GREEN SYNTHESIS OF COPPER OXIDE

NANOPARTICLES

A PROJECT SUBMITTED TO
FAKIR MOHAN AUTONOMOUS COLLEGE
IN FULFILLMENT OF THE REQUIRMENT
FOR THE BACHELOR DEGREE OF SCIENCE

SUBMITTED BY
NAME: - MANORANJAN NAYAK
EXAM ROLL NO.: -22S81034
REGD. NO.: -
UNDER THE SUPERVISION OF
SHRI TUSHARKANTA ICHAGUTU
ASSISTANT PROFESSOR
P.G DEPARTMENT OF CHEMISTRY
PLACE OF RESEARCH
P.G DEPARTMENT OF CHEMISTRY
FAKIR MOHAN AUTONOMOUS COLLEGE,
BALASORE

ACADEMIC SESSION-2022-25
 ACKNOWLEDGEMENT
I feel pleasure to submit the
Project On
A review study on Green Synthesis of Copper Oxide Nanoparticles.

This becomes possible only by the valuable guidance, constant

support and needful help of Shri Tusharkant Ichagutu, Assistant
Professor in the P.G. Department of Chemistry, Fakir Mohan
Autonomous College, Balasore.
So, I would like to express my sincere thanks and gratitude to him
and also my colleagues who helped me in my work a lot.

Signature of Candidate
Place-Balasore
Date: / /2025
Exam Roll No.: -22S81034
Regd. No.: -
 Certificate

This is to certify that the project report entitled “A review

study on Green synthesis of Copper Oxide Nanoparticles”
which is being submitted by MANORANJAN NAYAK, Exam
Roll No.: -22S81034, Regd. No.: - in fulfillment for the
award of The Bachelors Degree of Science from FAKIR
MOHAN AUTONOMOUS COLLEGE, BALASORE, is a
record of Bonafide work carried out by him under my guidance
and supervision.

Neither this project report nor any part of it has

been submitted for any degree or academic award. It is a routine
experiment only.

Signature of The Supervisor

Name: Shri Tusharkant Ichagutu
Designation: Assistant Professor
F.M AUTONOMOUS COLLEGE, BALASORE
 Declaration
I do hereby declare that the project entitled “A review
on Green Synthesis of Copper Oxide Nanoparticles” is being
submitted to Fakir Mohan Autonomous College, Balasore,
Odisha for the award of The Bachelor Degree of Science in an
original piece of work done by under me under the guidance of
Shri Tusharkant Ichagutu, Assistant Professor in P.G.
Department of Chemistry and the same has not been submitted
elsewhere for any other academic degree to this college or any
other college/ university.

Signature of the Guide Signature of the Candidate

Place: - Balasore
Date.: -
Exam Roll No.-22S81034
Regd. No.:

Signature of the Head of the Dept.

 CONTENTS
1.Introduction
2. Literature Review
3.Experimental Procedure
4. Characterization
5.Conclusion

Reference
1.Introduction
Nanotechnology has significantly advanced over the past few decades,
playing a crucial role in various industries, including medical, pharmaceutical,
and textile sectors. Metal nanoparticles such as silver, zinc, and gold have been
extensively utilized as therapeutic agents in medical institutions. Moreover,
transition metal oxides, including copper oxide (CuO), titanium dioxide (TiO₂),
iron oxide (Fe₃O₄), zinc oxide (ZnO), and nickel oxide (NiO) nanoparticles, have
emerged as advanced materials with broad applications in energy, biomedical,
and environmental fields. The strong adsorption capabilities exhibited by these
nanoparticles (NPs) significantly enhance their functionality and effectiveness in
various applications.
In recent years, there has been an increasing interest in the biological properties
of metal nanoparticles. Numerous studies have focused on evaluating the
biological activities of metal oxide nanoparticles, particularly copper oxide
nanoparticles (CuO NPs), which have demonstrated superior biological and
photocatalytic activities compared to conventional metal nanoparticles. CuO NPs
have been widely applied in gas sensing, wastewater treatment, catalysis, battery
technology, food preservation, high-temperature superconductors, solar energy
conversion, photovoltaic devices, dye removal, field emission emitters, and
agricultural advancements. Additionally, CuO NPs exhibit exceptional
anticancer, antimicrobial, and antioxidant properties, making them highly
promising for biomedical applications.
Several physical and chemical approaches have been explored for the synthesis
of CuO NPs, including microwave irradiation, thermal decomposition, sol-gel
techniques, colloidal thermal synthesis, sonochemical methods, hydrothermal
synthesis, and quick precipitation techniques. However, these conventional
methods often require labor-intensive processes, high energy input, and complex
procedures. To overcome these limitations, green synthesis methods have
emerged as an eco-friendly and sustainable alternative for nanoparticle
production.

2. Literature Review
This project report focuses on the green synthesis of CuO NPs, highlighting their
biomedical and environmental applications while emphasizing their advantages
over traditional synthesis methods. The study aims to explore the potential of
plant-mediated synthesis as a cost-effective, non-toxic, and environmentally
sustainable approach for the production of CuO NPs with enhanced properties for
diverse applications.

2. Synthesis of CuO Nanoparticles (CuO-NPs)

Various methods have been developed for synthesizing CuO nanoparticles (CuO-
NPs), including ultrasound irradiation, hydrothermal synthesis, biosynthesis,
electron beam lithography, solid-state reactions, sol-gel techniques, template-
assisted synthesis with surfactants, microwave-assisted protocols, copper acetate
decomposition, and sonochemical synthesis.

The choice of synthesis method significantly impacts the morphological

properties and toxicity of CuO-NPs. A flowchart illustrating these synthesis
approaches is presented in Figure 1.
3. Chemical Approach
The chemical approach involves the use of specific reagents to reduce copper ions
during the synthesis of CuO nanoparticles (CuO-NPs). This method is broadly
classified into two categories: the green chemical approach and the traditional
approach.

In the green chemical approach, organic compounds such as ascorbic acid are
used as reducing agents. In contrast, the traditional approach relies on inorganic
reducing agents like sodium borohydride and potassium borohydride. Studies
have demonstrated the effectiveness of ascorbic acid in CuO-NP synthesis, as
well as the role of potassium borohydride in nanoparticle formation.

However, several reports highlight significant limitations of the chemical

synthesis method, including high energy consumption, environmental pollution,
the need for high pressure and temperature, and the use of expensive and toxic
chemicals. These challenges also apply to the synthesis of other transition metal
oxide nanoparticles.

3.1. Physical Method

The physical method utilizes electric current as a source of electrons for
generating the required reduction process during CuO nanoparticle (CuO-NP)
synthesis. Common techniques in this approach include electro-spraying, laser
pyrolysis, laser ablation, and evaporation-condensation.

Among these techniques, pulsed laser-induced ablation has gained significant

attention due to its simplicity, eco-friendliness, and ability to produce uniformly
sized nanoparticles. In this method, a high-power pulsed laser is essential for
ablating the surface of the sample. Adjusting parameters such as pulse width,
wavelength, laser repetition rate, ablation time, and temperature allows precise
control over the morphological characteristics of the synthesized nanoparticles.

The physical method offers several advantages, including the production of CuO-
NPs with uniform size, controlled morphology, and high purity. However, its
limitations include high operational costs, the need for specialized skills, and the
significant power and energy requirements, which pose challenges to its
widespread application.

4. Biological Approach to Synthesis

The biological approach to nanoparticle synthesis involves the use of organisms
such as bacteria, yeast, fungi, and plant extracts as reducing agents for metal ions.
Several studies have reported the biosynthesis of CuO nanoparticles (CuO-NPs)
using bacteria such as Phormidium cyanobacterium, Morganella morganii,
Serratia sp., and Escherichia coli.

Despite the eco-friendly advantages of microbial nanoparticle synthesis, this

method has certain limitations, including the potential toxicity of some bacterial
strains, as well as challenges in isolation and incubation processes.

Plants, however, remain the most promising source for the synthesis of metal and
metal oxide nanoparticles. Their advantages include rapid reaction rates with low
energy requirements, the presence of multiple biomolecules, cost-effectiveness,
high stability, the absence of hazardous chemicals, and safe, straightforward
operational procedures.

Biomolecules found in plant extracts serve as both reducing and stabilizing agents
during CuO-NP synthesis. Compounds such as flavonoids, proteins, tannins,
phenols, and terpenoids have been identified as effective agents for CuO-NP
formation. Several plant species have been successfully utilized for CuO-NP
synthesis, as summarized in Tables 1 and 2. Additionally, Table 3 presents the
advantages and disadvantages of different synthesis methods.

4.1. Formation Mechanism of CuO-NP Synthesis from Plant Extracts

The green synthesis of CuO nanoparticles (CuO-NPs) using plant extracts as an

electron source for copper salt reduction offers several advantages over microbial
synthesis. Unlike microbial methods, it does not require cell culture maintenance
and can be easily scaled up for large-scale production.

During CuO-NP formation, an observable colour change occurs in the plant

extract upon the addition of copper salt, indicating nanoparticle synthesis. Studies
suggest that phytochemicals in plant extracts initially form complexes with metal
ions before reducing them to nanoparticles. Biomolecules present in the extract
interact with copper ions, facilitating their reduction and transformation into
CuO-NPs.

The probable mechanism for CuO-NP synthesis is represented by the following

reactions:

5. Effects of Experimental Parameters on CuO-NP Synthesis

5.1. Effect of pH

pH plays a crucial role in determining the acidity or basicity of a solution,

significantly influencing the synthesis of CuO nanoparticles (CuO-NPs) and
other metal oxide nanoparticles. Studies have shown that variations in pH impact
the size and texture of biosynthesized nanoparticles from plant extracts.
Additionally, pH adjustments have been utilized to control the shape and size of
synthesized nanoparticles. Optimal pH conditions for nanoparticle synthesis
using Aeromonas hydrophila extract have been reported to range between 7 and
9.

5.2. Effect of Plant Extract Type and Concentration

The efficiency of CuO-NP synthesis using plant extracts depends largely on the
types of biomolecules present and the extract concentration. The volume of plant
extract used influences the synthesis duration, with higher extract volumes
accelerating the reaction due to an increased availability of chemical constituents
that bind to the precursor, facilitating rapid bio-reduction and nanoparticle
stabilization. For optimal green synthesis conditions, the extract volume must
correspond appropriately to the concentration of the copper precursor. Studies
suggest that both the volume and type of plant extract used significantly impact
nanoparticle yield, morphology, and biological activity.

5.3. Effect of Incubation Time

The incubation time of nanoparticles synthesized using plant extracts affects their
morphological properties and overall quality. Factors such as storage conditions
and light exposure also influence the reaction time of CuO-NPs. Prolonged
incubation has been reported to cause particle aggregation and shrinkage,
potentially affecting their stability and functionality.

5.4. Effect of Temperature

Temperature is a key factor influencing the synthesis of metal oxide

nanoparticles. The recommended temperature range for CuO-NP synthesis using
plant extracts is between 25°C and 100°C. However, room-temperature synthesis
is more common due to the volatility of certain secondary metabolites found in
plant extracts. Studies indicate that higher temperatures promote rapid and
complete synthesis; however, excessively high temperatures may hinder
nanoparticle formation by inactivating biomolecules responsible for precursor
reduction.

This mechanism highlights the role of plant-derived biomolecules in reducing and stabilizing
CuO-NPs during synthesis.

5.5. Characterization of CuO-NPs

Since the applications of CuO nanoparticles (CuO-NPs) largely depend on their

physical, chemical, and structural properties, proper characterization is essential.
Various analytical techniques are employed to determine the morphology, size,
crystallinity, surface charge, and composition of CuO-NPs.
5.6. UV-Visible (UV-Vis) Spectroscopy

UV-Vis spectroscopy is a molecular spectroscopic technique based on the

Bouguer-Lambert-Beer law. It operates by measuring plasmon resonance and the
collective oscillations of conduction band electrons in response to
electromagnetic waves. This method is widely used for analyzing the absorption
properties of fluids and various materials.

During UV-Vis spectroscopy analysis, a beam of light is split into two: one
portion passes through the sample in a transparent cell, while the other serves as
a reference. The solution absorbs light at specific wavelengths, known as the
surface plasmon resonance (SPR), which is characteristic of the material being
analyzed. For CuO-NPs, the SPR typically falls within the range of 200–350 nm.

Studies indicate that factors such as extract type, pH, temperature, and synthesis
method can influence the SPR of CuO-NPs, subsequently affecting their
morphological properties. UV-Vis spectroscopy provides critical insights into
nanoparticle size, structure, stability, and aggregation behavior.

5.7. Fourier Transform Infrared (FTIR) Spectroscopy

FTIR spectroscopy is used to identify functional groups associated with

nanoparticles. The technique operates by detecting specific vibrational
frequencies of molecular bonds. When incident infrared light matches the
vibrational frequency of a functional group, absorption occurs, revealing the
chemical composition of the sample.

The unique arrangement of atoms in a molecule results in distinct FTIR spectra.

Prominent absorption peaks associated with CuO-NPs include:

• 3000–3350 cm⁻¹: Corresponding to N–H (amines) or O–H

(alcohols/phenols).
• 820–880 cm⁻¹: Attributed to aromatic C–H bending.
• 2900–3000 cm⁻¹: Linked to C–H stretching vibrations.
• 1600–700 cm⁻¹: Related to CuO vibrations.
• 1600–1790 cm⁻¹: Corresponding to C=O (carbonyl groups).

FTIR spectroscopy helps determine the biomolecules responsible for CuO-NP

stabilization during synthesis.

5.8. Size and Morphological Analysis

The size and morphology of CuO-NPs are critical parameters influencing their
applications. Various characterization techniques are commonly used for detailed
analysis:

5.9. Scanning Tunneling Microscopy (STM)

STM provides detailed surface information, including size estimation,

morphology, and topography. This technique operates based on quantum
tunneling, where variations in tunneling current create high-resolution surface
images. STM is widely used for studying metals and semiconductors.

5.10. Atomic Force Microscopy (AFM)

AFM is used to determine nanoparticle morphology, including size, surface

roughness, and texture. Unlike electron microscopy, AFM provides 3D
characterization of nanoparticles. It can also analyze the geometry and magnetic
properties of CuO-NPs. However, AFM has limitations, such as slow scanning
speed and potential inaccuracies in topography measurements when the probe is
not properly calibrated.

5.11. Transmission Electron Microscopy (TEM)

TEM is one of the most powerful techniques for analyzing CuO-NP morphology
and structure. It operates by transmitting a beam of energetic electrons through
an ultra-thin sample, forming high-resolution images. An advanced version, high-
resolution transmission electron microscopy (HRTEM), allows for imaging at the
atomic scale, providing detailed crystallographic information.

5.12. Scanning Electron Microscopy (SEM)

SEM is widely used for morphological characterization of CuO-NPs. However,

it has limitations, such as an inability to provide precise population and size
distribution data. Additionally, SEM can damage some nanopolymers, and the
nanoparticles must be vacuum-resistant for effective imaging. Despite its
advantages, SEM is expensive and time-consuming compared to other
techniques.

5.13. Dynamic Light Scattering (DLS)

Dynamic Light Scattering (DLS), also known as quasi-elastic light scattering, is

a widely used technique for determining nanoparticle size and aggregation
behavior. It is a fast and highly sensitive method capable of estimating particle
sizes across both the nano and macro scales.
One of the key advantages of DLS is its ability to provide rapid size distribution
measurements in a liquid medium. However, a major limitation is its inability to
differentiate between individual nanoparticles and aggregates. Additionally, the
accuracy of DLS measurements depends on factors such as sample concentration,
viscosity, and the presence of multiple particle populations.

The speed of DLS measurements is influenced by particle size: smaller particles

exhibit faster Brownian motion, resulting in higher scattering intensities, while
larger particles move more slowly. This technique is particularly useful for
studying nanoparticle stability and dispersity in colloidal solutions.

Here is the table without the reference column:

Functional
SPR Techniques for
Plant Group Size
S/N Plant Name Peak Morphological Shape
Part Prediction (nm)
(nm) Assessment
(cm⁻¹)
3976 (O–H),
Eupatorium UV, FTIR, XRD,
1 Leaf 301 2936 (C–H), Spherical -
odoratum TEM
1618 (C=O)
3406 (O–H),
Acanthospermum UV, FTIR, XRD,
2 Leaf 305 2838 (C–H), Spherical -
hispidum TEM
1520 (C=C)
Hylotelephium 3388 (O–H), UV, FTIR, SEM,
3 Flower 350 Spherical 83
telephium 1980 (C=O) TEM, XRD, EDX
3467 (N–H),
UV, FTIR, SEM, 26–
4 Kalopanax pictus Leaf 368 1584 (C=C), Spherical
TEM, EDX, XPS 67
1360 (C–N)
3420 (O–H),
Pterolobium XUV, FTIR, XRD, 10–
5 Leaf 274 2915 (C–H), Spherical
hexapetalum TEM, EDX 50
1625 (C=C)
Coriandrum XRD, FESEM,
6 Seed - - Irregular 18.2
sativum L. DLS
3415 (O–H), UV, FTIR, Quasi-
7 Oak Fruit - 34
1654 (C=O) FESEM, XRD cubic
SEM, EDS, UV,
8 Albizia lebbeck Leaf 413 - Spherical 100
XRD, TEM
3314 (O–H), UV, FTIR,
Eichhornia 15–
9 Leaf 310 1624 (N–H), FESEM, TEM, Spherical
crassipes 30
1217 (C–O–C) EDX
3000–3350,
Citrofortunella 820–880 (C– UV, FTIR, XRD, 54–
10 Leaf 305 Spherical
microcarpa H), 1357 SEM, FTIR, EDS 68
(C=O)
 Functional
SPR Techniques for
Plant Group Size
S/N Plant Name Peak Morphological Shape
Part Prediction (nm)
(nm) Assessment
(cm⁻¹)
3442 (O–H),
Verbascum UV, FTIR, XRD,
11 Leaf 350 2922 (C–H), Spherical -
thapsus SEM, FTIR
1616 (C=O)
Euphorbia 3384 (O–H), FTIR, XRD,
12 Flower 240 Cubic 19.2
pulcherrima 1595 (C=O) HRTEM
3439 (O–H), UV, FTIR, XRD,
13 Sida rhombifolia Leaf 471 Spherical 10
1658 (C=O) FESEM
3346 (O–H),
Seidlitzia Bark UV, FTIR, XRD,
14 385 2900 (C–H), - 26
rosmarinus Ashes FESEM
1500 (C=O)
Enicostemma UV, XRD, EDS,
15 Leaf 264 - - 30
axillare (Lam.) SEM, TEM
3209 (O–H),
Terminalia UV, XRD, FTIR, 103–
16 Leaf 372 2920 (C–H), Spherical
catappa L. SEM, TEM 29
1557 (C=O)
Fruit 3379 (O–H), UV, XRD, FTIR, 10–
17 Punica granatum 282 Spherical
Peel 1577 (C=O) SEM 100
3323 (O–H),
UV, XRD, FTIR,
18 O. cochinchinense Leaf - 1550 (C=C), - -
SEM, SEAD
1338 (C–N)
3550–3200
UV, XRD, FTIR,
(O–H), 1670 15–
19 Rosa canina Fruit 262 FESEM, WDX, Spherical
(C=O), 1405 25
EDX, TEM
(C=C)
3405 (O–H), UV-Vis, PL,
15–
20 Aloe barbadensis Leaf 285 1538 (C=C), FTIR, XRD, SEM, Spherical
30
944 (C–C) TEM
3300–3500
(O–H), 2299
21 Sambucus nigra Fruit 278 UV, XRD, FTIR - -
(C–H), 1621
(C=C)
3414 (O–H),
Calotropis UV, XRD, FTIR,
22 Leaf 355 2923 (C–H), Cylindrical 46
procera SEM
1598 (C=C)
5.14. Energy Dispersive X-ray (EDX)

EDX is a widely used technique for both qualitative and quantitative

identification of the elemental composition of CuONPs and other metal
nanoparticles. When a beam of electrons from EDX interacts with CuONPs, it
causes the emission of X-rays. These emitted X-rays are then analyzed to
determine the elemental composition.

For quantitative analysis, the concentration of specific elements in CuONPs is

measured based on the intensities of the peaks. Meanwhile, qualitative analysis
identifies elements by examining the positions of the X-ray peaks on the EDX
spectrum.

With advancements in technology, EDX can now be integrated with SEM and
TEM, allowing for enhanced identification and quantification of trace elements.
This technique is capable of detecting elements with atomic numbers ranging
from 4 to 92. The use of EDX in the characterization of biosynthesized CuONPs
has provided valuable insights into their elemental composition and purity.
 Figure 3. Probable mechanism of CuONPs toxicity against bacteria

Antibacterial Application

The inhibitory antibacterial potential exhibited by biosynthesized CuONPs

against both gram-positive and gram-negative bacterial strains has been studied.
The antibacterial activities of phytosynthesized CuONPs from the extract of
Tecoma castanifolia leaf displayed reliable bactericidal activity, which may be
useful in biomedical applications.

Studies have shown that the biomolecules in plant extracts used for CuONPs
synthesis enhance antibacterial efficacy against both Gram-positive and Gram-
negative bacterial strains. The increase in antibacterial activity of CuONPs has
been linked to the presence of biomolecules such as terpenoids, which play a role
during the capping process.

Antibacterial analysis of CuONPs obtained through the agar well diffusion

technique has demonstrated toxicity against both Gram-positive bacteria
(Streptococcus mutans and Staphylococcus aureus) and Gram-negative bacteria
(Pseudomonas aeruginosa, Klebsiella pneumoniae, and Escherichia coli),
effectively inhibiting their growth. The bactericidal effectiveness of CuONPs has
been attributed to the generation of highly reactive oxygen species on their
surface, such as hydroxyl radicals (OH), hydrogen peroxide (H₂O₂), and
superoxide anions (O₂⁻), which contribute to bacterial cell destruction.

Plant
S/N Plant Name Salt Applications Activities
Part
Eupatorium Copper
1 Leaf Antibacterial 12–30 mm
odoratum sulfate
Aloe barbadensis Copper Degradation, 98.89% removal of methylene
2 Leaf
Miller sulfate Antibacterial blue dye
Efficient catalyst for
Rheum palmatum Copper
3 Root Catalytic activity methylene blue and
L. chloride
rhodamine B reduction
Copper Strong activity against K.
4 Tea Leaf Antibacterial
nitrate pneumoniae and V. cholerae
Cell cycle disruption,
Eucalyptus Copper
5 Leaf Anticancer upregulation of pro-apoptotic
globulus sulfate
genes in MCF-7 cells
Eucalyptus Copper ROS generation for bacterial
6 Leaf Antibacterial
globulus sulfate cell interruption
Copper Effective degradation of
7 T. arjuna Bark Catalytic activity
nitrate methyl blue dye
Cupric Cytotoxic effect on breast
8 Camellia sinensis Leaf Anticancer
acetate cancer cells (MCF-7)
Copper Oxidative stress
9 Cistus incanus Leaf Enhanced stress resistance
nitrate improvement
Cissus Copper 86% inhibition against A.
10 Leaf Antifungal
quadrangularis acetate niger and A. flavus
Solar
Copper Fast degradation of methylene
11 Aloe Vera Leaf Photocatalytic
sulfate blue under solar irradiation
Activity
Platform for intracellular
Copper
12 Coleus aromaticus Leaf Anticancer miRNA delivery for lung
sulfate
cancer treatment
Copper Cytotoxic against cancer cells
13 Azadirachta indica Leaf Anticancer
acetate without affecting human cells
Hibiscus rosa- Copper Strong cytotoxicity against
14 Leaf Anticancer
sinensis acetate cancer cells
High cytotoxicity against
Copper
15 Murraya koenigii Leaf Anticancer cancer cells without harming
acetate
normal cells
Copper
16 Moringa oleifera Leaf Antioxidant Efficient antioxidant activity
acetate
 Plant
S/N Plant Name Salt Applications Activities
Part
Copper Promising antioxidant
17 Tamarindus indica Leaf Antioxidant
acetate potential
Aerial Copper Photocatalytic Higher catalytic activity
18 Euphorbia maculata
part sulfate activity compared to NiFe3O4 NPs
Copper Strong antimicrobial activity
19 Sida acuta Leaf Antibacterial
sulfate against pathogens
Copper Potential biomedical
20 Bauhinia tomentosa Leaf Antibacterial
sulfate applications
Copper High sensitivity towards Fe²⁺
21 Capparis spinosa Leaf Iron Sensing
sulfate and Fe³⁺ ions
Copper Sensing & Selective detection of
22 Fortunella japonica Fruit
sulfate Remediation analytes
Copper Cytotoxicity Cytotoxic against MCF-7
23 Acalypha indica Leaf
sulfate Activity breast cancer cells
Effective against P.
Copper
24 Zea mays Husk Antibacterial aeruginosa and B.
acetate
licheniformis
Eryngium Copper Nanoremediation for heavy
25 Leaf Remediation
campestre sulfate metal wastewater
Froriepia Copper Potential for soil and
26 Leaf Remediation
subpinnata sulfate groundwater remediation
Tabernaemontana Copper Maximum inhibition against
27 Leaf Antibacterial
divaricata sulfate urinary tract pathogens

Catalytic Application

Metallic and metal oxide nanoparticles have been reported to exhibit excellent
photocatalytic efficiency. The photocatalytic degradation assessment of green-
synthesized CuONPs on RB dye revealed a 94% degradation efficiency up to the
fifth cycle, demonstrating the durability of phytosynthesized CuONPs as a
reliable photocatalytic agent.

A comparative catalytic study of CuONPs and zinc oxide nanoparticles

(ZnONPs) for basic violet 3 degradation indicated that ZnONPs exhibited higher
catalytic activity than CuONPs. The degradation of basic violet 3 followed
pseudo-first-order kinetics.

CuONPs synthesized using Thymus vulgaris leaf extract have been reported as an
outstanding heterogeneous catalyst for N-arylation of amines and indoles,
achieving a remarkable percentage yield of N-arylated products. The catalyst also
demonstrated excellent recyclability and reusability without any loss in activity.
Additionally, the photocatalytic analysis of CuONPs derived from Aloe vera
leaves under solar simulator light irradiation showed complete degradation of
methylene blue within 10 minutes. This high activity has been attributed to the
phyto-constituents present in Aloe vera leaves. Further research has continued to
explore and confirm the catalytic efficiency of CuONPs.

Conclusion

The synthesis method of CuONPs significantly influences their ecological

identity, physiochemical, and morphological properties, which directly impact
their biological and catalytic applications. This discussion elaborates on the
biological synthesis of CuONPs from plants, offering promising opportunities for
medicinal institutions and various industries due to their bioactivity and eco-
friendly mode of synthesis.

The primary applications of CuONPs in biomedical and waste treatment sectors

are attributed to their antimicrobial efficiency, which is largely dependent on their
morphological properties. Recent advancements in characterization techniques
for analyzing CuONPs have been thoroughly highlighted. The effectiveness of
biosynthesized CuONPs in anticancer, antioxidant, antibacterial, and wastewater
treatment applications has been comprehensively examined.
Furthermore, the mechanisms of synthesis and toxicity have been well explained.
To enhance the biological applications of CuONPs, future research should focus
on minimizing their toxicity while maintaining and improving their biological
efficiency.

3.Experimental Procedure
1. Preparation of Neem Extract To prepare the neem extract, fresh neem
leaves were collected and thoroughly washed with distilled water to
remove any surface contaminants. The cleaned leaves were then chopped
into small pieces to enhance extraction efficiency. A measured quantity of
distilled water was added to the chopped leaves, allowing them to dissolve.
The mixture was continuously stirred for 2 hours to facilitate the release of
bioactive compounds from the neem leaves. After stirring, the extract was
filtered using standard filtration techniques to separate the liquid phase
from the solid residues. The obtained neem extract was then stored in a
freezer for further use.
 2. Preparation of CuONPs For the synthesis of CuO nanoparticles (CuONPs),
0.16 g of anhydrous CuSO₄ was accurately weighed and dissolved in 100
mL of distilled water. From this stock solution, 80 mL was taken for the
reaction.

To this 80 mL CuSO₄ solution, 20 mL of the previously prepared neem extract

was added dropwise under continuous stirring. Simultaneously, a small quantity
of NaOH solution was also introduced dropwise into the reaction mixture. The
reaction was carried out under continuous stirring for 3 hours, maintaining a
temperature range of 25–45°C to ensure proper interaction and formation of
nanoparticles.
After completion of the reaction, the product formed was subjected to
centrifugation to separate the solid precipitate. The centrifuged product was
washed multiple times with ethanol to remove any residual impurities.

3. Drying and Calcination The purified product was then transferred to a hot
air oven and dried at a controlled temperature for a minimum of 16 hours
to remove any remaining moisture. To further enhance the purity and
crystallinity of the synthesized CuONPs, the dried product was subjected
to calcination in a muffle furnace for 3 hours, resulting in the final
formation of CuO nanoparticles.
4. Characterization Studies To confirm the successful synthesis of CuONPs,
various characterization techniques were employed:
Electrochemical Study: The electrochemical behavior of the synthesized
CuONPs was analyzed, and the results indicated a good electrochemical
response.
UV-Visible Spectroscopy (UV-Vis): The UV study further confirmed the
formation of CuONPs, providing strong absorbance peaks, which verified the
successful synthesis of CuO nanoparticles.
5.Conclusion
The above experimental process led to the successful biosynthesis of CuO
nanoparticles using neem extract as a natural reducing and stabilizing agent. The
characterization results demonstrated the effectiveness of the synthesized
nanoparticles, making them suitable for potential applications in various fields
such as catalysis, electrochemistry, and biomedical sciences.
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