Nanoparticle

2. Biosynthesis and characterisation of
metal oxide nanoparticle from
pomegranate
Presented by
S.Vincy Jeba Malar
Under the guidance of
Dr.R.D.Femitha

3. Introduction
 In nanotechnology, a nanoparticle (10-9m) is defined as a small object that behaves as
a whole unit in terms of its transport and properties
 The science and engineering of nanosystems is one of the most challenging and
fastest growing sectors of nanotechnology.
 The first scientific description of the properties of nanoparticles was provide in 1857
by Michael Faraday in his famous paper “Experimental relations of gold to light”.
 The 1950’s and the 1960’s saw the world turning its focus towards the use of
nanoparticles in the field of drug delivery.
 The word pomegranate literaly means “seeded apple”.
 The word “pomegranate (Punica grannatum”) came from the Latin for “fruit many
seeds”.

4. Scope of the Research Work
 An important challenge in technology is to modify optical, electric and electronic
properties of nanoparticles by controlling their size and shape.
 Biomimetic synthesis of nanoparticles has opened its doors to a world of
nanoparticles with easy preparation protocols, less toxicity and a wide range of
applications according to their size and shape.
 Nanoparticles of desired size and shape have been obtained successfully using
plants extract.
 The field of nano biotechnology is still in its infancy and more research needs to be
focused on the mechanistics of nanoparticle formation which may lead to fine tuning
of the process ultimately leading to the biosynthesis of nanoparticles with a strict
control over the size and shape parameters.

5. Objective of the Research Work
 To biosynthesis nanoparticle by using plant extract
 To characterize the nanoparticles by UV-visible spectroscopy
 To find out wavelength of nanoparticles
 To characterize the nanoparticles by FTIR spectroscopy
 To characterize the nanoparticles by XRD
 To characterize the nanoparticles by AFM
 To characterize the nanoparticles by SEM
 To characterize the nanoparticles by TEM
 To study the anti microbial activity of the nanoparticles

6. Biosynthesis of copper oxide nanoparticle
Biosynthesis of copper oxide from seed extract
25gm of pomegranate seed crushed with 100 ml of double distilled water
4.9 gm of copper sulphate solution is made up to 200 ml SMF
10 ml of seed extract is added with 90 ml of copper sulphate solution
pink colour solution change to blue colour

7. Biosynthesis of copper oxide from peel extract
25gm of pomegranate peel crushed with 100 ml of double distilled water
4.9 gm of copper sulphate solution is made up to 200 ml SMF
10 ml of seed extract is added with 90 ml of copper sulphate solution
yellow colour solution change to green colour

8. Biosynthesis of iron oxide nanoparticle
Biosynthesis of Iron oxide from seed extract
25gm of pomegranate seed crushed with 100 ml of double distilled water
5.2 gm of ferric chloride solution is made up to 200 ml SMF
10 ml of seed extract is added with 90 ml of ferric chloride solution
yellow colour solution change to brown colour

9. Biosynthesis of Iron oxide from peel extract
25gm of pomegranate peel crushed with 100 ml of double distilled water
5.2 gm of ferric chloride solution is made up to 200 ml SMF
10 ml of peel extract is added with 90 ml of ferric chloride solution
pink colour solution change to brown colour

10. Characterization of copper oxide nanoparticle
UV-Vis spectroscopy
Fourier Transform Infra Red Spectroscopy(FTIR)
X-Ray diffraction
Transmission Electron Microscopy(TEM)
Scanning Electron Microscopy(SEM)
Atomic Force Microscopy(AFM)

11. UV visible spectroscopy
UV-Visible spectra of copper oxide nanoparticle from
pomegranate seed extract
300 400 500 600
0.5
1.0
1.5
2.0
2.5
Absorbance
Wavelength(nm)
370

12. UV-Visible spectra of copper oxide nanoparticle from
pomegranate peel extract
300 400 500 600
0.1
0.2
0.3
0.4
0.5
377
Absorbance
Wavelength(nm)

13. UV-Visible spectra of iron oxide nanoparticle
from pomegranate seed extract
100 200 300 400 500 600 700 800 900
0
1
2
3
4Absorbance
290
wavelength(nm)

14. UV-Visible spectra of iron oxide nanoparticle from
pomegranate peel extract
100 200 300 400 500 600 700 800
0.0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
251
282
Absorbance
wavelength(nm)

15. UV-Visible Spectral data λmax for copper and iron oxide from
seed and peel
Sample Material λmax
Copper oxide Seed 370
Peel 377
Iron oxide Seed 290
Peel 282

16. FTIR spectrum of pomegranate seed
4000 3500 3000 2500 2000 1500 1000 500
-5
0
5
10
15
20
25
30
35
40
910
23502950
3550 1620
1440
%Transmittance
wavelength(cm
-1
)

17. FTIR spectrum of pomegranate seed
Plant Characteristic
Absorption
(cm-1)
Bond Functional
group
1440 O-H Alcohol
Seed 1620 C=C Alkenyl
2950 C-H Alkyl
3550 O-H (strong) Alcohol

18. FTIR spectrum of copper oxide nanoparticle synthesized
by pomegranate seed extract
4000 3500 3000 2500 2000 1500 1000 500
0
5
10
15
20
25
30
7741440
32003550
2950
2350
2000
1620
1720
%Transmittance
wavelength(cm)
-1

19. FTIR spectrum of copper oxide nanoparticle from
pomegranate seed extract
Plant Characteristic
Absorption
(cm-1)
Bond Functional
group
1440 O-H Alcohol
Seed 1620 C=C Alkenyl
1720 C=O Aldehyde
2000 C=C=N Ketenimine
2950 C-H Alkyl
3200 O-H (weak) Alcohol
3550 O-H (strong) Alcohol

20. FTIR spectrum of pomegranate peel
4000 3500 3000 2500 2000 1500 1000 500
0
5
10
15
20
25
30
3700 1750
1440
1120
%Transmittance
wavelength(cm
-1
)

21. FTIR spectrum of pomegranate peel
Plant Characteristic
Absorption
(cm-1)
Bond Functional
group
1120 C=C Alkene
Peel 1440 S=O Sulfone
1750 O-H Carboxylic acid
3700 O-H Alcohol

22. FTIR spectrum of copper oxide nanoparticle synthesized
by pomegranate peel extract
4500 4000 3500 3000 2500 2000 1500 1000 500
-5
0
5
10
15
20
25
850
730
1120
1440
2500
3350
3700
2349
1750
1640
%Transmittance
wavelength(cm)
-1

23. FTIR spectrum of copper oxide nanoparticle
from pomegranate peel extract
Plant Characteristic
Absorption
(cm-1)
Bond Functional
group
730 C=C Alkene
Seed 1120 S=O Sulfone
1440 O-H Carboxylic acid
1750 C=O Ester
2349 O=C=O Carbon dioxide
2500 O-H (broad) Carboxylic acid
3350 N-H Secondary amine
3700 O-H Alcohol

24. FTIR spectrum of iron oxide nanoparticle synthesized
by pomegranate seed extract
4500 4000 3500 3000 2500 2000 1500 1000 500
-5
0
5
10
15
20
25
30
35
40
3400
2360
789
1030
1370
1500
1640
1720
3000
%Transmittance
wavelength(cm)
-1

25. FTIR spectrum of iron oxide nanoparticle from
pomegranate seed extract
Plant Characteristic
Absorption
(cm-1)
Bond Functional
group
1500 C=C Aromatic
Seed 1640 C=N Oxime
1720 C=O Carboxylic
acid
3000 N-H Amine salt
3400 N-H Primary amine

26. FTIR spectrum of iron oxide nanoparticle synthesized
by pomegranate peel extract
4500 4000 3500 3000 2500 2000 1500 1000 500
-5
0
5
10
15
20
25
30
35
3700
1750
1030
1440
1720
2370
2800
3400
%Transmittance
wavelength(cm)
-1

27. FTIR spectrum of iron oxide nanoparticle from
pomegranate peel extract
Plant Characteristic
Absorption
(cm-1)
Bond Functional
group
1030 S=O Sulfoxide
Peel 1440 O-H Carboxylic acid
1750 C=O Ester
2800 N-H Amine salt
3400 N-H Primary amine
3700 O-H Alcohol

28. XRD pattern of synthesized copper oxide
nanoparticle from Pomegranate Seed
The average diameter of the copper oxide nanoparticles is calculated
in the range 20nm by Scherrer formula.

29. XRD pattern of synthesized copper oxide
nanoparticle from Pomegranate Peel
The average diameter of the copper oxide nanoparticles is calculated
in the range 24 nm by Scherrer formula.

30. XRD pattern of synthesized iron oxide nanoparticle
from Pomegranate seed
The average diameter of the iron oxide nanoparticles is calculated
in the range 16 nm by Scherrer formula.

31. XRD pattern of synthesized iron oxide nanoparticle from
Pomegranate peel
The average diameter of the iron oxide nanoparticles is calculated in the
range 12nm by Scherrer formula.

32. TEM image of copper oxide nanoparticle from
Pomegranate Seed
The TEM monographs clearly show the distribution of spherical copper
oxide nanoparticles prepared by pomegranate seed extract.
The copper oxide nanoparticles were homogeneous and agglomerated
with a particle size of 20nm.

33. TEM image of copper oxide nanoparticle from
Pomegranate peel
The copper oxide nanoparticles are irregular and spherical in
morphology with a particle size of 24 nm.

34. TEM image of iron oxide nanoparticle from
Pomegranate Seed
The TEM image of iron oxide nanoparticles synthesized using
ferric chloride stabilized by pomegranate seed extract shows the size of iron
oxide nanoparticles is 16 nm.

35. TEM image of iron oxide nanoparticle from Pomegranate
peel
TEM micrograph of the synthesized iron oxide nanoparticle is suggest
that the particle are mostly spherical shape. The particle size 14nm.

36. SEM image of copper oxide nanoparticle from
pomegranate seed
Scanning electron microscope of the pomegranate seed extract treated
with copper oxide shows spherical and square like structure with diameter range
18 nm.

37. SEM image of copper oxide nanoparticle from
pomegranate peel
The formation of copper oxide nanoparticles as well as their
morphological dimensions in the SEM study demonstrated the size 24nm.

38. SEM image of iron oxide nanoparticle from pomegranate seed
Scanning electron microscope of the pomegranate seed extract treated with
iron oxide shows spherical and square like structure with diameter range 16nm.

39. SEM image of iron oxide nanoparticle from
pomegranate peel
SEM showed relatively spherical shape nanoparticle formed with
diameter range 15 nm.

40. 2D and 3D structure of copper oxide nanoparticle from
pomegranate seed
The surface topology of bio-synthesized copper oxide nanoparticles was
analyzed by AFM analysis. The average grain size was found to be 19.5nm.

41. 2D and 3D structure of copper oxide nanoparticle
from pomegranate peel
The topography of AFM micrographs clearly indicated that the formulated
copper oxide nanoparticle possess spherical shape, size range 24.5 nm.

42. 2D and 3D structure of iron oxide nanoparticle
from pomegranate seed
The average grain size was found to be 16.4 nm of 3D
profile image.

43. 2D and 3D structure of iron oxide nanoparticle from
pomegranate peel
The surface topology of bio-synthesized iron oxide nanoparticles was
analyzed by AFM analysis. The average grain size was found to be 14.5nm.

44. Sample Staphylococcus
albus
Bacillus
cereus
Pseudomonas proteus
Seed 11 13 11 R
peel 14 17 13 R
Control R R R R
Standard Disk
(AMIKACIN)
16 19 16 16
Copper oxide nanoparticle
Pomegranate seed exhibited potential antibacterial maximum ZOI
was found to be 13 mm for Bacillus cereus .whereas, the other three bacterial
strains of Pseudomonas, Staphylococcus albus , proteus showed ZOI of 11,11,0
mm.
Pomegranate peel exhibited potential antibacterial Maximum ZOI
was found to be 17mm for Bacillus Cereus whereas, the other three bacterial
strains of Pseudomonas, Staphylococcus albus. proteus showed ZOI of 14,13,0
mm.

45. Antimicrobial activity of copper oxide nanoparticle
using pomegranate

46. Anti microbial activity copper oxide nanoparticle
using pomegranate seed,peel extract
0
1
2
3
4
5
6
seed peel control standard
Series 1
Series 2
Series 3

47. Antibacterial activity of iron oxide nanoparticle using
pomegranate

48. Sample Staphylococcus
albus
Bacillus cereus Pseudomonas proteus
Seed 9 11 9 R
peel 11 13 10 R
Control R R R R
Standard Disk
(AMIKACIN)
16 19 16 16
Iron oxide nanoparticle
Pomegranate seed exhibited potential antibacterial maximum ZOI
was found to be 11 mm for Bacillus cereus. whereas, the other three
bacterial strains of Pseudomonas, Staphylococcus albus , proteus showed
ZOI of 9,9,0 mm.
Pomegranate peel exhibited potential antibacterial maximum ZOI
was found to be 13mm for Bacillus cereus whereas, the other three bacterial
strains of Pseudomonas, Staphylococcus albus, proteus showed ZOI of
11,10,0 mm.

49. Anti microbial activity iron oxide nanoparticle using
pomegranate seed, peel extract
0
1
2
3
4
5
6
Seed peel control standard
series 1
series 2
series 3

50. Conclusion
The metal oxide of copper and iron oxide nanoparticles were synthesized
using by pomegranate seed and peel extract.
The absorption peak of copper oxide and iron oxide nanoparticles formed in
the reaction media has absorbance peak around 380nm and 300nm.
XRD study the copper and iron oxide nanoparticles are crystalline in nature
and the size is around 24nm.
TEM analysis reveals that the copper and iron oxide nanoparticles were
spherical in shape and the size is around 24nm.
The surface morphology of the copper and iron oxide nanoparticle was
observed from the SEM analysis. The average size from 15 nm to24 nm.
Atomic force morphology reveals that the copper and iron oxide
nanoparticles are spherical shape and size ranging between 14 nm to 25 nm.
The copper and iron oxide nanoparticles were evaluated for antibacterial
activities against human pathogens viz. Bacillus cereus, pseudomonas and
staphylococcus albus, proteus. Bacillus cereus show good antimicrobial
activity compare with other human pathogens.