“Synthesis and Characterization of Copper Nanoparticles Using Plants

Extraction Method by Zea Mays”

1. Introduction:

Nanotechnology is concerned with nanostructures that are smaller than 100 nanometers in at
least one dimension and that can be altered at the atomic or molecular level. It is an
interdisciplinary research field that includes chemistry, biology, engineering, and medicine, and
it has great potential in early symptom diagnosis and treatment in clinical and environmental
research. The word ‘Nano’ means a one-billionth scale of physical magnitude, and one
nanometer is a one-billionth of a meter or 0.000000001 m. In recent years, nanoparticles (NPs),
mainly metal NPs, have expanded in biological sciences to be used for diagnosis and
therapeutics purposes. That extension in use and applications of NPs can be explained by their
distinctive small size, as well as their enormous surface-area-to-volume ratio, high
responsiveness to living tissues, stability at elevated temperatures, and cellular
transportation(Ahmad et al., 2022).

Their small dimensions do not only allow more surface functionality in a given volume, but also
lead to physical properties that often differ from their bulk counterparts in many aspects. Nano-
particles possess a much higher surface-to-mass ratio than bulk materials and, therefore, surface
atoms and surface energy strongly contribute to the material properties. These unique properties
render nanoparticles extremely attractive for a large range of applications, including catalysis,
gas and energy storage, photovoltaic, electrical and optical devices, and biological and medical
technologies. For this reason, nanoparticles are not only a growing topic of interest in research
settings, but they are also already widely used in consumer products(Modena, Rühle, Burg, &
Wuttke, 2019). The field of nanotechnology has grown exponentially during the last few
decades, due in part to the use of nanoparticles in many manufacturing processes, as well as their
potential as clinical agents for treatment of diseases and for drug delivery. This has created
several new avenues by which humans can be exposed to nanoparticles. Unfortunately,
investigations assessing the toxicological impacts of nanoparticles (i.e. Nano toxicity), as well as
their possible risks to human health and the environment, have not kept pace with the rapid rise
in their use. This has created a gap-in-knowledge and a substantial need for more
research(Missaoui, Arnold, & Cummings, 2018). Nanoparticles are considered magic bullets
because of their unique properties. Nowadays, the use of nanoparticles has emerged in almost
every field of science and technology, owing to its potential of revolutionizing specific fields. In
the field of food science and technology, the use of nanoparticles is being studied in diverse
areas, starting with the harvesting of crops up to final food consumption. With the increased
usage of nanoparticles in day-to-day life, concern over their safety has arisen in everyone’s mind.
There is an imbalance between the increase in research to identify new nanoparticle applications
and their safety, and this has triggered pressure on scientists to identify the possible effects of
nanoparticles on human health. There are numerous studies on the use of nanotechnology in food
and the effect of nanoparticles on human health, but there is a vacuum in the literature in terms
of the combined analysis of such studies. This review is an attempt to present and analyze
different studies on the use and the safety of nanoparticles in food(Naseer et al., 2018).

1.1 Importance of Nanoparticles:

Knowledge of the interactions between nanoparticles (NPs) and cell membranes is of great
importance for the design of safe and efficient Nano medicines. Extensive studies aimed at
understanding the correlation of NP properties with endocytosis have been carried out in the past
few years. Here, we review the recent progress of these studies and provide an overview of the
current state of knowledge on the influence of NP size, shape, stiffness and surface chemistry on
cellular uptake. Special attention was dedicated to the uptake of non-spherical nanoparticles.
Some general principles obtained from these fundamental studies will serve as guidelines for the
design of optimised NP for enhanced cellular uptake. Finally, the opportunities for polymer
chemists are discussed(J. Zhao & Stenzel, 2018).

NPs should typically be compatible with both the focused biological systems and properties
required for water solubility. Despite the fact that there are evolved strategies for coating NPs in
order to ensure water solubility and preferred features, perfect surface coating should permit for
particular criteria such as preventing unwanted NP accumulation during long-term storage,
retaining good water solubility, preserving NP features, and guaranteeing biocompatibility prior
to NP communication with targeted disciplines. In biology and medicine, successful applications
of NPs are sometimes required for entry into living cells, which means that NPs need to cross a
significant barrier, i.e., the cell membrane, a nm-scale lipid bilayer containing embedded or
peripherally bound proteins. NPs can be inserted into living cells through many well-known
methods: (a) non-specific endocytosis, in which NPs commonly end up in endocytic partitions;
(b) straightforward microinjection of nanoliter of NP scattering, which is a time-consuming
process and only applicable to a limited number of cells; (c) electroporation, which utilizes
charges to force NPs across the membrane; and (d) mediated/targeted ingestion based on NP
surface functionalization via biological interactions or promoters. Nanotechnology has the power
to play a critical role in a variety of customized drug applications. In recent years, there has been
a rise in the number of nanotechnology strategies developed to overcome the drawbacks of
targeted therapies, enhanced medical visualization and diagnosis of diseases, pest management,
improvement of crop production, theranostics operations incorporating diagnostic and
therapeutic innovations, and development of biosensors and biosensing gadgets. Nanoparticles
(NPs) are widely applied as nanomedicine and nanocarriers for drug delivery(Ahmad et al.,
2022).

1.2 Classification of Nanoparticles:

As there are many different sources of nanomaterials, nanomaterials must be classified and
categorized nanomaterials for better understanding them. This section presents the classification
of nanomaterials according to their dimensionality, origin, composition (Barhoum et al., 2022).

1.2.1 Nanomaterial Classification Based on Their Dimensionality:

Nanomaterials can be differentiated from the macroscale materials present on Earth based on
their dimensionality (size and morphology) into four main types of nanomaterials (0D, 1D, 2D,
and 3D) (Figure 8). Therefore, nanomaterials can be classified based on the number of
dimensions that are outside the nanometer scale(Barhoum et al., 2022).
Figure. Different morphologies observed in nanomaterials with different dimensionality

1.2.1.1 Zero-dimensional (0D) nanomaterials: 

These materials have all three dimensions in the nanometer scale (i.e., <100 nm). Some examples
are graphene quantum dots, carbon quantum dots, fullerenes, inorganic quantum dots, magnetic
nanoparticles, noble metal nanoparticles, up conversion nanoparticles, and polymer
nanoparticles. Due to their optical stability, wavelength-dependent photoluminescence, chemical
inertness, cell permeability, and biocompatibility, 0D nanomaterials are interesting for
optoelectronic and biomedical applications.recently reviewed the unique properties and emerging
applications of 0D nanomaterials(Wang, Hu, Liang, & Wei, 2020).

1.2.1.2 One-dimensional (1D) nanomaterials: 

These materials have only one dimension >100 nm. Metal, metal oxides, and carbon-based 1D
nanomaterials with a high aspect ratio (e.g., nanotubes, nanowires, and nanofibers) are excellent
electron sources that emit electrons in a low electric field. Polymer nanofiber mats, veils, and
webs are 1D NMs with large surface-to-volume ratio, elevated porosity and small pores that are
exploited for decontamination, catalysis, filtration, and also for super-absorbent and scaffold
nanomaterials for tissue engineering and wound dressings reviewed the unique properties,
emerging applications, and risks of 1D nanomaterials (Barhoum et al., 2021; Jeevanandam et al.,
2020; Jin et al., 2018; Salama et al., 2021).

1.2.1.3 Two-dimensional (2D) nanomaterials:

 These materials type has two dimensions >100 nm. They have plate-like shapes and have thin
layers with a thickness of at least one atomic layer. Graphene/graphene oxide/reduced graphene
oxide, silicate clays, layered double hydroxides, transition metal dichalcogenides, transition
metal oxides, black phosphorus, graphitic carbon nitride, hexagonal boron nitride, antimonite,
boron nanosheets, and tin telluride nanosheets are some examples of 2D nanomaterials. Their
uniform shapes, high surface-to-volume ratio (in contrast with the bulk material), and surface
charge are explained by the unique physical, chemical, optical, and biological characteristics(El-
Maghrabi et al., 2018).  

1.2.1.4 Three-dimensional (3D) nanomaterials: 

These materials are materials with three dimensions >100 nm. This class includes but is not
limited to box-shaped graphene nanostructured and bundles of nanowires, and nanotubes. In the
last decade, many researchers have been focusing on the design, production, and assessment of
3D nanostructures as electrodes for electrochemical energy conversion (fuel cells)(El-Maghrabi
et al., 2018; Gopalakrishnan et al., 2019). and storage (batteries and supercapacitors)(Nnaji et al.,
2018; H. Zhao & Lei, 2020), and water treatment(Ho, Ho, Babar, Kim, & Schwingenschlögl,
2020). Materials with complex 3D structures can display unique features, such as the critical
transition to bio stability of a square twist origami, and elevated mechanical strength. Complex
3D structures are important components of micro-electromechanical systems, biomedical
devices, robotics, and solar cells. The synergistic integration of nanomaterials with 3D printing
technologies enable the creation of architecture and devices with an unprecedented level of
functional integration(Wu, Yang, Huang, Chen, & Zheng, 2013)

1.2.2 Nanomaterial Classification in the Function of Their Origin:

Nanomaterials are part of the Earth system, and natural nanomaterials and living organisms have
been co-evolving harmoniously. Based on their origin, Nanomaterials can be classified into four
main types:

1.2.2.1 Natural Nanomaterials:

These nanoparticles and nanostructured materials are produced by natural (bio) geochemical or
mechanical processes, without any link with anthropogenic activities/processes. Some examples
are the foraminifera (primarily chalk) and virus (capsid, protein) structures, the wax crystals that
coat lotus or nasturtium leaves, spider and silk spider mites, the blue tarantula hues, the gecko
foot spatulas, some butterfly wing scales, natural colloids (milk, blood), horny materials (claws,
skin, feathers, hair), nacre, corals, and the human bone matrix. Natural inorganic nanomaterials
occur by crystal growth. For example, due to their crystal structure anisotropy, clays show
complex nanostructures. Moreover, volcanic activity may lead to the formation of opals, an
example of naturally occurring photonic crystals due to their nanoscale structure. Natural
nanomaterials sources include forest fires (combustion materials), volcanic ash, ocean spray,
radon gas decline, and weathering of metal- or anion-containing rocks and acid mine drainage
sites(Shalan, Barhoum, Elseman, Rashad, & Lira-Cantú, 2018).

 1.2.2.2 Incidental Nanomaterials:

These nanoparticles and nanostructured materials are unintentionally produced through direct or
indirect human influences or anthropogenic (e.g., mechanical or industrial) processes, such as
vehicle exhaust gases, welding gases, solid fuel heating (home heathers), and combustion during
cooking. Incidental atmospheric nanomaterials, inadvertently formed during a deliberate
procedure, might increase air pollution. Forest fires generate a wide range of nanomaterials (e.g.,
pigments, cement, fumed silica). It’s hard to say when human beings started making incidental
nanoparticles, but probably as soon as people started taming fire. Incidental nanomaterials,
byproducts of human activities, are generally have poorly controlled sizes and shapes. Incidental
nanomaterials have high environmental impacts and must be considered relative to engineered
nanomaterials.

1.2.2.3 Bioinspired Nanomaterials:

These are engineered nanomaterials whose properties mimic those of natural nanomaterials or
living matter. Using advanced nanofabrication technologies, many bioinspired nanomaterials
with specific functions can be fabricated by modulating their structures. For example,
chameleons can rapidly adjust their colors from a camouflaged state to a highly visible (excited)
state when fighting or courting(Lee et al., 2017). This color change occurs mainly by actively
tuning the lattice of guanine nanocrystals within iridophore cells. Mechanochromic elastomers
have been designed that mimic the photonic structure of the chameleon iridophore cells(Rasouli,
Barhoum, & Uludag, 2018). In these sensors, rigid silica nanocrystals are embedded in an
elastomers matrix to form non-close-packed crystals. These sensors display a color shift from red
to blue under stretching, and from red to green under compression. Similar to in chameleons, this
color change is reversible. These sensors could be used in wallpaper, signboards and optical
recording.

Figure. Bioinspired nanomaterials that respond to stimuli.

1.2.2.4 Engineered Nanomaterials: 

The engineered are nanoparticles and nanostructured materials produced for specific applications
based on their dimensionality and specific characteristics (e.g., nanostructured medical implants)
(Albalawi, Hussein, Fakurazi, & Masarudin, 2021; Samyn & Barhoum, 2018). These
nanomaterials must be extensively characterized to reduce or minimize their unplanned negative
effects(Buzea, Pacheco, & Robbie, 2007). The first commercial nanomaterials were engineered
by aerosol (fumed silica) produced in the 1940s. In this case, the first silica nanospheres were
synthesized from aqueous solutions in the 1960s. Both incidental and natural nanoparticles may
have regular or irregular shapes. Usually, engineered nanoparticles have regular shapes, such as
rings, spheres, tubes, etc. Engineered carbon nanostructures such as fullerenes, carbon
nanotubes, and graphene in which it has a more regular shape and structure than in carbon soot
(Incidental Nanomaterials).

1.2.2.5 Anthropogenic Nanomaterials:

This term refers to both incidental and engineered nanomaterials. Either the deliberate and
accidental release of anthropogenic nanomaterials in the environment is becoming a major public
issue.

1.2.3 Nanomaterial Classification as a Function of Their Chemical or

Elemental Composition:

Nanomaterials may contain one or two or more elements of the periodic table. In nature, they are
usually aggregates of different elements. In function of their chemical composition,
nanomaterials are differentiated into carbon, inorganic, organic, and hybrid nanomaterials.
Engineered nanomaterials are typically produced using a variety of methods and may include
one or more components(Gugulothu, Barhoum, Afzal, Venkateshwarlu, & Uludag, 2018).

1.2.3.1 Carbon Nanomaterials: 

They are made of sp2-bonded carbon atoms. They include nanodiamonds, fullerenes, graphene,
single- and multi-walled carbon nanotubes, carbon nanofibers, nano horns, nano-onions, and
nano-graphite(Thostenson, Ren, & Chou, 2001). The synthesis methods for carbon-based
materials are chemical vapor deposition (CVD), laser ablation, and arch discharge(Butler &
Sumant, 2008). Carbon-based nanomaterials are a specific nanomaterials class because of the
variety of allotropies, and they could also be considered to be organic nanomaterials because
they include C-C bonds. This class also includes non-sp 2 hybridized carbon atoms such as nano-
diamonds, carbon black, and activated carbon. Most of these nanomaterials found in the
environment reduce their size by milling or may be grown using a seeding growth technique
(e.g., CVD for nano-diamonds)(Barhoum et al., 2020). Carbon-based nanomaterials are crucial
for human activities for a long time ago (e.g., combustibles, composite materials, pigments,
reinforcement materials). In the sustainable energy field(Oschatz et al., 2016), graphite blocks
are part of nuclear reactors as reflectors and moderators. Carbon nanomaterials also act as
electrodes in lithium-ion rechargeable batteries and electric double-layer capacitors(Khan, Saeed,
& Khan, 2019).

Figure Carbon Nanoparticles

1.2.3.2 Organic Nanomaterials:

They have functional properties due to the chemical association of their principal constituent
(carbon) with other elements that confer specific functionalities and reactivity to the
nanostructured architecture. This class includes lipid and polymer nanoparticles. Usually, both
nanoparticles types are nanospheres or have a nano-encapsulated shape ranging from 10 to 1000
nm(Ramsden, 2011). Representative examples of organic nanoparticles are dendrimers, micelles,
liposomes, and ferritin. They are considered biodegradable and non-toxic nanoparticles. Lipid
bilayer films are self-assemblies formed by polar lipids found in cell membranes, some
microorganisms, and viruses. Langmuir–Blodgett films that mimic lipid bilayers are synthetic
self-assemblies composed of amphiphilic organic molecules. In these molecules, a polar
nanoblock joins with another polar block. The polar side is the head and the apolar side is the
tail; both have the same dimensions(Stevens & Etherington, 2019). These fabricated films
assemble nanoparticulated micelles, liposomes, and single or bilayer films by taking advantage
of the fact that the “head” is hydrophilic while the “tail” is hydrophobic. Micelles and liposomes
contain a hollow core(Stevens & Etherington, 2019).

Fig. Organic nanoparticles: a – Dendrimers, b – Liposomes and c – micelles.

1.2.3.3 Inorganic Nanomaterials: 

They form by non-carbon elements, including metals, metal oxides, metal salts. They have
different shapes (spheres, cylinders, oblates, ellipsoids, cubes, and stars) in the function of the
atom packing while retaining the crystallinity nature of metal-based compounds. However, there
are also amorphous inorganic nanoparticles. The surface is highly reactive and sensitive due to
dangling bonds of atoms at the surface. This problem overcomes by functionalization. Among
inorganic nanomaterials, metal-based quantum dots (1–10 nm) display exceptional properties
due to the transition stage between bulk and few atoms(Tang & Lo, 2013). Magnetic
nanoparticles are also fascinating due to the high coercive fields and superparamagnetic behavior
at the reduced nanoscale. Some examples of magnetic nanoparticles are magnetite (Fe3O4), γ-
Fe2O3, iron (Fe), cobalt (Co), and spinel-type ferromagnets(Essawy, El-Sabbagh, Tawfik, Van
Assche, & Barhoum, 2018). Nanoclays (2D silicates of 1 nm in thickness) are biocompatible and
have low toxicity(Rodríguez-Hernández, 2017). Their main application is polymer reinforcement
and barriers, membrane coatings, toxin adsorption, and antibacterial and sterilizing
materials(Siddiqui, Fatima, Tara, Rathi, & Chaudhry, 2019). Zeolite with nanosized cavities is
particularly interesting for wastewater treatment where they are the charged surface leading to
high ion-exchange capacity(Neitzel, Mochalin, & Gogotsi, 2012).
 Figure Inorganic Nanoparticles

1.2.3.4 Hybrid Nanocomposites: 

They include multiphase materials with at least one nanosized component (1–100 nm), or with
nanometric phase separation(Zhang, 2014). Hybrid nanocomposites are matrix-based, and the
matrix can be a polymer, ceramic, hydrogel, or metal. At nanocomposites, the polymers are the
matrix for organic or inorganic nanomaterials in different shapes. The organic or inorganic
nanomaterials used as reinforcement gives specific properties to the composite. For instance,
ceramic matrix composites are fiber-reinforced materials known as technical ceramics for exact
applications (e.g., aerospace industry). Hydrogels display bioactive properties adaptable to
precise environments, (e.g., tissue engineering), combined with the easy interaction between
polymeric chains and nanostructures. Metal nanocomposites combine two or more metals. For
example, intermetallic compounds, and alloys that contain nano-metals, core-shell nanoparticles,
or banded components. The metal matrix may contain carbonaceous materials that apply in the
aerospace industry (Barhoum et al., 2017; Rui; Saboori, Dadkhah, Fino, & Pavese, 2018).

Figure Hybrid Nanoparticles

1.3 Synthesis of Nanoparticles:

There are different methods for the synthesis of nanoparticles and these methods are divide into
two main classes(Ijaz, Gilani, Nazir, & Bukhari, 2020).

Figure Nanomaterial Synthesis

1.3.1 Top-down synthesis:

In this synthesis, destructive method is used. The larger molecule (bulk material) decomposed
into a smaller molecule and then these smaller molecules transform into the nanoparticles.
Grinding or milling, physical vapor deposition and other destructive approaches are the example
of Top-down synthesis(Iravani, 2011).

1.3.2 Bottom-up method:

Bottom-up method is also known as constructive method. It is the reverse of top-down method.
In this method, nanoparticles are formed from relatively simpler substances. Bottom-up method
includes Chemical vapor deposition (CVD), Sol–gel, spinning, pyrolysis and biological
synthesis(Ijaz et al., 2020).

1.4 Green Synthesis of Nanoparticles:

Traditional methods are used from past many years but researches have proved that the green
methods are more effective for the generation of NPs with the advantage of less chances of
failure, low cost and ease of characterization(Abdelghany et al., 2018). Physical and chemical
approaches of synthesizing NPs have posed several stresses on environment due to their toxic
metabolites. Plant-based synthesis of NPs is certainly not a troublesome procedure, a metal salt is
synthesized with plant extract and the response is completed in minutes to couple of hours at
typical room temperature. Generation of NPs from green techniques can be scaled up effortlessly
and they are fiscally smart too. In light of their exceptional properties the greenly orchestrated
NPs are currently favoured over the traditionally delivered NPs.  Use of more chemicals, which
are harmful and toxic for human health and environment, could increase the particle reactivity
and toxicity and might cause unwanted adverse effects on health because of their lack of
assurance and uncertainty of composition(Hussain, Singh, Singh, Singh, & Singh, 2016). Green
methods of synthesis are significantly attractive because of their potential to reduce the toxicity
of NPs. Accordingly, the use of vitamins, amino acids, plants extracts is being greatly
popularized nowadays(Baruwati, Polshettiwar, & Varma, 2009).

1.5 Characterization of Nanoparticles:

The nanoparticles can be categorized into quantitative and qualitative. These methods include
dynamic light scattering (DLS), scanning electron microscope (SEM), energy dispersive
spectroscopy (EDS), UV-Vis spectroscopy, X-ray diffraction (XRD), Fourier transform infrared
spectroscopy (FT-IR), surface-enhanced remain spectroscopy (SERS), Atomic force microscopy
(AFM), high angle annular dark field (HAADF), atomic absorption spectroscopy (AAS) and.ray
photoelectron spectroscopy (XPS)(Chanda, 2013; Mohammadlou, Maghsoudi, & Jafarizadeh-
Malmiri, 2016).
1.5.1 Qualitative analysis:

1.5.1.1 Fourier transform infrared spectroscopy (FT-IR):

Biomolecules those are responsible for capping, reduction and stabilizer, are identify by using
FT-IR.

1.5.1.2 UV–Vis spectrophotometry:

The nanoparticles of various metal ranges in size between 2 and 100 nm are characterized by
using UV–Vis. The wavelength of 300 to 800 nm is generally used. The formation and stability
of nanoparticles in aqueous solution are determined by using this technique.

1.5.1.3 Scanning electron microscope (SEM):

Output image is formed by using SEM in which electron is used instead of light. Shape, size,
morphology and distribution of synthesized nanoparticles are characterized by using SEM.

1.5.1.4 X-ray diffraction (XRD):

Atomic structure of materials is determined by using XRD. It is used for both qualitative and
quantitative analysis. It is used for the determination of crystal structure, calculation of
crystalline nanoparticles size and for confirmation of nanoparticles.

1.5.1.5 Atomic force microscopy (AFM):

AFM is used for study of shape, size and surface area of synthesized nanoparticles.

1.5.2 Quantitative analysis:

1.5.2.1 Transmission electron microscopy (TEM):

It is used for the study of particle size of material in nano-scale and crystal structure.

1.5.2.2 Annular dark-field imaging (HAADF):

The mechanism by which nanoparticles interact with bacteria can be studied by using HAADF.
HAADF image gave the information about the size distribution of nanoparticles interacting with
each type of bacteria.

1.5.2.3 Intracranial pressure (ICP):

ICP spectrometry is used to determine metal concentration in deionized and the original
nanoparticles solutions. Inductively coupled plasma mass spectrometry (ICP-MS) and
inductively coupled emission spectroscopy (ICP-ES) are used to measure the resulting metal
concentration.

1.6 Cu Nanoparticles:

The synthesis of the Cu nanoparticles via the physical and chemical approach usually makes use
of toxic reducing agents and organic solvents(Cheirmadurai, Biswas, Murali, & Thanikaivelan,
2014). As copper undergoes rapid oxidation, the preparation of stable Cu nanoparticles is rather a
complex process. They exhibit antibacterial properties as they have large surface area that helps
the bacteria to come in easy contact. They also exhibit nematicidal properties(Akhter et al.,
2020).
Only a few studies are reported related to Cu nanoparticle synthesis, where usually plant extracts,
microorganisms and ascorbic acid are used as reducing agents. Metallic copper usually exhibits
good conductivity but due to the presence of oxide layers the prepared Cu nanoparticles exhibit
low conductivity. Cu nanoparticles are widely used in electronic devices(Cheirmadurai et al.,
2014). Plant extracts like those of Aloe vera are helpful in the synthesis of Cu nanoparticles by
the reduction of copper ions(Jadoun, Arif, Jangid, & Meena, 2021). Copper oxide nanoparticles
with distinct sizes were prepared via colloidal heat combination process(Gour & Jain, 2019).
Figure SEM images of the Cu nanoparticles obtained at 100 C in the presence of SDBS

1.7 Synthesis of Cu Nanoparticles:

Preparation of metal nanoparticles should be done using appropriate method to obtain a

particular size of nanoparticle, as with use of particular method it reduces the size of particle and
stabilizes it. Copper nanomaterials are greatly in attention due its profuse amount, availability
and low cost in comparison to gold and silver, so large scale productions of copper nanoparticles
are using various physical and chemical method(Shah & Lu, 2018). The major methods used for
large scale production of nanoparticles is done through physical methods (Mechanical milling,
laser ablation and sputtering) and chemical methods (Solid state, liquid state, gas phase,
biological methods and other methods), terming them as Top down and Bottom up methods
respectively(Jamkhande, Ghule, Bamer, & Kalaskar, 2019). Top-down method for synthesis of
nanoparticles is a method where bulk material is the initial material which is catabolized to
reduce particle size, even though this methods are easy to perform but not suitable for preparing
uniform size of particles and due to this it can affect the surface chemistry of NP’s(Jamkhande et
al., 2019; Vickers, 2017). The mechanochemical method for synthesis of nanoparticles requires
precursor for copper, salts for dilution and as starting material which are ball milled at ambient
temperature, resulting into copper oxide (Ⅱ) nanoparticles enclosed into salt matrix further which
were washed by distilled water in ultrasonic bath(Harishchandra et al., 2020).
1.8 Cu Nanoparticle Formation by Green Plants:

Green synthesis of CuONPs using plant extracts as the source of electron generation for the
reduction of copper salt display some advantages over the use of microbes because it does not
require cell culture maintenance and it can be scaled up for large-scale synthesis. The formation
of CuONPs occurs with an observable change in the color of the extract when copper salt was
added. Several studies have revealed that the phytochemicals in the plant extracts first form
complexes with the iron salts and then reduce the ions to form nanoparticles. The biomolecules
in the plants extracts usually react with copper ion to cause reduction which subsequently
transform into CuONPs (Kerour, Boudjadar, Bourzami, & Allouche, 2018; Shayegan Mehr,
Sorbiun, Ramazani, & Taghavi Fardood, 2018).

1.9 Zea Maize:

Maize or Zea Mays, generally known as corn, is a special plant ever since American Indians used
it as a primary nourishment source. Maize was brought from western hemisphere at the start of
15th century to the Asia and Africa. Corn has an important position in the summer crops. It is
grown in the middle of reaping of spring and winter crops season. In present times, China,
Europe, Latin America and USA are the largest maize producers. China and U.S grow almost
60% of the world’s total maize(Sprague & Dudley, 1988). Corn basically originated from
teosinte, which is an old species of grass found in Latin America especially in countries such as
Guatemala and Mexico(Warman, 2003).

Figure Corn Plant

Maize is a yearly plant with approximate height of 4 meters. Every corn plant has both male
(tassels) and female (terminal flowers) organs. It has wide leaves with each node having single
leave at opposite ranks. The leaf arrangement in maize is distichous. Each leaf consists of a
covering around the stalk and a long blade joining it with the covering at the collar. A mature
maize plant usually has 30 leaves. The growth pattern of every maize type is same but place and
timing of planting can alter the height and count of leaves. A unique property of maize
contrasting to other plants is that the fruit or seed of maize are covered in the husk or shuck of
the plant. The pericarp (ovary wall) and testa (seed coat) are joined to make the fruit wall and
because of this tight grip between fruit and seed these two layers appear to be a single formation.
This formation is usually known by many names like fruit, kernel, grain and seed. The corn grain
is made of 3 major parts“embryo”,“endosperm” and the “fruit wall”. The quantity of grains
produced each ear and amount of ears which grow is generally confirmed when pollination is
done. Corn Plant has the ability to use sunlight better as compared to other grain producing crops
and it also develops quickly due to the size and distribution of its plant and leaves. Other than
corn oil it is also used to produce gluten feed, corn starch, syrup, dextrose(it is mostly used
pharmaceutical industry as the initial component for the production of Vitamin C and penicillin),
ethanol (which is used for beverage and fuel) syrup enriched in fructose (important ingredient
used in soft drink industry)(Sprague & Dudley, 1988). Maize is not only important for human
and animal food but also for commercial purposes. Edible portion of maize has different
minerals and vitamins in different quantity(Tabasum et al., 2019).

Figure. Zea Maize cultivation

2. Literature Review:

In 2018 One of the main subjects in nanoscience is the integration of green chemistry principles
to nanotechnology. Recently the ever-increasing need to develop eco-friendly metal
nanoparticle synthesis processes is observed. In this research, the ability of the fruit extracts
of Ziziphus spina-christi (L.) Willd. was examined as novel reducing agents for the green
synthesis of copper nanoparticles (Cu-NPs). Biosynthesized Cu-NPs were characterized by UV–
Vis spectroscopy, Fourier transform infrared spectroscopy (FT-IR), field emission scanning
electron microscopy (FESEM), transmission electron microscopy (TEM), and X-ray diffraction
(XRD). It was found that the as-prepared Cu-NPs can be used as an efficient
adsorptive nanomaterial to remove crystal violet (CV), from aqueous solution. The effects of
independent variables such as pH, initial dye concentration, adsorbent dosage and stirring time
on CV removal were studied using central composite design (CCD). Optimization of the
variables for maximum adsorption of target dye was performed using desirability function (DF)
combined response surface methodology (RSM). The results showed that 95% of CV with a high
adsorption capacity (37.5 mg g−1) was removed with a little amount of adsorbent (80 mg) in a
short time (7.5 min). In addition, antibacterial activity of the Cu-NPs was studied on two
different (g− and g+) bacteria; Escherichia coli and Staphylococcus aureus(Khani, Roostaei,
Bagherzade, & Moudi, 2018).

In 2018 Green synthesis of nanoparticles using biological molecules derived from the plant
sources in the form of extracts are exhibiting superiority over chemical and physical methods.
Development of green nanotechnology is generating interest of researchers towards eco-friendly
green synthesis of nanoparticles. In present study, green synthesis of stable copper nanoparticles
were done using Piper nigrum seed extract. The seed extract prepared by using deionised water
was mixed with 0.01M of copper sulphate (CuSO4.5H2O) solution and an alternative energy
source microwave irradiation used to get nanoparticles in a short incubation period. There is a
change in colour (Blue to Green) observed and that is indicating the formation of copper
nanoparticles. The plant based molecules present in the seed extract have highly controlled
assembly, which is suitable for synthesis of nanoparticles. These green synthesised copper
nanoparticles were characterised with the help of Uv-visible spectrophotometer, X-ray
diffraction analysis (XRD). It was observed that the Piper nigrum seed extract can reduce copper
ions to copper nanoparticles within 2 to 5 minutes of reaction time. Thus this method can be used
for rapid and eco friendly green synthesis of suitable copper nanoparticles (Sirisha & Asthana,
2018).

In 2018 Green synthesis of metal nanoparticles (NPs) has attracted considerable attention
because of its cheaper protocols and more environmentally friendly than standard synthetic
methods. In the present paper, Phoenix dactylifera L extract was used to synthesize copper oxide
nanoparticles with a differentvolume ratiobetween plant extract and metal (1mL of the plant
extract to 100mL of 1mM CuSO4.5H2O).The obtained copper oxide nanoparticles were
characterized by UV-Vis, FT-IR, XRD, SEM, and EDAX techniques. UV-Vis spectra showed a
maximum absorption at 275nm related to the copper oxide.FTIR spectra exhibit a weak peak at
554 cm−1 attributed to CuO vibration, confirming the formation of CuO nanoparticles. In
addition, a disappearance peaks at 3264cm-1 from the solution of as prepared (NPs) reveals that
polyphenols are responsible for thecopper oxide NPs formation. XRD confirmed the crystalline
natureof CuO and Cu2O NPs with average size ranged in 22-28 nm. SEM survey shows that the
obtained nanoparticles having in general aspherical shape. As established by EDAX to confirm
the presence of copper and oxygen, the weight percentage of the latterwas (72.39% Cu and
27.61% O), respectively(Berra et al., 2018).

In 2018 The current investigation involves the green synthesis of copper nanoparticles (CuNPs)
from an aqueous plant extract of Moringa oleifera Lam by two methods: (I) time-based approach
and (II) heat treatment of aqueous solution. Prepared CuNPs were characterised via Fourier
transform infrared spectroscopy, X-ray diffraction, scanning electron microscopy and
transmission EM. The study also reveals the potential bioactivity of the prepared CuNPs. In vitro
anti-microbial efficiency of CuNPs was estimated against bacterial and fungal strains by the agar
well diffusion method. Anti-oxidant capacity of CuNPs was determined using ferric reducing
ability of plasma (FRAP), lipid peroxidation (LPO) and peroxidase assays, while the antiplatelet
potential was determined by measuring two haemostatic parameters (PT & APTT assay). The
minimum inhibitory concentration was observed at 60 µg/ml against Streptomyces
griseus and Aspergillus niger when NPs were prepared by method II. CuNPs prepared by the
method I showed higher FRAP and LPO activities, while increased POX activity was found in
CuNPs prepared by method II. CuNPs prepared using method I also showed better anti-oxidant
and antiplatelet potential. It was observed that M. oleifera -derived CuNPs exhibits strong anti-
microbial, anti-oxidant and APTT potential. This indicates potential utilization of green
synthesized NPs for various industrial and therapeutic strategies(Saran, Vyas, Mathur, &
Bagaria, 2018).

In 2018 Cu nanoparticles (CuNPs) were biosynthesized by green nanotechnology and they

showed a inhibitory power on the growth of Gram (+) and Gram (−). CuNPs synthesis (0) was
performed at 65 °C from a solution of copper chloride and using as reducing cyanidine-O-3-
glucoside, selectively obtained from the skin of red plum (Pronus domestic). Their
characterization has shown typical UV–Vis bands, Cu nanoparticles refers to surface plasmon
resonance (SPR) at 550 nm. Structural analysis by HRTEM evidence growth is in the (1 1 1)
plane of the fcc phase of metallic Cu, with a size close to 10 nm, with a quasi-spherical
morphology, due to the role as surfactant of the cyanidine-O-3-glucoside for the preparation of
metal nanoparticles(Tovar-Corona et al., 2018).

In 2019 The scope of nanotechnology in the field of chemistry offers many applications and one
of major applications is the synthesis of metal and metal oxide nanoparticles in the green
manner. Copper (Cu) is nontoxic and have many medicinal and other importance. It is a cheaper
and good antibacterial and antifungal agent and widely used in the catalytic and sensors. Various
methods have been reported in literature for the synthesis of copper nanoparticles (CuNPs). The
biological/green methods taking part the copper salt and plant extracts are more efficient, very
low cost and better alternatives over such methods. The chacterization methods used for analysis
of CuNPS are very important in the evaluation of their functional aspects. The common
characterization techniques are Fourier transform infra-red (FTIR) spectroscopy, UV-Visible
spectroscopy (UVVis), Scanning electron microscopy (SEM), Transmission electron microscopy
(TEM), X-ray diffractometry (XRD), Dynamic light scattering (DLS) etc(Joshi, Kumar, &
Singh, 2019).

In 2019 The present work reports the extraction of phenolic compounds from
Polygonum minus using ionic liquid as extracting solvent. In this work, 1-Butyl-3-
methylimidazolium hydrogen sulfate [BMIM][HSO4] was used for the extraction of bioactive
compounds. Accordingly, ionic liquids based microwave-assisted extraction treatment for
separating of bioactive compounds from polygonum minus was first performed in the present
study. The results obtained in this work have high extraction yield in comparison with
conventional solvent. UV/Vis results showed that microwave synthesis was fast, well dispersed
and nanosized copper nanoparticle (CuNPs) in comparison with conventional synthesis. CuNPs
was characterised by X-Rays diffractometer (XRD), Fourier transform infrared (FTIR), dynamic
light scattering (DLS), field emission scanning electron microscopy combined with energy
dispersive x-rays (FESEM-EDX), and thermogravimetric analysis (TGA). All the instrumental
analyses confirmed the particles were nanosized. Furthermore, the antibacterial activity of as-
synthesised CuNPs showed effective inhibitory zone against three different bacteria. The
photocatalytic degradation of copper nanoparticles was studied using methylene blue (MB) and
methyl orange (MO) dyes under UV light and degraded 99.9% within short time 8 and 7 
min(Ullah, Wilfred, & Shaharun, 2019).

In 2019 In recent decades, nanotechnology is growing rapidly owing to its widespread

application in science and industry. The aim of the experiment was the green synthesis of copper
nanoparticles using Allium saralicum R.M. Fritsch aqueous extract and assessment of their
cytotoxicity, antioxidant, antibacterial, antifungal, and cutaneous wound healing effects under in
vitro and in vivo conditions. These nanoparticles were characterized by Fourier transformed
infrared spectroscopy (FT-IR), UV–visible spectroscopy, field emission scanning electron
microscopy (FE-SEM), transmission electron microscopy (TEM), and atomic force microscopy
(AFM). DPPH free radical scavenging test was done to assess the antioxidant properties, which
indicated similar antioxidant potentials for CuNPs Allium and butylated hydroxytoluene.
Minimum Inhibitory Concentration (MIC), Minimum Bactericidal Concentration (MBC), and
Minimum Fungicidal Concentration (MFC) were specified by macro-broth dilution
assay. CuNPs Allium indicated higher antibacterial and antifungal effects than all standard
antibiotics (p ≤ 0.01). Also, CuNPs Allium inhibited the growth of all bacteria at 1–8 mg/ml
concentrations and removed them at 2–8 mg/ml concentrations (p ≤ 0.01). In the case of
antifungal properties of CuNPs Allium, they prevented the growth of all fungi at 1–4 mg/ml
concentrations and destroyed them at 2–8 mg/ml concentrations (p ≤ 0.01). In vivo experiment,
after creating the cutaneous wound, the rats were randomly divided into six groups (n = 10):
untreated control, treatment with Eucerin basal ointment, treatment with 3% tetracycline
ointment, treatment with 0.2% CuSO4 ointment, treatment with 0.2% A. saralicum ointment, and
treatment with 0.2% CuNPs Allium ointment. Use of CuNPs Allium ointment in the treatment
groups substantially reduced (p ≤ 0.01) the wound area, total cells, neutrophil, macrophage, and
lymphocyte and remarkably raised (p ≤ 0.01) the wound contracture, hydroxyl proline,
hexosamine, hexuronic acid, fibrocyte, and fibrocytes/fibroblast rate compared to other groups.
The synthesized CuNPs Allium had high cell viability dose-dependently (Investigating the effect
of the plant on HUVEC cell line) and revealed this method was nontoxic. The results revealed
the useful non-cytotoxic, antioxidant, antibacterial, antifungal, and cutaneous wound healing
effects of CuNPs(Tahvilian et al., 2019).

In 2019 Facile green synthesis of copper nanoparticles from different biological procedures has

been indicated, but among all, biosynthesis of copper nanoparticles from medicinal plants is
considered as the most suitable method. The use of medicinal plant material increases the
therapeutical effects of copper nanoparticles. The aim of this study was green synthesis of copper
nanoparticles from aqueous extract of Falcaria vulgaris leaf (CuNPs) and assessment of their
cytotoxicity, antioxidant, antifungal, antibacterial, and cutaneous wound healing properties.
These nanoparticles were characterized by X-ray diffraction (XRD), fourier-transform infrared
spectroscopy (FT-IR), ultraviolet–visible spectroscopy (UV), transmission electron
microscopy (TEM), and field emission scanning electron microscopy (FE-SEM) analysis. The
synthesized CuNPs had great cell viability dose-dependently (Investigating the effect of the
CuNPs on human umbilical vein endothelial cell (HUVEC) line) and indicated this method was
nontoxic. Also, 2,2-diphenyl-1-picrylhydrazyl (DPPH) test was done to assess the antioxidant
activities, which indicated similar antioxidant potentials for CuNPs and butylated
hydroxytoluene. In part of cutaneous wound healing property of CuNPs, after creating the
cutaneous wound, the rats were randomly divided into six groups: treatment with 0.2% CuNPs
ointment, treatment with 0.2% CuSO4 ointment, treatment with 0.2% F. vulgaris ointment,
treatment with 3% tetracycline ointment, treatment with Eucerin basal ointment, and untreated
control. These groups were treated for 10 days. Treatment with CuNPs ointment remarkably
increased (p ≤ .01) the wound contracture, vessel, hexosamine, hydroxyl proline, hexuronic
acid, fibrocyte, and fibrocytes/fibroblast rate and substantially reduced (p ≤ .01) the wound area,
total cells, neutrophil, and lymphocyte compared to other groups. In antibacterial
and antifungal parts of this research, the concentration of CuNPs with minimum dilution and no
turbidity was considered minimum inhibitory concentration (MIC). To determine minimum
fungicidal concentration (MFC) and minimum bactericidal concentration (MBC), 60 μL MIC
and three preceding chambers were cultured on Sabouraud Dextrose Agar and Muller Hinton
Agar, respectively. The minimum concentration with no fungal and bacterial growth were
considered MFC and MBC, respectively. CuNPs inhibited the growth of all fungi at 2–4 mg/mL
concentrations and removed them at 4–8 mg/mL concentrations (p ≤ .01). In case of antibacterial
effects of CuNPs, they inhibited the growth of all bacteria at 2–8 mg/mL concentrations and
removed them at 4–16 mg/mL concentrations (p ≤ .01). The results of XRD, FT-IR, UV, TEM,
and FE-SEM confirm that the aqueous extract of F. vulgaris leaf can be used to yield copper
nanoparticles with notable amount of antioxidant, antifungal, antibacterial, and cutaneous wound
healing potentials without any cytotoxicity. Further clinical trials are necessary for confirmation
these therapeutical effects of CuNPs in human (Zangeneh et al., 2019).

In 2019 In the present study, we have developed a green, facile, eco-friendly and cost-effective
approach for the synthesis of copper nanoparticles (CuNPs) from copper salt (CuCl) solution by
using Camelia sinensis leaf extract. The biomolecules present in C. sinensis leaves extract were
responsible for the reduction and stabilization of CuNPs. The synthesized nanoparticles were
characterized by using FT-IR, SEM, TEM and EDX analysis. FT-IR analysis was used to ratify
the presence of biomolecules in C. sinensis leaves extract, while SEM and TEM analysis showed
that the synthesized nanoparticles are in agglomerated form and having an average size of 60 ± 6 
nm. The percentage weight of synthesized nanoparticles was measured through EDX analysis.
Moreover, the photocatalytic degradation of synthesized CuNPs was also evaluated by using
bromophenol blue (BPB) dye under sunlight irradiation. The maximum photo-degradation of
BPB dye was up to 83.7%. UV-Vis spectral analysis of CuNPs proved them as an efficient
photocatalyst in dye degradation(Ahmed et al., 2019).

In 2020 A facile environmentally friendly hydrometallurgical approach to recover copper as

nanoparticles from electronic waste (e-waste) has been presented. Selective leaching of copper
from powdered old computers waste printed circuit boards (WPCBs) was executed in
ammoniacal/ammonium salt solutions at room temperature. The effect of parameters controlling
copper recovery has given due consideration. Copper nanoparticles were prepared from leachant
solutions by reduction using L-ascorbic acid reductant and (CTAB) as a modifier at room
temperature. Characterization tests for the produced copper nanoparticles were performed to
confirm its structure. XRD analysis manifested the pure crystallinity phase of copper
nanoparticles and TEM images revealed its spherical shape with particle size in the range of 5–
32 nm. It is considered that the studied process is merely efficacious and environmentally
sustainable for preparing copper nanoparticles from WPCBs with the intention of recycling e-
waste to gain a higher valued product(El-Nasr, Abdelbasir, Kamel, & Hassan, 2020).

In 2020 Chromium (Cr) concentration has been increasing substantially in the environment due
to industrial and anthropogenic factors. Plants can absorb Cr and undergo unrestrained oxidation
cascades, resulting in cell injury. The ameliorative role of biogenic copper nanoparticles to
relieve wheat plants from Cr stress by supporting their growth is still unclear. The present work
aims at the biosynthesis and characterization of copper nanoparticles (CuNPs) from a
native Klebsiella pneumoniae strain, followed by assessment of wheat growth and physiological
responses to CuNPs mixed in Cr-rich soil. The taxonomic rank of K. pneumoniae SN35 was
established by the 16 S rRNA gene sequence analysis. The properties of biogenic CuNPs were
elucidated by using UV–vis spectroscopy, FTIR, XRD, SEM, and TEM. It was found that 19.01–
47.47 nm spherical shaped CuNPs were stabilized by different functional groups produced
extracellularly by the strain SN35. The XRD data revealed the crystalline nature of CuNPs as a
face-centered cubic structure. Different concentrations of CuNPs (0, 25, 50 and 100 mg kg−1 of
soil) were added into the soil mixed with 3.5 mg kg−1 K2Cr2O7 and the pots were placed in
a growth chamber for 30 days. The results revealed that the CuNPs, at 25 and 50 mg kg−1 of soil,
augmented plant growth, biomass, and cellular antioxidants contents, whereas decreased
the reactive oxygen species and Cr translocation from soil to roots and shoots as compared to
control plants. Overall, the results revealed that the soil amendment of CuNPs could immobilize
the Cr in the soil to prevent its translocation to the upper plant parts and support wheat growth by
relieving cellular oxidative stress (Noman et al., 2020).
In 2020 Green synthesis of nanoparticles has gained enormous attention in today's world due to
ongoing demand to develop safe, sustainable, cost-effective and environmentally friendly
process for synthesizing a wide variety of materials including metal/metal oxides nanoparticles,
hybrid and bioinspired materials. In the current study, we have carried out green synthesis of
copper oxide (CuO) nanoparticles (NPs) using psidium guajava leaf extract as capping agent and
copper acetate as metal precursor. The biosynthesized nanoparticles were characterized using
wide variety of techniques i.e. powder X-ray diffraction (PXRD), FTIR, TGA and HRTEM. The
PXRD result confirms the synthesis of copper oxide nanoparticles in pure phase having
monoclinic symmetry. The average particle size using PXRD was found out to be ∼33 nm
(Varughese, Kaur, & Singh, 2020).
In 2020 The synthesis of metal nanoparticles using plant extracts is a very promising method in
green synthesis. The medicinal value of Moringa oleifera leaves and the antimicrobial activity of
metallic copper were combined in the present study to synthesize copper nanoparticles having a
desirable added-value inorganic material. The use of a hydroalcoholic extract of M.
oleifera leaves for the green synthesis of copper nanoparticles is an attractive method as it leads
to the production of harmless chemicals and reduces waste. The total phenolic content in the M.
oleifera leaves extract was 23.0 ± 0.3 mg gallic acid equivalent/g of dried M. oleifera leaves
powder. The M. oleifera leaves extract was treated with a copper sulphate solution. A color
change from brown to black indicates the formation of copper nanoparticles. Characterization of
the synthesized copper nanoparticles was performed using ultraviolet-visible light (UV-Vis)
spectrophotometry, Fourier-transform infrared (FTIR) spectrometry, high-resolution
transmission electron microscopy (HRTEM), scanning electron microscopy (SEM), and X-ray
diffraction (XRD). The synthesized copper nanoparticles have an amorphous nature and particle
size of 35.8-49.2 nm. We demonstrate that the M. oleifera leaves extract and the synthesized
copper nanoparticles display considerable antioxidant activity. Moreover, the M. oleifera leaves
extract and the synthesized copper nanoparticles exert considerable anti-bacterial activity
against Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus, and Enterococcus
faecalis (MIC values for the extract: 500, 250, 250, and 250 µg/mL; MIC values for the copper
nanoparticles: 500, 500, 500, and 250 µg/mL, respectively). Similarly, the M. oleifera leaves
extract and the synthesized copper nanoparticles exert relatively stronger anti-fungal activity
against Aspergillus niger, Aspergillus flavus, Candida albicans, and Candida glabrata (MIC
values for the extract: 62.5, 62.5, 125, and 250 µg/mL; MIC values for the copper nanoparticles:
125, 125, 62.5, and 31.2 µg/mL, respectively). Our study reveals that the green synthesis of
copper nanoparticles using a hydroalcoholic extract of M. oleifera leaves was successful. In
addition, the synthesized copper nanoparticles can be potentially employed in the treatment of
various microbial infections due to their reported antioxidant, anti-bacterial, and anti-fungal
activities (Das, Abu-Yousef, Majdalawieh, Narasimhan, & Poltronieri, 2020).
In 2020 A biological extract of Cynomorium coccineum was used for the synthesis of
copper nanoparticles using copper sulphate as a starting matter. Fourier transform infrared
spectroscopy (FT-IR) showed the presence of the functional groups characteristics of biological
extract, i.e., triterpenes, phenolics, flavonoids, and other reducing agents. SEM analysis showed
that the surface of the particles were clusters and rough. The Elemental analysis by energy-
dispersive X-ray (EDX) proved the purity of the particles, showing the presence of
elemental copper oxide which was surrounded with some elements of the plant. X-ray diffraction
(XRD) pattern confirmed that the crystalline nature of the particles was affected by temperature.
The average crystallite size was calculated to be about 14.2 nm. The total weight loss registered
within the thermogravimetric (TG) analysis was 52.8%, suggesting that about 47.2% of the
metallic copper is present in the prepared copper nanoparticles which supported the data reported
within EDX analysis. The adsorption characteristics of the nanoadsorbents were examined
using methylene blue as adsorbate and the bio-sorption capacity reached 64 mg/g at room
temperature. The values of B and bt, calculated from Temkin equation, increased with the
increase in temperature (295–328 K), indicating endothermic adsorption and strong dye-
nanoadsorbent interaction. The mean free energy (E = 100–129.1 kJ/mol), calculated from
Dubinin-Radushkevich indicated that the chemisorption is the process involved for the
adsorption of methylene blue. In summary, the results obtained in this study deliver the design
and the synthesis of new materials for removing pollutants (Sebeia, Jabli, Ghith, & Saleh, 2020).
In 2021The disposal of solid waste containing toxic metals in the composition is a problem of
environmental and human contamination. The amount of waste as printed circuit boards has been
increasing year by year. New processes that aim at recycling and subsequent application
implementing a circular economy of this waste have been gaining importance. This study
proposes the printed circuit boards characterization for the copper recycling by acid leaching in
two steps, green synthesis of copper nanoparticles using ascorbic acid (vitamin C), and later
application of nanoparticles into surfactant degradation through the Fenton-like process. Acid
leaching proved a possible methodology for recycling copper, obtaining over 90% recovery. The
printed circuit boards show to be a copper secondary source with a weight percentage of about
26 and recovery of 97% in two leaching stages. The synthesis of CuNPs was inferred through
UV–Vis with absorption peak in 585 nm and modification of the coloration over time of
solutions to dark brown. The syntheses were performed at 70 °C, 80 °C, and 90 °C and in 0.17,
0.5, 0.83, 1, 2, and 3 h, and the stability of copper nanoparticles was verified in 168, 720, and
960 h after syntheses. Stable nanoparticles were produced with different diameters, and they
were applied as a catalyst in the Fenton reaction to degrade an amine-based surfactant used in the
mining industry. The total carbon degradation found was 57%. It was possible to observe that the
recycling of printed circuit boards is strategic to obtain copper from a secondary source, and the
synthesis of copper nanoparticles could be used as a product to applicate in the circular
economy using the Fenton process (Martins et al., 2021).

In 2021 Copper nanoparticles (CuNP) were obtained by a green synthesis method using cotton
textile fibers and water as solvent, avoiding the use of toxic reducing agents. The new synthesis
method is environmentally friendly, inexpensive, and can be implemented on a larger scale. This
method showed the cellulose capacity as a reducing and stabilizing agent for synthetizing
Cellulose–Copper nanoparticles (CCuNP). Nanocomposites based on CCuNP were characterized
by XRD, TGA, FTIR and DSC. Functional groups present in the CCuNP were identified by
FTIR analysis, and XRD patterns disclosed that nanoparticles correspond to pure metallic Cu°,
and their sizes are at a range of 13–35 nm. Results demonstrated that CuNPs produced by the
new method were homogeneously distributed on the entire surface of the textile fiber, obtaining
CCuNP nanocomposites with different copper wt%. Thus, CuNPs obtained by this method are
very stable to oxidation and can be stored for months. Characterization studies disclose that the
cellulose crystallinity index (CI) is modified in relation to the reaction conditions, and its
chemical structure is destroyed when nanocomposites with high copper contents are synthesized.
The formation of CuO nanoparticles was confirmed as a by-product, through UV spectroscopy,
in the absorbance range of 300–350 nm (Pérez-Alvarez et al., 2021).

In 2021 The aim of this work was the green synthesis of copper nanoparticles (Cu-NPs) using
aqueous extracts of (i) bilberry (Vaccinium myrtillus L.) waste residues from the production of
fruit juices and (ii) non-edible “false bilberry” fruits (Vaccinium
uliginosum L. subsp. gaultherioides). Different cupric salts (CuCl2, Cu(CH3COO)2 and
Cu(NO3)2) were used for the synthesis. The formation of stable nanoparticles (CuNPs) was
assessed by transmission electron microscopy and the oxidation state of copper in these
aggregates was followed by X-ray photoelectron spectroscopy. The polyphenol composition of
the extracts was characterized, before and after the synthesis, using spectrophotometric methods
(i.e. total soluble polyphenols and total monomeric anthocyanins) and high-performance liquid
chromatography coupled with tandem mass spectrometry (i.e. individual anthocyanins).
Polyphenol concentration in the extracts was found to decrease after the synthesis, indicating
their active participation to the processes, which led to the formation of Cu-NPs. The
antimicrobial activity of Cu-NPs, berry extracts, and cupric ion solutions were analysed by broth
microdilution and time-kill assays, on prokaryotic and eukaryotic models. The antimicrobial
activity of Cu-NPs, especially those derived from bilberry waste residues, appeared to be higher
for both Gram-negative and Gram-positive bacteria, and for fungi, compared to the ones of its
single components (cupric salts and berry extracts). Therefore, Cu-NPs from the green synthesis
here proposed can be considered as a cost-effective sanitization tool with a wide spectrum of
action (Benassai et al., 2021).
In 2021 Recently, nanocomposite photocatalysts based on semiconductors have attracted much
attention due to their suitable bandgap. Combination of tow of several semiconductors can slow
down the electron-hole recombination. In this regard, we have depicted an eco-friendly and
green fabrication technique to synthesize RGO/Cu nanocomposite by the reduction of graphene
oxide and Cu2+ ion utilizing spearmint extract as a reductant and capping agent. The sample was
identified by FTIR, XRD, FESEM, EDS, HRTEM, and CV. The results of photocatalytic
performance revealed that RGO/Cu is an efficient catalyst for degrading organic pollutants. This
compound can eliminate Rhodamine B (RhB) and Methylene blue (MB) 91.0% and 72.0%,
respectively (Safajou et al., 2021).
In 2021 In recent years, the green chemistry based-approach for the synthesis of nanoparticles
has shown tremendous promise as an alternative to the costly and environmentally unfriendly
chemically synthesized nanoparticles. In this study, copper oxide nanoparticles (CuO NPs) were
synthesized through a green approach using the water extract of papaya (Carica papaya L.) peel
biowaste as reducing as well as stabilizing agents, and copper (II) nitrate trihydrate salt as a
precursor. The structural properties, crystallinity, purity, morphology, and the chemical
composition of as-synthesized CuO NPs were analyzed using different analytical methods. The
analytical results revealed that the synthesized CuO was observed as spherical-like in particles
with measured sizes ranging from 85–140 nm and has monoclinic crystalline phase with good
purity. The Fourier transform infrared (FTIR) spectroscopic results confirmed the formation of
the Cu-O bond through the involvement of the potential functional groups of biomolecules in
papaya peel extract. Regarding photocatalytic activity, the green-synthesized CuO NPs were
employed as a photocatalyst for the degradation of palm oil mill effluent (POME) beneath the
ultraviolet (UV) light and results showed 66% degradation of the POME was achieved after 3 h
exposure to UV irradiation. The phytotoxicity experiment using mung bean (Vigna radiata L.)
seed also showed a reduction of toxicity after photodegradation (Phang et al., 2021).
In 2022 The green synthesis of nanoparticles from bioactive compounds have attracted a wide
range of application, due to increased drug efficacy and less toxicity in the nanosized mediated
drug delivery model. In this study, we have fabricated copper nanoparticles (CuNPs) from the
fruits of Prunus nepalensis (P. nepalensis) extract. Therefore, the aim of present study was to
investigate the anticancer ability of P. nepalensis fruit phytochemical copper nanoaprticles
(PNFPCuNP) on cancerous human breast cell line (MCF-7) and healthy (MCFA10) cell lines.
Crystalline CuNPs of P. nepalensis synthesis was confirmed by different physicochemical
analytical techniques such as UV–Visible Spectroscopy (UV–Vis); Fourier-Transform Infrared
Spectroscopy (FT-IR); Scanning Electron Microscopy (SEM); and Transmission Electron
Microscopy (TEM). Nanoparticle Size was found to be ranging from 35 to 50 nm with the
average size of 42.5 nm. Further, after synthesized compounds were tested anticancer activity on
human breast cancer cell lines. Following 72 h treatment to PNFPCuNP, the expression of
apoptotic marker genes (P21, p53, P14/P19, Caspase-3) were studied in MCF-7 cells treated at
100 to 200 μg of PNFP-CuNP. Our results showed that PNFP-CuNP increased the gene
expression of apoptotic genes in a dose-dependent manner. The real-time PCR data showed a
significant upregulation in p53, Bax, caspase-3, and caspase-9 and down regulation in the mRNA
expression of Ras and Myc genes in MCF-7 cells exposed to PNFP-CuNP. Collectively, the data
from this study stated that P. nepalennsis fruit extract phytochemical derived nanoparticles
induced apoptosis via the up regulation of tumour suppressor genes and down regulation of
oncogenes in MCF-7 cells. Finally, our study confirmed the CuNPs synthesis from P. nepalensis
fruit phytochemical, which showed environmental friendly anticancer activity (Biresaw &
Taneja, 2022).
In 2022 Copper nanoparticles (CuNPs) with antimicrobial and cytotoxic properties were
obtained using extracts from Hyptis suaveolens (L.) commonly called wild tulsi. Green synthesis
using plant extracts is a preferred means to obtain nanoparticles because of the simplicity,
efficiency, and clean process. Plant extracts contain phytochemicals that provide microbial and
other medical properties to the nanoparticles. In this study, Hyptis suaveolens (L.) was treated in
water and the extracts obtained were used to synthesize copper nanoparticles. Transmission
electron microscopy (TEM) images showed that the synthesized copper nanoparticles are
spherical in shape with an average diameter of 7.2 nm. X-ray diffraction (XRD) and selected
area electron diffraction (SAED) analysis showed that the particles were mostly of copper 92%
and about 8% of oxygen indicating partial conversion of the copper sulphate into copper oxide.
Plasmonic absorbance also indicated that the nanoparticles were predominantly made of copper.
The nanoparticles showed excellent antimicrobial activity against bacteria and fungi in the
lowest concentration tested. Both wild tulsi extract (WTE) and wild tulsi nanoparticle (WTN)
were non-toxic on NIH 3T3 cells at lower concentrations tested, however, WTN showed
cytotoxicity at higher concentrations and with extended incubation times (Shubhashree et al.,
2022).

In 2022 The emergence of antibiotic resistance of bacteria, fungi, and viruses recently, which led
to the interest in developing antimicrobial agents and making them more effective in the fields of
sterilization, public health, food preservation, etc., which led to the adoption of mineral elements
as antimicrobials. Bacterial and antiviral agents. Nickel and copper are among the options that
include antimicrobial because they are corrosion resistant and strong. Copper was recognized by
the US Environmental Protection Agency (EPA) in 2008 as an antimicrobial as the first metallic
agent. In this work, the green method successfully synthesized copper nanoparticles from ginger
extract within a short period and the samples were examined using multiple analytical techniques
such as scanning field emission electron microscopy (FE-SEM), X-ray diffraction
(XRD), Fourier transform infrared spectroscopy (FT-IR) and energy-dispersive X-ray (EDX)
analysis to study its structure and morphology. Then it was used as an antibacterial. Here, tests
were performed on two pathogenic strains of Gram-negative (G−ve) and Gram-positive (G+ve)
bacteria, Staphylococcus aureus, and Escherichia coli. At 37 °C, copper showed a response to
inhibition of the action of these two strains during 24 h of incubation. Although copper is
improved, this method can be extended to other types of metallic surfaces and other bacterial
strains (Abbas & Fairouz, 2022).

In 2022 Copper, a naturally occurring element, has medicinal potential and has recently gained in
the textile industry. This paper seeks to chart the way forward on the commercial production of
copper nanoparticles (Cu-NPs) for textile industry. The leaves and bark of Adenathera pavonina
linn were used to synthesize copper nanoparticles (Cu-NPs) which are utilized in a range of
applications as heterogeneous catalysts in textile industry. Copper nanoparticles were created
using a chemical and natural method (Leaf extraction). In the absence of any reducing or
stabilizing agents, the assynthesized samples were submitted to a range of analytical techniques
in order to learn more about the physical properties of the materials. The radius of the Cu-NP
from leave extract ranged from 56 nm – 89 µm with copper, carbon, and oxygen having a
composition of 73.31%, 6.40% and 20.29% respectively. Likewise, the radius of CuNP from
bark extract ranged from 56 nm – 293 µm with copper, oxygen, carbon, silicon, manganese, and
aluminum having a composition of 65%, 24%, 1.22%, 4.20%, 3.10%, and 2.12% respectively.
The impurities are largely from soil which the plant takes its nutrient. Carbon was found in both
extract as it is expected in organic compounds in plants. The crystalline characterization shows
that their exist a significant composition of Cu (I) species in both Cu-NP from the extracts that
makes it highly relevant to the catalytic procedure in the textile industries. Each feature of the
planes shows the copper atom represented as Cu-1 and Cu-2. Since Adenathera pavonina linn is
in abundant in the tropics, its sustainability and low-cost production and processing are highly
desired for textile industries(Owoeye, Akinlabu, Emetere, & ADEBOYE, 2022).

In 2022 The copper nanoparticles (CuNPs) synthesizing potential of Cynodon dactylon aqueous

leaf extract and their antibacterial as well as dye degradation potentials were investigated. The
synthesized CuNPs was initially characterized by gradual colour change from dark brown to blue
in colour and then found absorbance peak at 469 nm. Furthermore, the SEM and DLS analyses
showed that biosynthesized CuNPs were spherical in shaped and size ranging from 120 to
129 nm. The FTIR spectrum confirmed the presence of flavonoids, alkaloids, terpenoids, and
phenols, which involved in the reduction, capping, and stabilization of CuNPs. This green
synthesized CuNPs also demonstrated remarkable antibacterial activity against the
bacterial pathogens such as Escherichia coli, Bacillus subtilis and Staphylococcus
aureus and Klebsiella pneumoniae. This green synthesized CuNPs exhibited considerable dye
degrading potential in the following order as methyl organge > methyl red > Erichrome black T
dyes in the presence of sunlight through photocatalytic degradation process. These results
conclude that C. dactylon aqueous leaf extract mediated nanoparticles possess remarkable
antibacterial and dye degrading potential (Chompunut et al., 2022).

In 2023 The use of nanoparticles has increased significantly in domestic and industrial processes
in recent years. These particles show special physical and chemical behavior due to their high
surface-to-volume ratio, small size, and visual characteristics related to their size. Metal
nanoparticles are widely used in the fields of biotechnology, bioassays, clinical diagnosis and
treatment, food safety, and water and wastewater treatment. In the present study, copper
nanoparticles were green-synthesized using the aqueous extract of Foeniculum vulgare leaf
aqueous extract. The synthesized Cu NPs were characterized by analytical techniques including
energy dispersive X-ray analysis (EDX), field emission scanning electron microscopy (FE-
SEM), X-ray diffraction analysis (XRD), ultraviolet–visible (UV–vis) spectroscopy, and fourier
transform infrared spectroscopy (FTIR). The anti-human lung cancer activity of Cu NPs was
evaluated using MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. The
nanoparticles were formed in a spherical shape in the range size of 33.62 to 74.81 nm. In the
antioxidant test, the IC50 of Foeniculum vulgare, Cu NPs, and butylated hydroxytoluene (BHT)
against 2,2-diphenylpicrylhydrazyl (DPPH) free radicals were 99, 42, and 26 µg/mL,
respectively. In the cellular and molecular part of the recent study, the treated cells with Cu NPs
were assessed by MTT assay for 48 h about the cytotoxicity and anti-human lung cancer
properties on normal (human umbilical vein endothelial cells (HUVECs)) and lung cancer cell
lines i.e., NCI-H2126, NCI-H1299, and NCI-H1437. The IC50 of Cu NPs were 108, 168, and
122 µg/mL against NCI-H2126, NCI-H1299, and NCI-H1437 cell lines, respectively. The
viability of malignant lung cell line reduced dose-dependently in the presence of Cu NPs. The
IC50 of Foeniculum vulgare leaf aqueous extract were 594, 781, and 610 µg/mL against NCI-
H2126, NCI-H1299, and NCI-H1437 cell lines, respectively (Gu et al., 2023).
In 2023 The nano-enabled crop protecting agents have been emerging as a cost-effective, eco-
friendly, and sustainable alternative to conventional chemical pesticides. Here, the antibacterial
activity and disease-suppressive potential of biogenic copper nanoparticles (bio-CuNPs) against
bacterial fruit blotch (BFB), caused by Acidovorax citrulli (Ac), in watermelon (Citrullus
lanatus L.) is discussed. CuNPs are extracellularly biosynthesized using a locally isolated
bacterial strain Bacillus altitudinis WM-2/2, and have spherical shapes of 29.11–78.56 nm.
Various metabolites, such as alcoholic compounds, carboxylic acids, alkenes, aromatic amines,
and halo compounds, stabilize bio-CuNPs. Foliar application of bio-CuNPs increases the Cu
accumulation in shoots/roots (66%/27%), and promotes the growth performance of watermelon
plants by improving fresh/dry weight (36%/39%), through triggering various imperative
physiological and biochemical processes. Importantly, bio-CuNPs at 100 µg mL−1 significantly
suppress watermelon BFB through balancing reactive oxygen species system, improving
photosynthesis capacity, and modulating stomatal immunity. Bio-CuNPs show obvious
antibacterial activity against Ac by inducing oxidative stress, biofilm inhibition, and cellular
integrity disruption. These findings demonstrate that bio-CuNPs can suppress watermelon BFB
through direct antibacterial activity and induction of active immune response in watermelon
plants, and highlight the value of this approach as a powerful tool to increase agricultural
production and alleviate food insecurity (Noman et al., 2023).

In 2023 Using a simple greener and environmentally free method for the synthesis of cuprous
oxide nanoparticles (Cu2O NPs), the effect of Hydroxypropyl-β-cyclodextrin (HP-β-CD) as a
capping agent has also been analyzed in detail. In order to produce the NPs, the copper sulfate in
aqueous solutions is reduced with L-ascorbic acid and capped with HP-β-CD. Energy-dispersive
X-ray spectroscopy (EDX) and X-ray Photoelectron spectroscopy (XPS) are used for the
elemental analysis whereas field emission scanning electron microscopy (FE-SEM), and high-
resolution tunneling electron microscopy (HR-TEM) are carried out for the analysis of surface
morphology of the prepared NPs. In the typically selected area electron diffraction (SAED)
pattern along with the lattice fringes, the crystalline structure of NPs could be described as face-
centered cubic (FCC) with cu (111), (200), and (220) lattice planes. In addition, HR-TEM
images are evident for the Cu2O NPs and revealed that the spherical with an average size of 33.2
nm. In-vitro cytotoxic ability with the NPs has been tested on the MDA-MB-231 (human breast
cancer cell) cells and it was revealed to attain nearly 50% toxicity in 2.0 mg/ml (50% cell
survival) and other concentrations attained 90-98% survival and a similar pattern is expressed in
FACS analysis (Rajamohan, Ashokkumar, & Lee, 2023).

In 2023 Nanomaterials based on metal–doped fluorapatite (FAP) have attracted considerable

interest as potential next–generation antimicrobial agents. In this study, Cu2+–doped
FAP nanocrystals have been successfully synthesized by a neutralization method at room
temperature. Their structural, optical, antimicrobial, and hemcompatible properties have been
investigated. XRD, FTIR, FESEM, and N2 adsorption–desorption studies indicate the formation
of single–phase FAP mesoporous nanopowders, composed of rod–like particles. TEM images
confirmed the formation of nanorodes with a length of 60 nm and a width of about 18 nm.
Rietveld analysis shows that the Cu2+ ions preferentially substitute Ca2 (6 h) sites in the
hexagonal fluorapatite crystal structure. Fluorescence spectroscopy accompanied by MCR–ALS
method confirms substitution of Cu2+ ions in FAP crystal lattice with extracting additional d–
d band transition at green color from FAP broadband self–activated luminescence in violet–blue
color. Antimicrobial studies conducted on Staphylococcus aureus, Escherichia coli
and Micrococcus lysodeikticus showed that FAP nanopowder with the highest Cu2+ content have
strong bacteriostatic action on Staphylococcus aureus bacterial strain in mediums
containing nutrition matters. In addition, this sample in comparison to pure FAP achieved a high
percentage of relative reduction of bacterial population for all three species, being >90% in most
cases. Fungistatic action is noticed too, throwgh the slowing down mycelium growth of
fungus Aspergillus niger, Aspergillus flavus and Penicillium roqueforti and reduction
of sporulation of Aspergillus niger species. Cu2+ doped FAP nanocrystals shows a synergistic
antimicrobial effect with Cu2+ and F− ions. Concerning the potential biomedical applications, the
hemolysis ratios of the Cu2+ doped FAP samples were below 5%. The obtained results pointed
out the possible use of the synthesized nanocrystals as broad–spectrum antimicrobial agents for
various biomedical and health care preparations (Milojkov et al., 2023).
In 2023 Copper oxide nanoparticles (CuO NPs) can be synthesized by a one-pot synthetic
process using microwave irradiation and an extract of apple peel as a nontoxic reducing agent
and capping agent. This process has many advantages, including ease of operation, easy use of
natural and non-toxic precursors, and the absence of harmful reagents. FE-SEM, FE-SEM, HR-
TEM, XRD, XPS, Raman, and TGDTA are commonly used to characterize synthesized NPs.
Based on the XRD results, the structure is face-centered cubic (FCC) with orientation (111). In
addition, the synthesized green CuO NPs have been found to be highly effective in the reduction
of MB dyes as well as CV dyes. Recovering and reusing catalysts with almost no loss of activity
is possible (Rajamohan & Lee, 2023).
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