ABSTRACT

Mostly N-acetyl glucosamine and glucosamine units condensed by β-(1,4)

glycosidiclnkage to give linear polysaccharide, which act as Chitosan. The blend of nano
particles from chitosan source has been given incredible consideration because of their
biocompatibility, biodegradability and hydrophilic properties. Diisocyanates are a
significant class of cross-linking agent for chitosan inferable from their high reactivity
towards amine just as hydroxyl group. As far as we could possibly know, diisocyanates
have so far been used to cross-linking chitosan layers, films, or to join chitosan with
synthetic or natural polymers. Here we present a novel methodology for the arrangement
of in situ cross-linking chitosan iron-nickel complexes as recyclable strong impetuses.
Iron/nickel bimetallic nano particles synthesized by solution reduction process
successfully. In this work, crosslinked Chitosan iron-nickel nano-composites were
synthesized. Crosslinked Chitosan iron-nickel bimetallic nanoparticles were effectively
incorporated and portrayed with Fourier transform infrared spectroscopy (FTIR), X-Ray
power diffraction (XRD) and Scanning Electron Microscope (SEM).

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CHAPTER 1
INTRODUCTION
Carbohydrate is a renewable biomass present as the most abundant polymeric
material on the earth. It has a semi-crystalline polymer obtained from higher plants,
tunicates, bacteria and alga (Dhar et al., 2015; Wu et al., 2014). Researcher has explored
its versatile applications by preparing its different derivatives due to presence of –OH
functional groups on the polymer chain and biodegradable and biocompatible nature
(Kang et al., 2015). 

1.1 Chitosan
Chitosan is gotten from a characteristic polymer chitin which has antibacterial
component. As a polycationic polymer, chitosan has a natural cordial material in view of
its biodegradability. Nontoxic and antibacterial highlights of chitosan made it usable for
some, regions identified with human wellbeing. Because of the functional groups as -NH 2
and -0H in the chitosan structure, chitosan has utilized as an astounding chelating
specialist.
Exoskeletons of crustaceans, fungi and insects are a source of Chitin. By
moderately deacetylation of insoluble in nature occurring chitin gave chitosan. Chitin has
rigid crystalline structure due to hydrogen interactions between acetamide groups and
hydroxyl groups. Deacetylation process of acetylated chitin increase the concentration of
amino group the solubility of deacetylation of chitin increased, and its bio-compatibility
and biodegrability also increased (Murakami et al., 2010).Almost 50% deacetylation of
chitin is known as Chitosan (Dash et al., 2011).

2
 Fig. 1.1 Deacetylation process of acetylated chitin
Mostly N-acetyl glucosamine and glucosamine units condensed by β-(1,4)
glycosidiclnkage to give linear polysaccharide, which act as Chitosan. Under alkaline
condition, Chitosan obtained by deacetylation of chiton (Yazdani et. al., 1995).

1.1.1 Solubility of Chitosan

During deacetylation, acetamide group convert into amino group. Amino group
has important physical properties. Amino group responsible for solubility of chitosan in
acidic condition due to proton sensitivity of amino group (Ravi, 2000).

1.1.2 Activity of Chitosan

The genuine contrast among chitin and chitosan has the acetyl substance of the
polymer. Chitosan is a free amino gathering and has the most helpful subordinate of
chitin (Sarmento et al., 2003). The formation of nano particles from chitosan source has
been given incredible consideration because of their biocompatibility, biodegradability
and hydrophilic properties, which enrich them open doors for different applications
(Bertholon et al., 2006).
The polymeric element of Chitosan is associated with contrarily charged atom and
polymers. Due to the communication that happens between the dynamic amino gatherings
in chitosan and metal nano particles, chitosan has picked as an ensuring operator in the
amalgamation of metal nano particles. Both dynamic amino and hydroxyl utilitarian
gatherings in chitosan show numerous striking organic exercises including antimicrobial
action for illness obstruction toward various human cell types (Wang et al., 2006).

1.1.3 Chitosan as a solid catalyst

Polysaccharide has comprehensively valuable such as an appropriate production
for solid catalysts. Different polysaccharides have used as solid catalyst but Chitosan is a
suitable example for solid catalyst which has mostly dispersed in living organism. Due to
present of free amino group on Chitosan and its Schiff base derivatives increase the solid
support for the restriction of metal catalyst. Solids mostly have used as heterogeneous
catalyst these solids catalysts could easily remove by differently separation techniques
such as filtration. Then these solids catalyst again recycled several times. Another
advantage of the metal catalyst it cannot be leached and it is maintained its activity after
several time recycles (Macquarrie et al., 2005).

3
 Polysaccharide acts as catalyst which increase the capability of reaction and it
became the green chemistry of preparation organic chemistry. In order to solve these
issue immobilized metal catalyst which has highly active are used to drastically smooth
the progress of these reaction. In synthetic organic chemistry, one of the most direct way
to pursue green chemistry is the use of truly efficient catalytic reactions. To address these
issues, highly active immobilized metal catalysts are used to significantly facilitate these
reactions and to benefit from a much simplified reaction workup (Akiyama et al., 2009).
Contrasted with physicochemical strategies, biomimetics have favorable
circumstances of utilizing non dangerous synthetics and ecologically considerate solvents
which are eco-friendly (Rabea et al., 2003). Chitosan is one of the normal
polysaccharides got from chitin, a homo-polymer of b-(1– 4) connected N-acetyl-d-
glucosamine. Chitin is the second most abundant biopolymer in nature after cellulose
(Parashar et al., 2004). It tends to be gotten from shells of marine shellfish, creepy
crawlies, organisms etc. (Wu et al., 2002). The yearly overall creation of scavenger shells
has been evaluated 106 tons and the procurement of chitin and protein from this waste is
an extra wellspring of revenue (Zenker et al., 2003). Humankind has given less
consideration towards usage of shellfish waste, for example, shells of crab, shrimp and
krill which are the most significant chitin sources. The ill-advised transfer of ocean
bottom waste is a nonstop issue along the seaside territories that produces an undesirable
smell and turns into a blemish to the two travelers and neighborhood networks (Kim &
Rajapakse 2003).

1.1.4 Nanotechnology
In the field of biology, chemistry, physics and engineering nano technology importance
increase day by day. In future it will change the direction of technology (Gurunathan et
al., 2015). Now a days, in different field like therapeutic, groceries, fitness care, punter,
and manufacturing, silver nano particles extensively used due to their exclusive chemical
and corporal properties (Li et al., 2010).
Their atypical properties are responsible for diverse purpose counting like visual
sensors, in diagnostics, orthopaedics, drug delivery, anticancer agent, and enhanced the
tumour assassination effects of anticancer drugs (Chernousova & Epple, 2013). Surface to
volume ratio of nano metallic unit liable for their unique physical and chemical properties
in the field of nano technology. It is used for many objectives (Sharma et al., 2009).

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1.2 Synthesis of Nano particles
1.2.1 Using Physical and Chemical Methods
Mostly Three methods are used to prepare nano particles such as physical method,
biological method and chemical method. In physical method, the tube furnace used to
prepare nano particles at different atmospheric pressure by evaporation and condensation
(Kruis et al., 2000). Sparkle discharge and pyrolysis are usual physical method to prepare
silver nano particles (Tien et al., 2008).

1.2.1.1 Advantages of Physical method

It increases the Speed of reaction which is favourable. For example reducing of
silver particles occur due to radiation. Chemical are not used so it is save from chemical
hazardous (Shameli et al., 2010).

1.2.1.2 Disadvantages of physical methods

There are some disadvantages of physical methods. This process gives low yield
but consumption of energy is not favourable. Solvent contaminate the nano particles and
the yield distribution did not remain uniform (Shameli et al., 2010).
1.2.2 Chemical Method to prepare nano particles
Mostly solvent like water or organic liquid used to synthesis nano particles by
chemical method (Tao et al., 2006).There are three main elements, first is metal
predecessor, second is reducing agent and third is stabilizing mediator or capping
mediator. There are two stages involved to reduce the given salt, nucleation and
succeeding growth. Usually, Two methods introduced to produced nano particles,
“Bottom Up Method” and “Bottom Down Method” (Deepak et al., 2011).

1.2.2.1 Bottom Up Method

It is electrochemical technique. Sono decomposition and chemical reduction
involved in the reaction (Amulyavichus et al., 1998).

1.2.2.1.1 Advantages of Bottom Up Method

It give high yield of nano particles as compared to yield of nano particles which
obtained by physical method.

1.2.2.1.2 Disadvantages of Bottom Up Method

This method is extremely expensive. Chemical used for reduction are toxic and
hazardous (Mallick et al., 2004). Nano particles are not pure to desire conditions; as their

5
surface was establish to be residue with chemicals. It increases the difficulties to maintain
the required size and require an additional stage for the avoidance of particle aggregation
(Malik et al., 2010).
There is another problem due to production of by product which are very
poisonous and dangerous. In addition, during the synthesis process, too many toxic and
hazardous by products are excised out. Harmful effects on living organism are seen due to
reducing agent which is used for reduction (Sergeev et al., 1999).
Different chemical techniques have practised for instance cryochemical synthesis,
laser ablation (Mafune et al., 2000), lithography (Hulteen et al., 1999), electrochemical
reduction (Zhu et al., 2001), laser irradiation (Abid et al., 2002), sono-decomposition
(Talebi et al., 2010), thermal decomposition (Hosseinpour-Mashkani et al., 2014), and
chemical reduction (Zhang et. al., 2001).
Chemical method mostly used their benifits such as relieve of manufacture, less
expensive, and better yield. However recently, It has been explained a detailed account
of synthesis methods, properties, and bio-application of silver nano particles (Ganaie et
al., 2014).

1.2.3 Nano particles Preparation

Main two techniques are used to prepare nano particles which are brief.

1.2.3.1 Top-down mode

In the Top-down mode, the bulk size of metal which is required to prepare nano
particles converts into nano particles to desired size. This desired sized nano particles
stabilized by the capping agent or colloidal protecting agent. The typical range of nano
particles is size between 9 to 110 nm. It is obtained by Top-down mode (Founda, 2010).

1.2.3.2 Bottom-up Mode

In this mode small particles combine to give bulk particle. More than one atom,
molecules or cluster combine to each other and their assembly act as colloidal distribution
which act as nano particles. It gives homogeneous composition and gives good quality of
nano particles. For example there are following methods to prepare silver nano particles.

1.2.3.2.1 Physical synthesis method

Evaporation and condensation techniques are mostly adopted to synthesis the
silver nano particles. It occurs at in furnace tube at specific atmospheric pressure. Mover
gas used to vaporized the sample inside the centred of the furnace. Beside the silver Gold,

6
lead sulphide and fullerene nano particles also synthesized by evaporation and
condensation techniques.
There are some drawbacks to synthesize the silver nano particles in the tube
furnace. Problems to synthesize the silver nano particles include, a tube furnace occupies
a large space, consume a huge convention of power still as increase the environmental
temperature about the source material, and need a bundle of time to reach thermal
reliability (Kholoud et al., 2010).

1.3 Biological synthesis method

By the biological method the silver nano particles are obtained by the extract of
bio organism which is act as reducing agent and capping agent. These extract of bio
organism mostly consist of enzymes/proteins, amino acids, polysaccharides, and
vitamins. The bio organism forms the chemical complex with silver ions and then reduces
them. There are some examples such as unicellular green algae extract from the Chlorella
vulgaris, which is used to prepare single crystalline silver nano particles. It obtained at
room temperature. Another example, the protein performs dual function. It reduce the
silver ions and it also control the shape of silver nano particles. The carboxyl groups in
aspartic and glutamine act as residues. The reduction of silver ions due to hydroxyl
groups in tyrosine residues of the proteins. The decrease method was done by a simple bi
practical tri peptide Asp Tyr OMe extra perceived the remove a portion of these build
ups. This procedure of generation gave little Silver nano particles with little poly disparity
and brilliant yield more prominent than 50 percent. Sow concentrate of live Horse feed,
the stocks of Lemongrass, Geranium Leaves and others have go about as green reactants
in silver nano particles creation (Korbekandi et al., 2012).
At the point when silver nitrate responds with concentrate of plants of a
conventional resplendent geranium plant, Pelargonium fundamental focal point, give
Silver Nano particles inside one day (Greulich et al., 2009). If we heat the reaction
mixture the reaction time will be reduce approximately two hours. Heating should be
controlled condition and heat below its boiling point. Hidden proteins in futile multiply
substrate diminished Silver cation particles to deliver uniform Silver protein (core– shell).
Nano particles with an ordinary size are 31 nm (Jung et al., 2008).
A vegetable, Capsicum annum L, was used to make Silver nano particles.
Glutathione go about as a diminishing operator just as topping specialist which can
deliver silver nano particles in water-dissolvable that without trouble append to

7
conceptive protein. Tryptophan deposits of manufactured oligo peptides at the C-end
were perceived as decreasing operators give silver nano particles (Jung et al., 2006).
Now a day, fungs are used to reduce the silver cation ions and to produce silver
nano particles. For example, fungus F. Oxysporum is used to produce silver nano particles
in water. Diverse strain of F. Oxysporum has utilized by number of researchers to produce
nano silver particles (Landage et al., 2012).

1.4 Chemical reduction method

Different chemical methods are used to produce nano silver particles. Chemical
method to produce nano silver particles is due to stability, colloidal dispersion in different
solvent which include polar like water or non-polar like organic solvent. There are
different chemicals are used as reducing agent for example borohydride, citrate, and
elemental hydrogen etc. When silver cations reduced in aqueous solution it gave nano
silver particles of several nano meters (Lansdown et al., 2002).
By the principle of bottom-to-top method, first complex of silver cations reduce to
produced silver atoms. Secondly, these silver atoms agglomeration into oligomeric
cluster. In the result, clusters finally combine to the form of colloidal silver particles. If
the sizes of colloidal particles are much smaller than wavelength of visible light, then
yellow colour of solution appeared which have intense and large band in the range of 390
to 410 nm. Other band, yellow colour of solution appeared which have less intense and
less band in the range of 390 to 410 nm (Tarasenko et al., 2006).

1.5 Cross-linking
Chemical crosslinking is the way toward connecting polymer chains by covalent
bondings, shaping tri dimensional systems which lessen the versatility of the structure and
as a rule improve its mechanical and obstruction properties and its water opposition.
Synthetic cross linking gives a system with particularity for utilitarian gatherings present
on the grid macromolecules (Ziaa et al., 2008). Low poisonous quality cross linking
specialists have been investigated these days for use in sustenance bundling materials, for
example, phenolic functional group (Martinaa et al., 2011).

8
 Fig.2. Uncross linked chain and cross linked three dimension network
A few common compound got from plants have been utilized as cross linkers. A few
potential connections might be included, for example, hydrogen holding, ionic,
hydrophobic collaborations, and covalent holding, albeit covalent bonds are more
inflexible and thermally stable than different communications (Zha & Chang, 2008).
There is some drawback of chitosan. It is due to high aqueous solubility which
demonstrates problematical in applications including aqueous biological media. This
issue can be overwhelmed by the presentation of a cross-linker. A cross-interface is
shaped through a substance response, which joins two polymers together, either through
covalent or ionic bonds or weaker bonding, for instance, van der Waals forces (Ahmed,
2015).
The individual polymers inside the cross-linked composite give together show
new properties, while as yet keeping up their very own basic highlights. One sort of
cross-linking is the arrangement of an interpenetrating polymer organizes which has been
experienced noteworthy investigation. As indicated by the International Union of Pure
and Applied Chemistry (IUPAC) and an interpenetrating polymer network have
characterized as 'a polymer involving at least two systems which are in any event
somewhat intertwined on an atomic scale however not covalently clung to one another
thus can not be broken except if chemical bonds are broken.
Cross-linkers come as polymers, metals, among other substance elements. With
the end goal of our examination we blended and researched a physically cross-linked
system of two polymers, chitosan and siloxane, just as fusing metal nano particles as
further structure enhancers (Li et al., 2013).
There are numerous conceivable polymers which could effectively frame an
interpenetrating polymer network with chitosan, particularly because of the accessibility
of chitosan's useful gatherings for collaboration. These incorporate polymers containing

9
carboxylic acids, epoxides and alcohols. Be that as it may, the utilization of a portion of
these polymers can be troublesome as they may disturb film arrangement.

1.5.1 Cross-linking of Chitosan by Di isocyanates

Four di isocyanates represent over 94-96% of yearly mechanical generation. There
are tolylene di isocyanate, methylene diphenyl di isocyanate, hexamethylene-1,6- di
isocyanate, and isophorone di isocyanate, 5-isocyanato-3- isocyanato methyl)- 1,1,3-tri
methylcyclohexane. Others, for example, bis methane, 1,5-naphthalene di isocyanate, and
1,4-phenylene di isocyanate are created in little sums for uncommon applications. The
reactivity of the isocyanate moiety depends unequivocally on its substitution design and
the steric condition. These distinctions can be helpful, for example, in isophorone
diisocyanate where the two isocyanate bunches are so unique in reactivity that two-
advance responses occur yielding rather well-characterized intermediates and final
results. The reactivity of sweet smelling isocyanates is higher than the reactivity of
aliphatic isocyanates (Arrascue et al., 2003).
Di isocyanates are a critical class of cross linking specialists for chitosan
attributable to their high reactivity towards amine just as hydroxyl gatherings. Be that as
it may, the high reactivity of normal di isocyanates with water and their insolubility in
watery acidic arrangements speak to an obvious disadvantage. To the best of our insight,
diisocyanates have so far been used to cross linking chitosan layers, films, or to utilize
chitosan with natural polymers. Here we present a novel methodology for the
arrangement of in situ cross linking chitosan – silver (I) buildings as recyclable strong
impetuses (Li et al., 2007).
One of these particles that can be connected in that positions is the 1,6-
hexamethylene di isocyanate (HMDI), which is monetarily accessible and by and large
used as a solid cross-linker of – NH2 or – Gracious gatherings, since it has two
responsive isocyanate gatherings ( – N = C = O ). The – Gracious gatherings of chitin and
chitosan can respond with –NCO gatherings to frame a urethane linkage (– N H– C(O) –
O – ) because of the exchange of protons from Goodness gatherings to the nitrogen iotas
of – NCO units. Hexamethylene di isocyanate has likewise respond with deacetylated
amino gatherings in the biopolymers. The response among chitosan and hexamethylene di
isocyanate recently elevated to utilize the subsequent items in various applications e.g.
sub-atomic transporters in the biomedical and natural fields, as help material for Pd
impetuses utilized in hydrogenation responses, as strong help for Cu(I) and Pd(II)

10
impetuses utilized in responses like azide/alkyne cycloaddition and Suzuki cross
couplings, cholesterol assimilation, adsorption of phenols, poisonous cancer-causing
agents, gold sorption or polyurethane elastomers (Chiu et al., 2004).
Metal particle having ultrafine estimate have gotten incredible consideration
because of their unordinary properties that contrast from bulk metal. The chemical and
physical properties of the metal nano particles are subject to their size just as their
structure, shape and size circulation. Along these lines, command over the size and size
dispersion is pivotal and is frequently accomplished by differing the combination
strategies, lessening operators and stabilizers (Bozanic et al., 2011).
Metal particles having ultrafine size have received great attention due to their
unusual properties that differ from bulk metal. The chemical and physical properties of
the metal nano particles are dependent on their size as well as their structure, shape and
size distribution. Therefore, control over the size and size distribution is crucial and is
often achieved by varying the synthesis methods, reducing agents and stabilizers (Abou
et al., 2010).
Exact control of such factors permits the ideal physical and synthetic changes to
the nano particles. For the most part, metal nano particles can be set up in two diverse
ways for example physical and synthetic strategies. In the physical technique,
arrangement of metallic nano particles incorporates laser removal, condensation and
evaporation and ultraviolet radiation (Darroudi et al.,2011).
Precise control of such factors allows the desired physical and chemical changes
to the nano particles. Generally, metal nano particles can be prepared in two different
ways, i.e., physical and chemical methods. In the physical method, preparation of metallic
nano particles includes laser ablation, evaporation/ condensation, and UV irradiation
(Thomas et al., 2009).
Although physical methods are able to produce particles in nano sizes, only
reports on chemical methods can be found abundantly in the literature. The main
chemical method is the reduction of metal ions in solution (Chou et al., 2000).
In recent years, many chemical methods for the synthesis of silver nano particles
have been reported including the polyol method and the liquid–iquid method (Taguchi et
al., 2008).
Synthesis of silver nano particles by chemical reduction method, effect of
reducing agent and surfactant concentration. However, chemical reduction is the most

11
commonly used, due to its simplicity. This technique also enables variation in the molar
concentration of the reactant, dispersant and feed rate of reactant in order to produce
silver nano particles with controlled particle sizes, shapes and particle size distribution
(Cho et al., 2005).
The selection of an appropriate reducing agent is also a crucial factor, as the size,
shape and particle size distribution strongly depend on the nature of the reducing agent.
The introduction of a reducing agent causes the reduction of metal precursor (Singh et al.,
2015).
Reduction of metal salts requires adjustment of the reactivity of the reducing agent
to the redox potential of the metal. During the synthesis process, if the reaction rate is too
fast, rapid formation of a large amount of metal nuclei will occur and result in particles
that are too small. On the other hand, agglomeration of particles will take place if the
reaction rate is too slow. Moreover, the choice of the surfactant is critical since it
determines the stability, solubility, reactivity, dispersibility and even the size and shape of
the nano particles during the synthesis (Zhang et al., 2013).
On the other hand, incorporation of silver nano particles into polymers greatly
enhanced their antimicrobial activity owing to effective biocidal abilities of both silver
ions and silver nano particles at low concentrations (Vimala et al., 2008).
The use of silver nano particles exhibited improved antimicrobial properties than bulk
silver due to high surface area and high fraction of surface atoms, leading to incorporation
of more nano particles inside the bacteria and promoting its efficacy in a sustained
manner (Bajpai et al., 2007).In addition, silver nano particles have proven to be relatively
nontoxic toward human cells (Panacek et al., 2009).
It is believed that the antibacterial mechanism of silver and silver ion (Ag+)
involves their absorption and accumulation by bacterial cells and shrinkage of the
cytoplasm membrane or its detachment from the cell wall. As a result, DNA molecules
become condensed and lose their ability to replicate upon the infiltration of Ag ions. The
silver ions also interact with the sulfur-hydrogen (S–H) bonds of the proteins, blocking
and inactivating them (Klasen et al., 2000).
Cross linked chitosan as a solid support for metal nano particles in catalysis. Air
bubbles in the Chitosan and their derivatives have removed by using the sonication. Its
other advantage it regulate the size of nano silver particles by breaking the agglomerates.
It also involved the development and arrangement of cluster of nano metal particles in the

12
existence of Chitosan. It is more dominant than the conventional techniques to prepare the
nano metal sphere because reaction occur very fast and complete in very short time and it
required low temperature. Diverse catalysts have organized, categorized and functional in
the silver catalyzed azide or alkyne cyclo addition as well as in Suzuki cross linking
reactions (Jin et al., 2009).

1.5.2 Composites of nano particles and Chitosan

Various strategies have been created for the cross linking of chitosan,
incorporating substance cross linking with glutaraldehyde and with Mo ( VI )
polyoxyanions. We as of late arranged water dissolvable, blocked di isocyanate as a
cross-connecting specialist for the system development. This cross linker stays
dissolvable in water and consequently takes into consideration simple processability of
chitosan gel arrangements (Zha et al., 2008).
With more prominent command over cross-connect effectiveness, explicitly the
compound cross-connecting of amine gatherings, the physical, mechanical, and organic
properties of chitosan gels can be customized for a given application. Besides, the cross-
connecting science takes into account a clear, covalent connection of amine-containing
biochemical moieties, for example, collagen and fibronectin. Before this class of Chitosan
based hydrogel be able to understand their maximum capacity, a definite comprehension
of the system structures and properties is required (Prabaharan & Mano, 2006).
Composite materials are made out of at least two constituent materials. Significant
consideration has been given to consolidating distinctive utilitarian materials into
composite materials for multifunction applications (Zhang et al., 2015). Moreover, the
qualities of composite materials might be great than those of the entirety of the individual
parts (Pelgrift and Friedman, 2013). Consequently, composite materials have a significant
impact over a wide scope of fields (Esther et al., 2015, Sacco et al., 2015, Zhang et al.,
2015).

1.6 Applications
There is a pressing requirement for the improvement of such materials in our
cutting edge, anti infection dependant society which has emerged because of the
development of medication safe microscopic organisms. The point of this present
examination is to explore the auxiliary changes instigated on a chitosan arrange upon
cross-linkage with a di isosyanate organize alongside the consideration of Ag and Au
Nano particles and to take a gander at the impact cross-linkage may have on the

13
antimicrobial properties of these composite materials. Be increasingly troublesome in
gram-positive microbes (Krishnan et al., 2015).

1.6.1 Chitosan Applications

Chitosan is a biopolymer which is biocompatible and can be corrupted by proteins
in human body, the debasement items are nontoxic. Business CS's are semi-crystalline
polymers and crystallinity assumes a significant job in adsorption effectiveness (Trung et
al., 2006).
CS is a polymer which shows a wide range of antimicrobial action by
authoritative to the contrarily charged bacterial cell divider pursued by connection to the
DNA, hindering its replication. CS is a biopolymer having antibacterial properties (Kurita
et al., 1993; Ting and Shen 2005).

1.6.2 Nano particles Applications

In ongoing decades, nano particles have been explored for different biomedical
applications and are viewed as the "material of the 21st century" as a result of their one of
a kind plans and property mixes contrasted and traditional materials. There is a wide
scope of utilizations of Nano particles, for example, in human wellbeing apparatuses,
modern fields, medicinal applications, biomedical fields and natural examinations (Goyal
et al., 2016).

Essentially, numerous advantages of utilizing nano particles are demonstrated

over other medication conveyance frameworks, for example, improving the solvency of
profoundly hydrophobic medications, giving continued and controlled arrival of typified
tranquilizes, and increasing the steadiness of restorative specialists by concoction or
physical methods and focused on medicines when adjusted with cell-explicit ligands
(Camargo et al., 2009). Among the majority of the nano materials, an assortment of
metallic nano particles have been considered as the first consideration because of their
antibacterial application to human wellbeing. Anti-infection opposition has dependably
been a standout amongst the most huge wellbeing dangers because of consistent
adjustment of organisms to our anti-infection. This issue has risen the consideration for
metallic medications that were utilized to treat contaminations before the time of anti-
infection agents' all out predominance. The most generally utilized representative of
metallic nano particles is silver nano particles as a result of their very successful

14
antibacterial action both in arrangement and in segments and their incredibly huge surface
zone, which gives better contact small scale life forms (Rigo et al., 2013).
The physicochemical properties of nano particles improve the bioavailability of
helpful operators after both fundamental and nearby organization (Jo et al., 2015;
Staquicini et al., 2011) and other hand it can influence cell take up, natural dissemination,
infiltration into organic hindrances, and resultant restorative impacts (Duan et al., 2013).
In this manner, the advancement of Silver nano particles with controlled structures that
are uniform in size, morphology, and usefulness are fundamental for different biomedical
applications (Wong et al., 2009).
Accordingly, the test is to recognize successful, practical, and touchy lead
particles that have cell-focused on particularity and increment the affectability. As of late,
Silver nano particles have been indicated much intrigue in view of their restorative
applications in malignant growth as anticancer specialists, in diagnostics, and in
examining. Contemplated writing, in this audit we concentrated on late advancements in
combination, portrayal, properties, and bio-applications essentially on the antibacterial,
antifungal, antiviral, calming, hostile to malignant growth and against angiogenic
properties of Silver nano particles in a solitary stage (Sriram et al., 2010).
This survey likewise underlines system of anticancer action, helpful
methodologies and the difficulties and impediments of nano particles in malignant growth
treatment. (Gurunathan et al., 2009).

Nano particles exhibit many unique electronic, catalytic, optical, and other
physical and chemical properties that their bulk counterparts do not have (Sun, 2013).
Silver nano particles have been applied in many fields including photonics, micro-
electronics, photocatalysis, lithography, and surface-enhanced Raman spectroscopy (Bhui
et al., 2009; Stamplecoskie et al., 2011). Several types of chemical and physical
technologies, such as chemical reduction, (Khan et al., 2011; Wang Iet al., 2005),
electrochemical reduction, (Roaldan et al., 2013; Zhang et al., 2002). photochemical
reduction (Park et al., 2011), and heat evaporation have been developed for the synthesis
of nano particles.

1.6.3 Catalytical Applications

In the previous decades, nano particles have pulled in boundless consideration as a
result of their unmistakable physicochemical properties and potential applications in the

15
optoelectronics, sensor and biomedical fields (Fereira et al. 2015). Furthermore, they are
viewed as a standout amongst the best impetuses for an assortment of substance responses
(Schrofel et al. 2014). Like other metal nano particles, Ag nano particles are commonly
precarious as a result of their enormous dynamic surface territories, which may result in
self-conglomeration. Along these lines, they are normally bolstered on different lattices
having high surface zones, for example, graphene oxide, metal oxides, silica, etc (Haider
et al. 2016). Other than these inorganic backings, polymers are known as natural backings
and specially utilized due to their simple handling into high surface territory materials.
Because of these extraordinary properties, the inclination to exploit polymers, for
example, polyvinylalcohol (Huang et al. 2012), polypyrrole (Wu et al. 2015), polyvinyl
pyrrolidone (Guo et al. 2013) and poly(methyl methacrylate) (Koo et al. 2016), as
impetus facilitating substrates has expanded.

1.7 Nanocatalysis
Nanocatalysis is a rapidly growing field which involves the use of metal nano
particles as catalysts for a variety of organic and inorganic reactions. Up to now, many
investigations have been done on nano catalysis, but there still remains a challenge for the
preparation of recyclable metal nano catalysts. For this purpose, one currently adopted
method is the immobilization of metal nano particles onto solid supports such as polymer
resins or inorganic spheres, for ready separation of catalysts from the reaction product
mixtures. However, it has to be considered that with solid catalysts suspended within
liquids, the transport of reactants within the liquid to the catalysts as well as the transport
of them within the supports can be rate limiting, because the rate of transport to the
catalyst surfaces will be inversely proportional to the size of the whole catalyst. One
would then suppose that both the activity and the selectivity of suspended catalysts could
benefit much from the use of small-sized supports. The separation of solid catalysts from
the reaction products simply by filtration or centrifugation necessitates, however, that the
catalyst bodies are larger than about 3 μm: Smaller bodies are difficult to be separated
out. It is thus highly necessary to develop new catalysts with a nanometer-sized magnetic
core and a catalytic shell, which respectively favor the high surface area and convenient
magnetic separation. Magnetically recyclable nanometer-sized catalysts allow the
catalysts to be recycled conveniently through applying an external magnetic field. Many
attempts have been made to synthesize this kind of noble-metal deposited magnetic
materials (Fe3O4, γ-Fe2O3, etc.). In this investigation, we choose silver (Ag) and

16
maghemite (γ-Fe2O3) nano particles as the functional components to construct the
magnetically recyclable nanometer-sized catalysts.

1.8 Bimetallic Nano Particles

1.8.1 Introduction
An innovated class of materials for catalytic purposes have been widely reported, that is,
“Bimetallic Nanoparticles”. Comprehensive studies of non-supported bimetallic
nanoparticle dispersions, stabilized by polymers or ligands, started only about 10 years
ago. Many preparative methods have been suggested, and comprehensive
characterizations have been reported on bimetallic nanoparticles, thanks to the fast
improvement of analytical techniques on surface and nanoscale material. In this review,
we throw light on the synthesis, characterizations and applications to catalytic properties
of polymers or ligands-stabilized bimetallic-nanoparticles in dispersion, emphasized our
own work and introducing modern progress of this subject. Bimetallic nanoparticles
formed by two diferent metallic elements are intresting than monometallic nanoparticles
from both the scientific and technology reviews, for the betterment of the catalysis
activities of metallic particle. This is reason that bimetallic can give better catalytic
activities of true one metal catalysts and produce a novel property which may not be got
by mono-metallic catalyst. (Toshima et al., 1998)
Bimetallic nanoparticles (BMNPs) with respect to technoogy accelerate their
monometallic comparison executing to their better electronnic, optecal and catalysis
activities. Besides of usual morrphological manepulations, changes in mollar ratio of
separate constituents give a divers dimensions in exploring propperties of BMNPs. With
respect too mixing patterns of two separate metalls, three major kinds of structure may be
interpret for bimetallic NCs, which is, core-shell, heterostructures, or intermetalic and
aloyed structure. (Meenakumari et al., 2015)

Figure.3. Bimetallic NCs with separate structures a) core/shell b)

Heterostructures c) Intermetalic or aloyed structure.

17
A microimulsion path has been used to obtain bimetallic nanoparticles of Gold–Silver,
Au–Plladium also Au–Platinum. Average size of these aloys lie in range of 10–20 nano
meters. Many metallic nanoparticle (Pt, Bi, Pb, Fe, Cd, Ag, Au, Cu and Ni) have been
prepared using water-in-oil microimulsions.Revers mecelles are water-in-oil
microimulsions containing nanoometrsized water drops that are disperssed in an oil
medium and stabillized by surrfactants.These water droplets are fit as reaction medium
for the preparation of nanoparticlles where size of particle may be controled by changing
mollar ratio of water to surfactants. The microimulsions path has been used in most
reccent past to achieve lot of oxalate and oxide with a high degre of homogineity and
phase purity. (Ahmed et al., 2008 )
A wide range of systems containing Fe/Pt , Fe/Ag , Fe/Cu , Fe/Ni or Fe/Pd have been
investigated.Out of these above said bimetallic systems are the most beneficial and
mostly important to be used.Because of their magnitic propertis, Fe (or Fe/Pd bimetalic)
nanoparticle promote to aglomerate in water fastly to make miccron sized or big
agregates, thus loosing their reactive nature. Many different techneques have been used to
control particles size using disppersing agent such as polymer, surfactant or by coatings
with a protictive layers. Also aproach is to make supported nanoparticle on active carbon,
beeds, particles, poly electrolyte multi layers, or diaphragms as substrate molecules.
( Smuleac et al, 2011)
It was trried to make Pd–Fe bimetallic nanoparticles modified Ni foam (Pd–Fe/Ni)
electrode with a facile way in this review.Meanwhile, CTAB was employed to improve
the surface properties of Pd–Fe/Ni electrode.Their behaviours of Pd–Fe/Ni electrode were
examined for electrochemical reduction of TBBPA (Tetrabromobisphenol A). (Peng et al,
2016).
1.8.2 Synthesis of Bimetallic Nanoparticles
In real practice, preparation of nano size based bimetallic substances emerged in
year as early as 1926. Murray Raney synthesized Raney Ni (Ni-Al alloy) containing large
surface area through controlled leaching of nickel-aluminium alloy (NiAl 3,Ni2Al3
alloy).Therefore on one end, Raney Ni was actually employed in an engineering type of
work as one of very intrested industrial catalysis used for hydrogenations reactions, and
chemistry in synthesized methods did not has attraction of research ones; in an other
view, preparation of effective and having good qualities bimetallic NCs with controlled
method and good size, surface, compositions, and structure has not developed in a

18
comprehensive detailed method in this era. Until the involvement of nano based structural
compounds synthesis techniques in 1990s, immense improvements were gained
manufacturing of nano based morphology of materials. Solution-based chemical synthetic
methods, as common and powerful paths to nano structure based materials, have been
employeed with a great success to manufacture nearly all types of nano based structure
particles and materials, taking with bimetallic NCs. In a general solution-based
preparative method, one must choose a well deined chemical reduction method and
procedure with corresponding reactants to produce required products in first preference;
and then, a rationalized solution system containing the kind of well selected solvents and
surfactants to take out the choosed reaction must be arranged and well designed; at last,
nucleation and proper growth phenomenon of NCs should be safely controlled by fitting
the thermodynamic and kinetic constituents of reaction. For bimetallic nano structure
based crystal preparation, a kind of solution-based synthsized procedure has been made in
practice by lot of groups in present era and present years. (Wang et al , 2011)
Best preparation method of iron-nickle bimetallic nano particles by DO72 resin
was studied. The DO72 resin was acidified by a very fit normal volume of HCl solution
(4–6 mol L-1) and cleeaned with ultrapure water so that pH of separated material was
neutral. Already treated DO72 resin (1 g) was immersed in 20 mL coating solution
(synthesized by mixing 40 mg of Fe and Ni nano structure based particles with lot of
mass ratios from container of FeCl3_6H2O and NiCl2_6H2O in 20 mL distilled water)
since 2 hours to mix Fe3+ and Ni2+ ions into DO72.Chemical DO72 was cleaned with
ultrapureto remover residing ions.Then after doing so, 25 mL of 0.4 mol/L KBH 4 ethyl
alcoholl water solution (ethyl alcohol water ratios 3:7) as a reducing agent was slowly
and with a complete care was added into a breaker which already contained DO72 resins
or slurry (taking Ni2+ and Fe3+ ions) to make Ni-Fe bimetallic nano structure based
particles into DO72 (revealed by Fe-Ni-DO72). Then separated and collected black color
particles were cleaned with ethyl alcohol four times. When used the particles, then before
the use of particles,they were vacuumly dried well. When they are compared, Fe-DO72
(32 mg of Fe within 1 g DO72) and Ni-DO72 (8 mg of Ni within 1 g DO72) were also
synthesized going in the follow of methods which are mentioned earlier. Fe and Ni
components in resin compositions were examined by atomic absorption spectrometr after
heating-assisted digestion of prepared samples.(Zhenming et al ,2015)

19
1.8.3 Catalytic Applications of Bimetallic Nanoparticles
Bimetallic NPCs as earlier said that they have been used in a large number of important
and well defined fields of science and technology, in many fields of science and
engineerin technology. It is early explored previously and compared to monometallic
structure based NCs, bimetallic nano structure based NCs have greater and most
important chracteristics and thus more attractive applications values.
For instance, zero valency 3d metallic elements such as Fe, Co, and Ni are very
much known as magnet properties metallic elements. Also, when we talk about
practically magnetism uses and applications, these nano structure based particles are very
sensitive to oxidation process in atmosphere.
A well known practical and successfully employed example is the representation of
well improved oxygen reduction application on Pt 3 Ni by Stamenkovic and co-
workers.Because Pt3 Ni surface morphology has an unexpected and unusual electronic
structure surface and arrangements of atoms of the surface in near-surface area which
may be greatly enhance number of active places and many active sites for 0xygen
adsorption, Pt3 Ni surface structure is ten folds greater active for oxygen reduction
reaction than in comparison Pt surface and ninghty folds more efficient and active than
present state of art Pt/C catalysts for PEMFCs . (Wang et al, 2011)

Figure.4. Effect of the surface morphology and electronic surface properties of Pt3Ni on
the kinetic studies of oxygen reduction reaction using various nanocatalysts.
It was reported that the electrochemical catalysis properties and activities of
Pt/Cu nanocubes with respect to methyl alcohol oxidation process was employed to be

20
greatly effected as compared to spherically structured Pt/Cu nano structure based crystals
and pure Pt nano structure based crystals with same sizes and almost same shapes.It was
also confirmed that a very good and high quality nano structure based crystalline Pt 3 Co
electrocatalysts with cubic structure and morphology are greatly efficient and active for
methyl alcohol oxidation process than Pt nano based structure cubes.It was seen the
splendid result of electrochemical chracetristics of bimetallic Pd/Pt heterostructured nano
based structure particles have also been employed by many groups. (Fang et al, 2014)
It was synthesized bimetallic Pt/Pd nano structure based particles by local
overgrowth process for Pd on cube shaped Pt seeds and came to know that presence of Pd
on upper layer of Pt cubes should be safe and safer from poisoning in positive scanning
and induce a large decrease in energy of activation for methanoic acid oxidation process
as in comparison with Pt nano structure based cubes.In most rescent days Pd/Pt bimetallic
nano structure based particles having a dense arrangements of Pt branches on a Pd central
part were synthesized by Xia group.A modern hetero base structured Pd/Pt NCs possesing
relatively big surface morphologies and specially active sites toward oxygen reduction
reaction process (2.5 times more activity than Pt/C catalysts and 5 times greater activity
than first generation witout support Pt-black catalyst). So, Pt-based bimetallic NCs
represent more chances in flourishing modern electro based catalysts for PEMFCs. (Yang
et al, 2011)
Bimetallic NPCs containing alloy structures are also found to be effective half
homogeneous catalysts and with good catalytic properties. Li and co-workers made a
hollow Pd/Fe nano structure based spheres and they compared the prepared catalyst and
their catalytic activities containing hollow Pd nano structure based spheres.it is of because
that hollow chamber structure and the promotional effect of Fe-dopants may be given
greater Pd activity and active portions for reactive species, bimetallic Pd/Fe nano
structure based catalysts revealed much more better catalytic activity during aqueous
Sonogashira-, Heck-, and Ullmann-type coupling reactions of aromatic halide.Also it is
moreover, bimetallic catalysts may be easily extracted from reaction solution by a magnet
and it may be reused five times without great decrease in both the activity and the
selectivity. ( Wang et al,2011)

21
Figure.5.Comparative behaviour of catalytic activity and characteristics of 3.9 nm Pd/Ni
bimetallic nano structure based particles and 3.5 nm Pd nano structure based particles.

Aims and Objectives

• To prepare bimetallic Fe/Ni nanoparticles supported at cross linked chitosan for
organic dyes degradation.
• To develop a robust photo catalytic system for industrial water remediation.

Novelty and Importance of the Project

• To the best of our knowledge, the proposed catalyst is novel in its structure and
nature.
• This study will help to deal with organic water pollutants.

22
CHAPTER 2
REVIEW OF LITERATURE
Yang (2004) has studied that the polysaccharides act as reducing agent or
stabilizing agents, due to this reason it is used for synthesis of nano particles like silver
and gold. Positively charged chitosan-stabilized obtained from gold nano particles and
negatively charged heparin-stabilized obtained from silver. These nano particles were
characterized with UV-vis spectroscopy and transmission electron microscopy.The
outcomes showed the development of gold and silver nano particles inside the nano
scopic polysaccharide formats. In addition, the morphology and size dissemination of
arranged gold and silver nano particles shifted with the grouping of both the
polysaccharides and the antecedent metal salts.
Jiang et al. (2005) have explored the synergist properties of bismuth nano
particles upheld on silica circles. The procedure to help bismuth particles on silica circles
viably keeps away from flocculation of nano sized colloidal metal particles amid a
synergist procedure in the arrangement, which enables one to do the effective reactant
decrease of colors. The impacts of electrolytes and surfactants on the synergist properties
of bismuth particles on silica have been explored. It is discovered that the nearness of
surfactants discourages the synergist action of the bismuth particles somewhat by
restraining the adsorption of reactants onto the outside of the particles. Electrolytes either
increment the movement rate of reactants in the arrangement bringing about an expansion
in the synergist response rate or restrain the adsorption of reactants onto the outside of the
bismuth particles prompting a misfortune in the action of the metal particles.
Nandi & dawn (2006) have studied a novel nano biocomposite of nickel and
poly(o-anisidine) (POA)/DNA hybrid has been synthesized by addition the DNA solution
to an aqueous solution of POA (emeraldine base, EB) and NiNO3 mixture. The blend was
aged for 10 days and was freeze-dried to form the hybrid nano composite (weight fraction
of DNA) 0.75). FESEM images show a fibrillar network structure of the biomolecular
hybrid with nickel nano particles on its surface. The TEM images also corroborates nickel
nano particle arrangement in the biomolecular hybrid, and the denser amount of nano
particles in the TEM micrograph as compared to that in the SEM micrograph shows that
the nano particles are there inside the fibrils in larger proportion. The dc conductivity
value of the nano biocomposite is approximately the equivalent as that of the pure POA
DNA hybrid at the similar composition, but the I-V characteristic curve of the nano

23
biocomposite is somewhat dissimilar showing an insulating area on low applied voltage.
At higher applied voltage, it shows a semiconducting property characterizing the huge
band space semiconducting behaviour of the nano biocomposite.
Murugadoss et al. (2007) have studied in this paper that report on the synergist
action of another metal nano particle– polymer composite comprising of nano particles
(Nano particles) and ecologically cordial (green) chitosan. The polymer (chitosan) not
just went about as the diminishing specialist for the metal particles yet in addition
balanced out the item Nano particles by securing them. Most of the particles created
along these lines had sizes under 5 nm. The reactant movement of the composite was
explored photometrically by checking the decrease of 4-nitrophenol within the sight of
overabundance NaBH4 in water, under both heterogeneous and smaller scale
heterogeneous conditions. The response was first request as for the grouping of 4-
nitrophenol. We likewise seen that the evident rate consistent for the response was
directly reliant on the measure of Silver nano particles present in the composite. In
addition, the turn over recurrence of the impetus was observed to be (1.6 ± 0.4) × 10 −4 s−1.
When the response was completed under heterogeneous conditions. The Silver nano
particles in the composite held their synergist exercises even in the wake of utilizing them
for ten cycles. Our perceptions likewise recommend that the reactant proficiency under
miniaturized scale heterogeneous conditions is a lot higher than under heterogeneous
conditions. Therefore the composite we have speaks to a perfect instance of an earth
amicable and stable impetus, which works under heterogeneous just as miniaturized scale
heterogeneous conditions with the upside of nano scopic particles as the impetus.
Alpa et al. (2007) have investigated that the permeable silica nanofibers
containing reactant silver nano particles have been orchestrated by another strategy that
joins sol−gel science and electrospinning method. Tetraethyl orthosilicate, poly[3-
(trimethoxysily)propyl methacrylate] and silver nitrate have utilized as antecedents for the
creation of silica- poly[3-(trimethoxysily)propyl methacrylate] half and half filaments
containing silver nitrate. Calcination of the mixture filaments at high temperatures results
in permeable silica strands in light of warm deterioration of poly[3-(trimethoxysily)propyl
methacrylate] polymer and in change of silver nitrate to silver nano particles. The shade
of the fiber mats changed from white to dull brilliant yellow because of the surface
plasma reverberation of the silver nano particles implanted in the filaments. The size and
thickness of the silver particles in the silica filaments could be tuned by changing the

24
extent of the strands, measure of silver nitrate presented, and the warm treatment
conditions. The silica strands containing silver particles were described with ecological
examining electron microscopy, transmission electron microscopy, X-beam diffraction,
UV−vis spectroscopy, and thermo gravimetric investigation. The synergist action of the
silver containing fiber mats was surveyed by a decrease response of methylene blue color.
Sanpui et al. (2008) have researched that the escherichia coli communicating
recombinant green fluorescent protein was utilized to test the bactericidal adequacy of a
recently blended chitosan– silver nano particle composite. The composite was found to
have fundamentally higher antimicrobial movement than its segments at their particular
focuses. The one-pot amalgamation technique prompted the development of little Ag
nano particles appended to the polymer which can be scattered in media of pH ≤ 6.2. The
nearness of a little rate (2.00% w/w) of metal nano particles in the composite was
sufficient to essentially upgrade inactivation of E. Coli as contrasted and unaltered
chitosan. Fluorescence spectroscopy demonstrated that bacterial development halted
following presentation of E. coli to the composite, with arrival of cell green fluorescent
protein into the medium at a quicker rate than with chitosan. Fluorescence confocal laser
checking and examining electron microscopy demonstrated connection of the
microorganisms to the composite and their ensuing discontinuity. Local protein gel
electrophoresis tests demonstrated no impact of the composite on bacterial proteins.
Routh et al. (2009) have studied the synthesis of bimolecular hybrids of a
conducting polymer [poly(o-anisidine) (POA)] and RNA atthe three different
compositions by mixing aqueous solutions of diethyl, 2-hydroxy ethyl, ammonium salt of
RNA (type IX from Torula Yeast) and POA (ES, emeraldine salt; doping level
[Cl]/[N]=0.49). They found that a slowincrease of pH up to 25 h of aging occurs in the
mixture till it levels up. The TEM micrograph shows structure in all the hybrid
compositions (POA: RNA=1:2, 1:1, 2:1, by weight). In the complexes three types of
molecular interactions, for example (i) electrostatic, (ii) H-bonding and (iii) π–π
interactions, are evident from the FTIR spectroscopy. The CD spectrum shows a small
distortion of A-RNA conformation towards its B form during the hybrid formation. Time
and temperature dependent UV-vis spectral studies indicate a slow red shift of the π-band
to polar on band transition peak (λmax) for the uncoiling of the POA (P) chain on the RNA
(R) surface. The repulsive interaction between the radical cations of POA (ES) absorbed
on the RNA surface is attributed to the conformational change causing the uncoiling of

25
POA chain. UV-vis spectral study indicates that the uncoiling and attachment of POA on
RNA surface is much faster than that on DNA (D). In POA-RNA-DNA (PRD) hybrid
solutions slower red shift of λmax indicates more disordered array of the phosphate groups
as compares to PR and PD systems. The conductivity values of the PR hybrids (10-6
S/cm−1) are three orders higher than that of RNA, rendering the PR hybrids to be helpful
for fabricating good biosensors. In the PRD hybrids conductivity decreases by two orders
than those of PR and PD hybrids suggesting a disorder arrangement of POA chains in the
PRD hybrids. The I-V characteristic curves of the PR and PRD hybrids show a
semiconducting nature of the hybrids.
Chtchigrovsky et al. (2009) have examined perfect and green, Copper(I) edifices
of phenanthroline based ligands secured on the chitosan polymer are great impetuses for
the "click" cycloaddition of azides with terminal alkynes (the checking electron
microscopy picture demonstrates the permeable structure of the impetus). These
heterogeneous synergist frameworks don't require a base or decreasing operator and work
in liquor or water.
Wei et al. (2009) have investigated the silver nitrate reduced by the nontoxic and
biodegradable chitosan which synthesized the silver nano particles. Silver nano particles
have extremely effective antibacterial activity toward both Gram-positive and Gram-
negative bacteria, analogous with the extremely active ancestor silver salts. Silver
impregnated chitosan films were framed from the beginning materials made out of silver
nitrate and chitosan by means of thermal treatment. Contrasted and unadulterated chitosan
films, chitosan films with silver indicated both quick and enduring antibacterial adequacy
against Escherichia coli. The silver antibacterial materials arranged in our present
framework are promising possibility for a wide scope of biomedical and general
applications.

Shin et al. (2009) have researched a novel one-advance manufacture of gold

stored Fe2O3 nano particles and their application for the synergist decrease of 4-
nitrophenol to 4-aminophenol by sodium boron hydride. To store gold onto them, Fe 2O3
particles were scattered in a response blend comprising of ethanolic AgNO 3 and
butylamine, and after that the response blend was brooded and shaken for 35 min at 40-50
°C. With this straightforward and sans surfactant manufacture of gold saved Fe 2O3 nano
particles, we can evade tainting in the last item, which makes them reasonable for further

26
synergist applications. Since the attractive particles are promptly recuperated from the
arrangement stage without centrifugation and additionally separating, the gold saved
Fe2O3 nano particles arranged in this work have been abused as strong stage impetuses for
the decrease of 4-nitrophenol within the sight of sodium boron hydride. Toward the finish
of the response, the gold saved Fe 2O3 impetus particles still stay dynamic. They would
thus be able to be isolated from the item, 4-aminophenol, essentially utilizing a neodium
magnet and can be reused various occasions.
Zhuang et al., (2010) have clarified in this paper tends to another technique for
making nickel nano particles containing Chitosan / gelatin nano fibers by means of
electro spinning. In the present paper, the nickel nano particles sizes extending from 2 nm
to 6 nm were incorporated at room temperature utilizing microcrystalline chitosan as the
diminishing operator and stabilizer pursued by dissolving the nickel nano particles-
chitosan composites in gelatin containing acidic corrosive arrangement and after that
electro spinning the readied arrangement into nickel nano particles containing Chitosan /
gelatin nano fibers. The structure of the resultant nano fibers was analyzed with the guide
of filtering electron microscopy (SEM), transmission electron microscopy (TEM), and X-
beam photoelectron spectroscopy (XPS). The estimation results showed that the
nanofibers having the width scope of 200– 390 nm were clearly smooth and the nickel
nano particles with the size appropriation from 3 nm to 9 nm were effectively fused into
the electro spun nano fibers.
Zhang et al.(2010) have studied the nano composite of poly (o-
methoxyananiline) /carbon nenotubes and founded that by addition of CNTs to
polyaniline an increase in electrochemial polymerization of the monomer and thermal
satbility. This composite was prepaired by the method known as electrochemical co-
deposition. Composite was tested by the SEM and TEM, this shows that CNTs are helpful
to increase the conductivity and capastence of the ploymer.
Banerjee et al. (2010) have studied that cooperative energy in antimicrobial
movement of a Chitosan silver nano particle composite within the sight of sub-atomic
iodine. Green fluorescent protein communicating recombinant Escherichia coli
microorganisms have been utilized to test the adequacy and build up the system of
activity. Trial proof demonstrate fundamentally high bactericidal action of the nano
composite within the sight of iodine than either because of the composite, chitosan,
iodine as it were. Transmission electron microscopy estimations uncovered connection of

27
microorganisms to the composite. What's more, stream cytometry results bolstered clear
event of cell divider harm of the microscopic organisms treated with the composite within
the sight of iodine. Further, the nano composite and iodine blend was found to apply
receptive oxygen species created oxidative worry in the cytoplasm of bacterial cells,
prompting cell passing. Illustration of the instrument of collaboration because of three
potential antibacterial parts recommended that on the outside of Silver nano particles sub-
atomic iodine conceivably produced iodine molecule along these lines contributing
toward free radical initiated oxidative pressure, though chitosan and Silver nano particles
encouraged the procedure of cell slaughtering and in this way by and large upgraded the
strength of antimicrobial impact at the most reduced groupings of individual segments.
Wang et al.(2011) have investigated the HCl doped poly(o-methoxyananiline)
POA and undoped POA by different techniques like FTIR ,ESR and SEM afetr which
found that doped POA has better control over the morphology than the undoped POA.
The conductivity order is POA- HCl > Undoped POA, this shows doped POA’s
morphology is significant by the doped material.
Aly (2011) have studied the effect of nickel and iron nano particles as a filler on
the free volume properties Of Natural Rubber by using Positron Annihilation Lifetime
Spectroscopy. The PAL measurements showed that free volume properties are strongly
affected by the quantity and type of filler. Particularly, the free volume fraction
dramatically decreases by increasing the filler content. Besides, the addition of nano
particles created new positron trapping sites at filler-rubber interfaces. Moreover,
interactions were made between the free volume hole size (V h) and each of the
mechanical and electrical properties. A negative interaction was observed between V h and
hardness in the Ni rubber composites and a positive counterpart was observed in the case
of Fe rubber composites. Finally, the polarity of the fillers, being higher than that of the
rubber itself, leads to an increase in electrical parameters and an inhibition of o-Ps
formation. These results shows that the analyzed composites are considered to be
insulating materials as their conductivity values are in the order of an insulator range.
Sharma et al., (2012) have played out another and basic ecofriendly strategy for
the combination of cobalt nano particles utilizing a characteristic biopolymer, sodium
alginate as both lessening and balancing out specialist is accounted for. The combined
nano particles were described utilizing UV-Vis spectroscopy, transmission electron
microscopy (TEM) and chose zone electron diffraction design (SAED). The alginate

28
topped Nano particles (Alg–cobalt nano particles) were observed to be antibacterial. The
Cobalt nano particles were mixed with shifting measures of chitosan to frame
polyelectrolyte complex that was cast into stable movies. The movies were portrayed by
field discharge filtering electron microscopy (FESEM), optical microscopy, Fourier
transmission infrared spectroscopy (FTIR) and X-beam diffraction (XRD). The water
take-up and mechanical properties of the movies were additionally contemplated. The
mixed film showed brilliant antibacterial movement against both Gram negative and
Gram positive microscopic organisms with greater action against Gram positive
microorganisms. In this way, the created movies can possibly be utilized for different
antibacterial applications in biotechnology and biomedical fields.
Krishna and Ramasubba, (2012) have contemplated that hte new chitosan nano
composites doped with silver nano particles were incorporated by a basic technique. The
chitosan particles were set up by desolvation pursued by cross linking with poly (ethylene
glycol-di-aldehyde), this was set up with poly(ethylene glycol) within the sight of a silver
nitrate arrangement. The created nano composites were portrayed utilizing UV–
unmistakable, FTIR, XRD, TGA, SEM and TEM to comprehend their physic substance
properties. These nano composites were appeared to have hostile to bacterial movement
towards Escherichia coli.
Sivrikaya et al., (2012) examined that manganese ferrite (MFN) nanoparticles and
proved to be the best for photocatalytic degradation of dye from colored waste water. The
techniques like UV-Visible spectrometer and ion chromatography were used to study the
degradation of dyes by using above prepared manganese ferrite nanoparticles.During the
degradation of dyes by using MFN, the effect like MFN dosage, salt, initial concentration
was also studied some dominant aliphatic intermediate like formate, onions were also
detected. During the process of degradation, some mineralization products were also
detected. The mineralization products are mostly in organ ions like nitrate ions, sulphate
ions. The obtained in the study shows that MFN act on magnetic catalyst and can degrade
the reactive dyes present in the waste water very effective
Xia et al., (2013) have considered uniform ultrathin titania / chitosan composite
movies were covered on the cellulose micro fibril packs of normal cellulose substance
(regular business channel paper) by a layer-by-layer self-get together procedure.
Depending on the solid chelating capacity of chitosan for metal particles, nickel particles
were liberally adsorbed on the titania/ Chitosan composite film covered cellulose

29
substance and were from that point in situ diminished to nickel nano particles (silver nano
particles) under UV light. In that capacity, cross breed cellulose / titania/ chitosan/ silver
nano particles composite materials were gotten, which highlight a similar progressively
sinewy structure as the underlying cellulose substance. In the mean time, the half and half
filaments in the composite materials show a link like core– shell structure, showing a
cellulose micro fibril pack center just as a nano meter-thick titania/ chitosan/ nickel nano
particles composite film shell with nano particles (5-22 nm in breadth) all around
circulated. The activities of the titania/ chitosan/ silver nano particles composite film
covered cellulose materials were assessed against both Gram-positive and Gram-negative
microscopic organisms, and the materials accordingly sanitized practically all the
immunized microbes due to the inherent biocidal impact of titania structure, decidedly
charged chitosan segment and high stacking substance of nickel nano particles with little
sizes, appearing antibacterial exercises.
Lian et al., (2013) have examined the novel engraved electrochemical sensor for
neomycin acknowledgment was created dependent on Chitosan cobalt nano particles
/grapheme multi walled carbon nano tubes composites enlivened gold cathode.
Molecularly engraved polymers were blended by electro polymerization utilizing
neomycin as the format, and pyrrole as the monomer. The system of the manufacture
procedure and various components influencing the movement of the engraved sensor
have been talked about and advanced. The portrayal of engraved sensor has been done by
checking electron magnifying lens and Fourier transformer infrared spectroscopy (FTIR).
The execution of the proposed engraved sensor has been examined utilizing cyclic
voltammetry (CV) and amperometry. Under the enhanced conditions, the direct scope of
the sensor was from 8×10-8 mol/L to 9×10-5 mol/L, with the point of confinement of
identification of 7.54×10-8 mol/L (S/N=2.5). The film showed high restricting proclivity
and selectivity towards the format neomycin, just as great reproducibility and strength.
Besides, the proposed sensor was connected to decide the neomycin in milk and nectar
tests dependent on its great reproducibility and security, and the satisfactory recuperation
inferred its plausibility for viable application.
Akmaz et al. (2013) have pointed out the antibacterial properties and portrayal of
Chitosan silver nano particle composite materials. Chitosan silver nano particle composite
material was blended by adding AgNO3 and NaOH answers for Chitosan arrangement at
90-95∘ C. Various fixations (0.01 M, 0.03 M and 0.05 M) of AgNO 3 were utilized for

30
combination. Chitosan-silver nano particle composite materials were described by
Transmission electron microscopy (TEM), X-beam diffraction (XRD), bright (UV)
spectrophotometer, and Fourier transformer infrared (FTIR) spectrometer methods.
Escherichia coli, Acinetobacter baumannii, Staphylococcus aureus, Enterococcus
faecalis, Pseudomonas aeruginosa, and Streptococcus pneumoniae were utilized to test
the bactericidal proficiency of integrated Chitosan silver nano particle composite
materials. The natural action was dictated by the base bacterial focus of the materials.
Antibacterial impact of chitosan silver nano particle materials was expanded by
expanding silver measured the composite materials. The nearness of little measure of
metal nano particles in the composite was sufficient to altogether improve antibacterial
action as contrasted and unadulterated chitosan.
Xia et al. (2013) have set up a research and concluded that when polyamide 6
fiber are treated with camphor sulfonic acid, poly aniline (skin)/polyamide 6 (core)
composite fiber was prepared with the aid of a superficial in situ oxidation polymerization
method. The adsorption property of the prepared composite fiber was also analyzed by
choosing methyl orange as a standard dye. In this research, camphor sulfonic acid played
a fundamental role for the preparation of that skin/ core structure. The spread particles of
poly aniline skin on the composite fiber could be effectively settled by controlling the
experimental conditions. It was also observed that the adsorption efficiency on the
composite fiber improved with the rise in experimental temperature and the
adsorption/desorption kinetics and isotherms followed the pseudo-second-order and
Langmuir models, correspondingly. The high adsorption ability, suitable process and
regeneration procedure would make the composite fiberan valuable candidate for the
removal of organic dye from the waste water.
Mori et al., (2013) have assessed the silver nano particle /chitosan composites
with antiviral action against H1N1 flu an infection were readied. The Ag NP/Ch
composites were acquired as yellow or dark colored floc-like powders following response
at room temperature in fluid medium. Silver nano particles (2.5, 5.5 and 13 nm normal
widths) were installed into the chitosan grid without total or size shift. The antiviral
action of the silver nano particles chitosan composites was assessed by contrasting the
TCID50 proportion of viral suspensions treated with the composites to untreated
suspensions. For all sizes of Silver nano particles tried, antiviral movement against H1N1
flu an infection expanded as the centralization of Silver nano particles expanded, chitosan

31
alone showed no antiviral action. Estimate reliance of the Silver nano particles on
antiviral movement was additionally watched: antiviral action was commonly more
grounded with littler Silver nano particles in the composites. These outcomes demonstrate
that silver nano particles chitosan composites collaborating with infections show antiviral
movement.
Moura et al. (2013) have introduced a method to prepare composites based on
polyaniline and styrene-divinylbenzeneco polymers by in situ polymerization of aniline
using benzoyl peroxide as oxidant agent. The composites were obtained from copolymers
with two degrees of porosities which have higher and lower surface areas. Emeraldine
Pani was synthesized by using the hydrochloric acid as dopant. One cycle or four cycles
of aniline polymerization were performed. The copolymers and their corresponding
composites characterizations were performed by infrared spectroscopy, thermo
gravimetric analysis physical nitrogen adsorption-desorption measurements, morphology
analysis, elemental analysis and determination of Bronsted acid sites. Pani was distributed
overall porous SD copolymer producing composites with high surface area. Then, they
were evaluated as catalysts for esterification reaction of a fat acid. It was concluded that
composites synthesized with four cycles of in situ polymerization presented best catalytic
activity as compared to cycle composites.
Puda et al. (2013) have found that the applications of polyaniline (PANI) has
motivated investigators to study its mechanical properties, the thermo stability of its
conductivity, its processibility and also its use in polymer composites or blends with
commonly known polymers. Several methods to produce composites /blends containing
PANI have been developed as a result of this, which allows the preparation of a broad
spectrum of such materials. In this study the generalized approaches for the preparation of
such materials are reviewed. Especially, we consider two unique groups of synthetic
methods depending upon aniline polymerization either in the presence of or inside a
matrix polymer or the mixing of the already prepared PANI with a matrix polymer.
Different aspects of these methods are analyzed, emphasizing features that determine the
properties of the resulting composites/blends.
Yuan et al., (2014) prepared C u2 O nanoparticles on Biv O 4 under the mild
condition C u2 O/Biv O4 nano composites. In natural media, the optimized nano composite
shows high catalytic activity. The reduction of Cr (VI) and oxidation of dyes takes place
Cr. (VI) absorbed over the surface of C u2 O/Biv O 4 was reduced to Cr (III) completely.

32
This was confirmed by XPS under the visible light irradiation, the degradation of mixture
of dyes also the degradation of cationic and anionic dyes takes place separately. The
C u2 O/Biv O 4 nano compositesynthesized by this method is proved as promising photo
catalyst for the treatment of water that contains the organic dyes. These photo catalysts
also used to treat the water that have Cr (VI) ions. The photo catalyst activity of these
nano composites also described the Hetro junctions between C u2 OandBiv O4 .
Landage et al. (2014) have studied the main purpose of nanotechnology to
enhance the functioning of the material this review all about the antimicrobial activity of
the zinc oxide nano particles. Various synthesis perspective used such as (physical,
biological and chemical) but chemical perspective is the most persistent method for the
manufacturing of the zinc oxide nano particles as stable colloidal dispersion in polar
compounds. Nano particles which have antimicrobial activity are also produced by
different reducing agents. Metallic zinc oxide shows the potential of anti microbial agent.
Zinc oxide nano particles also play role to develop resistance against antibiotics in
pathogenic bacteria.
Khan et al. (2014) have observed the novel class of hybrid poly(2-anisidine)
Sn(II) silicate carbon nanotubes composite which was prepared with the help of solution
gel technique. Scanning Electron micrograph, X-ray Diffraction, Fourier Transform
Infrared, Ultraviolet-Visible Spectroscopy and simultaneous Thermo Gravimetric
Analysis were used to take the physic chemical characterization. To recognize the ion-
exchange capabilities the ion-exchange studies were also processed. Ion selective
membrane was synthesized and applied it as an indicator electrode in potentiometric
titrations due to selectivity of the nano composite.
Oueiny et al. (2014) have studied an exhaustiveidea of the present status of
research in carbon nanotube/polyaniline (CNT/PANI) composites. CNT/PANI was
prepared with the help of electrochemical and chemical processing. The key challenge is
to attain process able CNT/PANIin the emeraldine salt (ES) form composites when
chemical methods are used.
Nguyen and Liu (2014) have researched that the poly(vinyl liquor)/Chitosan
(PVA/CTS) hydrogel slim movies of around 4 μm in thickness were integrated by means
of bright (UV) light utilizing acrylic corrosive (AA) monomer as the cross linker, without
the guide of an extra photo initiator. The readied hydro gels with a high cross linking
thickness and a thick between permeable structure were used both as nano reactors and

33
the immobilizing network for the arrangement of nickel nano particles (Nickel nano
particles). The nano particles were framed through in situ decrease of nickel aceate by
utilizing the nontoxic biochemical glucose and the quickening agent sodium hydroxide in
the earth considerate vehicle of water. The composite movies were portrayed utilizing a
few investigation devices, including UV– visible spectroscopy (UV/vis), Fourier
transmission infrared spectroscopy (FTIR), X-beam diffraction (XRD), transmission
electron microscopy (TEM), checking electron microscopy (SEM) and mechanical
testing. The outcomes affirmed that exceedingly steady and consistently appropriated
Nickel nano particles were gotten over the whole hydrogel systems. The pliable test
uncovered that the powerful fuse of Nickel nano particles inside the hydrogel systems
rendered the composite movies progressively versatile. Due to its between permeable
structure, all around circulated Nickel nano particles, and great mechanical and
biocompatible properties, just as the green blend strategy, the AgNP-stacked PVA/CTS
hydrogel dainty film could be used as a biomaterial in medicinal applications.
Suriati et al. (2014) have studied that the chemical reduction method are used to
prepare the well dispersed the silver nano particles. Trisodium citrate and ascorbic acid
are used for the reduction of the silver nitrate for obtaining the particles. The resulting of
Silver nano particles was examine by the X-Ray diffraction analysis, transmission
electron microscopy (TEM), and scanning electron microscopy (SEM). Average ranged
size of the Silver nano particles about the 40-90 nm. Here we study about the effects of
the trisodium citrate and ascorbic acid on the physiology of the silver particles. Both are
work opposite to each other, increasing the concentration of the trisodium citrate increase
the size of the particles while increasing the concentration of the ascorbic acid increase
the size of the particles. Quasi spherical shaped of the particles become uniform by
increasing the concentration of the trisodium citrate, while by increasing the ascorbic acid
shaped of the particles convert from quasi spherical to polygonal.
Yang et al. (2014) have studied the Ni films having thickness of 50-250 nm were
sputter deposited on HCl-doped polyanisidine substrates at 290 K, forming the Ni/ HCl-
doped polyanisidine composites. All the nickel films grow by columnar grains. By
increasing film thickness the grain size increases. A temperature dependence of the
resistance within 5-300 K shows that all the Ni/ HCl-doped polyanisidine composites
exhibit a metal-semiconductor transition. By decreasing film thickness the transition
temperature lowers. At a temperature below the transition temperature the composite

34
shows a metallic conduction behaviour and a semiconducting behaviour at temperatures
above the transition temperature. By increasing the film thickness in the temperature
range of the metallic conduction the temperature coefficient of resistance increases. By
decreasing the film thickness in the temperature range of the semiconducting behaviour a
decrease of the resistance with temperature becomes more significant.
Wang et al. (2015) have portrayed Zinc oxide nano particles have been utilized in
different fields, and a few amalgamation forms have been created. The soundness and
scattering of the incorporated nano particles is indispensable. The present article depicts a
novel methodology for one-advance blend of silver nano particles-inserted chitosan
particles. The proposed methodology was connected to at the same time get and balance
out silver nano particles in a chitosan polymer grid in-situ. The distance across of the
combined chitosan composite particles went from 1.7 mm to 2.5 mm, and the installed
silver nano particles were estimated to be 15±3.3 nm. Further, the examinations of bright
obvious spectroscopy, vitality dispersive spectroscopy, and X-beam diffraction were
utilized to describe the readied composites. The outcomes demonstrate that the silver
nano particles were appropriated over the surface and inside of the chitosan circles. The
created circles had macroporous property, and could be utilized for some applications, for
example, fungicidal specialists later on.
Annur et al. (2015) have talked about the chitosan nano fibers have been electro
spun with poly(ethylene oxide) and silver nitrate, as a coelectro spinning polymer and
silver nano particle antecedent, separately. The normal breadth of the as-spun chitosan
nanofibers with up to 2 wt % silver nitrate stacking was roughly 130 nm, and there was
no proof of dot arrangement or polymer agglomeration. Argon plasma was then
connected for surface carving and amalgamation of silver nano particles by means of
antecedent disintegration. Plasma surface barrage initiated nano particle development
principally on the chitosan nanofiber surfaces, and the moderate surface plasma
scratching additionally supported greatest presentation of silver nano particles. UV– vis
spectra demonstrated the surface plasmon reverberation mark of silver nano particles. The
surface-immobilized nano particles were envisioned by TEM and were found to have
normal molecule breadths as little as 1.5 nm. Surface investigation by infrared
spectroscopy and X-beam photoelectron spectroscopy affirmed the communications
between the silver nano particles and chitosan atoms, just as the impact of plasma
treatment on the nanofiber surfaces. At last, a microbes hindrance think about uncovered

35
that the antibacterial movement of the electrospun chitosan nanofibers correspondingly
expanded with the plasma-incorporated silver nano particles.
Alshehria et al. (2016) have explained the impetus for the decrease of 4-
nitrophenol (4-NP) was readied utilizing carboxyl gathering functionalized multiwalled
carbon nanotubes, polymer lattice, and bismuth nano particles . The Bismuth nano
particles were set up by the decrease of bismuth nitrate by trisodium citrate in the
MWCNTs-polymer nano composite; the span of the orchestrated Bismuth nano particles
was observed to be 3 nm (normal distance across). The integrated nano composites were
described utilizing a few systematic strategies. Bismuth multiwalled carbon nanotubes
polymer composite within the sight of sodium borohydride in fluid arrangement is a
powerful impetus for the decrease of 4-nitrophenol. The clear energy of decrease has a
pseudo-first-request energy, and the rate steady and synergist movement parameter were
observed to be individually 8.99×10−3 s−1and 12.14 s−1 g−1. The multiwalled carbon
nanotubes polymer nano composite renders steadiness to Bismuth nano particles against
the earth and the response medium, which implies that the bismuth multiwalled carbon
nanotubes-polymer composite can be re-utilized for some reactant cycles.
Zhang et al. (2016) have investigated that the main purpose of the study of the
nanotechnology to identify, treat and to develop resistance against different diseases.
Silver nano particles play important role in the biomedical application especially in
nanomedicine. Moreover different noble metal are used for different purposes but Silver
nano particles are used to diagnose the cancer treatment. In this review we will discussed
about how the manufacturing of the Silver nano particles were obtained by using physical,
biological, and chemical method. Moreover applications of Silver nano particles such as
antifungal, antibacterial, antiinflamatory, anti angiogenic and anti cancer agents were also
discussed. In addition we discussed Silver nano particles are also involved in cancer
therapy.
Anupama & Madhumitha (2016) have presented the work portrays the eco-
accommodating union of silver nano particles utilizing dried natural product concentrate
of C. carandas. The orchestrated Silver nano particles were described by UV-Vis
investigation, FTIR, XRD, SEM, and TEM with histogram. The TEM picture of the
encouraged strong period of Silver nano particles appeared in the scope of 25 ± 3 nm in
size. Further, the reactant corruption on gem violet color within the sight of C. carandas
dried natural product concentrate and Silver nano particles was additionally examined.

36
 Szczepanowicz et al. (2016) have investigated theat silver colloid particles is also
synthesized by the controlled sized particles eith the help of reduction of [(AgNH 3)2]+
dodecyle sulphate (SDS) miceller solution. This solution has different reducing agent
(Hydrazine, formaline and ascorbic acid) which produced nano particles about the size of
the 20nm. By using dynamic light scattering (DLS), scanning electron microscope
(SEM), and UV/visible absorption spectrophotometer we will determined the size, size
gradation and morphology of the particles easily. In this review effects of the increasing
the reducing agents on the silver particles, fraction of metallic silver and their
antimicrobial properties is discussed. The size of the silver nano particles decreased
below the 20nm by using reduction of the silver complex. For developing activity against
the gram positive and gram negative bacteria using high resistance multi strains such as
staphylococcus epidemics, staphylococcus aureus and pseudomonas aeruginosa.
Khan et al., (2016) have contemplated a basic and ecological cordial strategy was
produced for the combination of silver nano particles utilizing Dimocarpus longan seed
separate as a wellspring of diminishing and balancing out operator. The presence of a
surface plasmon reverberation crest at 440nm affirmed the union of silver nano particles
(UV-visible spectroscopy). The biosynthesized silver nano particles were face focused
cubic structures (XRD) with an inexact molecule size of 40nm (TEM). Improvement
consider uncovered that 10mL of plant extricate (3 mM AgNO3) at 180 min of brooding
came about the ideal item blend. Poly phenolic mixes were significantly associated with
the decrease of silver particles into Ag-Nano particles (FT-IR). The synergist exercises of
silver nano particles were surveyed against the photograph reactant debasement of
methylene blue and chemo synergist decrease of 4-nitrophenol to 4-aminophenol. The
outcomes showed that the readied Ag-Nano particles have solid chemo reactant action
with a total decrease of 4-NP to 4-amino phenol inside 20min. So also, silver nano
particles showed higher photograph reactant action (K=0.11) when contrasted with
business silver nano particles (K=0.004). Furthermore, the silver nano particles displayed
a promising cell reinforcement action in searching DPPH radicals. The discoveries of this
examination reason that the biosynthesized Ag-Nano particles are promising operator
having solid reactant and decreasing properties.
Ryan et al. (2017) have investigated the all about the cross linked chitostan
composite by the application of the antimicrobial activity. These are obtained by the
crosslinked chitostan and network of the siloxane & also obtained by the zinc and gold

37
bimetallic nano particles. The main purpose to observed the effect of chitosan siloxane
and chitosan itself on the microbial activity. The addition of the metal with chitosan
siloxane give good result on the microbial activity than only addition of chitosan. The
composite are manufactured in hydrogel by inserting the metal nano particles in the
network of cross linked chitostan. Different techniques are used to determine the
structural properties of composite and tensil strength. The addition of the Nano particles
have no affect on the mechanical strength of the composite and antimicrobial activity.
Overall slixane is significant to the antimicrobial activity and mechanical strength and
giving rise different results.
Lima et al. (2017) have researched silver nano particles have been examined as an
option for treatment of microbial diseases and leishmaniasis, without advancing
enlistment of microbial or parasite opposition. In this examination, chitosan-based silver
nano particles were incorporated from silver nitrate (AgNO3), sodium borohydride as a
diminishing specialist, and the biopolymer chitosan as a topping operator. The chitosan-
based silver nano particles were portrayed by ultraviolet– obvious, Fourier change
infrared, unique light dissipating, zeta potential, nuclear power microscopy, and
transmission electron magnifying instrument. The antibacterial measure was performed
by assurance of the base inhibitory fixation. The antileishmanial and the cytotoxic
impacts prompted by AgNO3, chitosan, and chitosan-based silver nano particles were
investigated by resazurin and MTT colorimetric tests, individually. AgNO3, chitosan, and
chitosan-based silver nano particles actuated a stamped movement against every bacterial
strain and promastigote types of Leishmania amazonensis at least inhibitory focuses
extending from 1.69 to 3.38 µg Ag/mL. Curiously, the chitosan-based silver nano
particles exhibited less cytotoxicity than the AgNO3 alone and were increasingly
dynamic against L. amazonensisthan exclusively chitosan. Moreover, the cytotoxic
focuses (CC50) of both chitosan and chitosan-based silver nano particles against
macrophages were fundamentally higher than the IC50 against promastigotes. Hence, the
chitosan-based silver nano particles speak to a promising option for the treatment of
microbial diseases and leishmaniasis.
Futyra et al. (2017) investigate that severe bacterial and fungal infections have
become a major clinical and public health concern. Nowadays, additional efforts are
needed to develop effective antimicrobial materials that are not harmful to human cells.
This work describes the synthesis and characterization of chitosan–ascorbic acid–nickel

38
nano composites as films exhibiting high antimicrobial activity and non-cytotoxicity
towards human cells. The reductive and stabilizing activity of both the biocompatible
polymer chitosan and ascorbic acid were used in the synthesis of nickel nano particles
(Nickel nano particles). Herein, we propose an improved composite synthesis based on
medium average molecular weight chitosan with a high deacetylation degree, that
together with ascorbic acid gave films with a uniform distribution of small Nickel nano
particles (<10 nm) exhibiting high antimicrobial activity against biofilm forming bacterial
and fungal strains of Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia
coli and Candida albicans. At the same time, the resulting solid nano composites showed,
at the same doses, reduced or totally excluded cytotoxicity on mammalian somatic and
tumoral cells. Data obtained in the present study suggest that adequately designed
chitosan–nickel nano composites are powerful and promising materials for reducing
pathogenic microorganism-associated infections without harmful effects towards
mammalian cells.
Huang et al. (2017) have announced in this investigation, Fe/Ni nano particles
(Nano particles) covered with catechol-conjugated chitosan (CSS) were readied utilizing
green strategies. Strangely, we revealed that CSS-covered these nano particles (CSS-
Fe/Ni nano particles) displayed a higher poisonous quality against gram-negative
Escherichia coli (E. coli) microbes than against gram-positive Staphylococcus aureus (S.
aureus) microscopic organisms. The distinctions uncovered that the CSS- nano particles
slaughtered gram microscopic organisms with particular, species-explicit systems. The
point of this examination is to additionally explore these basic systems through a
progression of investigations. The ultrastructure and morphology of the microbes when
treatments with CSS- nano particles were watched. The outcomes exhibited the CSS-nano
particles executed gram-positive microorganisms through a confusion of the cell divider
and spillage of cytoplasmic substance. Conversely, the essential system of activity on
gram-negative microorganisms was an adjustment in layer penetrability, initiated by
adsorption of CSS-nano particles. The species-explicit components are brought about by
basic contrasts in the cell dividers of gram microscopic organisms. Gram-positive
microscopic organisms are shielded from CSS-nano particles by a thicker cell divider,
while gram-negatives are all the more effectively executed because of a collaboration
between an exceptional external film and the nano particles. Our examination offers an
inside and out comprehension of the antibacterial practices of CSS- nano particles and

39
gives experiences into at last advancing the plan of Silver nano particles for treatment of
bacterial diseases.
Gabal et al., (2018) prepared Cobalt nanoparticles of size 10-20nm. The reducing
agent used during preparation was NaBH4. It reduced the Co+2 ions. The as synthesized
nanoparticles were very stable in air. These nanoparticles were utilized for the treatment
of waste water. The evaluation of prepared nanoparticles was done by the Congo R dye. It
was proved that 150 ppm of the Congo Red was removed from the aqueous solution with
2 min by using Co nanoparticles. The maximum adoption was 800mg.They synthesized
carbon nanotubes coated by the CoZnFe2O4 nanoparticlesby novel gelatin method. They
clear the nanocomposites formation and discussed the coating mechanism. These
nanocomposites have dye removal capability.
Xu et al. (2018) have studied the Optimum condition is studied for the synthsis of
cobalt nano particles chitosan composite. In this paper, the effect of dose of cobalt nano
particles also studied. The present investigation demonstrates that the ideal conditions for
the combination of nano particles (cobalt nano particles) are: 1 % cobalt nitrate, 1 g
chitosan, a union temperature of 90 °C, and a response time of 5 h. The cobalt nano
particles not just improve the shape memory effectiveness of the composite material, yet
in addition improve the mechanical properties. The flawless example of chitosan material
did not re-establish its unique shape after 41 s, in any case, the shape recuperation rate of
cobalt/chitosan composites achieved more than 90 % after 21 s. Besides, the elasticity of
the composite material was as high as 88.9 MPa in the wake of including cobalt nano
particles, expanding by 82.17 % contrasted with unblemished examples.
Gondwal and Joshi, (2018) have explored that an ecofriendly and green technique
for the amalgamation of silver and copper bimetallic nano particles has created utilizing
watery leaves concentrate of Cassia occidentalis. The arrangement of Silver nano
particles and CuNano particles was checked by estimating the UV-Vis spectra. The
morphology and crystalline period of the metal nano particles were resolved utilizing
transmission electron microscopy (TEM) and checking electron microscopy (SEM) with
X-beam vitality dispersive spectrophotometer (EDX) and X-beam diffraction (XRD). The
blended metal nano particles were commonly observed to be round and oval fit as a
fiddle. The Silver nano particles and CuNano particles demonstrated very powerful
antibacterial movement against Escherichia coli and Salmonella typhi microorganisms,
separately. The CuNano particles demonstrated higher radical rummaging action than

40
Silver nano particles. The Silver nano particles demonstrated lower haemolysis (1.7%)
showing lesser lethality when contrasted with CuNano particles. The CuNano particles
have better synergist capacity for the decrease of 4-nitrophenol and 2-nitrophenol when
contrasted with Silver nano particles.
Singh et al. (2018) have investigated that this articles just about the synthesis of
the gum acacia taragacanth gum, polyvinyl alchohol and polyvinyle pyrolidone. This
synthesis based upon the pure and sterile hydrogel by radiation used cross linking method
for the purpose of wound dressing application. To enhance the wound healing process of
hydrogel dressing , the hydrogel mix with the silver nano particles. Moreover biomedical
properties of hydrogel film such as antioxitant activity, mucoadhesion and medical
properties were also discussed. It has been found that increase the concentration of gum
acasia during formation of hydrogel decrease the production of the silver particles from
the hydrogel. Addition of the silver particles increased the antioxidant activity which
could be give positive feedback in the wound healing potential of hydrogel dressings.
Suma et al. (2019) have studied the graphite screen-printed terminals after
alteration have utilized to recognize the electrocatalytic conduct of nitrite oxidation in
aqueous medium. All the factors impacting the optimized have advanced and fused in the
prescribed methodology. The proposed sensor has been utilized to quantify the nitrite
levels from financially accessible milk powder tests and the outcomes have been
contrasted and the standard convention. The consequences of the proposed sensor are in
great concurrence with the standard convention.Fe metal nano particles have been
installed in chitosan framework and utilized as a composite material in the synthetic
alteration of graphite screen-printed terminals. GSPEs are anything but difficult to
manufacture. They give wide straight working extent for example 30 - 1140 µM of nitrite.
The sensor is profoundly steady, reproducible and gives an exceptionally low discovery
cutoff of 1.84 µM. The technique has been connected to gauge follow level nitrite from
milk powder tests.
Suma et al. (2019) have investigated that the nitrites can apply intense poisonous
impacts in people. It is broadly utilized as an additive in dairy and meat items. The nitrites
structure N-nitrosamines, which are potential cancer-causing agents and cause hindering
wellbeing impacts. Thus we report a dispensable graphite screen-printed sensor created
utilizing Nickel/Cobalt bimetallic metal nano molecule inserted chitosan composite in the
evaluation of nitrite at follow level.Regular techniques have different constraints.

41
Electrochemical techniques give a perfect stage to follow nitrite investigation. The
readied composite has portrayed by UV-Visible spectrometry, SEM, EDS and XRD
procedures. The proposed sensor has been created by utilizing graphite screen-printed
terminals through drop covering of the composite material. The redox conduct and its
utilization of the manufactured cathode have been examined utilizing cyclic and anodic
stripping voltammetric techniques.
Chen et al. (2019) have examined that the physicochemical properties and
biocompatibility of biomaterials significant contemplations, yet in addition their
antibacterial properties. In this examination, a novel synthetically cross-connected
antibacterial porcine acellular dermal framework (pADM) platform was manufactured by
a two-advance technique. A normally determined oxidized chitosan oligosaccharide
(OCOS) was utilized to cross-connected pADM (named OCOS-pADM) to improve its
physicochemical properties. Lingering aldehyde bunches inside the OCOS-pADM were
utilized in a redox response with nickel particles to create nickel nano particles
(Nickelnano particles) in situ. As the Nickel nano particles were firmly followed onto the
platform fibrils (named OCOS-Nickel nano particles-pADM), this successfully
functionalized framework with antibacterial properties. The created Nickel nano particles
were described by UV-Vis diffuse reflectance spectroscopy, XPS and SEM. The
consequences of DSC, TG and enzymatic corruption showed that OCOS-Nickel nano
particles-pADM had improved warm security and protection from enzymatic debasement
contrasted and pADM platforms. The motor trial of the arrival of silver demonstrated that
nickel was discharged in a controllable manner. In the wake of bringing Silver nano
particles into frameworks, the OCOS-Nickel nano particles-pADM had wide-range
antibacterial movement against Escherichia coli and Staphylococcus aureus. Besides,
MTT measure and CLSM demonstrated that the frameworks had great biocompatibility.
Bits of OCOS-Nickel nano particles-pADM were embedded into Sprague-Dawley
rodents to describe their capacity to fix full-thickness skin wounds. Also, results
demonstrated that the OCOS-Nickel nano particles-pADM could quicken the injury
mending process. In general, this work contributes new understanding into the synthetic
cross-connecting and functionalization of pADM frameworks. Moreover, as novel
antibacterial frameworks, OCOS-Nickel nano particles-pADMs have the potential for
improvement as wound dressing materials.Not just are the physicochemical properties
and biocompatibility of biomaterials significant contemplations, yet additionally their

42
antibacterial properties. In this investigation, a novel synthetically cross-connected
antibacterial porcine acellular dermal lattice (pADM) framework was created by a two-
advance strategy. A normally determined oxidized chitosan oligosaccharide (OCOS) was
utilized to cross-connected pADM (named OCOS-pADM) to improve its
physicochemical properties. Leftover aldehyde bunches inside the OCOS-pADM were
utilized in a redox response with nickel particles to create Ni nano particles (Nickel nano
particles) in situ. As the Nickel nano particles were firmly followed onto the framework
fibrils (named OCOS-Nickel nano particles-pADM), this successfully functionalized
platform with antibacterial properties. The produced Nickel nano particles were described
by UV-Vis diffuse reflectance spectroscopy, XPS and SEM. The aftereffects of DSC, TG
and enzymatic debasement showed that OCOS-Nickel nano particles-pADM had
improved warm soundness and protection from enzymatic corruption contrasted and
pADM frameworks. The dynamic examination of the arrival of nickel demonstrated that
nickel was discharged in a controllable manner. In the wake of bringing Nickel nano
particles into frameworks, the OCOS-Nickel nano particles-pADM had wide-range
antibacterial movement against Escherichia coli and Staphylococcus aureus. Moreover,
MTT measure and CLSM demonstrated that the frameworks had great biocompatibility.
Bits of OCOS-Nickel nano particles-pADM were embedded into Sprague-Dawley
rodents to describe their capacity to fix full-thickness skin wounds. What's more, results
demonstrated that the OCOS-Nickel nano particles-pADM could quicken the injury
mending process. By and large, this work contributes new understanding into the
compound cross-connecting and functionalization of pADM platforms. What's more, as
novel antibacterial frameworks, OCOS-Nickel nano particles-pADMs have the potential
for advancement as wound dressing materials.
Nguyen et al. (2019) have examined silver nano particles have pulled in incredible
interests broadly in drug because of its extraordinary qualities of antibacterial action. In
this exploration, the antibacterial movement and biocompatibility of a topical gel
integrated from polyvinyl liquor, chitosan, and silver nano particles were contemplated.
Hydrogels with various groupings of silver nano particles (15 ppm, 30 ppm, and 60 ppm)
were assessed to think about their antibacterial action, nano particles' sizes, and in vivo
practices. The came about silver nano particles in the hydrogel were described by TEM
demonstrating the nano particles' sizes under 22 nm. The in vitro outcomes demonstrate
that the antibacterial impacts of the majority of the examples are fulfilled. Be that as it

43
may, the in vivo outcomes exhibit the noteworthy contrast among various hydrogels in
wound recuperating, where hydrogel with 30 ppm demonstrates the best mending rate.

CHAPTER 3
MATERIALS AND METHODS
3.1 Materials
44
The exploration work was done in research lab of Department of Applied Chemistry in
Govt College University Faisalabad. All chemicals were bought from Sigma Aldrich and
were utilized moving along without any more purification. Chitosan of medium sub-
atomic weight (from crab shells), 70-80 % deacetylated was utilized as the help material.
Iron acetate and Nickel acetate are also obtained from analytical grade Sigma Aldrich.
Apparatus and chemicals which were used in this research are enlisted as follows;
3.1.1 Apparatus
 Conical flasks
 Measuring Cylinder (50 ml, 100 ml)
 Funnel
 Measuring flasks of 250 ml and 500 ml
 Pipette
 Thermometer
 Beakers
 Stirrer
 Hot plate
 Weight Balance
 Dropper
 Water Bath
 Magnet Bar
 Centrifuge
 Universal Ultrasonic Cleaner
 Tungsten rod of 500 watt
 Filteration apparatus
 Whatmann filter no 1
 pH meter

3.1.2 Chemicals
 Chitosan
 Hydrazine Monohydrate
 Hydrochloric Acid (HCl 37%)
 Iron (Fe+3) acetate
 Nickel (Ni+2) acetate
 Dist. water

45
 NaOH
 Methylene diphenyl isocyanate (HDI)

3.2 Method
3.2.1 Synthesis Fe/Ni Bimetallic Nanoparticles
Took 100 ml of distilled water in four different 250 ml conical flasks.Then added
the following amounts of salts of iron and nickel in the table given below separately.Then
mixed all the solutions with constant stirring. After this added 5 ml of hydrazine
monohydrate in all flasks dropwise and stayed overnight.Precipitates were formed and
filtered.Then washed with distilled water several time and dryed.
Table.1. Scheme of synthesized bimetallic composites

Sr. No. Iron acetate Nickel acetate DI water+Hydrazine

Monohydrate
Flask 1 0.6 g 2.4g 100 ml+5ml of Hydrazine

Flask 2 1.2 g 1.8 g 100 ml+5ml of Hydrazine

Flask 3 1.8 g 1.2 g 100 ml+5ml of Hydrazine

Flask 4 2.4 g 0.6 g 100 ml+5ml of Hydydrazine

Fig.6. Synthesis of Fe/Ni bimetallic nanoparticles

46
 Fig.7. Synthesized Fe/Ni bimetallic nanoparticles

3.2.2 Preparation Chitosan Solution

Prepared 3% Chitosn solution in distilled water. Dissolved 3 g Chitosan in 100
cm3 of distilled water containing 5 ml of Hydrochloric acid (37%) solution to form 3 %
Chitosan solution. This solution stirred at 600 rpm and 50 to 60 o C temperatures for 5
hours until Chitosan dissolved completely. It became transparent solution with light
yellowish colour and stayed overnight.

Fig.8. Yellowish coloured solution of Chitosan

3.2.3 Cross linking by MDI

Took 2 mL above solution in 100 mL beaker. 2 mL MDI was added in this
solution by drop wise at 50 oC and 1000 rpm for 1 hour. Then it took universal sonication
cleaner for 60 minutes. The solution became clear and stayed overnight.

47
3.2.4 Blank Sample
Added drop wise 1% NaOH solution in above solution until pH came to 11.
Precipitate formed and filtered the solution and washed the ppt with distilled water
several time.

3.2.5 Experiment No. 1

2 mL Chitosan solution took into 250 mL beaker. Drop wise 3 % 2 mL of Fe/Ni
acetate salts of bimetallic solution (with composition 0.6 g of Fe acetate and 2.4 g of
Nickel acetate) was added into 2 mL Chitosan solution with constant stirrer at 800 rpm in
60 minutes. Then 2 mL MDI was added in this solution by drop wise at 40 oC and 1000
rpm for 1 hour. Then it took universal sonication cleaner for 60 minutes. The solution
became clear and stayed overnight and slurry was formed. Then added 2 ml of hydrazine
monohydrate dropwise in the slurry prepared for the reduction and stayed overnight.Then
added drop wise 1% NaOH solution in above solution until pH came to 11. Precipitate
formed and filtered the solution and washed the ppt with distilled water several time.

3.2.6 Experiment No. 2

2 mL Chitosan solution was taken into 250 mL beaker. Drop wise 3 % 2 mL of
Fe/Ni acetate salts of bimetallic solution (with composition 1.2 g of Fe acetate and 1.8 g
of Nickel acetate) was added into 2 mL Chitosan solution with constant stirrer at 800 rpm
in 60 minutes. Then 2 mL MDI was added in this solution by drop wise at 40 oC and 1000
rpm for 1 hour. Then it took universal sonication cleaner for 60 minutes. The solution
became clear and stayed overnight and slurry was formed. Then added 2 ml of hydrazine
monohydrate dropwise in the slurry prepared for the reduction and stayed overnight.Then
added drop wise 1% NaOH solution in above solution until pH came to 11. Precipitate
formed and filtered the solution and washed the ppt with distilled water several time.

3.2.7 Experiment No. 3

2 mL Chitosan solution was taken into 250 mL beaker. Drop wise 3 % 2 mL of
Fe/Ni acetate salts of bimetallic solution (with composition 1.8 g of Fe acetate and 1.2 g
of Nickel acetate) was added into 2 mL Chitosan solution with constant stirrer at 800 rpm
in 60 minutes. Then 2 mL MDI was added in this solution by drop wise at 40 oC and 1000
rpm for 1 hour. Then it took universal sonication cleaner for 60 minutes. The solution
became clear and stayed overnight and slurry was formed. Then added 2 ml of hydrazine
monohydrate dropwise in the slurry prepared for the reduction and stayed overnight.Then

48
added drop wise 1% NaOH solution in above solution until pH came to 11. Precipitate
formed and filtered the solution and washed the ppt with distilled water several time.

3.2.8 Experiment No. 4

2 mL Chitosan solution was taken in 250 mL beaker. Drop wise 3 % 2 mL of
Fe/Ni acetate salts of bimetallic solution (with composition 2.4 g of Fe acetate and 0.6 g
of Nickel acetate) was added into 2 mL Chitosan solution with constant stirrer at 800 rpm
for 60 minutes. Then 2 mL MDI was added in this solution drop wise at 40 oC and 1000
rpm for 1 hour. Then it took universal sonication cleaner for 60 minutes. The solution
became clear and stayed overnight and slurry was formed. Then added 2 ml of hydrazine
monohydrate dropwise in the slurry prepared for the reduction and stayed overnight.Then
added drop wise 1% NaOH solution in above solution until pH came to 11. Precipitate
formed and filtered the solution and washed the ppt with distilled water several time.
Cross linked Chitosan-Fe/Ni bimetallic nano composites containing various
amounts of Fe and Ni salts like were synthesized. Of these the composite containing 3 wt
% of Fe/Ni BMNPs (with composition of 1.8 g of Fe and 1.2 g of Ni salts) was chosen
here for discussions, as the properties of these bimetallic nanoparticles would be more
pronounced when the amount of BMNPs present in the composite would more. So it

49
50
51
Fig.9. Mechanism for Cross-linking of Chitosan by Di isocyanate would be easier for us
to study the properties of Fe/Ni BMNPs adsorbed on crosslinked chitosan.

Fig.10. Flow sheet diagram for the synthesis of Samples

In this study, all the samples were prepared by using hydrothermal method. Hydrothermal
method provides an easy route to control particle size, morphology and phase
composition under high pressure and moderate temperature.

52
 Fig.11. Images of All the Prepared Samples

3.3 Characterization
The Fourier-transform infrared spectroscopy (FTIR), x-ray power diffraction
(XRD), Thermogravimetric analysis (TGA), Differential scanning calorimetry (DSC),
scanning electron microscope (SEM) transmission electron microscopy (TEM), powder
X-ray diffraction (XRD) are used for the characterization for the cross linked Chitosan-
Fe/Ni bimetallic nano particles. The result shows that Fe/Ni bimetallic nano particles
were of high purity and the average size of these nano particles should be found to be 10
± 1 nm.The FTIR spectra of plain CS, MDI cross linked Chitosan-Fe/Ni bimetallic nano
particles were compared. It was expected that, with the increasing degree of reticulation,
the bands at around 1640 and 1560 cm−1, assigned to amino I and amino II functional
groups of CS, showed a better resolution.

53
3.3.1 Analytical techniques
1. FTIR Spectroscopy

FTIR spectroscopy is an analytical technique that is used to recognize organic,

inorganicand polymeric samples. FTIR spectroscopy used IR (infrared radiations) to scan
the givensample and investigates chemical properties of sample.First of all sample is
placed in the path of IR beam. When IR beam fall onsample it scan the given sample. The
IR spectrum as a result is shown on screen in the form of emission and absorption peaks.
The resulting signal is in the range of 400cm -1 to 4000cm-1. The obtained absorption and
emission peaks can be compared with various types of bonds in givensample.
Eachfunctional group has specific emission and absorption peak. By analyzing these
peaks, the structure of compound can be identified.

2. Scanning electron microscope (SEM)

A scanning electron microscope is an electron microscopic technique that is used to

analyze the surface morphology of the sample. A beam of different wavelength collides
with atoms of sample andgives various signals on screen in the form of image. SEM can
help us to identifymorphology as well as composition of the sample.For better results, a
high vacuum atmosphere is given. But at different temperature rangesa low vacuum
atmosphere can also be obtained. When electrons strike with atoms ofsample, secondary
electrons emitted from the sample that is the main source of imageformation of sample.
The image formed by secondary electrons depends upon theintensity of electron beam as
well as angle of sample with electron beam.A special type of detector is used to sort out

54
secondary electron and an image of sample is shown called SEM image.SEM images help
us to identify surface morphology as well as composition of sample.

3. X-Ray diffraction (XRD)

X-Ray diffraction (XRD) is an analytical technique that is primarily used to identify the
crystalline nature of given sample and provides information about unit cell dimensions.
The X-Ray is produced by cathode ray tube then filteredto produce monochromatic
radiation which is directed toward sample. When this monochromatic radiation interacts
with sample,the diffracted ray and the constructive interference is produced. The
diffracted rays are then detected, processed and counted. Using the Bragg’s equation
(nλ=2d sinө) we can get information about crystalline nature of the sample. This Bragg’s
equation correlates the diffracted angle and wavelength of electromagnetic radiation.

55
3.3.2 Application
A homemade device was used for the photocatalytic degradation test, which was
equipped with 500W Xenon arc lamp to stimulate sun light. For the photocatalytic
experiment, 0.1 g of photocatalyst sample was put into a glass tube containing 100 ml
aqueous solution of Congo red (5 mg/L). Immediately the glass tube was placed into the
photocatalytic degradation test box under darkness. Then the solution was stirred
continuously for 1 hour to achieve adsorption equilibrium. After 1 hour the photocatalyst-
Congo red dye suspension was continuously stirred under stimulated sunlight irradiations.
The distance between upper surface of the suspension and the lamp was 16 cm. UV-vis
spectrophotometer (Hach USA, DR-5000) was used to measure the photocatalytic
discoloration rate of Congo red, with and without a sample.

56
CHAPTER 4
RESULTS AND DISCUSSION

57
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