INTRODUCTION

NANOPARTICLES-

Nanotechnology refers to an emerging field of science that includes synthesis and development of
various nanomaterials which have a wide variety of potential applications in biomedical, optical and
electronic field. Nanoparticles can be defined as objects ranging in size from 1-100nm that due to their
size may differ from the bulk material. The properties of bulk materials are different from that of the
nanoparticles due to their large surface. Nanoparticles are of great scientific interest as they are
effectively a bridge between bulk material and atomic or molecular structure. [1]

Nanoparticles are not simple molecules itself and therefore composed of three layers i.e. (a) The
surface layer, which may be functionalized with a variety of small molecules, metal ions, surfactants and
polymers. (b) The shell layer, which is chemically different material from the core in all aspects, and (c)
The core, which is essentially the central portion of the nanoparticles and usually refers the nanoparticles
itself. [2]

TYPES OF NANOPARTICLES:-

Nanoparticles are broadly divided into various categories depending on their morphology, size and
chemical properties. Based on physical and chemical characteristics, some of the well known classes of
nanoparticles are given as below—

1. Carbon-based nanoparticles :- Fullerenes and carbon nanotubes (CNTs) represent two major
classes of carbon-based nanoparticles. Fullerenes contain nanomaterials that are made of globular hollow
cage such as allotropic forms of carbon. [3]
2. Metal nanoparticles: - Metal nanoparticles are purely made of the metals precursors. Due to well-
known localized surface plasmon resonance (LSPR) characteristics, these nanoparticles possess unique
optoelectrical properties. Nanoparticles of the alkali and noble metals i.e. Cu, Ag and Au have broad
absorption band in the visible zone of the electromagnetic solar spectrum. The facet, size and shape
controlled synthesis of metal nanoparticles is important in present day cutting-edge materials. Due to their
advanced optical properties, metal nanoparticles find applications in many research areas. [4]

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 3. Ceramic nanoparticles :- Ceramics nanoparticles are inorganic nonmetallic solids, synthesized via
heat and successive cooling. They can be found in amorphous, polycrystalline, dense, porous or hollow
forms. [5]
4. Semiconductor nanoparticles :- Semiconductor materials possess properties between metals and
nonmetals and they possess wide band gaps and therefore showed significant alteration in their properties
with band gap tuning. [6]
5. Polymeric nanoparticles :- These are normally organic based nanoparticles and in the literature a
special term polymer nanoparticle (PNP) collective used for it. They are mostly nanospheres or
nanocapsular shaped [7]. Nanospheres are matrix particles whose overall mass is generally solid and the
other molecules are adsorbed at the outer boundary of the spherical surface. In nanocapsular, the solid
mass is encapsulated within the particle completely. [8]
6. Lipid-based nanoparticles :- These nanoparticles contain lipid moieties and effectively using in
many biomedical applications. Generally, a lipid nanoparticles is characteristically spherical with
diameter ranging from 10 to 1000 nm. Like polymeric nanoparticles, lipid nanoparticles possess a solid
core made of lipid and a matrix contains soluble lipophilic molecules. Surfactants or emulsifiers stabilized
the external core of these nanoparticles. [9]

USES OF NANOPARTICLES :-
Nanoparticles can be used in variety of applications. Some important of these are given below—
1. Applications in drugs and medications :- Nanoparticles have drawn increasing interest from every
branch of medicine for their ability to deliver drugs in the optimum dosage range often resulting in
increased therapeutic efficiency of the drugs, weakened side effects and improved patient compliance
[10]. Most of the semiconductor and metallic nanoparticles have immense potential for cancer diagnosis
and therapy on account of their surface plasmon resonance (SPR) enhanced light scattering and
absorption. [11]
2. Applications in manufacturing and materials :- Nanoparticles focus on the characterization,
designing and engineering of biological as well as non-biological structures less than 100 nm, which
show unique and novel functional properties. The potential benefits of nanotechnology have been
documented by many manufacturers at high and low level and marketable products are already being
mass-produced such as microelectronics, aerospace and pharmaceutical industries. The presence of
nanoparticles in commercially available products is becoming more common. [12]

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 3. Applications in the environment :- Most of environmental applications of nanotechnology fall into
three categories:
i. Environmentally benign sustainable products (e.g. green chemistry or pollution prevention).
ii. Remediation of materials contaminated with hazardous substances and
iii. Sensors for environmental stages. [13]
4. Applications in electronics:- There has been growing interest in the development of printed
electronics in last few years because printed electronics offer attractive to traditional silicon techniques
and the potential for low cost, large area electronics for flexible displays, sensors. Printed electronics with
various functional inks containing NPs such as metallic nanoparticles, organic electronic molecules,
CNTs and ceramics nanoparticles have been expected to flow rapidly as a mass production process for
new types of electronic equipment. [14]
5. Applications in energy harvesting :- Recent studies warned us about the limitations and scarcity of
fossil fuels in coming years due to their nonrenewable nature. Therefore, scientists shifting their research
strategies to generate renewable energies from easily available resources at cheap cost. They found that
nanoparticles are the best candidate for this purpose due to their, large surface area, optical behavior and
catalytic nature. Especially in photocatalytic applications, Nanoparticles are widely used to generate
energy from photoelectrochemical (PEC) and electrochemical water splitting. [15]
6. Applications in mechanical industries:- Nanoparticles can offer many applications in mechanical
industries especially in coating, lubricants and adhesive applications. Besides, this property can be useful
to achieve mechanically stronger nanodevices for various purposes. [16]

SILVER NANOPARTICLES-

Silver nanoparticles are formed when Ag+ ions dissociate from a silver compound when it is
dissolved and gain an electron in an oxidation-reduction reaction with a reducing agent. The interest in
the study of AgNPs with respect to their various different behaviors has recently increased because of
their unique and attractive physical, chemical, and biological properties. AgNPs are also known to have
unique properties in terms of toxicity, surface plasmon resonance, and electrical resistance. Based on
these, intensive works have been conducted to investigate their properties and potential applications for
several purposes such as antimicrobial agents in wound dressings, anticancer agents, electronic devices,
and water treatment. [17]

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TYPES OF SYNTHESIS OF Ag-NANOPARTICLES :-
Ag nanoparticles can be synthesized using three methods, namely physical, chemical, and
biological.
1. Physical approach :- Generally, the physical approach used to synthesize Ag nanoparticles
employs the evaporation–condensation method. It is commonly performed using a tube furnace at
atmospheric pressure, which is reliable to synthesize various sizes. [18]

2. Chemical approach :- The chemical approach is widely used for synthesizing Ag nanoparticles
using water or organic solvents. It is an easy way to synthesize Ag nanoparticles in solution [19].
However, a certain amount of toxic material may be produced as residue. Some reducing agents such as
borohydride, citrate, ascorbate, and glucose have been used to address this problem. [18]

3. Biological approach :- There are some evidences that the above methods are harmful to the
environment. To avoid this harmfulness in the environment, we can opt or develop the plant-mediated
green synthesis method which includes the nontoxic and very environmentally friendly process of the
preparation and the production of the silver nanoparticles. The advancement of green synthesis over the
other synthetic methods is because of its eco-friendly and cost-effective characteristics and it can easily
scale up for the large-scale synthesis. The very important factor is there is no need to use the high
temperature, pressure, energy, and any toxic chemicals for the production of silver nanoparticles. [20]
It had been known that the synthesis of Ag nanoparticles using the chemical approach requires three
main ingredients: a silver salt, a reducing agent, and a stabilizer or capping agent. In the biological
approach, the reducing agent and the stabilizer are replaced using molecules obtained from living
organisms such as plants, bacteria, fungi, yeast, and algae. The use of the plants for the production of
silver nanoparticles is the single-step technique for the process of green synthesis. Plant extracts can work
as reducing agents to synthesize Ag nanoparticles and provide an alternative solution that is
environmentally friendly. [17]

USES OF Ag NANOPARTICLES :-
Silver has long been recognized as having an inhibitory effect toward many bacterial strains and
microorganisms commonly present in medical and industrial processes [21]. The most widely used and
known applications of silver and silver nanoparticles are in the medical industry. These include topical
ointments and creams containing silver to prevent infection of burns and open wounds [22]. Other widely
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used applications are medical devices and implants prepared with silver-impregnated polymers [23]. In
addition, silver-containing consumer products such as colloidal silver gel and silver-embedded fabrics are
now used in sporting equipment.

ORGANIC DYES :-
Organic dyes are used in numerous industries like food, pharmacy, cosmetics, paints, plastics,
paper and textiles. These synthetic dyes are harmful for human health and they are hazardous to
environmental as well. Various conventional treatments like coagulation, filtration, adsorption and
reverse osmosis have been used for dye removal. Nevertheless, it is difficult to remove these dyes from
water, because of their aromatic structural stability. Nano-catalysts are one of the promising agents for the
reduction of synthetic dyes. Thus, in the present work, we have been investigating
the degradation of Methylene Blue (MB) and Methyl Orange (MO) dye using the green-synthesized silver
nanoparticles (AgNPs) as catalyst in the presence of sodium borohydride (NaBH4). [24]

AIM AND OBJECTIVE

1. To biosynthesize silver nanoparticles using leaf extract of Azadirachta indica (neem).
2. To characterize the synthesized nanoparticles by UV-visible spectroscopy, Fourier transform
infrared spectroscopy and Scanning Electron Microscopy.
3. To study the catalytic action of the synthesized silver nanoparticles in reduction of dyes viz.
methylene blue and methyl orange.

EXPERIMENTAL SECTION

i. CHEMICALS AND REAGENT :-

All the chemicals used are of reagent grade. Silver nitrate (AgNO3) was purchased from Fisher Scientific,
Sodium borohydride (NaBH4) was purchased from EmpluraR Merck Life Science Pvt. Limited.
Methylene blue and methyl orange was purchased from Ranbaxy Laboratories Limited. The leaves of
neem were collected from campus.

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 ii. METHODS :-
a. Preparation of plant extract ---
Green tender neem (Azadirachta indica) leaves were used to prepare the aqueous plant extract. 10-15
number of neem leaves were taken and thoroughly washed in distilled water, cut into fine pieces and
were added to a beaker containing 20mL distilled water. This was boiled for a few minutes until the water
level decreased to 10mL and then filtered through Whatmann No.1 filter paper. The filtrate obtained was
the neem leaf extract. [25]
b. Synthesis of Ag nanoparticles ---

0.1 M of AgNO3 solution was prepared in 40mL distilled water. 20mL of the AgNO3 solution was taken
in a glass beaker at room temperature. 5-6 drops of the plant extract was added dropwise to the AgNO3
solution with continuous stirring. The mixture was kept still for few minutes to observe the colour change
to reddish brown, change in colour indicated the synthesis of Ag-nanoparticles. [25]

c. Reduction of dye ---

Methylene blue(MB) and methyl orange(MO) solutions were prepared by dissolving in using deionised
water. 1.5 mL sodium borohydride solution was added to 30mL MB and MO separately. The solutions
were then vigorously stirred. 1 mL of Ag-nano was added to each of the solutions and stirred for 1min.
The degradation of dyes is indicated by the decolorisation of the solution.

d. Characterization of the Ag-nanoparticles ---

 UV-Visible Spectroscopy:-

Preliminary identification of the silver nanoparticles was carried out using UV-visible
spectroscopy. The UV-Vis spectra are recorded on a LABINDIA ANALYTICAL UV-3200 UV-Vis
Spectrometer. The formation of silver nanoparticles was confirmed by measuring the UV-visible spectra
of the solutions after diluting a small portion of the Ag-nano sample. The distilled deionized water was
used as blank.

 Fourier Transform Infrared Spectroscopy :-

To determine the functional groups of the aqueous plant extract and their possible involvement in
the synthesis and stabilization of silver nanoparticles, Fourier transform infrared analysis was carried out
of the test sample (plant extract after reaction with AgNO3). The FTIR spectra were recorded on a
Perkin Elmer RXI FTIR system.
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  Scanning Electron Microscopy :-
The morphology of the synthesized nanoparticles from neem extract were investigated by
Scanning Electron Microscopy(SEM) images. The images were recorded in Zeiss FESEM SIGMA 300
METEK EDAX scanning electron microscope at an acceleration voltage of 5kV.

RESULTS AND DISCUSSION

When neem leaf extract is added to silver salt (AgNO3) solution it results into a colour change
from colourless to brown colour, as shown in Fig.1. The change in colour of the solution is due to the
presence of silver nanoparticles formed by the reduction of silver salt. The reduction of silver salt to
elemental silver occurs due to the presence of reducing agents that are present in the neem leaves such as
terpenoids and flavanones. AgNPs show brown colour in aqueous solution due to the excitation of surface
plasmon vibrations. It is noticed that the complete colour change took after about 5min as shown in Fig.1.

addition of neem extract

~ 5mins

(AgNO3 before addition of extract) (Formation of Ag-nano)

Fig.1: Silver nanoparticle synthesis

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 UV-Vis Analysis –

The UV-visible spectrum of the synthesized silver nanoparticles showed absorption peak at 440
nm which is the characteristic peak for AgNPs (Fig.2).

1.0

0.8

Absorbance
0.6

0.4

0.2

0.0
300 450 600 750 900
Wavelength (nm)

Fig.2: UV/Visible spectrum of AgNPs synthesised from Azadirachta Indica (neem) extract.

 Fourier transform infrared spectroscospy analysis -

FTIR analysis on the biosynthesized Ag NPs was carried out to identify the functional groups
responsible for the reduction of metal precursors (AgNO3). The spectral (Fig.3) analysis of the extract
shows IR bands at 3465, 1645 and 1372 cm-1. The band at 3465 cm -1 corresponds to O-H stretching of
H-bonded alcohols and phenols. The peak found at 1645 cm -1 showed a stretch for C=O and C=C bond
and the peak at 1372 cm -1 can be attributed to the OH bending. Therefore the synthesized nanoparticles
were surrounded by proteins and metabolites such as terpenoids obtained from the neem leaf extract
which could have possibly served the purpose of capping agents and reducing agents for the synthesis
of the AgNPs.

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 Fig.3: FTIR spectrum of the Ag-nanoparticles synthesized by Neem extract

 Scanning Electron Microscopy (SEM) –

SEM analysis provides the information about the morphology of the synthesized AgNPs. It also
gives the idea about the approximate size of particles and features. From the SEM images the AgNPs
were found to be uniform and spherical in shape (Fig.4).

Fig.4 : SEM image of the synthesized Ag-nanoparticle

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 Catalytic activity of Ag nanoparticles –

The catalytic activity of biosynthesized AgNPs was investigated using the reduction reaction of
methylene blue and methyl orange by NaBH4 as a model reaction.

Catalytic reduction of Methylene blue :

MB is a basic aniline dye also known as methylthioninium chloride, which has many uses in
diverse fields including biology and chemistry, as a stain and as a medicine. However there are some of
its side effects and the common side effects include abnormal urine or stool color.
Methylene blue (MB) has blue color in oxidized state and loses its color upon reduction forming
colorless leucomethylene blue. The degradation of MB by NaBH4 was carried out in presence of AgNPs
at different time intervals. The absorption peak of MB was found at 670 nm and hence the degradation
studies were monitored via UV-Visible spectroscopy. The absorption peak of MB was found to decrease
with time as shown in Fig.5. This decrease in peak indicated the reduction of the dye by NaBH4 in
presence of the green synthesized AgNPs as catalyst. The fading of the blue colour of MB during the
process also proved its reduction (Fig.6). After 35mins complete degradation was observed.

Fig.5 : UV-Vis absorption spectra for reduction of MB by NaBH4 in the presence of synthesized AgNPs

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 NaBH4 + AgNP

(methylene blue solution) (reduced methylene blue)

Fig.6 : Reduction of the methylene blue dye

Catalytic reduction of Methyl Orange :

Methyl orange is a toxic and stable dye which is dangerous to the environment. MO is an organic
sulfosalt dye, which can be reduced by reductants like NaBH4 to form small organic molecules firstly and
non toxic species finally, but the reduction rate is very slow.

The absorption band of MO appeared at 470 nm and hence its reduction studies were performed by
UV-visible spectroscopy. The absorption spectra of reduction of MO by NaBH4 in presence of AgNPs as
catalyst at regular time intervals are shown in Fig.7. The peak at 470 nm is found to decrease gradually
with varying time intervals and eventually disappearing. It was obvious from the decrease in the peak that
the degradation of the dye had occurred. After 45mins complete degradation was observed.

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 Fig.7 : UV-Vis absorption spectra for reduction of MO by NaBH4 in the presence of synthesized
AgNPs

CONCLUSION

Silver nanoparticles were synthesized by using Azadirachta Indica (neem). Characterization of

synthesized nanoparticles was studied by using UV-Visible spectra, Fourier transform infrared
spectroscopy and Scanning Electron Microscopy(SEM). With the help of UV-Visible spectra the
maximum absorbance of silver nanoparticles was found to be at 440nm. From the analysis of FTIR
studies it was confirmed that the carbonyl groups from the amino acid residues and proteins has the
stronger ability to bind metal indicating that the proteins could possibly form the metal nanoparticles (i.e.;
capping of silver nanoparticles) to prevent agglomeration and thereby stabilize the medium. By using
SEM images the morphology of the AgNPs were observed. Synthesized silver nanoparticles were used
for catalytic reduction of the dyes methylene blue and methyl orange. Their reduction was confirmed by
using UV-Visible spectroscopy where the absorption peak decreased gradually with regular intervals of
time.

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