bionanotech-presentation1.pptx

bionanotech-presentation1.pptx

• 1.
  BETA VULGARIS MEDIATEDBIO-INSPIRED COPPER NANOPARTICLE and ITs APPLICATION AS ANTIBACTERIAL AGENT AYESHA ANDLEEB
• 2.
  Why copper nanoparticles..?? Biosynthesis of silver and gold nanoparticles as antibacterial agents have been reported but as Both the silver and gold are expensive metals, and lower resistant towards the migration of ions. • So, copper nano-materials, are an area of attraction; as copper is: • Cost effective, • highly abundant in nature, • has a high melting point, • has low electrochemical migration behavior, • has chemical stability and possesses catalytic and antimicrobial properties. • In our bodies; it is important for various metabolic activities. • acts as a cofactor for various enzymes • cell signaling and defense mechanism. • It is one of the important elements for plant growth as well.  So, Cu nanoparticles can be a substitute for noble metals.
• 3.
  “Green Nanoparticles” • Useof plant or plant parts for the bio-reduction of metal ions into their elemental form in the size range 1–100 nm. • So, plants parts have extensively been used for the biosynthesis of NPs as secondary metabolites are abundant in the plants. • The process is more efficient, simpler, economical, and can easily be scaled up. • Aqueous extract of plant materials can reduce size and provide stability. • Choice of solvent, eco-friendly, reducing agent and stabilizing agents are essential considerations. • In the reference paper, green technology was adopted for the biosynthesis of Cu nanoparticles. • Aqueous extract of Beta Vulgaris to explore its antimicrobial properties against three bacterial strains. • Beta vulgaris is enriched with flavonoids, carotenoids, limonene, sugars etc. that has the capability to reduce and stabilize the metallic Cu to its NPs.
• 4.
  Material and reagents •Analytical grade chemicals were used for the synthesis of beta vulgaris mediated nanoparticle synthesis including: • CuSO4. 5H2O, • ethanol, • peptone, • yeast powder, • Muller agar and • NaCl.
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  Beta vulgaris extract •Beta Vulgaris was crushed in a mortar and pestle. • To the groundmass, about 70 ml of double distilled water was added and stirred at 40°C on a magnetic stirrer for about 10-15 min. • Then, it was filtered and centrifuged for about 15 min. The obtained supernatant was used as plant extract throughout the experiment.
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  • 20 mlof copper salt solution (1 mM) was mixed with 5 ml of Beta Vulgaris extract. • It was stirred on a magnetic stirrer for 1 hr. • A change in color of the reaction mixture is the primary confirmation of nanoparticles. • The formation of Cu nanoparticles was indicated by the color change of Beta Vulgaris extract from blood red to deep brown. • The spectroscopic studies were made in the UV-Vis Spectrophotometer. Color Change Showing the Visual Formation of Cu NPs Cuso4 solution Beet root+ CuSO4 CU-NPS
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  Antibacterial Activity • Thedisk diffusion method was adopted to assess the antibacterial efficacy of nanoparticles. • The nutrient media was prepared in double distilled water. • Approximately 20 ml of sterile molten and cooled media was poured into pre-sterilized Petri dishes. • Plates were left overnight at 35°C to check if any contamination. • After inoculation of 3 different target bacteria, sterile paper discs of 3 mm diameter were impregnated with 50μl copper nanoparticles placed on each plate. • Chloramphenicol and tetracycline were used as the standard. • The experimental plates were then incubated at 35°C for 24 hours and 48 hours along with the control.
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  Result interpretation UV-Vis SpectralAnalysis Scanning Electron Microscopy (SEM) EDAX (electron dispersive X-ray spectroscopy) Fourier Transform Infrared Spectroscopy (FT-IR) Observation of Antibacterial activity
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  UV-Vis Spectral Analysis •The preliminary detection was studied by UV-Vis spectrophotometer. • It is an indirect method to study the bio reduction of an aqueous solution of copper salt to copper NPs. • The UV-Visible spectrum showed an absorption maximum band (SPR, λspr) in the range of 550- 575 nm as a result of collective electron oscillation around the surface mode of the particles and confirmed the formation of Cu nanoparticles (Cu NPs). UV-Visible Spectrum of Cu-NPs
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  Scanning Electron Microscopy(SEM) • The morphology and size of synthesized copper NPs using Beta Vulgaris extract were studied by Scanning Electron Microscopy. • The SEM micrograph showed the mono-dispersed nature of Cu NPs mostly having a cubical shape and few are spherical with the size ranging from 35 to 60 nm SEM Image of Biologically Synthesized Copper Nanoparticles
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  EDAX (electron dispersive X-rayspectroscopy) • The elemental analysis of the synthesized sample was done using electron dispersive X-ray spectroscopy. • EDAX image showed the characteristic peaks of metallic copper along with peaks of C and O which might be due to the presence of biomolecules (betalains) that are involved in the stabilization of Cu NPs. • It suggests that nanoparticles were synthesized successfully from an aqueous solution of Beta Vulgaris. EDAX of Copper Nanoparticle
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  Fourier TransformInfraredSpectroscopy • FT-IRis an important technique to identify the biomolecules that are involved in the reduction and stabilization of the synthesized Cu NPs. • It was recorded in the range from 4,000 to 400 cm-1. Broad band observed at 3400 cm-1 was assigned to the O-H stretching vibration that represents the high concentration of alcoholic groups present in Beta Vulgaris. • The hydroxyl groups present in the biomolecules have a stronger ability to bind with copper ions and are involved in the green synthesis of Cu NPs. The peaks at 2900 cm-1 and 2976 cm-1 showed symmetric & asymmetric C-H stretching vibrations. The carbonyl group band appeared at 1654 cm-1 indicating the presence of aldehydes, ketones or carboxylic groups. FT-IR Spectrum of Copper NPs
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  Observation of Antibacterialactivity •The biologically synthesized copper nanoparticles were tested against three bacterial strains; Bacillus subtilis (gram +ve), Escherichia Coli (gram -ve), and Promicromonospora. • Figure shows that the zone of inhibition was maximum for E. Coli and minimum for Bacillus subtilis . • The exact mechanism of the antibacterial activity of Cu NPs is uncertain. • It is believed that the Cu NPs get attached to Sulphur and -COOH group of amino acids that lead to the deactivation of the enzymes essential for DNA biosynthesis. Zone of Inhibition Diameter (mm) Promicrospora 18±2 E. coli 20±2 Bacillus subtilis 7±2
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  Conclusions • An environmentallybenign method was proposed to synthesize Cu NPs using Beta Vulgaris aqueous extract and without using any external stabilizer. • Copper is prone to oxidation and therefore, the synthesis of copper NPs is generally carried out in a non-aqueous medium. • Plenty of betalains in Beta Vulgaris might be imparting additional stability to the synthesized Cu NPs. • The synthesized product was confirmed by FT-IR, UV-Visible spectrophotometer, SEM and EDAX spectroscopic techniques. • The particles of the size range 35-60 nm were obtained and could be further controlled by adopting suitable reaction conditions. • The growth of E. coli was suppressed significantly with copper NPs. • Biologically synthesized copper NPs showed good antibacterial activity.