The document discusses the biosynthesis of CuO/Fe2O3 nanocomposites using Raphanus sativus and explores their potential in dye adsorption and antioxidant activities. It highlights the harmful effects of Congo Red dye in water pollution and the advantages of using green synthesis methods with Raphanus sativus due to its rich phytochemical profile. The findings indicate the effectiveness of the synthesized nanocomposites in removing Congo Red dye and suggest their non-toxic application in addressing environmental issues.

1. BIOSYNTHESIS OF CuO/Fe2O3
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
Fe2O3 and CuO exhibit intrinsic
antioxidant properties without need for
functionalization (Valgimigli et al. 2018)
(Valgimigli et al. 2018)
• Fe2O3 and CuO are two biocompatible
and inexpensive metal oxide whose
nancomposite was prepared and
analyzed for antioxidant activity and dye
removal.
Mechanism of Antioxidant Action in Fe2O3 (Imam et al. 2017)

6. METAL OXIDE NANOPARTICLES
PROPERTIES OF METAL OXIDE NANOPARTICLES
• Increased surface area leading 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
Depend 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
Milder reaction conditions
One pot synthesis
Easier
implementation
in batches as
well as on a
larger scale
Easily 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=
Me+
Coalescence and Ostwald Ripening for
sizeable particles
Determines the final shape of the
nanoparticles
Plant 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
• Possesses 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, Cu2O, 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
• Essential 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
CuSO4
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℃) for
2 hours and Grind
Color Change
from Green to
Red to Brown

15. SYNTHESIS of HEMATITE (α-Fe2O3) NPs
FeCl3
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℃) for
2 hours and Grind
Color Change
from Light
Brown to Dark
Brown

16. SYNTHESIS of NANOCOMPOSITES
Calcination of α-
Fe2O3
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 Fe2O3 CuO Total weight
(Fe2O3)0.75(CuO)0.25
0.375g 0.125g 0.500g
(Fe2O3)0.25(CuO)0.75
0.125g 0.375g 0.500g
(Fe2O3)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-Fe2O3 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-Fe2O3.
NANOCOMPOSITES:
XRD
CuO/α-Fe2O3 nanocomposites crystallize in the same crystal
system (cubic) and space group (Fd-3 m)

20. A shift in 2 θ towards lower values indicates an increase in grain
size on nanocomposite formation.
Shannon-Prewitt
data for Effective
Ionic Radii:
Cu2+ = 57 pm
Fe3+ = 54 pm

21. minor peak at 398nm:
Fe2O3
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
absorbance was determined at 517 nm with respect to DPPH
[(Ao - An)/Ao] * 100
(Ao = absorbance of DPPH
; An = 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%
0
10
20
30
40
50
60
70
80
90
100
25% CuO 75% CuO 50% CuO
%Antioxidant Activity
25% CuO
75% CuO
50% CuO

23. MECHANISM OF ANTIOXIDANT ACTION
Mostly ROS-mediated
CuO and Fe2O3 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 Fe2O3/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- Fe2O3 and CuO NPs have negative surface charge that
attracts cationic form of Congo Red by electrostatic
attraction
α-Fe2O3 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.
(Fe2O3)0.50(CuO)0.50 demonstrated finest applicability at neutral pH and at pH
4 while ((Fe2O3)0.25(CuO)0.75) exhibited best properties at all other pH values
α-Fe2O3 / 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