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  1. 1. Biosynthesis and characterisation of metal oxide nanoparticle from pomegranate Presented by S.Vincy Jeba Malar Under the guidance of Dr.R.D.Femitha
  2. 2. 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”.
  3. 3. 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.
  4. 4. 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
  5. 5. 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
  6. 6. 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
  7. 7. 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
  8. 8. 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
  9. 9. 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)
  10. 10. 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
  11. 11. 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)
  12. 12. 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)
  13. 13. 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)
  14. 14. 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
  15. 15. 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 )
  16. 16. 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
  17. 17. 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
  18. 18. 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
  19. 19. 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 )
  20. 20. 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
  21. 21. 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
  22. 22. 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
  23. 23. 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
  24. 24. 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
  25. 25. 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
  26. 26. 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
  27. 27. 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.
  28. 28. 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.
  29. 29. 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.
  30. 30. 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.
  31. 31. 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.
  32. 32. 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.
  33. 33. 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.
  34. 34. 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
  35. 35. 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.
  36. 36. 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.
  37. 37. 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.
  38. 38. SEM image of iron oxide nanoparticle from pomegranate peel SEM showed relatively spherical shape nanoparticle formed with diameter range 15 nm.
  39. 39. 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.
  40. 40. 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.
  41. 41. 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.
  42. 42. 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.
  43. 43. 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.
  44. 44. Antimicrobial activity of copper oxide nanoparticle using pomegranate
  45. 45. 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
  46. 46. Antibacterial activity of iron oxide nanoparticle using pomegranate
  47. 47. 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.
  48. 48. 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
  49. 49. 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.

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