaller ones.
In this study, Banerjee et al.[124], proposed a Leaf extract mediated green synthesis of silver
nanoparticles from widely available Indian plants: Musa balbisiana (banana), Azadirachta
indica (neem) and Ocimum tenuiflorum (black tulsi) and their synthesis, characterization,
antimicrobial property and toxicity analysis. This study investigates an efficient and
sustainable route of AgNP preparation from 1mM aqueous AgNO3 using leaf extracts of
three indian plants. The Ag NPs were characterized by UV-visible spectrophotometer,
particle size analyzer (DLS), scanning electron microscopy (SEM), transmission electron
microscopy (TEM) and energy-dispersive spectroscopy (EDS), Fourier transform infrared
spectrometer (FTIR) to determine the nature of the capping agents in each of these leaf
extracts. Ag NPs showed significantly higher antimicrobial activities against Escherichia coli
(E. coli) and Bacillus sp. in comparison to both AgNO3 and raw plant extracts. Additionally,
a toxicity evaluation of these Ag NPs containing solutions was carried out on seeds of Moong
Bean (Vigna radiata) and Chickpea (Cicer arietinum). Results depicts the treatment of seeds
with Ag NPs solutions exhibited better rates of germination and oxidative stress enzyme
activity (nearing control levels), though detailed mechanism of uptake and translocation are
yet to be analyzed.
Figure 15: UV–Vis absorption spectrum of silver nanoparticles. From (A) banana, (B)
neem and (C) Tulsi leaf extracts.
Figure 16: Graphs obtained from FTIR analysis of AgNPs. Curve: (A)- Banana, (B)-
Neem and (C)- Tulsi leaf extracts respectively.
Figure 18: TEM images of silver nanoparticles formed by the reaction of 1 mM silver
nitrate and 5% leaf extract of (A) banana. (B) neem and (C) tulsi leaves respectively.
CONCLUSION
As metal nanoparticles seems to fascinate for the future diverse industry due to their enrich
chemical, electrical and physical properties. Metal nanoparticles are synthesized
predominantly by wet chemical methods, where the chemical used are toxic and flammable.
So there is need of eco friendly nanoparticles synthesis approach. Green nanotechnology
accomplish this need by synthesize the metal nanoparticles without using any toxic chemical
as a reducing agent. Plant extract mediated biological synthesis of nanoparticles is known as
Green Synthesis or Green Nanotechnology and these nanoparticles are known as biogenic
nanoparticles. Green synthesis provides benefits over chemical and physical method as it is
cost effective. eco friendly, more stable and there is no need any high energy, pressure,
temperature and toxic chemicals.
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