A REVIEW ON GREEN SYNTHESIS AND CHARACTERIZATION OF SILVER NANOPARTICLES AND THEIR APPLICATIONS: A...

7/25/2019 A REVIEW ON GREEN SYNTHESIS AND CHARACTERIZATION OF SILVER NANOPARTICLES AND THEIR APPLICATIONS: A

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www.wjpps.com Vol 5, Issue 7, 2016. 730

Rawat et al. World Journal of Pharmacy and Pharmaceutical Sciences

A REVIEW ON GREEN SYNTHESIS AND CHARACTERIZATION OF

SILVER NANOPARTICLES AND THEIR APPLICATIONS: A GREEN

NANOWORLD.

Jagpreet Singh1, Gurleen Kaur

1, Pawanpreet Kaur

1, Rajat Bajaj

1and Mohit Rawat

1*

1Department of Nanotechnology, Sri Guru Granth Sahib World University, Fatehgarh

Sahib, Punjab.140406.

ABSTRACT

Nanoparticles are the spearheads of the rapidly expanding field of

nanotechnology. An array of physical and chemical methods is used

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

challenging obstructions in the field of nanotechnology. The use of

toxic chemicals and non-polar solvents in synthesis leads to the

inability to use nanoparticles in clinical fields. Therefore, development

of clean, non-toxic, biocompatible and eco-friendly method for

synthesis of nanoparticles deserves recognition. Silver nanoparticles

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.

WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES

SJIF Impact Factor 6.041

Volume 5, Issue 7, 730-762 Review Article ISSN 2278 4357

*Corresponding Author

Mohit Rawat

Assistant Professor,

Department of

Nanotechnology, Sri Guru

Granth Sahib World

University, Fatehgarh

Sahib, Punjab.140406.

Article Received on

09 May 2016,

Revised on 29 May 2016,

Accepted on 19 June 2016

DOI: 10.20959/wjpps20167-7227


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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 1100 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 fabricationof 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.


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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.

Figure 2: Different approaches of synthesis of silver nanoparticles.

Green Chemistry of the Synthesis of Nanomaterials

The growing demand in the chemical industries is due to the ample benefits of nanomaterials

and preparation of nanomaterials include chemicals such as solvents, raw materials, reagents,

and template materials. Such chemicals have engendered the noxious intermediates and

products. To deminsh the undesirable products, the concept of green chemistry was brought

in the chemical industries.[1] The advantages of green synthesis of nanoparticles over the

physical and chemical methods are:

a)

Clean and eco-friendly approach, as toxic chemicals are not used;

b)

the active biological component itself act as reducing and capping agent , thereforereduction the overall cost of synthesis process;


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c) can be used at large scale production of nanoparticles;

d)

external experimental conditions like high energy and high pressure are not required,

leads to energy saving process.

Elemental Silver and Silver Nanoaprticles

Elemental or metallic silver (Ag) is a chemical element which is malleable and ductile

transition metal with a white metallic luster appearance. Of all metals, silver has.[2,3]

highest electrical conductivity(higher than copper that is currently used in many electrical

applications)

thermal conductivity

lowest contact resistance

high optical reflectivity.[4]

Silver is stable in pure air and water. The presence of ozone or hydrogen sulfide or sulfur in

the air or water may result in silver tarnishing[5]

due to the formation of silver sulfide. Silver

can be present in four different oxidation states: Ag0, Ag2+, Ag3+. The former two are the

most abundant ones, the latter are unstable in the aquatic environment.[6]

Silver has many

isotopes with 107 Ag being the most common. Although acute toxicity of silver in the

environment is dependent on the availability of free silver ions, investigations have shown

that these concentrations of Ag+ ions are too low to lead toxicity.[7]Metallic silver appears to

pose minimal risk to health, whereas soluble silver compounds are more readily absorbed and

have the potential to produce adverse effects. [8] The reduction in the size of silver to

nanosized silver increases its ability to control bacteria and fungi. Due to the large surface

area of nanomaterials leads to increased contact with bacteria and fungi which increases its

effectivity. Nanosilver, when in contact with bacteria and fungus, adversely affects the

cellular metabolism of the electron transfer systems and the transport of substrate in the

microbial cell membrane. Bacteria and fungi causes itchiness, infection, odor, sores, the use

of nanosilver repress the proliferation of bacteria and fungi. Nanosilver have been widely

used due to its antibacterial microbial activity for the development of products containing

siver include food contact materials (such as cups, bowls and cutting boards), odor-resistant

textiles, electronics and household appliances, cosmetics and personal care products, medical

devices, water disinfectants, room sprays, childrens toys, infant products and health

supplements.


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Properties of Silver Nanoparticles

Surface effects and quantum effects are two factors which differentiate nanomaterials from

bulk.[9]

Due to these effects there significant change in chemical, mechanical, optical,

electrical and magnetic properties of materials. Therefore nanosilver has unique optical and

physical properties that are not present in bulk silver, and which are very useful in medical

applications. Mainly the properties of silver nanoparticles are antibacterial, antifungal[10]

Antiviral[11]., anti-inflammatory.[12]etc.

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

patients 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, Staphylococcusand 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 forantibacterial 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.


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ACTION OF SILVER NANOPARTICLES ON MICROBES

Silver nanoparticles are able to physically interact with the cell surface of various bacteria.

This is particularly in the case of Gram- negative bacteria where numerous studies have been

observed the adhesion and accumulation of AgNPs to the bacteria surface. Silver

nanoparticles have the ability to anchor to the bacterial cell wall and subsequently penetrate

it, thereby causing structural changes in the cell membrane which renders bacteria more

permeable. AgNPs accumulate on the membrane cell creates gaps in the integrity of the

bilayer which predisposes it to a permeability increase and finally bacterial cell death. There

have been electron spin resonance spectroscopy studies that suggested that there is formation

of free radicals by the silver nanoparticles when in contact with the bacteria and these free

radicals have the ability to damage the cell membrane and make it porous which can

ultimately lead to cell death.[20]It is likely that a combined effect between the activity of the

nanoparticles and the free ions contributes in different ways to produce a strong antibacterial

activity of broad spectrum. It has also been proposed that there can be release of silver ions

by the nanoparticles[21]and these silver ions can interact with the thiol groups of many vital

enzymes and inactivate them.[22]

The bacterial cells in contact with silver take in silver ions,

which inhibit several functions in the cell and damage the cells. Then, there is the generation

of reactive oxygen species(ROS), forming free radicals with a powerful bactericidal action,

which are produced possibly through the inhibition of a respiratory enzyme by silver ions and

attack the cell itself. Silver ions have the tendency to enter the microbial body causing

damage to its structure. As a consequence, ribosomesmay be denatured with inhibition of

protein synthesis, as well as translation and transcription can be blocked by the binding with

the genetic material o the bacterial cell. Protein synthesis has been shown to be altered by

treatment with AgNPs and proteomic data have shown an accumulation of immature

precursors of membrane proteins resulting in destabilization of the composition of the outer

membrane. Silver is a soft acid, and there is a natural tendency of an acid to react with a base,

in this case, a soft acid to react with a soft base. The cells are majorly made up of sulfur and

phosphorus which are soft bases. The action of these nanoparticles on the cell can cause the

reaction to take place and subsequently lead to cell death. Another fact is that the DNA has

sulfur and phosphorus as its major components; the nanoparticles can act on these soft bases

and destroy the DNA which would definitely lead to cell death.[23]


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Figure 3: Various modes of action of silver nanoparticles on bacteria

Applications of Silver Nanoparticles

Scientific Applications

Due to the surface Plasmon resonance (SPR)[24,25] and surface enhanced raman scattering

(SERS) properties of silver nanoparticles, they have many applications such as sensing

applications including detection of DNA sequences.[26] laser desorption/ionization mass

spectrometry of peptides[27], colorimetric sensors for Histidine[28], enhanced IR absorption

spectroscopy, biolabeling and optical imaging of cancer.[29]

biosensors for detection of

herbicides[30]and glucose sensors for medical diagnostics.[31]

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 dai ly basis is

measured in the millions.


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Table 1: Emerging applications of nanosilver in medical products. (Reprinted from

Nanotoxicology, Vol. 3 (2), Wijnhoven, S.W.P., Peijnenburg, W.J.G.M., Herberts, C.A.,

Hagens, W.I., Oomen, A.G., Heugens, E.H.W., Roszek, B., Bisschops, J., Gosens, I., van

de Meent, D., Dekkers, S., de Jong, W.H., van Zijverden, M., Sips, A.J.A.M., Geertsma,

R.E., Nanosilver a review of available data and knowledge gaps in human and

environmental risk assessment, pp109-138, Copyright 2009 with permission from

Informa Healthcare).

Medical domains Examples References

AnesthesiologyCoating of breathing mask and endotracheal tube

for mechanical ventilatory supportPatent -

DentistrySilver-loaded SiO2nanocomposite resin filler as

aadditive in polymerizable dental materials[32]

Diagnostics

Nanosilver pyramids for enhanced biodetectionUltrasensitive and ultrafast platform for clinicalassays for diagnosis of myocardial infarction

Fluorescence-based RNA sensing Magnetic

core/shell Fe3O4/Au/Ag nanoparticles with turnable

plasmonic properties

[33]

Drug deliveryRemote laser light-induced opening of

microcapsules[34]

Eye care Coating of contact lens

Imaging

Silver dendrimer nanocomposite for cell labeling

Fluorescent core-shell Ag@SiO2nanoballs for

cellular imaging Molecular imaging of cancer cells

[36]

Neurosurgery Coating of catheter for cerebrospinal fluid drainage

Orthopedics

Additive in bone cement Implantable material using

clay-layers with starch-stabilized silvernanoparticles Coating of intramedullary nail for

long bone fractures Coating of implant for jointreplacement Orthopedic stockings

[38]

Patient care Superabsorbent hydrogel for incontinence material

Pharmaceutics

Treatment of dermatitis Inhibition of HIV-1

replication Treatment of ulcerative colitis Treatment

of acne

[40]

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.


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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.

Plant Extract Mediated Green Synthesis of Silver Nanoparticles

There is growing imposition of silver nanoparticles in the field of medicine, optics,

biotechnology, microbiology, environmental remediation and material science has lead to

increasing demand in chemical industry. For the production of silver nanoparticles, various

reducing agents are reported such as H2 gas[44], sodium borohydride[45], hydrazine[46],

ethanol[47]

, ethylene glycols[48]

, Tollens reagent[49]

, ascorbic acid[50]

and aliphatic amines.[51]

Depending on the strength of the reducing agents the particle size can be controlled. Hence

umpteen number of toxic chemicals have been utilized to blend the silver nanoparticles as a

reducing and stabilizing agent and poly (ethylene glycol) block copolymers are used to

reduce the Ag+

ions into Ag nanoparticles in aqueous or non- aqueous solution. Although

these methods may successfully manufacture the well defined nanoparticles but they are quiet

expensive and dangerous to environment. Thus, the concept of green chemistry was

introduced to nanoparticles synthesis with the elimination or decline tradition of toxic

chemicals. In previous study, plants and herbs antioxidant are present as a phytochemical

constituents in seeds, stems, fruits and in leaves. The plant-based phytochemicals in the

synthesis of nanoparticles creates a meaningful symbiosis between natural/plant science and

nanotechnology. This connection provides a green approach to nanotechnology, referred to asgreen nanotechnology or Green Synthesis.The major foredeal of using plant extracts for

silver nanoparticle preparation is that they are easily available, safe and nontoxic in most

cases, have a broad variety of metabolites that can aid in the reduction of silver ions and

quicker than microbes in the synthesis. The main mechanism considered for the route is

plant-assisted reduction due to phytochemicals. The main phytochemicals involved are

terpenoids, flavones, ketones, aldehydes, amides and carboxylic acids. Flavones, organic

acids and quinones are water-soluble phytochemicals that are responsible for the instant

reduction of the ions and formation of nanoprticles. Studies have revealed that xerophytes


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contain emodin, an anthraquinone that undergoes tautomerization, leading to the formation of

the silver nanoparticles.

Mechanism of Plant Mediated Synthesis of Silver nanoparticles

In plants the abundant compunds are Ascorbic acid (C6H8O6) and polyphenols. The acid has

functions in photosynthesis as an enzyme cofactor (including synthesis of ethylene,

gibberellins and anthocyanins) and in the control of cell growth. Also in plants the widely

chemicals are polyphenol or hydroxyphenol are found which are used as a reducing agents

for one pot synthesis of silver nanoparticles nature, polyphenol is one of the most important

chemicals in many reductive biological reactions widely found in plants and animals.

Figure 4: Ascorbic acid reduction mechanism of gold and silver ions to obtain Ag0and

Au0nanoparticles.

Figure 5: Mechanism of Plant mediated synthesis of Silver nanoparticles


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Table 2: Plants extract mediated synthesis of silver nanoparticles

Plants Size(nm) Plantss part Shape References

Alternanthera dentate 50100 Leaves Spherical

Acorus calamus 31.83 Rhizome Spherical

Boerhaavia diffusa 25 Whole plant SphericalTea extract 2090 Leaves Spherical

Tribulus terrestris 1628 Fruit Spherical

Cocous nucifera 22 Inflorescence Spherical

Abutilon indicum 717 Leaves Spherical

Pistacia atlantica 1050 Seeds Spherical

Ziziphora tenuior 840 Leaves Spherical

Ficus carica 13 Leaves -

Cymbopogan citrates 32 Leaves -

Acalypha indica 0.5 Leaves -

Premna herbacea 1030 Leaves Spherical

Calotropis procera 1945 Plant SphericalCentella asiatica 3050 Leaves Spherical

Argyreia nervosa 2050 Seeds -

Psoralea corylifolia 100110 Seeds -

Brassica rapa 16.4 Leaves -

Coccinia indica 1020 Leaves -

Vitex negundo 5 & 1030 Leaves Spherical & fcc

Melia dubia 35 Leaves Spherical

Portulaca oleracea 80 Leaves -Trachyspermum ammi 87, 99.8 Seeds

Swietenia mahogany 50 Leaves

Musa paradisiacal 20 Peel

Moringa oleifera 57 Leaves

Garcinia mangostana 35 Leaves

Eclipta prostrate 3560 Leaves Triangles, pentagons, hexagons

Nelumbo nucifera 2580 Leaves Spherical, triangular

Acalypha indica 2030 Leaves Spherical

Allium sativum 422 Leaves Spherical

Aloe vera 50350 Leaves Spherical, triangular

Citrus sinensis 1035 Peel Spherical

Eucalyptus hybrid 50150 Peel

Memecylon edule 2050 Leaves Triangular, circular, hexagonal

Nelumbo nucifera 2580 Leaves Spherical, triangular

Datura metel 1640 Leaves Quasilinear superstructures

Carica papaya 2550 Leaves

Vitis vinifera 3040 Fruit


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Table 3: Antimicrobial activities of silver nanoparticles synthesized using plant extracts.

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,

Aloeveraplant 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 370

C using aqueous solution of 5mM silver nitrate. Cymbopogan citratus

(DC) stapf (commonly known as lemon grass) a native aromatic herb from India and also

Biological entity Test microorganisms Method References

Alternanthera dentate

Escherichia coli, Pseudomonas

aeruginosa, Klebsiella pneumonia

and Enterococcus faecalis

[93]

Boerhaavia diffusa

Aeromonas hydrophila, Pseudomonas

fluorescens and Flavobacterium

branchiophilum

[94]

Tea E. coli

Tribulus terrestris

Streptococcus pyogens, Pseudomonas

aeruginosa, Escherichia coli, Bacillus

subtilis and Staphylococcus aureus

Kirby-Bauer [96]

Cocous nucifera

Klebsiella pneumoniae, Bacillus

subtilis, Pseudomonas aeruginosa and

Salmonella paratyphi

[97]

Aloe vera E. coli Standard plate count

Solanus torvumP. aeruginosa, S. aureus, A. flavusand Aspergillus niger

Disc diffusion[99]

Trianthema decandra E. coli and P. aeruginosa Disc diffusion

Argimone mexicanaEscherichia coli; Pseudomonas

aeruginosa; Aspergillus flavus

Disc diffusion for

bacteria and food

poisoning for fungi

[101]

Abutilon indicumS. typhi, E. coli, S. aureus and B.

substilus[102]

Cymbopogan citratus

P. aeruginosa, P. mirabilis, E. coli,

Shigella flexaneri, S. somenei andKlebsiella pneumonia

Disc diffusion[103]


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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 (110 -3mol dm-3, 210-3mol dm-3, 310-3 mol dm-3, 410-3 mol dm-3and 510 -3mol

dm-3AgNO3) 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.


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Figure 6: Colour change of AgNO3solution 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 AgNO3solution; (1) 110-3

mol dm-3

,

(2) 210-3 mol dm-3, (3) 310-3 mol dm-3, (4) 410-3 mol dm-3, (5) 510-3 mol dm-3; (d) UV-

Vis absorption spectra of AgNP2 using different concentrations of aqueous AgNO3

solution; (1) 110-3

mol dm-3

, (2) 210-3

mol dm-3

, (3) 310-3

mol dm-3

, (4) 410-3

mol dm-

3, (5) 510-3mol dm-3

Figure 7: (a) XRD pattern of AgNP1 (b) XRD pattern of AgNP2


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Figure 8: (a) TEM images of AgNP1 and (b) AgNP2

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.

I n another study, Sunlight-induced rapid and efficient biogenic synthesis of silver

nanoparticles using aqueous leaf extract of Ocimum sanctum Linn. With enhanced

antibacterial activity has been reported by Goutam Brahmachari et.al.[122]In this study the

O.sanctum leaf extracts of varying concentrations (10%, 7%, 5% and 3%) transferred (5mL

each) into four different 100-mL conical flasks containing 45mL of 1030M silver nitrate

solution leads to final volumes of 50mL each. The resulting solutions were kept under direct

sunlight; gradual colour change was then noticed as an indication of silver nanoparticle

formation (Figure 9) and confirmed by UV-Vis spectrophotometer studies at a regular

interval of time. The nanoparticles were characterized with the help of UV-visible

spectrophotometer and transmission electron microscopy (TEM). The prepared silver

nanoparticles exhibited considerable antibacterial activity.


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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 103M 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 103 M AgNO3measured at 60 min.


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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 () for treated]. All

values are means of three sets of experimental data.

I n one more study,the Green Synthesis of Silver nanoparticles byMulberry Leaves Extract

has been reported by Akl M. Awwad1, Nid 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.


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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.


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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,


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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: UVVis 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.


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Figure17: SEM 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.

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.


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Table 4:Showing zone of inhibitions found in Bacillus and E. coli cultures

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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123.Akl M. Awwad, Nid M. Salem Green Synthesis of Silver Nanoparticles by Mulberry

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124.Priya Banerjee, Mantosh Satapathy, Aniruddha Mukhopahayay and Papita Das Leaf

extract mediated green synthesis of silver nanoparticles from widely available Indian

plants: synthesis, characterization, antimicrobial property and toxicity analysis.

Bioresources and Bioprocessing, 2014; 1: 3.

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