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ISSN No: 2456
International
Research
Biosynthesis and Characterization o
Nanoparticles u
N. Tensingh Baliah
Post Graduate and Research Department
of Botany, Ayya Nadar Janaki Ammal
College, Sivakasi, Tamil Nadu, India
ABSTRACT
The wide application of nanoparticles stimulates the
need for synthesizing them but, the conventional
methods are usually hazardous and energy
consuming. This leads to focus on “green synthesis”
of nanoparticles which seems to be easy efficient and
ecofriendly approach. In this study, the plant mediated
synthesis of zinc oxide nanoparticles (ZnO NPs) was
carried out using bulb extract of Allium cepa as a
reducing agent. The optimized nano zinc thus
obtained was quantified and characterized using UV
Visible spectroscopy, Fourier Transform Infrared
Spectroscopy (FT-IR), X-Ray diffraction,
Electron Microscope (SEM), EDAX
potential analyses. Further, the synthesized ZnO NPs
were tested for antimicrobial activity.
Keywords: Nanoparticles, zinc oxide,
Allium cepa
I. INTRODUCTION
Nanotechnology is a emerging field with its
application in science and technology for the purpose
of manufacturing new materials at nano scale level
(Albrecht et al., 2006). Recent advance in the field of
nanotechnology, particularly the ability to prepare
highly ordered nanoparticles of any size and shape
(Sangeetha et al., 2012). Nanotechnology is a
multidisciplinary scientific field undergoing explosive
development. The nanometer-sized particles offer
novel structural, optical and electronic properties that
are not attainable with individual molecules or bulk
@ IJTSRD | Available Online @ www.ijtsrd.com | Volume – 2 | Issue – 2 | Jan-Feb
ISSN No: 2456 - 6470 | www.ijtsrd.com | Volume
International Journal of Trend in Scientific
Research and Development (IJTSRD)
International Open Access Journal
Biosynthesis and Characterization of Zinc Oxide
Nanoparticles using Onion Bulb Extract
Post Graduate and Research Department
of Botany, Ayya Nadar Janaki Ammal
College, Sivakasi, Tamil Nadu, India
S. Lega Priyatharsini
Post Graduate and Research Department
of Botany, Ayya Nadar Janaki Ammal
College, Sivakasi, Tamil Nadu, India
application of nanoparticles stimulates the
need for synthesizing them but, the conventional
methods are usually hazardous and energy
consuming. This leads to focus on “green synthesis”
of nanoparticles which seems to be easy efficient and
oach. In this study, the plant mediated
synthesis of zinc oxide nanoparticles (ZnO NPs) was
carried out using bulb extract of Allium cepa as a
reducing agent. The optimized nano zinc thus
obtained was quantified and characterized using UV-
Fourier Transform Infrared
Ray diffraction, Scanning
EDAX and Zeta
. Further, the synthesized ZnO NPs
Nanoparticles, zinc oxide, biosynthesis,
Nanotechnology is a emerging field with its
application in science and technology for the purpose
of manufacturing new materials at nano scale level
., 2006). Recent advance in the field of
ogy, particularly the ability to prepare
particles of any size and shape
., 2012). Nanotechnology is a
multidisciplinary scientific field undergoing explosive
sized particles offer
ral, optical and electronic properties that
are not attainable with individual molecules or bulk
solids (Anitha et al., 2011). The characters of metal
and metal oxide nanoparticles have been of great
interest due to their distinctive feature such as
catalytic activity, optical, magnetic and electrical
properties (Garima et al, 2010). Nanoparticles
interaction with biological materials and established a
series of nanoparticle / biological interfaces that
depend on colloidal forces as well as dynamic
biophysicochemical interactions. These interactions
lead to the formation of new nanomaterial with
control size shape, surface chemistry, roughness and
surface coatings.
The use of plants for the synthesis of nanoparticles is
novel and provides a cost
environmentally friendly alternative to chemical and
physical synthesis. In addition, the use of plants can
be easily scaled up for large-
the use of toxic chemicals or the need for high
pressures, energy and temperature
2017). Plant-mediated biosynthesis of zinc oxide
nanoparticles has been achieved in
(Sangeetha et al., 2011), Physalis alkekengi
2011), Parthenium hysterophorus
Zingiber officinale (Bhuyan et al
indica (Wang et al., 2014),
Medicago sativa (Nagaiyothi
Anisochilus carnosus (Bala et al
of Calotropis procera and fruit juice of
aurantifolia (Samat et al., 2013)
2018 Page: 36
6470 | www.ijtsrd.com | Volume - 2 | Issue – 2
Scientific
(IJTSRD)
International Open Access Journal
f Zinc Oxide
sing Onion Bulb Extract
S. Lega Priyatharsini
Post Graduate and Research Department
of Botany, Ayya Nadar Janaki Ammal
College, Sivakasi, Tamil Nadu, India
., 2011). The characters of metal
and metal oxide nanoparticles have been of great
interest due to their distinctive feature such as
catalytic activity, optical, magnetic and electrical
, 2010). Nanoparticles
ical materials and established a
series of nanoparticle / biological interfaces that
depend on colloidal forces as well as dynamic
biophysicochemical interactions. These interactions
lead to the formation of new nanomaterial with
e chemistry, roughness and
The use of plants for the synthesis of nanoparticles is
novel and provides a cost-effective and
environmentally friendly alternative to chemical and
physical synthesis. In addition, the use of plants can
-scale synthesis without
the use of toxic chemicals or the need for high
sures, energy and temperature (Bhainsa et al.,
mediated biosynthesis of zinc oxide
nanoparticles has been achieved in Aloe barbadensis
Physalis alkekengi (Qu et al.,
Parthenium hysterophorus (Rajiv et al., 2013),
et al., 2015), Azadirachta
., 2014), Ocimum basilicum,
(Nagaiyothi et al., 2013),
et al., 2015), milky latex
and fruit juice of Citrus
., 2013) etc.
International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456-6470
@ IJTSRD | Available Online @ www.ijtsrd.com | Volume – 2 | Issue – 2 | Jan-Feb 2018 Page: 37
It is known that the plant mediated synthesis of ZnO
nanoparticles is much safer and environmentally
friendly as compared to chemical synthesis. The size
of the synthesized ZnO nanoparticles was in the range
of 60-70 nm. The larger nanoparticles of ZnO resulted
from the agglomeration of smaller nanoparticles.
Highly ionic nanoparticulate metal oxides such as
zinc oxide nanoparticles are unique in that they can be
produced with high surface areas and with unusual
crystal structures (Anandraj et al., 2017). Compared
to organic materials, inorganic materials such as ZnO
possess superior durability, greater selectivity and
heat resistance (Padmavathy and Vijayaraghavan,
2008).
II. MATERIAL AND METHODS
SYNTHESIS OF ZINC OXIDE
NANOPARTICLES
The onion bulbs were washed with sterile distilled
water and the outer covering of the bulb was manually
peeled off and the fleshy part of the onion was
rewashed with sterile distilled water. The onion bulb
was cut into small pieces and 10g of bulb was ground
using mortar and pestle with distilled water. The
extraction was filtered using muslin cloth and then
Whatmann No.1 filter paper and used as reducing
agent and stabilizer. Zinc nitrate was used as
precursor for the synthesis of zinc oxide
nanoparticles. For ZnO NPs synthesis, the reaction
mixture was prepared by mixing of 10 ml of leaf
extract and 90 ml of 1mM zinc nitrate solution in a
250 ml Erlenmeyer conical flask. This mixture was
incubated in dark conditions at 37°C. Then, the
mixture was centrifuged at 10,000 rpm for 20
minutes. The pellet was taken after centrifugation and
air dried. The pellet was used for further studies.
RECOVERY OF ZnO NANOPARTICLES
The nanoparticles thus obtained were purified by
repeated centrifugation at 10000 rpm at 25°C for
10minutes. It was followed by re-dispersion of the
pellet in deionized water to get rid of any
uncoordinated biological molecules. The process of
centrifugation and re-dispersion were repeated with
sterile distilled water to ensure better separation of
free entities from the nanoparticles.
CHARACTERIZATION OF ZnO
NANOPARTICLES
The synthesized nanoparticles were characterized by
UV-Visible spectroscopy, Scanning Electron
Microscopy (SEM), Energy Dispersive X-ray
Spectroscopy Analysis (EDAX), Fourier Transform
Infrared Spectroscopy (FTIR), X-Ray Diffraction
(XRD) and Zeta potential analyses.
UV-VISIBLE SPECTROSCOPY ANALYSIS
The reduction of pure metal ions was confirmed by
measuring the absorption of reaction mixture by UV-
Vis Spectrophotometer (UV 1700 SHIMADZU) from
200 to 800 nm (Roy et al., 2013).
FOURIER TRANSFORM INFRARED (FTIR)
ANALYSIS
FTIR has become an important tool for understanding
the involvement of functional groups in interactions
between metal particles and biomolecules. FT-IR
spectra were recorded at 1 cm-1
resolution by FTIR
spectrophotometer (FTIR-8400S SHIMADZU) using
KBr pellet technique (Prathra et al., 2011). The
frequency range was measured as wave numbers
typically over the range 4000-400 cm-1
.
X-RAY DIFFRACTION (XRD) ANALYSIS
To determine the nature and size of the ZnO
nanoparticles, X-ray diffraction (XRD) was
performed. The pellet was dissolved in deionized
sterile water and washed thrice in the same by
centrifugation. The pellet was retained and air dried.
The powder from of the sample was coated on the
XRD grid, the spectra were recorded 40 kV and a
current of 30 mA with CuKα radiation using XRD
(Philips PW1050/37 model). The diffracted intensities
were recorded from 20°C to 80°at 2θ angles. The
crystalline nature of synthesized nanoparticles was
calculated from the width of the XRD peaks, using the
Debye-Scherrer formula (D= Kλ/β cosθ).
SCANNING ELECTRON MICROSCOPY (SEM)
ANALYSIS
The synthesized nanoparticles were dispersed in water
and the resultant suspensions were homogenized
using ultra sonicator for one to two hours. A drop of
the nanoparticles suspension was placed on a piece of
micro glass slide attached to a metal grid coated with
carbon film and dried it gradually at room
International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456-6470
@ IJTSRD | Available Online @ www.ijtsrd.com | Volume – 2 | Issue – 2 | Jan-Feb 2018 Page: 38
temperature. The sample was then sputter coated with
gold and visualized with a JEOL JSM-6480 LV SEM
to assess the particle size, shape and percentage
(Forough and Farhadi, 2010).
ENERGY DISPERSIVE X-RAY
SPECTROSCOPY (EDAX) ANALYSIS
A drop of the nanoparticles suspension was placed on
a piece of microglass slide attached to a metal grid
coated with carbon film, and dried it gradually at
room temperature. X-ray spectrometer (EDAX)
operated at an accelerating voltage at 10 KeV. The
sample was then sputter coated with gold and
visualized with a BRUKER to assess the particle size,
shape and percentage of synthesized particles.
ZETA POTENTIAL ANALYSIS
The supernatant was filtered and then sonicated for 5
minutes. The solution was centrifuged for 15 minutes
at 25°C with 5000 rpm and the supernatant was
collected. Then, the supernatant was diluted for 4 to 5
times and then set for Zeta-Potential analysis
(Mohammed et al., 2014).
ANTIMICROBIAL ACTIVITY
The ZnO nanoparticles were tested against the
bacterial pathogens such as E.coli (ATCC25922),
Staphylococcus aureus (ATCC9144), Pseudomonas
aeruginosa (ATCC25619) and fungal pathogens
Aspergillus niger (MTCC 1344) and Aspergillus
flavus (MTCC 1973). Agar disc diffusion method
(Baur et al., 2012) was employed for the study of
antimicrobial activity of the synthesized zinc oxide
nanoparticles.
STATISTICAL ANALYSIS
The XRD values were inferred through JCPDS file no
89-3722. Further, the UV-Visible spectrum, FTIR,
XRD values were interpreted by using ORIGIN
VERSION-8 (data analysis and graphing work space).
The data were reported as mean ± SE and in the figure
parentheses represent the per cent activity.
III. RESULTS & DISCUSSION
VISUAL OBSERVATION OF ZINC OXIDE
NANOPARTICLES
The synthesized zinc oxide nanoparticles were
initially confirmed by visual observation by colour
change in the reaction medium. The metal ions were
reduced during the exposure to aqueous extract of
onion within 24 hours of incubation period. It was
observed that the colour of the reaction medium was
changed to pale yellow to pale brown colour (Fig. 1).
UV-VISIBLE SPECTROSCOPY ANALYSIS
The reduction of zinc oxide ions in the onion extract
was further confirmed by UV-Vis Spectrophotometer.
UV-Vis absorption spectra of nanoparticles were
shown in (Fig. 2). The UV-Vis absorption spectrum of
ZnO NPs was as the peak maxima in 354 nm which
corresponding to the absorbance of zinc oxide
nanoparticles. After incubation period, the colour was
changed because of the excitation of Surface Plasmon
Vibrations in the ZnO nanoparticle. The reduction of
zinc was subjected to analysis by using the UV-Vis
Spectrophotometer. Absorption spectra of ZnO NPs
formed in the reaction media has absorbance peak at
310nm, broadening of peak indicated that the particles
are polydispersed. The frequency and width of the
surface plasmon absorption depends on the size and
shape of the metal nanoparticles as well as on the
dielectric constant of the metal itself and the
surrounding medium (Melvin et al., 2009).
The aqueous extract of Punica granatum and zinc
nitrate solution, the colour of the reaction medium
changed rapidly from colour less to brownish yellow.
The colour indicated surface plasmon vibration
typical of zinc oxide (ZnO) nanoparticles. The
maximum intensity at 364nm was observed indicating
complete reduction of zinc ions. The UV-Vis optical
absorption spectrum of ZnO nanoparticles showed a
sharp absorbance at 345 nm, which indicated an
almost uniform size of the nanoparticles. However,
upon change in particle size or particle shape, a slight
shift in the absorption was observed (Guo et al.,
2001). The UV-Vis absorption spectrum of ZnO
nanoparticles using flower extract of Nyctanthes
arbor-tristis showed a sharp absorbance at 369 nm
(Jamdagni et al., 2016).
FOURIER TRANSFORM INFRARED
SPECTROSCOPY (FTIR) ANALYSIS
FTIR analysis was carried out to identify the
biomolecules which were responsible for the
reduction of metal ions into ZnO nanoparticles in the
presence of onion extract (Fig. 3). The phytochemical
found in the onion extract were responsible for the
formation of various nanoparticles. The FT-IR
spectrum of onion extract showed several absorption
International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456-6470
@ IJTSRD | Available Online @ www.ijtsrd.com | Volume – 2 | Issue – 2 | Jan-Feb 2018 Page: 39
peaks ranged from 3421cm-1
to 677 cm-1
. The region
of band was phenols, alkanes due to N-H stretching of
proteins and O-H stretching, > C=O stretching of
esters, aromatics, ring C-C stretching of phenyl,
alkanes, C-O stretch in vibration combined with the
ring stretch of phenyl, aliphatic amines, alcohols,
carboxylic acids, ester, ether, functional groups
mainly from carbohydrate and alkyl halides. FTIR
measurements of zinc oxide nanoparticles with onion
extract showed the presence of bands at 3438.84,
2925.81, 1633.59, 1383.83, 1122.49, 1104.17 and
600.78 cm-1
. These bands were indicting the presence
of N-H bend primary amines, C-O stretching alcohols,
carboxylic acids, ester and ethers.
FTIR spectrum of ZnO nanoparticles revealed that the
peak at 417.52cm-1
was the characteristic absorption
of ZnO bond and the broad absorption peak at
3438cm-1
attributed to the characteristic absorption of
hydroxyl group (Khan et al., 2011). The FTIR
spectrums of ZnO NPs derived from Corymbia
citriodora leaf extract showed the strong absorption
peaks observed at 3300 and 1620cm−1
and were
assigned to O–H stretch and N–H bend functional
groups. Weaker bands observed at 2955, 777 and
633cm−1
were assigned to –C–H stretch (alkanes), C–
H (aromatics) and –C=C–H (alkynes). The absorption
peaks at 1520 and 1431cm−1
were attributed the C=C
bending and C–C stretching of aromatic rings,
respectively. The peak at 1053 cm−1
assigned to the
C–N stretching mode in aliphatic amines. The
formation of ZnO NPs may attribute to the chemicals
presence in the C. citriodora such as citronellal,
linalool, catechin, gallic acid, courmaric acid and
protocatechuric acid, which acted as reducing agents
as well as stabilizing agents (Tamuly et al., 2013).
The FTIR spectrum of ZnO nanoparticles derived
from Ocimum tenuiflorum showed the fundamental
mode of vibration at 3458.04cm−1
which
corresponded to the O-H stretching vibration,
1625.35cm−1
which correspond to the N-H bend,
1418.86cm−1
which corresponded to C-C stretching
vibration of alcohol, carboxlic acid, ether and ester
were confirmed. The band at 1148.10cm−1
corresponds to C-N symmetric stretching vibration
and O-H bending of the hydroxyl group at
3458.04cm−1
was observed. The absorption at
875.23cm−1
was due to the formation of tetrahedral
coordination of Zn. The bond at 835.61cm−1
was due
to the C-Cl stretching vibration. The peak at
668.29cm−1
indicated the stretching vibrations of ZnO
nano particle which was consistent with that reported
before (Ravichandrika et al., 2012). From FTIR
analysis of leaves extract of Ocimum tenuiflorum all
functional groups play important role in the
preparation of zinc oxide nanoparticles.
Green synthesis of ZnO nanoparticles using
Phyllanthus embilica stem extract revealed The FT-IR
spectra showed the presence of bonds due to O-H
stretching around 3423 cm-1
. Peak at 1405 cm-1
may
be assigned to symmetric stretching of the carbonyl
side groups in the amino acid residues of the protein
molecules. The band at 1022 cm-1
was corresponding
to C-N stretching vibration of amine. The peak at
around 1340 cm-1
present in green ZnO signified
amide III band of the random coil of protein (Joel and
Sheik, 2017).
X-RAY DIFFRACTION PATTERN ANALYSIS
The various peaks in the XRD pattern could be
assigned to the crystalline zinc oxide phase with the
hexagonal wurzite structure with the lattice
parameters a=3.2475 nm and c= 3.2501 nm. The XRD
pattern showed different intensity peaks in the whole
spectrum of 2θ values ranging from 20° to 80° for the
onion. The Allium cepa extract-mediated synthesized
ZnO nanoparticles were indexed as (100), (002),
(101), (102), (110), (103), (200), (112) and (201). The
zinc was indexed as (100) and (101). The average
crystalline size was determined using Scherrer’s
equation. The average crystalline size of zinc oxide
nanoparticles was 20-100 nm (Fig. 4). Zinc oxide
nanoparticles synthesized using Corymbia citriodora
leaf extract showed the peaks at 32.1°, 34.6°, 36.1°,
47.7°, 56.4°, 63.1° and 68.1° and indexed to
hexagonal wurtzite ZnO (JCPDS 36–1451). Further,
no other peaks have been detected in the sample and
confirmed the biosynthesis of ZnO NPs owning a high
purity. The mean crystalline size was found to be 21
nm (Yuhong Zheng et al., 2017). Biological synthesis
of zinc oxide nanoparticles from Catharanthus roseus
leaf extract showed strong diffraction peaks at 20, 32,
35 and 40 degrees of 2q which corresponds to 111,
200, 220 and 311 crystal planes, which were
significant agreement with the JCPDS file 36145 (a =
b = 3.249A0
, C = 5.206 A0
) and indexed as the
hexagonal wurtezite structure of ZnO. High purity and
crystallinity of the synthesized ZnO NPs were also
confirmed.
International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456-6470
@ IJTSRD | Available Online @ www.ijtsrd.com | Volume – 2 | Issue – 2 | Jan-Feb 2018 Page: 40
SCANNING ELECTRON MICROSCOPE (SEM)
ANALYSIS
The surface morphology of the nanoparticles was
characterized using Scanning Electron Microscopy.
The onion extract mediated ZnO nanoparticles were
found as nano-rod shape (Fig. 5). The biological
approach using milky latex of Calotropis procera has
been used for the first time as a reducing material as
well as surface stabilizing agent for the synthesis of
spherical-shaped ZnO NPs. The structure, phase and
morphology of synthesized product were investigated
by the SEM analysis (Sangeetha et al., 2011). Raut et
al. (2017) done the SEM analysis for zinc oxide
nanoparticles synthesized by Ocimum tenuiflorum leaf
extract. The SEM analysis showed the hexagonal
nanoparticle formed with diameter range 11-25nm.
ZnO nanoparticles synthesized using Trifolium
pratense flower extract revealed that SEM images of
the ZnO nanoparticles were agglomerated with a
particle size ranging from 100–190nm (Dobrucka and
Dugaszewska, 2015).
EDAX ANALYSIS
EDAX analysis is very much useful for further
confirmation of presence of nanoparticles. The
EDAX analysis showed the confirmative peaks for
ZnO NPs. The ZnO nanoparticles were confirmed by
typical absorption peak at 1KeV (Fig. 6). Sharma et
al. (2014) reported that the size of the nanoparticle is
more than that calculated from the Debye -Scherrer
formula indicating the agglomeration of crystallites in
ZnO nanoparticles. The aggregation of particles
should have been originated from the large specific
surface area and high surface energy of ZnO
nanoparticles. The aggregation occurred probably
during the process of drying (Raoufi, 2013). The
green synthesis of zinc oxide nanoparticles using
Moringa oleifera leaf extract the strong and weak
peaks are observed from Zn and O atom and weak
peaks are observed form S, K, C, P, Ca element
(Mishra et al., 2015). Biosynthesis and
characterization of ZnO nanoparticles using the
aqueous leaf extract of Imperata cylindrica the EDS
spectrum shows the high value of zinc (80.3%) and
oxygen (19.65%).
ZETA POTENTIAL ANALYSIS
Zeta potential measures the potential stability of the
nanoparticles in the colloidal suspension. From the
Zeta potential analysis, onion extract mediated ZnO
nanoparticles carried a charge of ±26.5 mV (Fig. 7).
The zeta potential of the biologically synthesized NPs
was determined in water as dispersant. The zeta
potential was found to be -20 to -30 mV for ZnO
nanoparticles. The high negative value approved the
repulsion among the particles and thereby increases in
stability of the structure of nanoparticles (Jafarirad et
al., 2016). It was suggested that the surface of the
nanoparticles was negatively charged and dispersed in
the medium. The negative value confirms the
repulsion among the particles and proves that they are
very stable (Anandalakshmi et al., 2016). The
biosynthesis of zinc oxide nanoparticles using Ixora
coccinea leaf extract zeta potential was found to be
−49.19 mV (Yedurkar et al., 2016). The synthesis and
characterisation of zinc oxide nanoparticles using
terpenoid fractions of Andrographis paniculata leaves
the zeta potential of synthesized ZnO nanoparticles
was 17.6 mV.
ANTIMICROBIAL ACTIVITY
Antimicrobial activity of onion extract mediated zinc
oxide nanoparticles were tested against bacterial
pathogens such as E.coli and Staphylococcus aureus,
Pseudomonas aeruginosa and two fungal pathogens
like Aspergillus niger and Aspergillus flavus. The
antibacterial activity (inhibition zone) of zinc oxide
nanoparticles was higher against Pseudomonas
aeruginosa followed by Staphylococcus aureus and
E.coli. In the case of fungal pathogens, the maximum
inhibition zone was observed against A. niger (Table
1).
Nanoparticles possess the antimicrobial properties
against several microorganisms including bacteria,
fungi and certain extremophiles (Haeed et al., 2008).
The properties of ZnO nanoparticle is strongly
influenced by the particle size and mechanism of cell
inhibition. The properties include disruption of cell
membrane, altering the permeability, electrostatically
binding to the cell surface and accumulation of
nanoparticle in cytoplasm (Stoimenov et al., 2002).
Antimicrobial activity of zinc oxide nanoparticles
were known from the very distant past and has many
applications in disinfecting medical devices, water
purification, and wound healing, creams, lotions and
antibacterial creams. The mechanism of action of
antimicrobial activity zinc oxide is similar to other
nanoparticles, but it acts mostly through destruction of
bacterial walls. Zinc oxide nanoparticles have been
widely used against Gram-positive and Gram negative
bacteria (Emami et al., 2013).
International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456-6470
@ IJTSRD | Available Online @ www.ijtsrd.com | Volume – 2 | Issue – 2 | Jan-Feb 2018 Page: 41
The mechanisms of antibacterial activity of ZnO NPs
were direct contact of ZnO NPs with cell walls,
resulting in destructing bacterial cell integrity,
liberation of antimicrobial ions mainly Zn+2
ions.
However, the toxicity mechanism varies in various
media as the species of dissolved Zn may change
according to the medium components besides the
physicochemical properties of ZnO NPs (Yedurkar et
al., 2016). Nano-sized ZnO exhibited varying
morphologies and showed a significant antibacterial
activity over a wide spectrum of bacterial species
explored by a large body of researchers. ZnO is
currently being investigated as an antibacterial agent
in both microscale and nanoscale formulations. ZnO
exhibited significant antimicrobial activities when
particle size is reduced to the nanometer range, then
nano-sized ZnO interacted with bacterial surface
and/or with the bacterial core where it entered inside
the cell and subsequently exhibited distinct
bactericidal mechanisms (Seil and Webster, 2012).
CONCLUSIONS
The biological production of metal nanoparticles is
becoming a very important field in chemistry, biology
and materials science. Metal nanoparticles have been
produced chemically and physically for a long time;
however, their biological production has only been
investigated very recently. The biological reduction of
metals by plant extracts has been known since the
early 1900s; however, the reduction products were not
studied. The rapid biological synthesis of zinc
nanoparticles using bulb extract of Allium cepa
provides an environmental friendly, simple and
efficient route for synthesis of nanoparticles. The use
of plant extracts avoids the usage of harmful and toxic
reducing and stabilizing agents. The synthesized nano
particles of ZnO are in the range of 20-100 nm. Zinc
nanoparticles can exist in ions only in the presence of
strong oxidizing substances. The synthesis of ZnO
nano particles is still in its infancy and more research
needs to be focused on the mechanism of nanoparticle
formation which may lead to fine tuning of the
process ultimately leading to the synthesis of
nanoparticles with a strict control over the size and
shape parameters.
ACKNOWLEDGEMENT
The authors are thankful to the Management and the
Principal of Ayya Nadar Janaki Ammal College,
Sivakasi, Tamil Nadu, India for providing laboratory
facilities to carry out the experiment.
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International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456-6470
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Onion Extract ZnNO3 ZnO NPs
Fig. 1: Visual observation of synthesis of Zinc
oxide nanoparticles
Fig. 2: UV-Visible spectrum of Zinc oxide
nanoparticles
Fig. 3: FTIR spectrum of Zinc oxide nanoparticles
Fig. 4: XRD spectrum of the Zinc oxide
nanoparticles
Fig. 5: SEM micrograph of Zinc oxide
nanoparticles at different magnifications
Fig. 6: EDAX spectrum of Zinc oxide
nanoparticles
Fig. 7: Zeta potential spectrum of Zinc oxide
nanoparticles
Table 1: Antimicrobial activity of Zinc Oxide nanoparticles
Microorganism
Diameter of inhibition zone (mm)
Onion Extract Zinc oxide
nanoparticles
E. coli 0.91
±0.02
1.9
±0.04
P. aeruginosa 1.72
±0.03
2.5
±0.04
S. aureus 1.6
±0.04
2.4
±0.07
A. niger 1.2
±0.09
2.9
±0.03
A. flavus 1.6
±0.09
2.2
±0.02

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Biosynthesis and Characterization of Zinc Oxide Nanoparticles using Onion Bulb Extract

  • 1. @ IJTSRD | Available Online @ www.ijtsrd.com ISSN No: 2456 International Research Biosynthesis and Characterization o Nanoparticles u N. Tensingh Baliah Post Graduate and Research Department of Botany, Ayya Nadar Janaki Ammal College, Sivakasi, Tamil Nadu, India ABSTRACT The wide application of nanoparticles stimulates the need for synthesizing them but, the conventional methods are usually hazardous and energy consuming. This leads to focus on “green synthesis” of nanoparticles which seems to be easy efficient and ecofriendly approach. In this study, the plant mediated synthesis of zinc oxide nanoparticles (ZnO NPs) was carried out using bulb extract of Allium cepa as a reducing agent. The optimized nano zinc thus obtained was quantified and characterized using UV Visible spectroscopy, Fourier Transform Infrared Spectroscopy (FT-IR), X-Ray diffraction, Electron Microscope (SEM), EDAX potential analyses. Further, the synthesized ZnO NPs were tested for antimicrobial activity. Keywords: Nanoparticles, zinc oxide, Allium cepa I. INTRODUCTION Nanotechnology is a emerging field with its application in science and technology for the purpose of manufacturing new materials at nano scale level (Albrecht et al., 2006). Recent advance in the field of nanotechnology, particularly the ability to prepare highly ordered nanoparticles of any size and shape (Sangeetha et al., 2012). Nanotechnology is a multidisciplinary scientific field undergoing explosive development. The nanometer-sized particles offer novel structural, optical and electronic properties that are not attainable with individual molecules or bulk @ IJTSRD | Available Online @ www.ijtsrd.com | Volume – 2 | Issue – 2 | Jan-Feb ISSN No: 2456 - 6470 | www.ijtsrd.com | Volume International Journal of Trend in Scientific Research and Development (IJTSRD) International Open Access Journal Biosynthesis and Characterization of Zinc Oxide Nanoparticles using Onion Bulb Extract Post Graduate and Research Department of Botany, Ayya Nadar Janaki Ammal College, Sivakasi, Tamil Nadu, India S. Lega Priyatharsini Post Graduate and Research Department of Botany, Ayya Nadar Janaki Ammal College, Sivakasi, Tamil Nadu, India application of nanoparticles stimulates the need for synthesizing them but, the conventional methods are usually hazardous and energy consuming. This leads to focus on “green synthesis” of nanoparticles which seems to be easy efficient and oach. In this study, the plant mediated synthesis of zinc oxide nanoparticles (ZnO NPs) was carried out using bulb extract of Allium cepa as a reducing agent. The optimized nano zinc thus obtained was quantified and characterized using UV- Fourier Transform Infrared Ray diffraction, Scanning EDAX and Zeta . Further, the synthesized ZnO NPs Nanoparticles, zinc oxide, biosynthesis, Nanotechnology is a emerging field with its application in science and technology for the purpose of manufacturing new materials at nano scale level ., 2006). Recent advance in the field of ogy, particularly the ability to prepare particles of any size and shape ., 2012). Nanotechnology is a multidisciplinary scientific field undergoing explosive sized particles offer ral, optical and electronic properties that are not attainable with individual molecules or bulk solids (Anitha et al., 2011). The characters of metal and metal oxide nanoparticles have been of great interest due to their distinctive feature such as catalytic activity, optical, magnetic and electrical properties (Garima et al, 2010). Nanoparticles interaction with biological materials and established a series of nanoparticle / biological interfaces that depend on colloidal forces as well as dynamic biophysicochemical interactions. These interactions lead to the formation of new nanomaterial with control size shape, surface chemistry, roughness and surface coatings. The use of plants for the synthesis of nanoparticles is novel and provides a cost environmentally friendly alternative to chemical and physical synthesis. In addition, the use of plants can be easily scaled up for large- the use of toxic chemicals or the need for high pressures, energy and temperature 2017). Plant-mediated biosynthesis of zinc oxide nanoparticles has been achieved in (Sangeetha et al., 2011), Physalis alkekengi 2011), Parthenium hysterophorus Zingiber officinale (Bhuyan et al indica (Wang et al., 2014), Medicago sativa (Nagaiyothi Anisochilus carnosus (Bala et al of Calotropis procera and fruit juice of aurantifolia (Samat et al., 2013) 2018 Page: 36 6470 | www.ijtsrd.com | Volume - 2 | Issue – 2 Scientific (IJTSRD) International Open Access Journal f Zinc Oxide sing Onion Bulb Extract S. Lega Priyatharsini Post Graduate and Research Department of Botany, Ayya Nadar Janaki Ammal College, Sivakasi, Tamil Nadu, India ., 2011). The characters of metal and metal oxide nanoparticles have been of great interest due to their distinctive feature such as catalytic activity, optical, magnetic and electrical , 2010). Nanoparticles ical materials and established a series of nanoparticle / biological interfaces that depend on colloidal forces as well as dynamic biophysicochemical interactions. These interactions lead to the formation of new nanomaterial with e chemistry, roughness and The use of plants for the synthesis of nanoparticles is novel and provides a cost-effective and environmentally friendly alternative to chemical and physical synthesis. In addition, the use of plants can -scale synthesis without the use of toxic chemicals or the need for high sures, energy and temperature (Bhainsa et al., mediated biosynthesis of zinc oxide nanoparticles has been achieved in Aloe barbadensis Physalis alkekengi (Qu et al., Parthenium hysterophorus (Rajiv et al., 2013), et al., 2015), Azadirachta ., 2014), Ocimum basilicum, (Nagaiyothi et al., 2013), et al., 2015), milky latex and fruit juice of Citrus ., 2013) etc.
  • 2. International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456-6470 @ IJTSRD | Available Online @ www.ijtsrd.com | Volume – 2 | Issue – 2 | Jan-Feb 2018 Page: 37 It is known that the plant mediated synthesis of ZnO nanoparticles is much safer and environmentally friendly as compared to chemical synthesis. The size of the synthesized ZnO nanoparticles was in the range of 60-70 nm. The larger nanoparticles of ZnO resulted from the agglomeration of smaller nanoparticles. Highly ionic nanoparticulate metal oxides such as zinc oxide nanoparticles are unique in that they can be produced with high surface areas and with unusual crystal structures (Anandraj et al., 2017). Compared to organic materials, inorganic materials such as ZnO possess superior durability, greater selectivity and heat resistance (Padmavathy and Vijayaraghavan, 2008). II. MATERIAL AND METHODS SYNTHESIS OF ZINC OXIDE NANOPARTICLES The onion bulbs were washed with sterile distilled water and the outer covering of the bulb was manually peeled off and the fleshy part of the onion was rewashed with sterile distilled water. The onion bulb was cut into small pieces and 10g of bulb was ground using mortar and pestle with distilled water. The extraction was filtered using muslin cloth and then Whatmann No.1 filter paper and used as reducing agent and stabilizer. Zinc nitrate was used as precursor for the synthesis of zinc oxide nanoparticles. For ZnO NPs synthesis, the reaction mixture was prepared by mixing of 10 ml of leaf extract and 90 ml of 1mM zinc nitrate solution in a 250 ml Erlenmeyer conical flask. This mixture was incubated in dark conditions at 37°C. Then, the mixture was centrifuged at 10,000 rpm for 20 minutes. The pellet was taken after centrifugation and air dried. The pellet was used for further studies. RECOVERY OF ZnO NANOPARTICLES The nanoparticles thus obtained were purified by repeated centrifugation at 10000 rpm at 25°C for 10minutes. It was followed by re-dispersion of the pellet in deionized water to get rid of any uncoordinated biological molecules. The process of centrifugation and re-dispersion were repeated with sterile distilled water to ensure better separation of free entities from the nanoparticles. CHARACTERIZATION OF ZnO NANOPARTICLES The synthesized nanoparticles were characterized by UV-Visible spectroscopy, Scanning Electron Microscopy (SEM), Energy Dispersive X-ray Spectroscopy Analysis (EDAX), Fourier Transform Infrared Spectroscopy (FTIR), X-Ray Diffraction (XRD) and Zeta potential analyses. UV-VISIBLE SPECTROSCOPY ANALYSIS The reduction of pure metal ions was confirmed by measuring the absorption of reaction mixture by UV- Vis Spectrophotometer (UV 1700 SHIMADZU) from 200 to 800 nm (Roy et al., 2013). FOURIER TRANSFORM INFRARED (FTIR) ANALYSIS FTIR has become an important tool for understanding the involvement of functional groups in interactions between metal particles and biomolecules. FT-IR spectra were recorded at 1 cm-1 resolution by FTIR spectrophotometer (FTIR-8400S SHIMADZU) using KBr pellet technique (Prathra et al., 2011). The frequency range was measured as wave numbers typically over the range 4000-400 cm-1 . X-RAY DIFFRACTION (XRD) ANALYSIS To determine the nature and size of the ZnO nanoparticles, X-ray diffraction (XRD) was performed. The pellet was dissolved in deionized sterile water and washed thrice in the same by centrifugation. The pellet was retained and air dried. The powder from of the sample was coated on the XRD grid, the spectra were recorded 40 kV and a current of 30 mA with CuKα radiation using XRD (Philips PW1050/37 model). The diffracted intensities were recorded from 20°C to 80°at 2θ angles. The crystalline nature of synthesized nanoparticles was calculated from the width of the XRD peaks, using the Debye-Scherrer formula (D= Kλ/β cosθ). SCANNING ELECTRON MICROSCOPY (SEM) ANALYSIS The synthesized nanoparticles were dispersed in water and the resultant suspensions were homogenized using ultra sonicator for one to two hours. A drop of the nanoparticles suspension was placed on a piece of micro glass slide attached to a metal grid coated with carbon film and dried it gradually at room
  • 3. International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456-6470 @ IJTSRD | Available Online @ www.ijtsrd.com | Volume – 2 | Issue – 2 | Jan-Feb 2018 Page: 38 temperature. The sample was then sputter coated with gold and visualized with a JEOL JSM-6480 LV SEM to assess the particle size, shape and percentage (Forough and Farhadi, 2010). ENERGY DISPERSIVE X-RAY SPECTROSCOPY (EDAX) ANALYSIS A drop of the nanoparticles suspension was placed on a piece of microglass slide attached to a metal grid coated with carbon film, and dried it gradually at room temperature. X-ray spectrometer (EDAX) operated at an accelerating voltage at 10 KeV. The sample was then sputter coated with gold and visualized with a BRUKER to assess the particle size, shape and percentage of synthesized particles. ZETA POTENTIAL ANALYSIS The supernatant was filtered and then sonicated for 5 minutes. The solution was centrifuged for 15 minutes at 25°C with 5000 rpm and the supernatant was collected. Then, the supernatant was diluted for 4 to 5 times and then set for Zeta-Potential analysis (Mohammed et al., 2014). ANTIMICROBIAL ACTIVITY The ZnO nanoparticles were tested against the bacterial pathogens such as E.coli (ATCC25922), Staphylococcus aureus (ATCC9144), Pseudomonas aeruginosa (ATCC25619) and fungal pathogens Aspergillus niger (MTCC 1344) and Aspergillus flavus (MTCC 1973). Agar disc diffusion method (Baur et al., 2012) was employed for the study of antimicrobial activity of the synthesized zinc oxide nanoparticles. STATISTICAL ANALYSIS The XRD values were inferred through JCPDS file no 89-3722. Further, the UV-Visible spectrum, FTIR, XRD values were interpreted by using ORIGIN VERSION-8 (data analysis and graphing work space). The data were reported as mean ± SE and in the figure parentheses represent the per cent activity. III. RESULTS & DISCUSSION VISUAL OBSERVATION OF ZINC OXIDE NANOPARTICLES The synthesized zinc oxide nanoparticles were initially confirmed by visual observation by colour change in the reaction medium. The metal ions were reduced during the exposure to aqueous extract of onion within 24 hours of incubation period. It was observed that the colour of the reaction medium was changed to pale yellow to pale brown colour (Fig. 1). UV-VISIBLE SPECTROSCOPY ANALYSIS The reduction of zinc oxide ions in the onion extract was further confirmed by UV-Vis Spectrophotometer. UV-Vis absorption spectra of nanoparticles were shown in (Fig. 2). The UV-Vis absorption spectrum of ZnO NPs was as the peak maxima in 354 nm which corresponding to the absorbance of zinc oxide nanoparticles. After incubation period, the colour was changed because of the excitation of Surface Plasmon Vibrations in the ZnO nanoparticle. The reduction of zinc was subjected to analysis by using the UV-Vis Spectrophotometer. Absorption spectra of ZnO NPs formed in the reaction media has absorbance peak at 310nm, broadening of peak indicated that the particles are polydispersed. The frequency and width of the surface plasmon absorption depends on the size and shape of the metal nanoparticles as well as on the dielectric constant of the metal itself and the surrounding medium (Melvin et al., 2009). The aqueous extract of Punica granatum and zinc nitrate solution, the colour of the reaction medium changed rapidly from colour less to brownish yellow. The colour indicated surface plasmon vibration typical of zinc oxide (ZnO) nanoparticles. The maximum intensity at 364nm was observed indicating complete reduction of zinc ions. The UV-Vis optical absorption spectrum of ZnO nanoparticles showed a sharp absorbance at 345 nm, which indicated an almost uniform size of the nanoparticles. However, upon change in particle size or particle shape, a slight shift in the absorption was observed (Guo et al., 2001). The UV-Vis absorption spectrum of ZnO nanoparticles using flower extract of Nyctanthes arbor-tristis showed a sharp absorbance at 369 nm (Jamdagni et al., 2016). FOURIER TRANSFORM INFRARED SPECTROSCOPY (FTIR) ANALYSIS FTIR analysis was carried out to identify the biomolecules which were responsible for the reduction of metal ions into ZnO nanoparticles in the presence of onion extract (Fig. 3). The phytochemical found in the onion extract were responsible for the formation of various nanoparticles. The FT-IR spectrum of onion extract showed several absorption
  • 4. International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456-6470 @ IJTSRD | Available Online @ www.ijtsrd.com | Volume – 2 | Issue – 2 | Jan-Feb 2018 Page: 39 peaks ranged from 3421cm-1 to 677 cm-1 . The region of band was phenols, alkanes due to N-H stretching of proteins and O-H stretching, > C=O stretching of esters, aromatics, ring C-C stretching of phenyl, alkanes, C-O stretch in vibration combined with the ring stretch of phenyl, aliphatic amines, alcohols, carboxylic acids, ester, ether, functional groups mainly from carbohydrate and alkyl halides. FTIR measurements of zinc oxide nanoparticles with onion extract showed the presence of bands at 3438.84, 2925.81, 1633.59, 1383.83, 1122.49, 1104.17 and 600.78 cm-1 . These bands were indicting the presence of N-H bend primary amines, C-O stretching alcohols, carboxylic acids, ester and ethers. FTIR spectrum of ZnO nanoparticles revealed that the peak at 417.52cm-1 was the characteristic absorption of ZnO bond and the broad absorption peak at 3438cm-1 attributed to the characteristic absorption of hydroxyl group (Khan et al., 2011). The FTIR spectrums of ZnO NPs derived from Corymbia citriodora leaf extract showed the strong absorption peaks observed at 3300 and 1620cm−1 and were assigned to O–H stretch and N–H bend functional groups. Weaker bands observed at 2955, 777 and 633cm−1 were assigned to –C–H stretch (alkanes), C– H (aromatics) and –C=C–H (alkynes). The absorption peaks at 1520 and 1431cm−1 were attributed the C=C bending and C–C stretching of aromatic rings, respectively. The peak at 1053 cm−1 assigned to the C–N stretching mode in aliphatic amines. The formation of ZnO NPs may attribute to the chemicals presence in the C. citriodora such as citronellal, linalool, catechin, gallic acid, courmaric acid and protocatechuric acid, which acted as reducing agents as well as stabilizing agents (Tamuly et al., 2013). The FTIR spectrum of ZnO nanoparticles derived from Ocimum tenuiflorum showed the fundamental mode of vibration at 3458.04cm−1 which corresponded to the O-H stretching vibration, 1625.35cm−1 which correspond to the N-H bend, 1418.86cm−1 which corresponded to C-C stretching vibration of alcohol, carboxlic acid, ether and ester were confirmed. The band at 1148.10cm−1 corresponds to C-N symmetric stretching vibration and O-H bending of the hydroxyl group at 3458.04cm−1 was observed. The absorption at 875.23cm−1 was due to the formation of tetrahedral coordination of Zn. The bond at 835.61cm−1 was due to the C-Cl stretching vibration. The peak at 668.29cm−1 indicated the stretching vibrations of ZnO nano particle which was consistent with that reported before (Ravichandrika et al., 2012). From FTIR analysis of leaves extract of Ocimum tenuiflorum all functional groups play important role in the preparation of zinc oxide nanoparticles. Green synthesis of ZnO nanoparticles using Phyllanthus embilica stem extract revealed The FT-IR spectra showed the presence of bonds due to O-H stretching around 3423 cm-1 . Peak at 1405 cm-1 may be assigned to symmetric stretching of the carbonyl side groups in the amino acid residues of the protein molecules. The band at 1022 cm-1 was corresponding to C-N stretching vibration of amine. The peak at around 1340 cm-1 present in green ZnO signified amide III band of the random coil of protein (Joel and Sheik, 2017). X-RAY DIFFRACTION PATTERN ANALYSIS The various peaks in the XRD pattern could be assigned to the crystalline zinc oxide phase with the hexagonal wurzite structure with the lattice parameters a=3.2475 nm and c= 3.2501 nm. The XRD pattern showed different intensity peaks in the whole spectrum of 2θ values ranging from 20° to 80° for the onion. The Allium cepa extract-mediated synthesized ZnO nanoparticles were indexed as (100), (002), (101), (102), (110), (103), (200), (112) and (201). The zinc was indexed as (100) and (101). The average crystalline size was determined using Scherrer’s equation. The average crystalline size of zinc oxide nanoparticles was 20-100 nm (Fig. 4). Zinc oxide nanoparticles synthesized using Corymbia citriodora leaf extract showed the peaks at 32.1°, 34.6°, 36.1°, 47.7°, 56.4°, 63.1° and 68.1° and indexed to hexagonal wurtzite ZnO (JCPDS 36–1451). Further, no other peaks have been detected in the sample and confirmed the biosynthesis of ZnO NPs owning a high purity. The mean crystalline size was found to be 21 nm (Yuhong Zheng et al., 2017). Biological synthesis of zinc oxide nanoparticles from Catharanthus roseus leaf extract showed strong diffraction peaks at 20, 32, 35 and 40 degrees of 2q which corresponds to 111, 200, 220 and 311 crystal planes, which were significant agreement with the JCPDS file 36145 (a = b = 3.249A0 , C = 5.206 A0 ) and indexed as the hexagonal wurtezite structure of ZnO. High purity and crystallinity of the synthesized ZnO NPs were also confirmed.
  • 5. International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456-6470 @ IJTSRD | Available Online @ www.ijtsrd.com | Volume – 2 | Issue – 2 | Jan-Feb 2018 Page: 40 SCANNING ELECTRON MICROSCOPE (SEM) ANALYSIS The surface morphology of the nanoparticles was characterized using Scanning Electron Microscopy. The onion extract mediated ZnO nanoparticles were found as nano-rod shape (Fig. 5). The biological approach using milky latex of Calotropis procera has been used for the first time as a reducing material as well as surface stabilizing agent for the synthesis of spherical-shaped ZnO NPs. The structure, phase and morphology of synthesized product were investigated by the SEM analysis (Sangeetha et al., 2011). Raut et al. (2017) done the SEM analysis for zinc oxide nanoparticles synthesized by Ocimum tenuiflorum leaf extract. The SEM analysis showed the hexagonal nanoparticle formed with diameter range 11-25nm. ZnO nanoparticles synthesized using Trifolium pratense flower extract revealed that SEM images of the ZnO nanoparticles were agglomerated with a particle size ranging from 100–190nm (Dobrucka and Dugaszewska, 2015). EDAX ANALYSIS EDAX analysis is very much useful for further confirmation of presence of nanoparticles. The EDAX analysis showed the confirmative peaks for ZnO NPs. The ZnO nanoparticles were confirmed by typical absorption peak at 1KeV (Fig. 6). Sharma et al. (2014) reported that the size of the nanoparticle is more than that calculated from the Debye -Scherrer formula indicating the agglomeration of crystallites in ZnO nanoparticles. The aggregation of particles should have been originated from the large specific surface area and high surface energy of ZnO nanoparticles. The aggregation occurred probably during the process of drying (Raoufi, 2013). The green synthesis of zinc oxide nanoparticles using Moringa oleifera leaf extract the strong and weak peaks are observed from Zn and O atom and weak peaks are observed form S, K, C, P, Ca element (Mishra et al., 2015). Biosynthesis and characterization of ZnO nanoparticles using the aqueous leaf extract of Imperata cylindrica the EDS spectrum shows the high value of zinc (80.3%) and oxygen (19.65%). ZETA POTENTIAL ANALYSIS Zeta potential measures the potential stability of the nanoparticles in the colloidal suspension. From the Zeta potential analysis, onion extract mediated ZnO nanoparticles carried a charge of ±26.5 mV (Fig. 7). The zeta potential of the biologically synthesized NPs was determined in water as dispersant. The zeta potential was found to be -20 to -30 mV for ZnO nanoparticles. The high negative value approved the repulsion among the particles and thereby increases in stability of the structure of nanoparticles (Jafarirad et al., 2016). It was suggested that the surface of the nanoparticles was negatively charged and dispersed in the medium. The negative value confirms the repulsion among the particles and proves that they are very stable (Anandalakshmi et al., 2016). The biosynthesis of zinc oxide nanoparticles using Ixora coccinea leaf extract zeta potential was found to be −49.19 mV (Yedurkar et al., 2016). The synthesis and characterisation of zinc oxide nanoparticles using terpenoid fractions of Andrographis paniculata leaves the zeta potential of synthesized ZnO nanoparticles was 17.6 mV. ANTIMICROBIAL ACTIVITY Antimicrobial activity of onion extract mediated zinc oxide nanoparticles were tested against bacterial pathogens such as E.coli and Staphylococcus aureus, Pseudomonas aeruginosa and two fungal pathogens like Aspergillus niger and Aspergillus flavus. The antibacterial activity (inhibition zone) of zinc oxide nanoparticles was higher against Pseudomonas aeruginosa followed by Staphylococcus aureus and E.coli. In the case of fungal pathogens, the maximum inhibition zone was observed against A. niger (Table 1). Nanoparticles possess the antimicrobial properties against several microorganisms including bacteria, fungi and certain extremophiles (Haeed et al., 2008). The properties of ZnO nanoparticle is strongly influenced by the particle size and mechanism of cell inhibition. The properties include disruption of cell membrane, altering the permeability, electrostatically binding to the cell surface and accumulation of nanoparticle in cytoplasm (Stoimenov et al., 2002). Antimicrobial activity of zinc oxide nanoparticles were known from the very distant past and has many applications in disinfecting medical devices, water purification, and wound healing, creams, lotions and antibacterial creams. The mechanism of action of antimicrobial activity zinc oxide is similar to other nanoparticles, but it acts mostly through destruction of bacterial walls. Zinc oxide nanoparticles have been widely used against Gram-positive and Gram negative bacteria (Emami et al., 2013).
  • 6. International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456-6470 @ IJTSRD | Available Online @ www.ijtsrd.com | Volume – 2 | Issue – 2 | Jan-Feb 2018 Page: 41 The mechanisms of antibacterial activity of ZnO NPs were direct contact of ZnO NPs with cell walls, resulting in destructing bacterial cell integrity, liberation of antimicrobial ions mainly Zn+2 ions. However, the toxicity mechanism varies in various media as the species of dissolved Zn may change according to the medium components besides the physicochemical properties of ZnO NPs (Yedurkar et al., 2016). Nano-sized ZnO exhibited varying morphologies and showed a significant antibacterial activity over a wide spectrum of bacterial species explored by a large body of researchers. ZnO is currently being investigated as an antibacterial agent in both microscale and nanoscale formulations. ZnO exhibited significant antimicrobial activities when particle size is reduced to the nanometer range, then nano-sized ZnO interacted with bacterial surface and/or with the bacterial core where it entered inside the cell and subsequently exhibited distinct bactericidal mechanisms (Seil and Webster, 2012). CONCLUSIONS The biological production of metal nanoparticles is becoming a very important field in chemistry, biology and materials science. Metal nanoparticles have been produced chemically and physically for a long time; however, their biological production has only been investigated very recently. The biological reduction of metals by plant extracts has been known since the early 1900s; however, the reduction products were not studied. The rapid biological synthesis of zinc nanoparticles using bulb extract of Allium cepa provides an environmental friendly, simple and efficient route for synthesis of nanoparticles. The use of plant extracts avoids the usage of harmful and toxic reducing and stabilizing agents. The synthesized nano particles of ZnO are in the range of 20-100 nm. Zinc nanoparticles can exist in ions only in the presence of strong oxidizing substances. The synthesis of ZnO nano particles is still in its infancy and more research needs to be focused on the mechanism of nanoparticle formation which may lead to fine tuning of the process ultimately leading to the synthesis of nanoparticles with a strict control over the size and shape parameters. ACKNOWLEDGEMENT The authors are thankful to the Management and the Principal of Ayya Nadar Janaki Ammal College, Sivakasi, Tamil Nadu, India for providing laboratory facilities to carry out the experiment. REFERENCES 1. Albrecht, M. A., Evan, C. W. & Raston, C. L. (2006). Green chemistry and the health implications of nanoparticles. Green Chemistry, 3(8), 417-432. 2. Anand Raj, L.F.A. & Jayalakshmy, E. (2017). Biosynthesis and characterization of zinc oxide nanoparticles using root extract of Zingiber officinale. J.Chem, 31, 51-56. 3. Anandalakshmi, K., Venugobal, J. & Ramasamy, V. (2016). Characterization of zinc oxide nanoparticles by green synthesis method using Pedalium murex leaf extract and their antibacterial activity. Appl. Nanosci. 6(2), 399. 4. Anitha, S. Kiruba, D., Singhal, G. (2011). Bhavesh, R., Kunalkasaraiya, M., AshishRajanan, S. & Singh R. Biosythesis of zinc oxide nanoparticles using Ocimum sanctum (Tulsi) leaf extract and screening its antimicrobial activity, J. Nanopart. Res, 13(9), 2981-2988. 5. Bala, N., Saha, S., Chakraborty, M., Maiti, M., Das, S., Basub, R. & Nandyc, P. (2015). Green synthesis of zinc oxide nanoparticles using Hibiscus subdariffa leaf extract: Effect of temperature on synthesis, anti-Bacterial activity and anti-diabetic activity. Int. J. Mol. Sci, 5, 4993- 5003. 6. Baur, R.W., Kirby, M. D. K., Sherris, J. C. & Turck, M. (2012). Antibiotic susceptibility testing by standard single disc diffusion method. Americ. J. Clin. Pathol. 45, 493-496. 7. Bhainsa K. C. & Souza, D. (2017). Extracellular biosynthesis of zinc oxide nanoparticles using the fungus Aspergillus fumigates. Biosurfaces, 47(2), 160-164. 8. Bhuyan, T., Mishra, K., Khanuja, M. & Prasad, R. (2015). Biosynthesis of zinc oxide nanoparticles from Azadirachta indica for antibacterial and photocatalytic applications. Mat. Sci. Semiconduct. Process, 32, 55-56. 9. Dobruckaand, R. & Dugaszewska, J. (2015). Biosynthesis and antibacterial activity of ZnO nanoparticles using Trifolium pratense flower extract. Saudi J. Biol. Sci. 23, 517-523. 10. Emami, S., Akya, A., Hossain Zadeh, A. & Barkhordar, S. (2013). Bacterial contamination of traditional ice creams in Kermanshah. Iran J. Med. Microbiol. 7(2), 59–62. 11. Forough, M. & Farhadi, K. (2010). Biological and green synthesis of zinc oxide nanoparticles. Turkish J. Eng. Env. Sci. 34, 281 – 287.
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  • 8. International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456-6470 @ IJTSRD | Available Online @ www.ijtsrd.com | Volume – 2 | Issue – 2 | Jan-Feb 2018 Page: 43 35. Sharama, G., Jasuja, N. D., Ali, M. I. & Joshi, S. C. (2014). A review on nanomedicinal and nanosensing potential of nanoparticles. Int. J. Biol. Chem, 8 (2), 58-84. 36. Stoimenov P. K., Klinger R. L., Marchin G. L., & Klabunde K. J. 2002. Metal oxide nanoparticles as bactericidal agents. Langmuir 18(17), 6679- 6686. 37. Tamuly, M., Hazarikaa, S., Borah, M. R., Das & M. P. Boruah. (2013). In situ biosynthesis of Ag, Au and bimetallic nanoparticles using Piper pedicellatum: Green chemistry approach. Colloids Surf. B Biointerf. 102, 627–634. 38. Wang, X.X., Wu, L., Zhou, P., Li, C., Zhao, L.B., An. W. and Chen, Y. (2014) Effect of ZnO nanoparticles on Medicago sativa at the germination stage. Appl. Mechan. Mat. 665, 583- 586. 39. Yedurkar, S., Maurya, C. & Mahanwar P. (2016). Biosynthesis of Zinc Oxide nanoparticles using Ixora coccinea leaf extract - a green approach. Open J. Syn. Theo. Appli.5, 1-14. 40. Yuhong Zheng, Li Fu, Fuigui Han, Aiwu Wang, Wen Cai, Jinping Yu, Jun Yang & Feng Peng (2017). Green biosynthesis and characterization of zinc oxide nanoparticles using Corymbia citriodora leaf extract and their photocatalytic activity. Green Chem. Lett. Rev. 8(2), 59–63. Onion Extract ZnNO3 ZnO NPs Fig. 1: Visual observation of synthesis of Zinc oxide nanoparticles Fig. 2: UV-Visible spectrum of Zinc oxide nanoparticles Fig. 3: FTIR spectrum of Zinc oxide nanoparticles Fig. 4: XRD spectrum of the Zinc oxide nanoparticles
  • 9. Fig. 5: SEM micrograph of Zinc oxide nanoparticles at different magnifications Fig. 6: EDAX spectrum of Zinc oxide nanoparticles Fig. 7: Zeta potential spectrum of Zinc oxide nanoparticles Table 1: Antimicrobial activity of Zinc Oxide nanoparticles Microorganism Diameter of inhibition zone (mm) Onion Extract Zinc oxide nanoparticles E. coli 0.91 ±0.02 1.9 ±0.04 P. aeruginosa 1.72 ±0.03 2.5 ±0.04 S. aureus 1.6 ±0.04 2.4 ±0.07 A. niger 1.2 ±0.09 2.9 ±0.03 A. flavus 1.6 ±0.09 2.2 ±0.02