project presentation on nanoparticles biosynthesis

Nanoparticles

1. BIOSYNTHESIS OF CuO /Fe 2 O 3 NANOCOMPOSITE USING Raphanus Sativus AND ITS ADSORPTION STUDIES

2. STEPS UNDERTAKEN DURING PROJECT Choice of Biomaterial Choice of Oxides to be Synthesized Choice of Solvent for Extract Determining Reaction Conditions Synthesis of Nanoparticles and Nanocomposite Choice of Dye for Adsorption Antioxidant activity

3. PROBLEM STATEMENT: DYE REMOVAL FROM WATER BODIES Congo Red is readily absorbed in the human body and has genotoxic, mutagenic and carcinogenic effects  ( Oladoye et al.2022) “ A River Turns Red: Pollution Bleeds Hinton” TOI:31-05-2023 Safe removal of Congo Red dye is of significance in tackling water pollution . Requires inexpensive, easily synthesized adsorbent.

4. HARMFUL EFFECTS OF CONGO RED DYE Toxic/Inhibitory Effect Affected Targets Reference Causing Infertility Water flea ( Ceriodaphnia dubia ) (Zamora et al. 2016) Increases COD Water bodies and aquatic flora and fauna (Rani, K.C et. al. 2017) It makes surface water unaesthetic Water bodies ( Oladoye , P.O et al. 2022) Allergic Humans ( Litefti , K et al. 2019) Phytotoxicity Plants (Kumar, V. 2020) Molecular Structure of Congo red Dye

5. PROBLEM STATEMENT: NEED FOR SAFE ANTIOXIDANTS Nanoantioxidants are defined as nano materials capable of slowing the overall rate of autoxidation by trapping the chain carrying radicals, or by decreasing the initiation events . Inorganic metal oxide nano particles like Fe 2 O 3 and CuO exhibit intrinsic antioxidant properties without need for functionalization ( Valgimigli et al. 2018) ( Valgimigli et al. 2018) Fe 2 O 3 and CuO are two biocompatible and inexpensive metal oxide whose n ancomposite was prepared and analyzed for antioxidant activity and dye removal. Mechanism of Antioxidant Action in Fe 2 O 3 (Imam et al. 2017)

6. METAL OXIDE NANOPARTICLES PROPERTIES OF METAL OXIDE NANOPARTICLES I ncreased surface area l eading to enhanced adsorptive properties and faster diffusivities Enhanced optoelectronic, optical, electrical, thermal properties due to quantum confinement effect Tunability of properties : size , shape, porosity and cellular penetration capability depending on synthesis conditions. Ability to be synthesized under controlled conditions with reproducibility in size and morphology D epend upon their composition, crystallographic structure, morphology, surface stoichiometry, surface functionalities and interactions of the phases ( Chavali and Nikolova , 2019)

7. TOP-DOWN APPROACHES BOTTOM-UP APPROACHES SYNTHESIS METHODS HYDROTHERMAL SYNTHESIS SOLVOTHERMAL SYNTHESIS SOL-GEL SYNTHESIS SOLID STATE METHODS THERMAL DECOMPOSITION MICROWAVE ASSITED SYNTHESIS WET CHEMICAL SYNTHESIS CHEMICAL PRECIPITATION GREEN SYNTHESIS VAPOR DEPOSITION ELECTROCHEMICAL DEPOSITION MILLING MACHINING LITHOGRAPHY LASER ABLATION SPUTTERING CHEMICAL ETCHING PHYSICAL METHOD CHEMICAL METHOD BIOLOGICAL METHOD

8. ADVANTAGES OF GREEN SYNTHESIS of NANOPARTICLES Inexpensive High Yield, good morphology, easier control of synthesis conditions No toxic chemicals required M ilder reaction conditions O ne pot synthesis E asier implementation in batches as well as on a larger scale E asily available biomaterial Sustainable with greener synthesis design (Toress,2020; Kumar et al. 2017)

9. GREEN SYNTHESIS USING PLANT PARTS Does not require elaborate maintenance of cultures that micro-organisms require Lower toxicity concerns Phytochemicals (proteins , amino acids, carbohydrates, steroids, coenzymes and secondary metabolites such as saponins , tanins , flavonoids phenolics , vitamins, trepenoids , alkaloids) act as reducing agents, as well as capping agents

10. Me + Me + Me + Growth phase Activation Phase Termination phase Metal nanoparticle synthesis in plant parts involves three phases : MECHANISM OF NP SYNTHESIS WITH PLANT PARTS Reduction of metal ions occurs after reaction with phytochemicals to form smaller NP. OH Me Me Me O= O= O= Me + C oalescence and Ostwald Ripening for sizeable particles Determines the final shape of the nanoparticles P lant reducing agent Nucleation: organic coat act as stabilizers Reduced metal ions

11. USING Raphanus sativus for GREEN SYNTHESIS In India, leaves of Raphanus sativus are often disposed away or used as fodder Thus, is easily available and inexpensive P ossesses anticancer, antioxidant and antimicrobial properties by the virtue of its bioactive constituents Rich in phytochemicals that can act as bioreducing and capping agents Raphanus sativus extracts have been used in the past to synthesize Co, Ag, Cu 2 O, ZnO and NiO nanoparticles utilizing either roots or leaves for the process ,( Perveen R et al. 2020, Tej Sigh 2016) ( Balu et al. 2020) ( Umamaheswari et al. 2021) ( Haq S et al. 2021) Raphanus sativus : Genus : Raphanus Family : Brassicacae

12. Major bioreductants and capping agents; polyphenols and flavonoids are present in the highest concentration in leaves E ssential biomolecules were preserved in distilled water. PHYTOCHEMICAL PROFILE of Raphanus sativus Phytochemical Aqueous Extract Flavanoid + Alkaloids + Tannins + Phenols + Glycosides + Proteins and Amino Acids + Steroids - Carbohydrates - ( Gamba et al. 2022) ( Gamba et al. 2022) ( Gamba et al. 2022) Major Phytochemicals in parts of Radish

13. MECHANISM OF SYNTHESIS of NP by Raphanus sativus (Al Awadh et al. 2022)

14. SYNTHESIS of TENORITE ( CuO ) NPs CuSO 4 Leaf Extract prepared by boiling Shade Dry Raddish Leaves Grind into Paste and Boil in Distilled Water NaOH dropwise (till pH=12) 2 hours Centrifuge Drying and Grinding Calcine (500℃) f or 2 hours a nd Grind Color Change from Green to Red to Brown

15. SYNTHESIS of HEMATITE ( α -Fe 2 O 3 ) NPs FeCl 3 Leaf Extract prepared by boiling Shade Dry Raddish Leaves Grind into Paste and Boil in Distilled Water NaOH dropwise (till pH=12) 2 hours Centrifuge Drying and Grinding Calcine (500℃) f or 2 hours a nd Grind Color Change from Light Brown to Dark Brown

16. SYNTHESIS of NANOCOMPOSITES Calcination of α -Fe 2 O 3 Calcination of CuO Post-Calcination Grinding Post-Calcination Grinding Dissolution in ethanol in different ratios Ultrasonication Heating below BP of ethanol till solvent evaporates Calcination Post-Calcination grinding NANO-COMPOSITE FORMATION Nano-Composite Fe 2 O 3 CuO Total weight (Fe 2 O 3 ) 0.75 ( CuO ) 0.25 0.375g 0.125g 0.500g (Fe 2 O 3 ) 0.25 ( CuO ) 0.75 0.125g 0.375g 0.500g (Fe 2 O 3 ) 0.50 ( CuO ) 0.50 0.250g 0.250g 0.500g

17. CHARACTERIZATION PURE HNP: XRD (JCPDS card no: 33-0664) Phase pure rhombohedral α-Fe 2 O 3 without impurities with average crystallite size 4.36 nm by Debeye Scherrer equation

18. Formation of phase pure monoclinic CuO without impurities with average crystallite size 4.848 nm according to Debeye Scherrer equation PURE TNP: XRD (JCPDS card no: 00-048-1548)

19. The XRD pattern for all three nanocomposites show diffraction peaks ascribed to both CuO and α-Fe 2 O 3 . NANOCOMPOSITES: XRD CuO /α-Fe 2 O 3 nanocomposites crystallize in the same crystal system (cubic) and space group (Fd-3 m)

20. A shift in 2 towards lower values indicates an increase in grain size on nanocomposite formation.   Shannon-Prewitt data for Effective Ionic Radii : Cu 2+ = 57 pm Fe 3+ = 54 pm

21. minor peak at 398nm : Fe 2 O 3 These peaks are observed as a result of scattering by the metal oxide nanoparticles minor peak at 255nm : CuO NANOCOMPOSITES: UV-Vis Spectroscopy

22. Antioxidant activity was examined through DPPH ASSAY where a bsorbance was determined at 517 nm with respect to DPPH [( A o - A n )/ A o ] * 100 ( A o = absorbance of DPPH ; A n = absorbance of DPPH@Nanocomposite mixture) . APPLICATIONS ANTIOXIDANT APPLICATION RESULT: values for 0.01mg/ml nanocomposite were calculated to be 83.99%, 72.37% and 91.24%

23. MECHANISM OF ANTIOXIDANT ACTION M ostly ROS-mediated CuO and Fe 2 O 3 are supposed to be CAT-mimic or catalase mimic

24. ADSORPTION OF CR DYE ADSORPTION TAKING CHANGE IN pH & CONC. Of CR DYE Batch adsorption performed :50 ppm solution of Congo red added to 0.001g of Fe 2 O 3 / CuO Nanocomposite. Four such solutions with pH 4,6,8 and 10 were prepared A UV-Vis spectrum of each supernatant was recorded

25. ADSORPTION OF CR DYE

26. ADSORPTION OF CR DYE

27. ADSORPTION OF CR DYE MECHANISM Congo Red is a dipolar molecule which exists as anionic form (deep red color) at neutral or alkaline pH and cationic form at acidic pH exhibiting a dark blue color α- Fe 2 O 3 and CuO NPs have negative surface charge that attracts cationic form of Congo Red by electrostatic attraction α-Fe 2 O 3 NP and CuO NP adsorbs Congo Red dye at acidic pH by H-bonding and electrostatic interactions

28. Biosynthesis implemented using Raphanus sativus leaves yielded phase pure nanomaterials with high crystallinity and small grain size. ( Fe 2 O 3 ) 0.50 ( CuO ) 0.50 demonstrated finest applicability at neutral pH and at p H 4 while ( (Fe 2 O 3 ) 0.25 ( CuO ) 0.75 ) exhibited best properties at all other pH values α-Fe 2 O 3 / CuO nanocomposite has potential for removal of other hazardous environmental pollutants via adsorption. Being non-toxic in moderate amounts, this nanocomposite has untapped potential as an antioxidant for use in industry CONCLUSION

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30. I would like to express my sincere gratitude to Prof. Nahid Nishat for her constant support and encouragement throughout the duration of this project. I am forever indebted to our Assistant Professor, Dr. Zeba Haque for providing her able guidance and direction through every step of the way. This project wouldn’t have been possible without her vision and planning. I would also like to thank our senior research scholars and my batchmates I would like to thank CIF, Jamia Millia Islamia and AMU, Aligarh ACKNOWLEGMENT