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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.
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”.
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).
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.
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.
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.
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.
• 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.
• 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.
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
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.
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
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.
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
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
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

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Mugunthan 2nd Msc physics Bio-Fabrication of Zinc Oxide NPs from Aegle marmelos leaf extract and their Antibacterial activity

  • 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