The document describes the bio-fabrication and characterization of zinc oxide nanoparticles synthesized from Aegle marmelos leaf extract, and their antibacterial activity. Zinc oxide nanoparticles were synthesized using a green chemistry approach involving the reduction of zinc ions to zinc oxide nanoparticles using the leaf extract. The nanoparticles were characterized using XRD, FTIR, and UV-Vis spectroscopy. XRD showed the nanoparticles had a hexagonal wurtzite crystal structure with an average size of 19nm. FTIR confirmed the formation of zinc oxide. UV-Vis showed high absorbance in the UV region. Disc diffusion assays found the nanoparticles had antibacterial activity against E. coli and Bacillus cereus.

1. Bio-Fabrication of Zinc Oxide NPs
from Aegle marmelos leaf extract and their
Antibacterial activity
SUBMITTED BY:
K. MUGUNTHAN,
MSc PHYSICS II YEAR,
BHARAT INSTITUTE OF HIGHER EDUCATION AND
RESEARCH
GUIDED BY:
Dr. S. ANNADHI, MSc, PhD,
HEAD OF THE DEPARTMENT,
DEPARTMENT OF PHYSICS,
BHARAT INSTITUTE OF HIGHER EDUCATION AND
RESEARCH.

2. INTRODUCTION
NANOSCIENCE
• Nanoscience is the study of structures and materials on an ultra-small scale, and the unique and
interesting properties these materials demonstrate.
• Nanoscience cover those field of physics, biology, medicine, chemistry, are studying and using for
those application.
NANOTECHNOLOGY
• Nanotechnology as “a science, engineering, and technology conducted at the nanoscale (1 to 100
nm), where unique phenomena enable novel applications in a wide range of fields, from chemistry,
physics and biology, to medicine, engineering and electronics”.

3. NANOPARTICLES
• The word nano is comes from ancient greek language and means dwarf. the prefix nano is
one billionth and its denoted by the factor 10-9 m or 0.000000001m.
• Nanoparticles are incredibly tiny material between 1 to 100 nm.
• 1nm = 10-9 m = 0.000000001m (1 billionth).

4. CLASSIFICATION OF NANOPARTICLES
ZERO DIMENSIONS: zero-dimensional (0d) nanomaterials, including graphene quantum dots
(gqds), carbon quantum dots (cqds), fullerenes, inorganic quantum dots (qds), magnetic
nanoparticles (mnps), noble metal nanoparticles, up conversion nanoparticles (ucnps) and polymer
dots (p dots), have attracted extensive research
1-D NANOMATERIALS 1-D nanomaterials have thin films or surface coatings and are used in the
circuitry of computer chips and for anti-reflective properties and hard coatings on eyeglasses.

5. 2D - NANOMATERIALS
2D nanomaterials have fixed and long nanostructures with thick membranes. They are used to prepare nanopore filters used
for small particle separation and filtration.
3D - NANOMATERIALS
Three-dimensional nanomaterials (3D) are materials that are not confined to the nanoscale in any dimension. This class can
contain bulk powders, dispersions of nanoparticles, bundles of nanowires, and nanotubes as well as multi-nanolayers.

6. SYNTHESIS APPROACHES
TOP-DOWN APPROACH:
Top-down approach begin with larger
particles which are reduced to nanoparticles after a
sequence of operations performed over them.
BOTTOM-UPAPPROACH
bottom - up approaches of production of
nanomaterials comprise the miniaturization of
materials constituents to the atomic.

7. MATERIALS AND METHODS
• MATERIALS:
All chemicals used in this study were of ar grade with 99% purity (merck and sd fine chemicals) and
were used without further purification. To synthesize zno and transition metals, all glassware used in
this experimental work was washed with acid to obtain, zinc nitrate hexahydrate (zn(no3)2.6h2o),
and acetone.
• SYNTHESIS OF PURE ZNO NANOPARTICLES:
The Zno nanoparticles were synthesized using chemical a precipitation method.
• PREPARATION OF (AEGLE MARMELOS) LEAF EXTRACT:
Fresh (aegle marmelos) (0.25%) 20 g of leaf was added to 100 ml sterile distilled water, magnetically
stirred at 800 c for 1 h, and then cooled at room temperature. finally, the extract was filtered through
whatman filter no.1 paper and stored at 40 c for further synthesis.

8. • X-RAY DIFFRACTION (XRD)
X-ray diffraction is a rapid analytical technique primarily used for the phase identification of crystalline materials. it
can provide information on the unit cell dimensions.
• FOURIER TRANSFORM INFRARED SPECTROSCOPY (FTIR)
Fourier transform infrared spectroscopy relies on the fact that most molecules absorb light in the
infrared region of the electromagnetic spectrum. this absorption corresponds to the bonds present in the
molecule. the frequency range is typically measured as wave numbers over the range of 4000 – 600
cm-1.

9. • UV-SPECTROSCOPY (UV-VIS)
Ultraviolet (uv) spectroscopy is a type of spectroscopy that uses light in the ultraviolet region of the
electromagnetic spectrum to study the absorption, transmission, and reflection of materials. The
principles of uv spectroscopy are based on the fact that molecules absorb light in the uv region of the
electromagnetic spectrum owing to the excitation of electrons from their ground state to higher energy
states.

10. ANTIFUNGALACTIVITY ASSAY
AGAR DISC DIFFUSION METHOD:
The antifungal activity of the extracts was
determined by the disc diffusion method on the
sabouraud dextrose agar (sda) medium. sabouraud
dextrose agar (sda) medium was poured into a petri plate.
after the medium solidified, the inoculums were spread
on solid plates with sterile swabs moistened with the
fungal suspension. The Samples were then placed over a
plate. amphotericin-b was used as the positive control.
the plates were then incubated at 37 ºc for 24 h. the
antifungal activity was determined by measuring the
diameter of the inhibition zone.
Zn(NO3)26H2O
Stirring 2 hours at 800 c
(600 rpm)
Annealing at 5000C for 4
hours
Precipitate of Zno NPs
40c 5 hours
NaOH
Stirring 2 hours in
deionized water
100 ml of Deionized water
&20 grams of leaf
800c 1 hour heat
Adding Drop by drop of leaf extract

11. RESULT AND DISCUSSION
THE X-RAY DIFFRACTION ANALYSIS:
The powder x-ray diffraction view of Zno is shown in fig. 1. a certain line
sharpening of xrd peaks indicated that the synthesized material particles were good
crystal structure.
Fig. 1. X-ray diffraction spectrum of synthesized Zinc
oxide nanoparticles.

12. FOURIER TRANSFORM INFRARED SPECTROSCOPY
FT-IR gives the composition and formation of functional groups of the synthesized ZnO
nanoparticles. it also suggests that the formation of ZnO nanoparticles is due to the interaction of the
phenolic compounds, alkynes, terpenoids, and flavonoids. fig. 2 represents FT-IR spectra of the
synthesized ZnO nanoparticles in the range 400–4000 cm-1. the functional groups were responsible for
reducing zinc ions to ZnO, which was observed as bands. each of the bands corresponds to various
stretching modes.
FT-IR SPECTRUM OF SYNTHESIZED ZINC OXIDE NANOPARTICLES.
RESULT AND DISCUSSION

13. ULTRAVIOLET–VISIBLE SPECTROSCOPY
A UV-VIS spectroscopic study of the absorption spectrum in the range of wavelengths of 200–800 nm was
used to determine the optical characteristics of ZnO nps made by using an extract from (aegle marmelos), as
shown in figure 3. fig. 3 shows that the dominant (strong) feature of all ZnO nps produced at high absorbance
in the uv region below 375 nm and a low absorbance rate in the visible range
RESULT AND DISCUSSION
UV–spectra for Zinc oxide nanoparticles Tauc’s plot of green synthesized ZnO-NPs.

14. ANTI BACTERIAL ASSAY
STATISTICAL ANALYSIS
The results of the 5 trials were given as mean± sd. one/two-way anova was followed by using pad prism
V.8.0 software version. the antimicrobial activity was statistically analyzed to know the level of significance at
P<0.001, P<0.01 and P<0.05. P value less than 0.05, 0.01, 0.001 was considered statistically significant.
RESULT AND DISCUSSION
Bacillus cereus E.coli
0
5
10
15
20
Concentrations
Zone
of
inhibition
in
mm
125(µg/ml)
250(µg/ml)
500(µg/ml)
Standard (5µg/ml)
E.coli Bacillus cereus
Antibacterial activity for different pathogens

15. The zinc-oxide nanoparticle was synthesized by using the biological approach and the usage of aegle
marmelos leaf extract effectively. The green synthesis method is eco-friendly, in comparison to the alternative
methods along with physical and chemical. The XRD examinations of synthesized ZnO-nps showed the formation
of a hexagonal-wurtzite shape. the common crystalline size of the nanoparticles was found to be 19 nm. the band
at 962 cm−1 in FT-IR-spectrum confirms the formation of ZnO-nps. the UV-spectrum is the dominant (strong)
feature of all ZnO-nps, produced at high absorbance inside the UV-region 375 nm, and the received price of the
optical band gap is 3.08 ev. Similarly, ZnO has a large hobby against e. coli and bacillus cereus with a quarter of
inhibition values reported. Biosynthesized ZnO nps prepared from aegle marmelos are expected to have
remarkable packages in the pharmaceutical and biomedical fields together with drug delivery and in cosmetic
industries.
CONCLUSION

16. 1. FOWSIYA, I.V. ASHARANI, SIMRON MOHAPATRA, AKHILA ESHAPULA, PRAGYA MOHI, NISHANT THAKAR, SUPRIYO
MONAD AND MADHUMITHA, AEGLE MARMELOS PHYTOCHEMICAL STABILIZEDSYNTHESIS AND CHARACTERIZATION OF
ZNONANOPARTICLES AND THEIR ROLE AGAINSTAGRICULTURE AND FOOD PATHOGEN. FEBRUARY 21, 2019
2. MOHAMED TAHA YASSIN, ASHRAF ABDEL-FATTAH MOSTAFA, ABDULAZIZ ABDULRAHMAN AL-ASKAR AND FATIMAH O.
AL-OTIBI, FACILE GREEN SYNTHESIS OF ZINC OXIDE NANOPARTICLES WITH POTENTIAL SYNERGISTIC ACTIVITY WITH
COMMON ANTIFUNGAL AGENTS AGAINST MULTIDRUG-RESISTANT CANDIDAL STRAINS. MAY 26, 2022
3. T.KAVIYARASI, B.MUTHULAKSHMI, C. KAVITHAGREEN SYNTHESIS AND CHARACTERIZATION OF ZINC NANOPARTICLE
USING AEGLE MARMELOS LEAF EXTRACT,2017
4. C.JOEL1 AND M. SHEIK MUHIDEEN BADHUSHA, GREEN SYNTHESIS OF ZNO NANOPARTICLES USING PHYLLANTHUS
EMBILICA STEM EXTRACT AND THEIR ANTIBACTERIAL ACTIVITY, 2016
5. M.S. GEETHA A, H. NAGABHUSHANA B, H.N. SHIVANANJAIAH, GREEN MEDIATED SYNTHESIS AND CHARACTERIZATION OF
ZNO NANOPARTICLES USING EUPHORBIA JATROPA LATEX AS REDUCING AGENT, JUNE 23,2016
REFERENCE