GREEN SYNTHESIS OF COPPER NANOPARTICLES FROM

CITRULLUS COLONCYNTHIS SEED EXTRACT AND THEIR

APPLICATION IN DYE WASTEWATER TREATMENT

By

Fakiha Safdar

Arsalna Naseer

Under the Supervision of

Dr. Azhar Ali

MSc.(Sweden), Ph.D. (pb)

A thesis submitted to the University of Punjab in the partial fulfillment of requirements

for the degree of BS Environmental Sciences

COLLEGE OF EARTH AND ENVIRONMENTAL SCIENCES

UNIVERSITY OF THE PUNJAB, LAHORE-PAKISTAN

SESSION: 2019-2023
GREEN SYNTHESIS OF COPPER NANOPARTICLES FROM
CITRULLUS COLOCYNTHIS SEED EXTRACT AND THEIR
APPLICATION IN DYE WASTEWATER TREATMENT

BS Environmental Sciences
Session: 2019-2023

COLLEGE OF EARTH AND ENVIRONMENTAL SCIENCES

THE UNIVERSITY OF THE PUNJAB,
LAHORE-PAKISTAN
 DEDICATION

This thesis is dedicated to our parents, siblings, and teachers.

 CERTIFICATE OF APPROVAL

This thesis by Fakiha Safdar (12M) and Arsalna Naseer (30M) is hereby approved for
submission to the University of the Punjab, Lahore in partial fulfillment of the
requirements for the degree of BS in Environmental Sciences.

SUPERVISOR PRINCIPAL

Dr. Azhar Ali Prof. Dr. Sajid Rashid Ahmed

Assistant Professor, Principal,
College of Earth and College of Earth and
Environmental Sciences, Environmental Sciences,
The University of Punjab, University of the Punjab,
Lahore, Pakistan. Lahore, Pakistan.
 Table of Contents

ABSTRACT .............................................................................................................................. i

ACKNOWLEDGEMENT ...................................................................................................... ii

LIST OF FIGURES ............................................................................................................... iii

LIST OF TABLES ................................................................................................................. iv

LIST OF ABBREVIATIONS ................................................................................................ v

Chapter 1

INTRODUCTION................................................................................................................... 1

1.1 Overview ............................................................................................................... 1

1.2 Water pollution ..................................................................................................... 1
1.3 Textile industry in Pakistan .................................................................................. 1
1.4 Textile dyes ........................................................................................................... 1
1.5 Methylene blue...................................................................................................... 2
1.6 Impact of dye on the environment and human beings ......................................... 2
1.7 Textile wastewater treatment ............................................................................... 2
1.8 Nanotechnology ................................................................................................... 2
1.9 Classification of nanoparticles ............................................................................. 2
1.10 Nanoparticles Synthesis ....................................................................................... 3
1.10.1 Top-Down approach ....................................................................................... 3

1.10.2 Bottom-up approach........................................................................................ 3

1.11 Green Method ........................................................................................................ 3

1.12 Citrullus colocynthis ............................................................................................. 3
1.13 Characterization of Nanoparticles ......................................................................... 4
1.14 Application in wastewater treatment ..................................................................... 4
1.15 Aims and Objectives ............................................................................................. 5
Chapter 2

LITERATURE REVIEW ...................................................................................................... 6

Chapter 3

MATERIALS AND METHODS ......................................................................................... 10

3.1 Sample collection ................................................................................................ 10

3.2 Preparation of seed extract .................................................................................. 11
 3.3 Preparation of standard salt solution ................................................................... 11
3.4 Preparation and collection of Cu-NPs................................................................. 11
3.5 Characterization of nanoparticles ....................................................................... 11
3.6 UV-Vis Spectroscopy.......................................................................................... 12
3.7 FTIR Spectroscopy.............................................................................................. 12
3.8 Scanning Electron Microscopy ........................................................................... 12
3.8 EDX ..................................................................................................................... 12
3.10 X-ray Diffraction ................................................................................................. 12
3.11 Photocatalytic Activity ........................................................................................ 12
3.12 Chemical Analysis............................................................................................... 13
3.13 Degradation study ............................................................................................... 13
3.14 Statistical analysis ............................................................................................... 14
Chapter 415

RESULTS AND DISCUSSION ........................................................................................... 15

4.1 UV-Visible Spectroscopy………………………………………………………15

4.2 FTIR spectroscopy .............................................................................................. 16

4.3 Scanning Electron Microscope (SEM) ............................................................... 16
4.4 EDX .................................................................................................................... 19
4.5 XRD .................................................................................................................... 20
4.6 Photocatalytic activity......................................................................................... 20
4.7 Calibration curve ................................................................................................. 21
4.8 Catalytic dose ...................................................................................................... 21
4.9 Contact time ........................................................................................................ 23
4.10 pH........................................................................................................................ 24
4.11 Temperature ........................................................................................................ 26
Chapter 5

CONCLUSION AND RECOMMENDATIONS ................................................................ 28

5.1 Conclusion................................................................................................................ 28
5.2 Limitations ............................................................................................................... 28
5.2.1 Synthesis Variability: .................................................................................... 28

5.2.2 Particle Size and Shape: ................................................................................ 29

5.2.3 Stability and Aggregation:............................................................................. 29

5.2.4 Long-Term Performance: .............................................................................. 29

 5.2.5 Scale-up Challenges: ..................................................................................... 29

5.2.6 Eco toxicity and Environmental Impact: ....................................................... 29

5.2.7 Complex Wastewater Matrices: .................................................................... 29

5.2.8 Regulatory and Safety Considerations: ......................................................... 30

5.2.9 Comparative Studies: .................................................................................... 30

5.3 Recommendations ............................................................................................... 30

5.3.1 Optimization of Synthesis Parameters: ......................................................... 30

5.3.2 Standardized Characterization:...................................................................... 30

5.3.3 Real Wastewater Studies: .............................................................................. 30

5.3.4 Mechanistic Studies:...................................................................................... 30

5.3.5 Long-Term Performance Assessment: .......................................................... 31

5.3.6 Environmental Impact Assessment: .............................................................. 31

5.3.7 Hybrid Systems: ............................................................................................ 31

5.3.8 Regulatory and Safety Considerations: ......................................................... 31

Chapter 6

REFERENCES ...................................................................................................................... 32
 i

ABSTRACT
Dye-mediated textile wastewater has a major contribution to environmental pollution.
Nanotechnology is showing effective results in dye degradation such as methylene blue.
The current research focuses on a green route for the production of Copper Nanoparticles
(Cu NPs). Leaf extract solution (15%) of Citrullus colocynthis (Desert Gourd) is mixed in
a ratio of 1:3 with 0.01M CuSO, solution for the production of Cu NPs. The method
developed is green, environment-friendly, and cost-effective. The plant extract is used as
both a reducing and capping agent. The change in color of the mixture indicates the
formation of Cu NPs.UV-Visible spectroscopy, Fourier Transform Infrared (FTIR)
spectroscopy, Scanning Electron Microscopy (SEM), Energy dispersive X-ray (EDX)
spectroscopy, and X-ray diffraction (XRD) spectroscopy were used to characterize the
synthesized Cu NPs. The particles show maximum absorption (ms) at 560 nm using UV-
Vis spectroscopy. The particles show an average size of 173 nm using XRD and they are
spherical in shape characterized by SEM and EDX. These nanoparticles are used as
photocatalysts when used for the degradation of methylene blue (MB) dye. Different
parameters such as dye concentration, catalytic dose, pH, and contact time were studied.
Each of the parameters was varied independently while keeping the other constant. When
irradiated under UV radiations, up to 96 % of MB dye degraded in the presence of these
particles. Biologically synthesized copper nanoparticles can effectively be utilized for the
treatment of textile wastewater.
 ii

ACKNOWLEDGEMENT
We extend our sincere gratitude to all individuals who have supported us during the
course of this thesis endeavor. We express our heartfelt thanks to Prof. Dr. Sajid Rashid
Ahmad, the esteemed Principal of our College, for his guidance, and unwavering support
throughout our academic pursuit. Our research team expresses sincere gratitude to
Dr.Azhar Ali, our respected supervisor, for his invaluable mentorship, steadfast
encouragement, and exceptional proficiency throughout our research work. The guidance
provided by the supervisor has played a crucial role in enhancing our concepts and
motivating us to achieve greater accomplishments. We express our utmost gratitude for
his unwavering dedication and the significant influence he has exerted on our research
work.

Furthermore, we express our gratitude to Dr. Bilal Shakoor and Dr. Abdul Qadir for
helping us in devising the methodology and providing us with the apparatus to execute
the experiment. Ultimately, we express our gratitude to our esteemed Lab Attendants for
their guidance throughout our research.

Fakiha Safdar

Arsalna Naseer
 iii

LIST OF FIGURES
Figure 3.1:- Flow chart of sample collection…………………………………………….10
Figure 3.2:- Stepwise Preparation and extraction of copper nanoparticles………………11
Figure 3.3:- Photocatalytic activity of Cu NPs…………………………………………...13
Figure 4.1:- UV-Vis scan of CuNPs between wavelength in nm and absorbance……….15
Figure 4.2:- FTIR spectrum of CuNPs between wavelength in cm-1 and transmittance...16
Figure 4.3:- SEM images of Cu NPs at various resolutions (a) at 1um (b) 2 µm (c) at 500
nm (d) at 200 nm…………………………………………………………………………18
Figure 4.4:- EDX Image of Cu NPs (a) Peak View of elements (b) Weight % of
elements…………………………………………...……………………………………...19

Figure 4.5:- XRD graph of Cu NPs between intensity (a.u) and angle (2θ)……………..20
Figure 4.6:- Graph showing Standard curve of calibration for MB dye…………………21
Figure 4.7:-Graph indicating Influence of adsorbent dose on the exclusion of MB dye...22
Figure 4.8:- Graph between Adsorbent dos (g) and Removal capacity ( q)……………...23
Figure 4.9:- Graph indicating Influence of Contact Time in the exclusion of MB dye….24
Figure 4.10:- Graph between Time (min) and Removal capacity ( q)…………………...24
Figure 4.11:- Graph showing Influence of pH in the exclusion of MB dye……………...25
Figure 4.12:- Graph between pH and Removal capacity ( q)…………………………….26
Figure 4.13:- Influence of Temp (C) in the exclusion of MB dye……………………….27
Figure 4.14:- Graph between Temp(°C) and Removal capacity ( q)…………………….27
 iv

LIST OF TABLES
TABLE 1.1:- SCIENTIFIC CLASSIFICATION OF CITTRULUS COLSYNTHIS
PLANT ......................................................................................................................... 4
TABLE 4.1:- CALIBRATION CURVE DATA FOR MB DYE ...................................... 21
TABLE 4.2:- ADSORPTION DOSE DATA FOR MB DYE ........................................... 22
TABLE 4.3:- CONTACT TIME DATA FOR MB DYE .................................................. 23
TABLE 4.4:- PH DATA FOR MB DYE ........................................................................... 25
TABLE 4.5:- TEMPERATURE DATA FOR MB DYE ................................................... 26
 v

LIST OF ABBREVIATIONS
NPs Nanoparticles
UV Ultraviolet
Vis Visible
FTIR Fourier Transform Infrared
SEM Scanning Electron Microscope
EDX Energy dispersive X-ray
XRD X-ray Diffraction
LSPR Localized Surface Plasmon Resonance
FCC Face centered cubic
JCPDS Joint Committee on Powder Diffraction Standards
FWHM Full width at half maximum
nm Nanometer
mm Micrometer
cm Centimeter
Cu Copper
MB Methylene Blue
TEM Transmission electron microscopes
DLS Dynamic light scattering
LSPR Localized Surface Plasmon Resonance
Chapter 1 1 Introduction

Chapter 1

INTRODUCTION

1.1 Overview

This chapter focuses on synthetic dye-mediated textile wastewater pollution and the
degradation of methylene blue. Nanotechnology is coupled with environmental
remediation to give eco-friendly and sustainable solutions. Different types of nanoparticle
routes for their generation are discussed along with the special focus on the green
synthesis of copper nanoparticles by Citrullus colosynthis seeds extract.
1.2 Water pollution

Water pollution is when human activities pollute water. It is adversely effecting

human health, aquatic ecosystems, and the economy. Water pollution is an important
environmental issue caused by various factors such as agricultural runoff, industrial
discharges, and poor waste management (Lin et al., 2022)
1.3 Textile industry in Pakistan

Pakistan's major manufacturing sector is the textile industry. The production of

textiles and clothing contributes significantly to Pakistan's economy, yet the sector is also
a major contributor to water pollution. (Ahmad, 2009). Textile effluent is wastewater
from the textile industry. It contains a variety of pollutants that can harm the environment,
aquatic habitats, and human health. Textile wastewater is frequently contaminated with
harmful substances such as hazardous metals, dyes, dissolved solids, and suspended
solids (Senthil Kumar & Saravanan, 2017).
1.4 Textile dyes

Textile dyes are used to color fabrics. Textile dyes can be natural or can be
synthesized in laboratories. Textile dyeing materials can be harmful to the environment if
not treated properly (Singh et al., 2017). According to the nature following are the types
of dye.
 Acid dye: Natural fibers such as wool and silk are dyed.
 Basic dyes: Used in synthetic fibers such as nylon and polyester.
 Direct Dyes: Used to dye cellulose fibers such as cotton.
Chapter 1 2 Introduction

 Reactive dyes: One of the most popular textile dyes. It can be used to dye a
variety of yarns.
 Vat dyes: For cotton and other cellulose fibers. It is not water-soluble but can be
converted to water-soluble.
 Sulfur dye: Used in the manufacture of cotton and other cellulose fibers. It is
inexpensive and easy to use, but not very stable (benkhaya said et al., 2017).
1.5 Methylene blue

Methylene Blue is an organic, synthetic, and reactive dye with a complex chemical
structure. A variety of industries uses Methylene blue dye, but it is also considered toxic
and potentially harmful to both environment and human beings (Khan et al., 2022). In this
study, we will carry out the dye methylene blue degradation.
1.6 Impact of dye on the environment and human beings

Textile dyes can have adverse effects on human health; Textile dyes cause birth
deformities, cancer, respiratory issues, and skin irritation. The severity of these effects
depends on the type of drug, how it is used, and how much it is exposed to.(Tang et al.,
2018)
1.7 Textile wastewater treatment

Traditional methods of textile wastewater treatment, such as chemical treatment, can

be harmful to the environment and cause secondary pollution, New methods of textile
wastewater treatment, such as biological and electrochemical methods, are
environmentally friendly (Azanaw et al., 2022).
1.8 Nanotechnology

Chemical treatment is one of the traditional textile wastewater treatment technologies

that can be hazardous to the environment and result in secondary contamination.
Environmentally friendly new techniques for treating textile effluent include biological
and electrochemical processes.(Amjad et al., 2021).
1.9 Classification of nanoparticles

Based on their size, shape, and other characteristics, nanoparticles are categorized
into different categories. The following list includes some significant nanoparticle classes
Chapter 1 3 Introduction

such as Polymeric Nanoparticles, Ceramic Nanoparticles, Metal nanoparticles,

Semiconductor Nanoparticles, and Carbon-based nanoparticles (Gao et al., 2022).

1.10 Nanoparticles Synthesis

The top-down approach and the bottom-up approach are two methods for creating
nanoparticles.
1.10.1 Top-Down approach
Bulk materials are separated in top-down methods to create nanostructured
materials(Baig et al., 2021). It includes Laser ablation, Mechanical milling,
Sputtering, Etching, and Electro explosion.
1.10.2 Bottom-up approach
The bottom-up strategy incorporates key elements into larger structures via
nanoscale physical and chemical processes (J. P. Singh et al., 2020). The most
popular bottom-up techniques for creating nanoparticles include Synthesis of
chemicals, Template-assisted synthesis, and self-assembly.
1.11 Green Method

The use of green-synthesized copper nanoparticles (CuNPs) has gain immense

attention due to their novel catalytic properties and minimal environmental impact and
reducing the number of processes used (Hajizadeh et al., 2022). Green synthesis is a
method which focuses on synthesizing nanoparticles using environmentally friendly
methods. This can be done using natural resources, such as plants, bacteria, or algae.
Green synthesis has several advantages over traditional methods of synthesis (J. Singh et
al., 2018).
1.12 Citrullus colocynthis

This Research focuses on developing green copper nanoparticles in this study

using a substance obtained from Citrullus colocynthis, a plant renowned for its abundance
of medicinal chemicals and reductive properties. This group's perennial herbaceous plant
is Citrullus colocynthis. Citrullus colocynthis, a desert plant with a fruit known as "bitter
apple," has been used medicinally for millennia and has a variety of pharmacological
properties, including purgative, anti-diabetic, laxative, anthelmintic, anti-inflammatory
and anti-cancerous (Li et al., 2022). With this study, the aim is to understand the efficacy
of green-synthesized CuNPs as catalytic agents for the degradation of methylene blue.
Chapter 1 4 Introduction

Table 1.1:- Scientific classification of Cittrulus colsynthis plant

Classification Name
Kingdom Plantae
Sub-Kingdom Viridiplantae –green-colored plants
Infra-Kingdom Streptophyta –land-living plants
Super-division Embryophyta
Division Tracheophyta –vascular plants
Sub-division Spermatophytina –seed plants
Class Magnoliosida
Superorder Rosanae
Order Cucurbitales
Family Cucurbitaceae –gourds, citrouilles
Genus Citrullus, watermelon
Specie Citrullus colocynthis (L.) Schrad. -colocynths

1.13 Characterization of Nanoparticles

There are many methods for characterizing nanoparticles. These techniques can
be used to study size, shape, composition, and surface properties. Common methods
include TEM, DLS, XRD, UV-Vis, and Raman spectroscopy. Each technique has
benefits and drawbacks. The method selected will rely on the unique characteristics of the
nanoparticles (Mourdikoudis et al., 2018).
1.14 Application in wastewater treatment

Nanoparticles have potential applications in wastewater treatment. It can be used to

purge water of impurities like heavy metals, bacteria, and viruses. It can be used to
improve wastewater treatment. They have the potential to reduce the environmental
impact of wastewater treatment. Nanoparticles are still being developed, but have the
potential to revolutionize wastewater treatment.
Chapter 1 5 Introduction

1.15 Aims and Objectives

 To couple the nanotechnology and environmental remediation in providing a

solution to synthetic dye pollution.
 To synthesize copper nanoparticles by green route and test their potential in
methylene blue dye degradation.
Chapter 2 6 Literature Review

Chapter 2

LITERATURE REVIEW
Nanoparticles are a fascinating area of scientific research because of their very
small size, and unexpected findings, a wide range of possible applications have been
made. But it also presents difficulties for scientists, who must develop new methods of
synthesis, characterization, and modeling to understand and harness their properties. This
chapter discusses the research that has been done on nanoparticles, focusing on their
synthesis, characterization, and applications in different fields of technology.
In a study conducted by (Amjad et al., 2021), Copper nanoparticles (CuNPs) were
produced using the environmentally friendly approach of mint (Mentha longifolia L.)
extraction. SEM, XRD and FTIR were used to characterize the CuNPs. The effects of
CuNPs on wheat germination and growth were assessed using a germination test and a
growth assay. According to the results, CuNPs at a concentration of 50 mg/L greatly
increased the rate of wheat seed germination and the growth of wheat seedlings. Further
investigation is needed to determine how CuNPs impact wheat growth. According to the
study's findings, mint extract-derived green CuNPs may be able to promote wheat
germination and growth.
(Mali et al., 2020) prepare the copper nanoparticles (CuNPs) using the leaf extract
of the plant Celastrus paniculatus. UV-Vis, SEM, XRD and FTIR were used to
characterize the CuNPs. The dye methylene blue was effectively photocatalyzed by the
CuNPs, with a 90% degradation rate occurring in just one hour. With a 12 mm inhibitory
zone, the CuNPs also demonstrated strong antifungal efficacy against the mold Fusarium
oxysporum. The outcomes demonstrated that CuNPs produced using Celastrus
paniculatus leaf extract have effective photocatalytic and antifungal activities.
The study by (Abbas et al., 2020), Aloe barbedensis, Azadirachta indica,and
Coriandum setivum aere used to prepare Silver and copper nanoparticles. SEM, X-ray
diffraction (XRD), and UV-visible spectroscopy were all used to characterize the
nanoparticles. The naphthalene was adsorbed from an aqueous solution using the
nanoparticles. With a maximum removal effectiveness of 98.81% for Azadirachta indica
and 95.29% for Coriandrum sativum, the nanoparticles demonstrated significant
adsorption capacity for naphthalene. The findings demonstrated that plant-derived silver
Chapter 2 7 Literature Review

and copper nanoparticles can function as efficient adsorbents for naphthalene

decontamination.
(Ghosh et al., 2020) reported a novel biological technique for producing
environmentally friendly CuNPs from the leaf extract of Jatropha carcass. Fourier
transform infrared spectroscopy (FTIR), SEM, X-ray diffraction (XRD), and UV-visible
were used to characterize the CuNPs. The CuNPs were spherical with size from 10 to 12
nm. The CuNPs displayed good optical characteristics and had a 3.6 eV band gap. The
CT-DNA was well-bound by the CuNPs, with a binding constant of 1.024 102 M-1. The
photocatalytic activity of the CuNPs under UV light was good with the methylene blue
degradation rate of 90%.
CuNPs were produced utilizing a green technique in this study by (Mali et al.,
2020), using the leaf extract of the curative herb Celastrus paniculatus. After
characterizing by various spectroscopy techniques, a panel of bacteria and fungi was used
to test CuNPs' photocatalytic activities. CuNPs with size range of 10-15 nm were created.
The CuNPs were well-dispersed and spherical in shape in the leaf extract, according to
the SEM pictures. The XRD pattern demonstrated that CuNPs are crystalline. Functional
groups like hydroxyl, carbonyl, and carboxyl groups, were detected by FTIR analysis.
CuNPs had strong photocatalytic activity against the dye methylene blue, degrading 90%
of it in just one hour.
In this study by (Pérez-Alvarez et al., 2021), Cotton textile fibers were used green
synthesis of CuNPs. FTIR, SEM, XRD, and UV-Vis were used to characterize the
CuNPs. The CuNPs were spherical in size range from 13 to 35 nm. The CuNPs were
stable in aqueous solution for at least six months. The degradation of the dye methylene
blue was successfully catalyzed by the CuNPs. The outcomes demonstrated that cotton
textile fibers might be used in a green way to create CuNPs and their Potential use as
catalyst in dye degradation.
(Rasool et al., 2022) showed that the effectiveness of silver nanoparticles (AgNPs)
produced from Citrullus colocynthis (bitter melon) fruit extracts in treating human
infection and breast cancer was examined. The silver nitrate solution was incubated with
the fruit extracts while being sonicated to create the AgNPs. The generated AgNPs were
characterized by TEM, dynamic light scattering (DLS), and UV-Vis spectroscopy. TEM
scans showed that the AgNPs were spherical and had an average size of 10 nm.
According to DLS measurements, the AgNPs' size distribution was condensed. AgNPs
contain a sizable visible spectrum absorption band, according to measurements obtained
Chapter 2 8 Literature Review

using UV-VIS spectroscopy. The antiproliferative effect of AgNPs was evaluated in the
breast cancer cell line MCF-7. MCF-7 cells were discovered to be efficient against three
pathogens, including E. coli. AgNPs created from C. colocynthis fruit extracts, according
to the authors, have the potential to be brand-new treatments for infections and breast
cancer. MICs for P. aeruginosa and E. coli are respectively 10 g/mL and 20 g/mL.
In this study, by (Nieto-Maldonado et al., 2022) ,Withania somnifera root extract
(WSRE), Rosa 'Andeli' double pleasure fresh petals (RAFE), and Gardenia jasminoides
Ellis fresh leaves (GJLE) were some of the extracts used to make CuNPs in a green way.
FTIR, SEM, XRD, and UV-Vis were used to characterize the CuNPs. A panel of
microorganisms, including Staphylococcus aureus (S. aureus), Escherichia coli (E. coli),
and Pseudomonas aeruginosa (P. aeruginosa), were used to test CuNPs' antibacterial
activity. The sizes and forms of the CuNPs that were created using the various plant
extracts varied. The tiniest particles were the RAFE-NPs, which had an average size of 10
nm. The GJLE-NPs, with an average size of 20 nm, were the biggest. The WSRE-NPs
had an average size and were of intermediate size. All of the CuNPs had a spherical form.
The CuNPs were well-dispersed in the plant extracts, as evidenced by the SEM pictures.
The crystalline character of CuNPs was confirmed by the XRD patterns. Functional
groups known to be important to CuNPs' antibacterial action.
This study by (Ying et al., 2022) illustrates that Plant extracts can be used to
produce nanoparticles Biomolecules found in plant extracts can revert metal ions to their
elemental state and regulate the size and structure of nanoparticles. Numerous methods
can be used to characterize the nanoparticles' size, shape, and surface characteristics.
There are numerous possible uses for the nanoparticles created by plant-based green
synthesis, including pharmaceuticals, catalysis, environmental clean-up, agriculture, and
cosmetics. A fascinating new technology with many potential uses is plant-based green
synthesis. This technique is sustainable and friendly to the environment because it uses
plant extracts. The synthesis of nanoparticles using this approach needs to be optimized,
and new uses for these nanoparticles need to be developed. Plant extracts, which are
renewable resources, are used in plant-based green synthesis. In general, plant extracts are
not harmful to people. Plant extracts are reasonably priced. The green production of
nanoparticles using plants also has significant drawbacks. Plant extract contaminants may
contaminate the nanoparticles created by plant-based green synthesis. Uncertainty
surrounds the mechanism through which plant-based green synthesis generates
nanoparticles. Carbonyls, and carboxyl groups, were detected by FTIR analysis. The
Chapter 2 9 Literature Review

CuNPs created utilizing the various plant extracts showed various levels of antibacterial
activity. The most potent antibacterial action was demonstrated by the RAFE-NPs, which
exhibited a minimum inhibitory concentration (MIC) of 10 g/mL. With a MIC of 100
g/mL, the GJLE-NPs exhibited the least bactericidal activity. The MIC of the WSRE-NPs
was 50 g/mL, indicating moderate antibacterial activity.
(Amaliyah et al., 2017) introduced a green route to prepare Copper nanoparticles
(CuNPs) using the plant Piper retrofractum Vahl. FTIR, SEM, XRD, and UV-Vis were
used to characterize the CuNPs. A panel of microorganisms, including Staphylococcus
aureus (S. aureus), Escherichia coli (E. coli), and Pseudomonas aeruginosa (P.
aeruginosa), were used to test the antibacterial activity of CuNPs. The Piper retrofractum
Vahl extract was used to create CuNPs, with an average size of 10-15 nm. Functional
groups known to be involved in the antibacterial activity of CuNPs, including hydroxyl,
carbonyl, and carboxyl groups, were detected by FTIR analysis.
This study by (Ying et al., 2022) illustrates that Plant extracts can be used to
produce nanoparticles Biomolecules found in plant extracts can revert metal ions to their
elemental state and regulate the size and structure of nanoparticles. There are numerous
possible uses for the nanoparticles created by plant-based green synthesis, including
pharmaceuticals, catalysis, environmental clean-up, agriculture, and cosmetics. A
fascinating new technology with many potential uses is plant-based green synthesis. This
technique is sustainable and friendly to the environment because it uses plant extracts.
The synthesis of nanoparticles using this approach needs to be optimized, and new uses
for these nanoparticles need to be developed. Plant extracts, which are renewable
resources, are used in plant-based green synthesis. In general, plant extracts are not
harmful to people. Plant extracts are reasonably priced. The green production of
nanoparticles using plants also has significant drawbacks. Plant extract contaminants may
contaminate the nanoparticles created by plant-based green synthesis. Uncertainty
surrounds the mechanism through which plant-based green synthesis generates
nanoparticles.
Chapter 3 10 Materials and Methods

Chapter 3

MATERIALS AND METHODS

The aim of this study was to provide an effective method for the green production of
copper nanoparticles (Cu-NPs) and the photo-catalytic degradation of methylene blue
dye. The experimental work was done in the College of Earth and Environmental
Science, University of the Punjab, Lahore, Pakistan. First, the copper nanoparticles (Cu-
NPs) were synthesized using Citrullus colocynthis seed extract and then used for the
methylene blue dye degradation. Following is the experimental procedure of this study.
3.1 Sample collection

Citrullus colocynthis plant seeds were collected from the desert of Cholistan
(Bahawalpur, Pakistan). To eliminate the dust and undesirable particles, the seeds were
washed with deionized water after washing with tap water. The dried seeds were then
ground to the powder in a mortar pestle and stored in an air-sealed jar for further use
(Sinha & Ahmaruzzaman, 2015)

Figure 3.1:- Flow chart of sample collection

Chapter 3 11 Materials and Methods

3.2 Preparation of seed extract

A 15% by-weight plant extract solution was made by mixing ground dry seeds of
citrullus colocynthis with deionized water. This mixture was heated for two hours at 80°C
in the oven. The mixture was cooled at room temperature before filtering twice using
watmann-42 filter paper to get a clear solution. This solution was stored in the freezer at
-4°C for further use. To avoid any kind of contamination, the solution was used within a
week.
3.3 Preparation of standard salt solution

A 0.01 M solution of Cu (II) ion was obtained by dissolving a sufficient amount of

anhydrous CuSO4 in deionized water and sonication the solution for full salt breakdown.
3.4 Preparation and collection of Cu-NPs

Cu-NPs were made by combining 1:3 of plant extract solution and standard solution
of cu (II) ions. The mixture is heated with constant stirring for 1 hour at 60°C in a hot
magnetic plate the solution is then placed for 48 hours for the formation of nanoparticles.
Green precipitates were formed at the bottom of the beaker indicating the formation of
Cu-NPs. Nanoparticles were washed with deionized water and absolute ethanol numerous
times to remove all the organic and inorganic impurities from the particles. The particles
were later obtained by centrifuging them at 6000 RPM in a Ke-Cheng L2-6K centrifuge
machine. Particles were dried in the oven at about 80°C for 1 day.

Figure 3.2:- Stepwise Preparation and extraction of copper nanoparticles

3.5 Characterization of nanoparticles

To check the synthesis, shape, size, and morphology of nanoparticles, several

spectroscopic methods such as UV-Vis spectroscopy, Fourier transform infrared (FTIR)
spectroscopy, scanning electron microscopy (SEM), and X-ray diffraction technique
(XRD) were utilized.
Chapter 3 12 Materials and Methods

3.6 UV-Vis Spectroscopy

Absorption spectroscopy is a useful tool for determining Cu (II) ion reduction to

Cu NPs. To examine the absorption spectra of the synthesized Cu NPs, Schimadzu-1800
double-beam spectrophotometer was used after they had been dissolved in water. The
200-800 mm scanning range was used for the study.
3.7 FTIR Spectroscopy

FTIR spectroscopy is used to identify the surface functional groups of Cu NPs. An

Agilent Technology Cary-630 FTIR spectrophotometer was used for analysis. The NPs
were dispersed onto a KBr pellet, and their spectrum was recorded within the range of
650-4000 cm.
3.8 Scanning Electron Microscopy

To examine the size and surface shape of Cu NPs, Scanning electron microscopy
(SEM) is utilized. A high-energy electron beam is used in scanning in SEM, and when it
interacts with the sample's atoms, it produces a variety of signals. These electron-atom
interactions provide important details about the surface shape, structure, chemical
makeup, and whether the material is crystalline or amorphous.
3.8 .EDX

An energy-dispersive X-ray analyzer (EDX) is employed in conjunction with the

SEM. EDX imaging employs secondary electrons. For these studies, a specific Nova
Nano SEM-450 device running at 10 kV is used. For best results, samples are coated with
a coating of Au and dehydrated at 373 K for 240 minutes before analysis.
3.10 X-ray Diffraction

To assess the sample's structural characteristics, X-ray Diffraction (XRD) analysis

was used. The Rigaku D/Max equipment was used to conduct XRD on the prepared
sample.
3.11 Photocatalytic Activity

A 50 mL dye solution (50 ppm) was prepared in distilled water, and the pH was
adjusted to 5 by adding 1 M NaOH / 0.5 M H2SO4. 0.1 g of Copper nanoparticles were
then added to the solution. The mixture-containing reaction vessel was shacked for 30
minutes in the dark to achieve adsorption-desorption equilibrium.2ml sample was
Chapter 3 13 Materials and Methods

obtained from the reaction vessel Several times after 15 minutes to check the progress of
the reaction. A spectrophotometer was used to measure the absorbance of these samples
at λ max.
Cu NPs concentration (Adsorption dose) was varied from 0.1g to 0.9g, and pH
from 1 to 9. Temperature was varied from 10 °C to 50 °C and the Contact Time from 05
minutes to 45 minutes. To optimize the experimental parameters, a systematic method
was used in which each parameter was changed separately while the other values
remained constant. The ideal conditions for reaction and the influence of each parameter
on the response could be assessed separately by this technique.

Figure 3.3:- Photocatalytic activity of Cu NPs

3.12 Chemical Analysis

All the experiments were performed thrice. UV spectrophotometer was used to

detect the residual dye concentration in the filtered solution at 660 nm. The following
formula was used to calculate the deterioration efficiency (%):

Degradation (%) = (Co - C) / Co x 100……………….. (i)

Where Co is the dye solution's initial absorbance and C is the absorbance at the
given time throughout the photocatalytic process.

3.13 Degradation study

The assessment of methylene blue dye degradation involved a sequential analysis,

encompassing bond cleavage and the subsequent generation of the novel compounds.
Chapter 3 14 Materials and Methods

3.14 Statistical analysis

The data was gathered in triplicate, and the values were estimated as the average
of triple measurements with the standard error of the means taken into account.
Chapter 4 15 Results and discussion

Chapter 4

RESULTS AND DISCUSSION

This chapter describes the results and discussions of the generated Cu NPs'
characterization and potential application in methylene blue degradation. Several
analytical methods, such as UV-Visible Spectroscopy, have been utilized to characterize
synthesized Cu NPs. FTIR, SEM, EDX, and XRD are all employed. UV-Vis
spectroscopy was used to verify the initial production of the green-manufactured Cu NPs.
The surface binding relationships of different green chemicals present in plant extract
solution with Cu NPs were investigated using FTIR spectroscopy. SEM, EDX, and XRD
were used to investigate the form and size of the green synthesized Cu NPs.
4.1 UV-Visible Spectroscopy
Cu NPs' conduction electrons swing in response to electromagnetic radiation
(EMR). These oscillations provide the basis of the absorption band of Cu NPs in UV-Vis
spectrophotometry. Localised Surface Plasmon Resonance (LSPR) describes the
oscillations created by NPs when they interact with EMR. The absorption band reveals
details about the composition of Cu NPs. The plant extract and the particles to be
generated determine the maximal absorption detected by NPs. Cu NPs' oscillation band is
claimed to be 560-600 nm. The UV-Vis spectrum of the LSPR displayed by the Cu NPs
at 560 nm is shown in the figure below.

Figure 4.1:- UV-Vis scan of CuNPs between wavelength in nm and absorbance

Chapter 4 16 Results and discussion

4.2 FTIR spectroscopy

FTIR spectroscopy is used to detect the presence of green substance employed as a

reducing agent. This green substance also serves as a capping agent for the Cu NPs. The
transmitted peaks in the FTIR measurements of green synthesized Cu NPs were 3278,
2924, 2854, 2321, 1741, 1637, 1542, 1377, and 1237 cm-1.. The peak at 1377 cm-1
corresponds to the Methyl C ̶ H asymmetrical/symmetrical bend. The peak at 2924 cm-1 is
due to the starching of asymmetrical/symmetrical Methylene C ̶ H. The stretching Normal
Polymeric Alcohol occurs at 3278 cm-1. The peak at 1237 cm-1 is due to Aromatic ethers,
aryl ̶ O stretch. Primary amines, NH bends at 1637cm-1. The peak at 1741 cm-1 shows
Alkyl carbonates. Aliphatic nitro compounds at a transmittance of 1542 cm -1 fall in the
Nitrogen-oxy group. The peak transmittance at 2321 cm-1 and 2854 cm-1 shows Thiols
(S ̶ H Stretch). (Coates, 2000)

Figure 4.2:- FTIR spectrum of CuNPs between wavelength in cm-1 and

transmittance

4.3 Scanning Electron Microscope (SEM)

SEM analysis is employed to evaluate the shape and size of the green
manufactured Cu NP. SEM images were taken at a variety of resolutions, including 1 μm,
Chapter 4 17 Results and discussion

2 μm, 500 nm, and 200 nm. The particles in the images are irregularly spaced, spherical
in shape, and form clusters. Particles are in a close connection to one another.
Chapter 4 18 Results and discussion

Figure 4.3:- SEM images of Cu NPs at various resolutions (a) at 1um (b) 2 µm (c) at
500 nm (d) at 200 nm
Chapter 4 19 Results and discussion

4.4 EDX

EDX analysis confirmed the presence of other components in addition to Cu NPs.

Because of the presence of proteins and other green material coupled to the Cu NPs,
additional elements like C, H, and S are present. These bio-molecules are connected to
the Cu NPs as capping agents. Figure 4.4 depicts the presence of numerous components
containing Cu NPs, as well as their weight percentage.

Figure4.4:- EDX Image of Cu NPs (a) Peak View of elements (b) Weight % of
elements
Chapter 4 20 Results and discussion

4.5 XRD

The metallic and crystalline character, as well as the size, of green Manufactured Cu
NPs are determined by powder XRD spectroscopy. The results reveal that face-centered
cubic (fee) crystals with planes (111), (200), and (220) may be created. These findings
corroborate the standard JCPDS Card number 85-132. (Biçer & Şişman, 2009). The
following equation shows the Scherrer formula to find the particle size of the Cu NPs.

Particle size L (mm) = K λ / β. Cos θ ………………….(1)

Where k is constant having a value of 0.9, λ is the wavelength of the X-ray which
is 0.15406 nm, B is the full width at half maximum (FWHM) value in radians, and is the
half of the diffraction angle of the center of the peak in radians. Using the Scherer
equation, the size of the Cu NPs is calculated to be 174.2 nm. This result agrees with the
SEM study and validates the XRD analysis. Figure shows the XRD results.

Figure 4.5:- XRD graph of Cu NPs between intensity (a.u) and angle (2θ)

4.6 Photocatalytic activity

To study the photocatalytic activity, dye solution was treated against biologically
synthesized cuNPs. Various factors were investigated, including contact time, catalytic
dosage, temperature, and pH. Only one parameter was changed at a time while the other
remained constant.
Chapter 4 21 Results and discussion

4.7 Calibration curve

Spectrophotometer was calibrated to produce the standard linear curve with

absorption at different dye concentrations.
Table 4.1:- Calibration curve data for MB dye

Calibration curve data for Methylene Blue Dye

Concentration
(ppm) Absorbance Equilibrium constant Slope Y Intercept
2 0.404 2.45407369
4 0.908 4.711987545
6 1.228 6.372599896
8 1.599 8.29787234

10 1.985 10.30098599 0.19265 0.0995

2.5
y = 0.1927x + 0.0689
R² = 0.995
2

1.5
Absorbance

Absorbance
1

0.5

0
0 5 10 15
Concentration (ppm)

Figure 4.6:- Graph showing Standard curve of calibration for MB dye

4.8 Catalytic dose

While keeping all other parameters constant, CuNP concentration was varied. The
following table shows the Adsorption dose data. The trend line of the graph shows that the
With the increase in the concentration of catalytic dose, the removal percentage also
increases.
Chapter 4 22 Results and discussion

Table 4.2:- Adsorption dose data for MB dye

Adsorption dose data for Methylene blue

Volume Volume Mass C˳ Cₑ q Removal
(ml) (L) (g) (mg/L) Absorbance (mg/L) (mg/g) (%)
50 0.05 0.1 50 0.670 3.49 23.26 93.02
50 0.05 0.2 50 0.596 3.10 11.72 93.79
50 0.05 0.3 50 0.564 2.94 7.84 94.13
50 0.05 0.4 50 0.510 2.66 5.92 94.69
50 0.05 0.5 50 0.467 2.43 4.76 95.14
50 0.05 0.6 50 0.458 2.39 3.97 95.23
50 0.05 0.7 50 0.453 2.36 3.40 95.28
50 0.05 0.8 50 0.447 2.33 2.98 95.34
50 0.05 0.9 50 0.430 2.24 2.65 95.52

96.00

95.50

95.00

94.50
Removal (%)

94.00

93.50
Removal (%)
93.00

92.50

92.00

91.50
0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9
Mass (g)

Figure 4.7:-Graph indicating Influence of adsorbent dose on the exclusion of MB

dye
Chapter 4 23 Results and discussion

25.00

20.00

15.00
q(mg/g)

10.00 q(mg/g)

5.00

0.00
0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9
Mass (g)

Figure 4.8:- Graph between Adsorbent dos (g) and Removal capacity ( q)
4.9 Contact time

The contact time data is shown below. The graph shows that up to a certain time, the
removal efficiency increases with contact time but then remains constant.

Table 4.3:- Contact Time data for MB dye

Contact Time data for Methyle Blue Dye

Volume Volume Mass Time C˳ Cₑ q Removal
(ml) (L) (g) (min) (mg/L) Absorbance (mg/L) (mg/g) (%)
50 0.05 0.1 5 50 0.697 3.630 23.185 92.740
50 0.05 0.1 10 50 0.671 3.495 23.253 93.010
50 0.05 0.1 15 50 0.558 2.906 23.547 94.188
50 0.05 0.1 20 50 0.494 2.573 23.714 94.854
50 0.05 0.1 25 50 0.461 2.401 23.799 95.198
50 0.05 0.1 30 50 0.460 2.396 23.802 95.208
50 0.05 0.1 35 50 0.460 2.396 23.802 95.208
50 0.05 0.1 40 50 0.461 2.401 23.799 95.198
50 0.05 0.1 45 50 0.462 2.406 23.797 95.188
Chapter 4 24 Results and discussion

95.500

95.000

94.500
Removal (%)

94.000

93.500

93.000 Removsl (%)

92.500

92.000

91.500
5 10 15 20 25 30 35 40 45
Time (min)

Figure 4.9:- Graph indicating Influence of Contact Time in the exclusion of MB dye

23.900
23.800
23.700
23.600
23.500
q(mg/g)

23.400
23.300
q(mg/g)
23.200
23.100
23.000
22.900
22.800
5 10 15 20 25 30 35 40 45
Time (min)

Figure 4.10:- Graph between Time (min) and Removal capacity ( q)

4.10 pH

By varying the pH from 1-9 it is found that the degradation happened maximum at 6.
The table shows the absorption data at different pHs while the graph shows the trend line
of the maximum absorption.
Chapter 4 25 Results and discussion

Table 4.4:- pH data for MB dye

Solution pH data for Methylene Blue Dye

Volume Volume Mass C˳ Cₑ q Removal
(ml) (L) (g) pH (mg/L) Absorbance (mg/L) (mg/g) (%)
50 0.05 0.1 1 50 0.691 3.599 23.201 92.802
50 0.05 0.1 2 50 0.635 3.307 23.346 93.385
50 0.05 0.1 3 50 0.548 2.854 23.573 94.292
50 0.05 0.1 4 50 0.473 2.464 23.768 95.073
50 0.05 0.1 5 50 0.455 2.370 23.815 95.260
50 0.05 0.1 6 50 0.295 1.536 24.232 96.927
50 0.05 0.1 7 50 0.324 1.688 24.156 96.625
50 0.05 0.1 8 50 0.388 2.021 23.990 95.958
50 0.05 0.1 9 50 0.457 2.380 23.810 95.240

98.000

97.000

96.000

95.000
Removal (%)

94.000
Removal (%)
93.000

92.000

91.000

90.000
1 2 3 4 5 6 7 8 9
pH

Figure 4.11:- Graph showing Influence of pH in the exclusion of MB dye

Chapter 4 26 Results and discussion

24.400
24.200
24.000
23.800
q (mg/g)

23.600
23.400
q (mg/g)
23.200
23.000
22.800
22.600
1 2 3 4 5 6 7 8 9
pH

Figure 4.12:- Graph between pH and Removal capacity ( q)

4.11 Temperature

It is noted that the removal percentage increases with the increase in temperature. The
table shows the absorption data while the graph shows the trend line at different
temperatures.
Table 4.5:- Temperature data for MB dye

Volume Volume Mass Temp C˳ Cₑ q Removal

(ml) (L) (g) (°C) (mg/L) Absorbance (mg/L) (mg/g) (%)
50 0.05 0.1 10 50 0.612 3.188 23.406 93.625
50 0.05 0.1 20 50 0.579 3.016 23.492 93.969
50 0.05 0.1 30 50 0.530 2.760 23.620 94.479
50 0.05 0.1 40 50 0.457 2.380 23.810 95.240
50 0.05 0.1 50 50 0.396 2.063 23.969 95.875
Chapter 4 27 Results and discussion

96.500

96.000

95.500

95.000
Removal (%)
94.500
Removal (%)
94.000

93.500

93.000

92.500
10 20 30 40 50
Temp (°C)

Figure 4.13:- Influence of Temp (C) in the exclusion of MB dye

24.100
24.000
23.900
23.800
23.700
q (mg/g)

23.600
23.500 q (mg/g)
23.400
23.300
23.200
23.100
10 20 30 40 50
Temp (°C)

Figure 4.14:- Graph between Temp(°C) and Removal capacity ( q)

Chapter 5 28 Conclusion and Recommendations

Chapter 5

CONCLUSION AND RECOMMENDATIONS

5.1 Conclusion

This study aimed to formulate copper nanoparticles with some eco-friendly and
economical techniques and to use them as photocatalytic agents. For this purpose green
method was selected in which some plants or microbes could be used for the formulation
of nanoparticles. Here in this research, extract of Citrullus colocynthis plant seeds was
used which act as both capping as well as reducing agent. As Citrullus colocynthis is an
herbaceous plant and can be easily found in deserts without any cost no solvents,
templates, or reducing agents were used in this technique, so the process is green, eco-
friendly, and cost-effective. The nanoparticles formulated were characterized by various
spectroscopic techniques like UV-Vis, FTIR. XRD, SEM, and EDX. The results of these
techniques showed that the particles formulated were irregularly spherical in shape and
their particle size was 17 nm. These Cu NPs when used as photocatalysts for the
degradation of MB dye they show good catalytic effects and degraded dye. Up to 96 %
removal efficiency was recorded.

5.2 Limitations

Research on the green synthesis of copper nanoparticles and their

application in dye wastewater treatment is an interesting and important field of study.
However, like any scientific research, it has its limitations. Here are some potential
limitations to consider
5.2.1 Synthesis Variability:
Green synthesis methods often involve natural extracts or biomolecules as
reducing agents. The composition and properties of these agents can vary significantly
due to factors such as seasonal changes, geographical location, and extraction methods.
This can lead to variability in the size, shape, and stability of the synthesized copper
nanoparticles, impacting their performance in dye wastewater treatment.
Chapter 5 29 Conclusion and Recommendations

5.2.2 Particle Size and Shape:

The effectiveness of nanoparticles in dye degradation depends on their size, shape,
and surface area. Controlling these parameters during synthesis can be challenging, as
they can affect the catalytic activity and stability of the nanoparticles. Variations in these
factors can result in inconsistent treatment outcomes.
5.2.3 Stability and Aggregation:
Copper nanoparticles synthesized using green methods may have limited stability
over time, leading to aggregation or precipitation. This can reduce their efficacy in
wastewater treatment and result in uneven treatment results.
5.2.4 Long-Term Performance:
This study focuses on short-term performance evaluations of synthesized
nanoparticles in wastewater treatment. However, the long-term stability and effectiveness
of these nanoparticles under real-world conditions, including repeated use, continuous
exposure to pollutants, and potential interference from other wastewater components,
might differ from initial laboratory results.
5.2.5 Scale-up Challenges:
Research conducted at the laboratory scale might not consider the challenges
associated with scaling up nanoparticle synthesis and application processes. Factors like
cost-effectiveness, scalability, and feasibility for industrial applications can pose hurdles
that are not evident in small-scale experiments.
5.2.6 Eco toxicity and Environmental Impact:
While green synthesis methods aim to reduce the environmental impact compared
to conventional methods, the nanoparticles themselves could still pose potential
environmental risks. The release of copper nanoparticles into the environment could have
unintended ecological consequences, such as bioaccumulation, toxicity to aquatic
organisms, and disruption of ecosystems.
5.2.7 Complex Wastewater Matrices:
Real-world dye wastewater is often complex, containing a variety of dyes,
additives, and other contaminants. The effectiveness of copper nanoparticles might vary
depending on the specific composition of the wastewater. Research conducted using
simplified or synthetic wastewater might not fully capture the challenges posed by real
wastewater matrices.
Chapter 5 30 Conclusion and Recommendations

5.2.8 Regulatory and Safety Considerations:

The use of nanoparticles in wastewater treatment raises concerns about their
potential release into the environment and their effects on human health and safety.
Researchers must address these considerations and ensure that the benefits of the
technology outweigh the potential risks.
5.2.9 Comparative Studies:
To evaluate the true benefits of copper nanoparticles in dye wastewater treatment,
it's important to compare their performance with other existing treatment methods. Some
studies might lack comprehensive comparisons with conventional methods, making it
difficult to assess their practical advantages.
While research on the green synthesis of copper nanoparticles for dye wastewater
treatment holds promise, researchers should aware of these limitations and work to
address them in order to develop a comprehensive understanding of the technology's
potential and limitations.
5.3 Recommendations

Following are the recommendations that can be considered for further research.

5.3.1 Optimization of Synthesis Parameters:

Investigate the effects of different synthesis parameters (such as precursor
concentration, temperature, pH, and reaction time) on nanoparticle properties. Optimizing
these parameters can lead to nanoparticles with enhanced catalytic activity and stability.
5.3.2 Standardized Characterization:
Develop standardized protocols for characterizing the size, shape, composition,
and surface properties of the synthesized copper nanoparticles. This will improve the
reproducibility of results across different studies.
5.3.3 Real Wastewater Studies:
Conduct studies using real dye wastewater samples to evaluate the nanoparticles'
performance in a more realistic context. This will provide a better understanding of their
efficiency and challenges in practical applications.
5.3.4 Mechanistic Studies:
Investigate the mechanisms involved in the dye degradation process catalyzed by
copper nanoparticles. Understanding the underlying chemistry can aid in optimizing the
nanoparticles' design and performance.
Chapter 5 31 Conclusion and Recommendations

5.3.5 Long-Term Performance Assessment:

Study the stability and efficacy of the synthesized nanoparticles over extended
periods and after multiple treatment cycles. This will provide insights into their potential
for sustainable and continuous wastewater treatment.
5.3.6 Environmental Impact Assessment:
Evaluate the environmental impact of releasing copper nanoparticles into the
environment after their use in wastewater treatment. This includes assessing their
potential for bioaccumulation and toxicity.
5.3.7 Hybrid Systems:
Explore the possibility of combining copper nanoparticles with other treatment
methods, such as adsorption, biological degradation, or other advanced oxidation
processes, to enhance overall treatment efficiency.
5.3.8 Regulatory and Safety Considerations:
Address regulatory and safety concerns related to the use of nanoparticles in
wastewater treatment. Collaborate with experts in environmental and health sciences to
ensure that the technology meets safety standards.
Incorporating these recommendations into future research can help overcome the
limitations and advance the understanding of green-synthesized copper nanoparticles for
dye wastewater treatment, ultimately contributing to more effective and sustainable water
treatment solutions.
Chapter 6 32 References

Chapter 6

REFERENCES

Abbas, S., Nasreen, S., Haroon, A., & Ashraf, M. A. (2020). Synhesis of Silver and
Copper Nanoparticles from Plants and Application as Adsorbents for Naphthalene
decontamination. Saudi Journal of Biological Sciences, 27(4), 1016.
https://doi.org/10.1016/J.SJBS.2020.02.011
Ahmad, Z. (2009). Research Report on Pakistan’s Textile Industry Analysis. SSRN
Electronic Journal. https://doi.org/10.2139/SSRN.1651789
Amaliyah, S., Pangesti, D. P., Masruri, M., Sabarudin, A., & Sumitro, S. B. (2017). Green
synthesis and characterization of copper nanoparticles using Piper retrofractum Vahl
extract as bioreductor and capping agent. Heliyon, e04636.
https://doi.org/10.1016/j.heliyon.2020.e04636
Amjad, R., Mubeen, B., Ali, S. S., Imam, S. S., Alshehri, S., Ghoneim, M. M., Alzarea, S.
I., Rasool, R., Ullah, I., Nadeem, M. S., & Kazmi, I. (2021). Green Synthesis and
Characterization of Copper Nanoparticles Using Fortunella margarita Leaves.
Polymers, 13(24). https://doi.org/10.3390/POLYM13244364
Azanaw, A., Birlie, B., Teshome, B., & Jemberie, M. (2022). Textile effluent treatment
methods and eco-friendly resolution of textile wastewater. Case Studies in Chemical
and Environmental Engineering, 6, 100230.
https://doi.org/10.1016/J.CSCEE.2022.100230
Baig, N., Kammakakam, I., Falath, W., & Kammakakam, I. (2021). Nanomaterials: a
review of synthesis methods, properties, recent progress, and challenges. Materials
Advances, 2(6), 1821–1871. https://doi.org/10.1039/D0MA00807A
benkhaya said, Elherfi, A., & Elherfi, S. (2017). (PDF) Classifications, properties and
applications of textile dyes: A review. Applird Journal of Environmental Engineering
Science.
Biçer, M., & Şişman, İ. (2009). Controlled synthesis of copper nano/microstructures
using ascorbic acid in aqueous CTAB solution.
https://doi.org/10.1016/j.powtec.2009.11.022
Coates, J. (2000). Interpretation of infrared spectra, a practical approach. 10815–10837.
https://www.researchgate.net/profile/Chandana-Chandrasekher/post/How-to-
interpret-my-FTIR-spectrum-Is-there-any-online-software-available-to-interpret-the-
Chapter 6 33 References

spectrum/attachment/5ac4a7094cde260d15d6bc94/AS%3A611637941780481%401
522837257071/download/joan+coates.pdf
Gao, Z., Lee, S., & Huang, J. (2022). The Application of Nanoparticle Ceramic Materials
in the Design of Modern Handicrafts. Journal of Nanomaterials, 2022.
https://doi.org/10.1155/2022/1998985
Ghosh, M. K., Sahu, S., Gupta, I., & Ghorai, T. K. (2020). Green synthesis of copper
nanoparticles from an extract of Jatropha curcas leaves: characterization, optical
properties, CT-DNA binding and photocatalytic activity. RSC Advances, 10(37),
22027–22035. https://doi.org/10.1039/D0RA03186K
Hajizadeh, Y. S., Harzandi, N., Babapour, E., Yazdanian, M., & Ranjbar, R. (2022).
Green Synthesize and Characterization of Copper Nanoparticles Using Iranian
Propolis Extracts. Advances in Materials Science and Engineering, 2022.
https://doi.org/10.1155/2022/8100440
Khan, I., Saeed, K., Zekker, I., Zhang, B., Hendi, A. H., Ahmad, A., Ahmad, S., Zada, N.,
Ahmad, H., Shah, L. A., Shah, T., & Khan, I. (2022). Review on Methylene Blue: Its
Properties, Uses, Toxicity and Photodegradation. Water (Switzerland), 14(2).
https://doi.org/10.3390/W14020242
Li, Q. Y., Munawar, M., Saeed, M., Shen, J. Q., Khan, M. S., Noreen, S., Alagawany, M.,
Naveed, M., Madni, A., & Li, C. X. (2022). Citrullus colocynthis (L.) Schrad (Bitter
Apple Fruit): Promising Traditional Uses, Pharmacological Effects, Aspects, and
Potential Applications. Frontiers in Pharmacology, 12, 791049.
https://doi.org/10.3389/FPHAR.2021.791049/BIBTEX
Lin, L., Yang, H., & Xu, X. (2022). Effects of Water Pollution on Human Health and
Disease Heterogeneity: A Review. Frontiers in Environmental Science, 10(June).
https://doi.org/10.3389/fenvs.2022.880246
Mali, S. C., Dhaka, A., Githala, C. K., & Trivedi, R. (2020). Green synthesis of copper
nanoparticles using Celastrus paniculatus Willd. leaf extract and their photocatalytic
and antifungal properties. Biotechnology Reports, 27, e00518.
https://doi.org/10.1016/J.BTRE.2020.E00518
Mourdikoudis, S., Pallares, R. M., & Thanh, N. T. K. (2018). Characterization techniques
for nanoparticles: comparison and complementarity upon studying nanoparticle
properties. Nanoscale, 10(27), 12871–12934. https://doi.org/10.1039/C8NR02278J
Nieto-Maldonado, A., Bustos-Guadarrama, S., Espinoza-Gomez, H., Z. Flores-López, L.,
Ramirez-Acosta, K., Alonso-Nuñez, G., & Cadena-Nava, R. D. (2022). Green
Chapter 6 34 References

synthesis of copper nanoparticles using different plant extracts and their antibacterial
activity. Journal of Environmental Chemical Engineering, 10(2), 107130.
https://doi.org/10.1016/J.JECE.2022.107130
Pérez-Alvarez, M., Cadenas-Pliego, G., Pérez-Camacho, O., Comparán-Padilla, V. E.,
Cabello-Alvarado, C. J., & Saucedo-Salazar, E. (2021). Green Synthesis of Copper
Nanoparticles Using Cotton. Polymers 2021, Vol. 13, Page 1906, 13(12), 1906.
https://doi.org/10.3390/POLYM13121906
Rasool, S., Tayyeb, A., Raza, M. A., Ashfaq, H., Perveen, S., Kanwal, Z., Riaz, S.,
Naseem, S., Abbas, N., Ahmad, N., & Alomar, S. Y. (2022). Citrullus colocynthis-
Mediated Green Synthesis of Silver Nanoparticles and Their Antiproliferative
Action against Breast Cancer Cells and Bactericidal Roles against Human
Pathogens. Nanomaterials 2022, Vol. 12, Page 3781, 12(21), 3781.
https://doi.org/10.3390/NANO12213781
Senthil Kumar, P., & Saravanan, A. (2017). Sustainable wastewater treatments in textile
sector. Sustainable Fibres and Textiles, 323–346. https://doi.org/10.1016/B978-0-
08-102041-8.00011-1
singh, karuna, kumar pankaj, & srivastava rajani. (2017). (PDF) An overview of textile
dyes and their removal techniques: Indian perspective. Pollution Research.
Singh, J., Dutta, T., Kim, K. H., Rawat, M., Samddar, P., & Kumar, P. (2018). ‘Green’
synthesis of metals and their oxide nanoparticles: applications for environmental
remediation. Journal of Nanobiotechnology 2018 16:1, 16(1), 1–24.
https://doi.org/10.1186/S12951-018-0408-4
Singh, J. P., Kumar, M., Sharma, A., Pandey, G., Chae, K. H., Lee, S., Singh, J. P.,
Kumar, M., Sharma, A., Pandey, G., Chae, K. H., & Lee, S. (2020). Bottom-Up and
Top-Down Approaches for MgO. Sonochemical Reactions.
https://doi.org/10.5772/INTECHOPEN.91182
Sinha, T., & Ahmaruzzaman, M. (2015). Green synthesis of copper nanoparticles for the
efficient removal (degradation) of dye from aqueous phase. Environmental Science
and Pollution Research, 22(24), 20092–20100. https://doi.org/10.1007/S11356-015-
5223-Y/FIGURES/7
Tang, A. Y. L., Lo, C. K. Y., & Kan, C. wai. (2018). Textile dyes and human health: a
systematic and citation network analysis review. Coloration Technology, 134(4),
245–257. https://doi.org/10.1111/COTE.12331
Ying, S., Guan, Z., Ofoegbu, P. C., Clubb, P., Rico, C., He, F., & Hong, J. (2022). Green
Chapter 6 35 References

synthesis of nanoparticles: Current developments and limitations. Environmental

Technology & Innovation, 26, 102336. https://doi.org/10.1016/J.ETI.2022.102336