Environmental Technology Reviews

ISSN: 2162-2515 (Print) 2162-2523 (Online) Journal homepage: www.tandfonline.com/journals/tetr20

A review on green synthesis and characterisation

of copper nanoparticles using plant extracts for
biological applications

Anjali Banger, Ankita Kumari, Nirmala Kumari Jangid, Sapana Jadoun,

Anamika Srivastava & Manish Srivastava

To cite this article: Anjali Banger, Ankita Kumari, Nirmala Kumari Jangid, Sapana
Jadoun, Anamika Srivastava & Manish Srivastava (2025) A review on green synthesis and
characterisation of copper nanoparticles using plant extracts for biological applications,
Environmental Technology Reviews, 14:1, 94-126, DOI: 10.1080/21622515.2025.2453950

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Published online: 16 Feb 2025.

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ENVIRONMENTAL TECHNOLOGY REVIEWS
2025, VOL. 14, NO. 1, 94–126
https://doi.org/10.1080/21622515.2025.2453950

A review on green synthesis and characterisation of copper nanoparticles using

plant extracts for biological applications
Anjali Bangera, Ankita Kumaria, Nirmala Kumari Jangida, Sapana Jadoun b
, Anamika Srivastavaa and
Manish Srivastava c
a
Department of Chemistry, Banasthali Vidyapith, Vanasthali, India; bSol-ARIS, Departamento de Química, Facultad de Ciencias, Universidad de
Tarapacá, Arica, Chile; cDepartment of Chemistry, Central University of Allahabad, Allahabad, India

ABSTRACT ARTICLE HISTORY

The ‘green’ synthesis of metal nanoparticles has emerged as a prominent and environmentally Received 31 May 2024
friendly method in materials science for synthesising various nanoparticles. Among all, copper Accepted 6 January 2025
nanoparticles, with their versatile applications across multiple industries, have garnered
KEYWORDS
significant attention. Plant-derived components, including fruits, leaves, flowers, roots, shoots, Sustainable synthesis;
seeds, and bark, contain active compounds that facilitate the reduction and stabilisation of copper nanoparticles; plants
copper ions to form copper nanoparticles. Harnessing the reducing power of these plant extract; sustainability;
extracts offers a sustainable and cost-effective alternative to conventional chemical methods, antimicrobial activity
minimising environmental impact. The synthesis process yields copper nanoparticles with
unique properties suitable for diverse biomedical, agricultural, and ecological applications.
Characterisation techniques such as UV-visible spectroscopy, X-ray diffraction, transmission
electron microscopy, Fourier-transform infrared spectroscopy, and many more discussed in
detail provide insights into their physicochemical properties. Moreover, these nanoparticles’
biological activities and potential applications in drug delivery, antimicrobial agents, catalysis,
and plant growth promotion are discussed. This review highlights significant progress in
utilising green synthesis for copper nanoparticle fabrication, underscoring their promising role
in advancing various fields of science and technology, including biomedical applications.

Highlights catalysis, and environmental remediation has driven

significant advancements in nanomaterial synthesis.
. Sustainable synthesis of copper nanoparticles.
However, conventional methods often rely on harsh
. Use of plant extract for the green synthesis.
chemicals and energy-intensive processes and use
. In-depth characterisation techniques are discussed.
complex equipment, raising environmental and
. Diverse applications in biological fields.
health concerns [1]. To address these challenges,
‘green synthesis’ has emerged as a sustainable and
environmentally friendly approach to nanoparticle
1. Introduction production. Green synthesis involves using organic,
In recent years, the widespread acceptance of nano­ eco-friendly, and biocompatible materials, such as
particles in industries like electronics, biomedicine, plant extracts, to produce nanoparticles [2]. This

CONTACT Nirmala Kumari Jangid [email protected] Department of Chemistry, Banasthali Vidyapith, Vanasthali, Rajasthan 304022, India;
Sapana Jadoun [email protected] Sol-ARIS, Departamento de Química, Facultad de Ciencias, Universidad de Tarapacá, Avda. General,
Velásquez, Arica 1775, Chile
© 2025 Informa UK Limited, trading as Taylor & Francis Group
 ENVIRONMENTAL TECHNOLOGY REVIEWS 95

technology aims to reduce the negative environ­ copper nanoparticles were synthesised through
mental impacts of traditional synthesis methods green methods using plant materials such as
while ensuring that the nanoparticles exhibit the flowers, stems, leaves, and fruits [13,14]. Commonly
desired characteristics and enhanced used precursors in the synthesis of copper nanoparti­
biocompatibility. cles include copper sulphate pentahydrate [15],
During green synthesis, biological entities such as copper acetate [16], copper chloride [17], copper
plants, bacteria, fungi, algae, etc., serve as excellent nitrate trihydrate [18]. The completion of the reaction,
reducing and stabilising agents in the synthesis of hence the formation of nanoparticles, is initially
inorganic nanoparticles [3,4]. Additionally, green syn­ confirmed via colour change based on the precursor
thesis facilitates the utilisation of various organic used: yellow to brown [19], blue to brown [20],
materials as stabilising and reducing agents, including yellow to green [13]. The UV absorption range
algae, plant extracts, and microorganisms. These ranges from 200–580 nm based on the energy state
agents prevent nanoparticle aggregation and of the nanoparticles [12,13].
accumulation and contribute to their stability. Utilising copper nanoparticles presents an
Examples of bioactive substances found in plant approach that is cost-effective, scalable, compatible
extracts, such as flavonoids, polyphenols, terpenoids, with existing procedures, and aligns with environmen­
and alkaloids, play a vital role in nanoparticle stabilis­ tally friendly practices. Incorporating plant extracts
ation [1,5]. Flavonoids and polyphenols like tannins, enhances the reliability of eco-conscious methods,
by donating electrons to metal ions, reduce the yielding nanoparticles with exceptional biocompat­
metal ions to form nanoparticles. [6]. These com­ ibility crucial for various biological applications, such
pounds reduce nanoparticles’ oxidation state and as medical practices, imaging, and diagnostics.
act as antioxidants [3]. The concentration, reaction This review paper delves into the synthesis of
time, temperature, and pH vary depending on the copper nanoparticles via green synthesis, covering
extraction method of different plant entities during experimental methodologies, characterisation tech­
the synthesis of the nanoparticles [1,7]. niques, and potential biological applications. By
Reports indicate that approximately 40–50% of embracing environmentally benign strategies, we
global food waste originates from plant sources, can foster the development of advanced nanomater­
including roots, flowers, fruits, vegetables, peels, ials that harmonise technological innovation with
leaves, and seeds, all rich in bioactive compounds. ecological responsibility.
So, they can be utilised as natural oxidising agents
to synthesise inorganic nanoparticles. This technique
2. Synthesis of nanoparticles
of synthesising nanoparticles via plant-based entities
directly correlates with the United Nations’ Sustain­ In general, the nanoparticles can be synthesised using
able Development Goals for the 2030 Agenda and the bottom-up and top-down methods, as shown in
clearly defines the principles of circular economy, Figure 1. The top-down method converts the bulk-
which emphasises the recycling, recovering, and reu­ sized materials into nano-sized materials correspond­
tilising of compounds [8]. ing to finer structures. They are simple to use but
Among various nanoparticles, copper nanoparti­ ineffective to produce tremendously small and irregu­
cles are particularly noteworthy due to their versatility larly shaped particles. The major drawback of this
in multiple applications. [9,10]. Copper nanoparticles method is that it is difficult to obtain the required
are considered one of the cheapest transition size and shape of particles. In the bottom-up
metals. They are receiving significant attention due method, small-sized materials are converted into
to their excellent chemical, physical, optical, catalytic, bulk-sized materials. This method is also known as
electrical, mechanical large surface area to volume the constructive method. Nanomaterials’ shape, size,
ratio, stability, magnetic, and biological properties and composition are compared to the top-down
[11,12], making them useful in a wide range of appli­ method. The bottom-up and top-down methods are
cations. Essential parameters for outstanding per­ further classified into distinctive techniques based on
formance are related to the nanoparticles’ sizes, the mechanism involved in the formation of nanopar­
shapes, compositions, and structures. In this review, ticles, the energy intake, i.e. high and low energy
96 A. BANGER ET AL.

Figure 1. Bottom-up and top-down approaches as general synthesising methods for nanoparticles.

processes like milling (top-down) and self-assembly Further, the micro-emulsion technique [29], synthesis
(bottom-up), the medium of synthesis, i.e. solid-, by hydro-thermal technique [30], synthesis by polyol
liquid – or gas- phase like mechanical, CVD, sol–gel technique [31], synthesis by chemical vapour tech­
or green respectively and the environmental par­ nique [32], and synthesis by chemical vapour trans­
ameters that affect during the synthesis of nanoparti­ port method enhanced with plasma [33]. For
cles. These conditions are responsible for their size, instance, CuSO4 and PEG solutions were combined
shape, and application in ecological remediations, bio­ to produce copper nanoparticles by chemical
medical diagnostics, and other research areas. reduction [34]. During this process, ascorbic acid
The physical, chemical, and biological processes was used as a protective agent to prevent the oxi­
that are used in the production of nanoparticles are dation of nanoparticles.
described further: The use of highly concentrated reducing agents,
stabilisers, and surfactants during the fabrication of
nanoparticles involves physical and chemical routes
2.1. Physical methods
of synthesis and high-beam radiation, directly impact­
Physical processes use high-intensity radiations, thermal ing human health and the ecosystem. Therefore,
energy, mechanical pressure, or electrical energy to green nanoparticle synthesis should be favoured
melt, evaporate, condense, and abrade materials to since it involves a one-step biological reduction
create nanoparticles. Highly intensive ball milling method that consumes less energy and uses inexpen­
energy technique [21], pulse-laser deposition and laser sive materials.
ablation [22], electrospray ionisation [23], condensation
of inert gases [24], electron-beam physical vapour trans­
2.3. Biological methods
port [25], devolatilization via laser and flame spray tech­
nique [26], and melt-mixing are few of the most widely Green nanotechnology creates nanoparticles with
used physical processes for synthesising nanoparticles specialised functionalities by combining biological
[27]. These techniques are tedious, labour intensive, concepts with chemical and physical processes. Its
and require high maintenance. foundation is the reduction of risk associated with
manufacturing and using nanomaterials. Creating
‘green chemistry’ processes, i.e. safe, biodegradable,
2.2. Chemical methods
less hazardous, and suitable to the environment,
The sol–gel technique is a widely used chemical tech­ could be advantageous for nanoparticle synthesis.
nique for the preparation of nanoparticles [28]. They might consist of living things, including fungi,
 ENVIRONMENTAL TECHNOLOGY REVIEWS 97

Figure 2. The biological synthesis of copper nanoparticles using plant extract. Initially, the plant extract is added to the precursor salt
solution. In most cases, the plant extract acts as a capping and reducing agent. The synthesised nanoparticles are lastly filtered to
obtain the desired product.

bacteria, algae, and plants. Figure 2 represents the can utilise various plant parts, each discussed in sep­
biological synthesis procedure of nanoparticles. arate sections.

3. Synthesis of copper nanoparticles using 3.1. Using leaves extract

plant entities A.M. Alahdal and others have bio-synthesised copper
Recent studies [5,35,36] have suggested that nanoparticles using Phragmanthera austroarabica leaf
extracts from plant entities could be employed as extract. The results obtained from TEM, SEM, and XRD
a precursor for creating ecologically friendly nano­ revealed that the particle size of copper nanoparticles
particles. Biomolecules like polysaccharides, pro­ was found to be 44.6 ± 2.7 nm, spherical, and crystal­
teins, enzymes, vitamins, organic acids, and amino line in nature. These fluorescent nanoparticles were
acids like citrates are found in plants. Additionally, used to detect hexavalent chromium ions and were
the appropriate function of phytochemicals in the highly efficient for removing dyes. They proved
bio-reduction of metal oxide nanoparticles was
examined [37]. These chemical compounds are
active and cap, stabilise, and reduce metal ions to
metallic nanoparticles. The effect on the plant
parts in the presence and absence of copper has
been described in Figure 3. Its application as an
electron carrier, antimicrobial agent, etc., and how
its deficiency could lead to wilting, diseases, and
stunted growth.
A standard procedure for synthesising copper
nanoparticles includes selecting a part (leaves,
stems, flowers, and fruits) of the desired plant
species and washing it with double-distilled water
to remove dust particles. Then, cut it into smaller
pieces and dry under sunlight. The plant part is
placed in distilled water at 30°–40°C [14,19] and at
higher temperatures >60°C [35,36]. It is further
cooled and filtered to get its extract. The extract can Figure 3. Effect on plant parts in the presence and absence of
be stored at 4° C until use. The synthesis process copper.
98 A. BANGER ET AL.

highly antioxidant in nature [38]. Bouzekri and others peels. The extraction is then mixed with the precursor
have fabricated silver-doped copper nanoparticles (metal salt) to form nanoparticles. Salas et al. have
from the plant extract (leaf) of Hammada scoparia. synthesised ultrasound-assisted nanoparticles of
Their potential was evaluated to prevent the develop­ copper metal using fruit extract of Vaccinium corym­
ment of micro-organisms and the removal of dyes bosum, as shown in Figure 4 (diagrammatic represen­
from the aqueous medium. The characterisation tation), and incorporated them into face masks. The
results discovered that the photocatalytic activity nanoparticles obtained were semi-spherical in
increased the addition of silver to nanoparticles [39]. shape. The TEM data confirmed the size of nanoparti­
Mali and others used leaf extract of Celastrus pani­ cles in the range of 3–12 nm. A characteristic Surface
culatus plant for the reduction of copper ions. The size Plasmon Resonance band was observed at 535 nm,
of the copper nanoparticles was in the 2–10 nm range confirming copper nanoparticle formation. The XRD
and were spherical in shape, exhibiting antifungal and data interpreted the FCC crystalline structure of the
some photocatalytic activity [13]. Wu and co-workers nanoparticles. They served as effective anti-bacterial
used the aqueous leaf extract of the Cissus Virginia additives after they were incorporated into face
plant for the reduction of copper ions. The nanoparti­ masks [41].
cles lay in the range of 5–20 nm and were spherical. Johnson and co-researchers synthesised eco-
The synthesised copper nanoparticles were used to friendly copper nanoparticles via peel extract of
treat urinary infection pathogens [20]. Sharma and P. granatum to check their electrical conductivity.
others synthesised copper nanoparticles using the The SPR absorption band was observed around 570
Tinospora cardifolia leaf extract. In this research, Tinos­ nm, indicating the presence of copper nanoparticles,
pora cardifolia was used as a reducing agent. The syn­ and the electrical conductivity was found to be 3.14
thesised copper nanoparticles exhibited resistance mS/cm [42]. Kausar and co-researchers synthesised
against certain bacterial species and were further copper nanoparticles from Mentha’s aqueous extract
used to impregnate cotton fabrics [17]. to investigate the growth and germination of
Gu and others fabricated copper nanoparticles T. aestivum [43]. Fragaria ananassa fruit extract was
from the aqueous leaf extract of Foeniculum vulgare. used by Hemmati and co-researchers to synthesise
The UV, FTIR, FESEM, and XRD data inferred that the eco-friendly copper nanoparticles [44]. TEM scans
nanoparticles were spherical and between 33.62 and revealed a uniformly spherical shape with sizes
74.81 nm in size. They studied their antioxidant between 10 and 30 nm. The 2,2-diphenyl-1-picrylhy­
effects and evaluated them for cytotoxicity and anti- drazylhydrate free radical scavenging assay was used
human lung cancer characteristics [40]. to assess the antioxidant properties of synthesised
copper nanoparticles. The spherical nanoparticles’
antifungal activity was examined against
3.2. Using fruit extract
C. guilliermondii, C. albicans, C. parapsilosis, C. Kruse,
The fruit extract is prepared by collecting different and C. glabrata. Mohamed used an extract of dates
fruit samples like oranges, papaya, lemon, etc. The without seeds as a reductant to make copper nanopar­
extract can be prepared with juice, pulp, or fruit ticles [45]. The spherical form of copper nanoparticles

Figure 4. Diagrammatic representation of the preparation of copper nanoparticles (ultrasound-assisted) from plant extract of Vacci­
nium corymbosum [41].
 ENVIRONMENTAL TECHNOLOGY REVIEWS 99

and SPR peak at 576 nm were detected by UV-vis spec­ washed with deionised water multiple times, and
troscopy. The produced nanoparticles were 78 nm in then it is cut into pieces and dried under sunlight.
size and had +41 mV zeta potential, according to DLS Further, it is boiled in water to form its extract.
analysis, indicating high particle stability. An absorp­ Finally, it is filtered and dried in the oven. The
tion peak at 560 nm was visible in copper nanoparti­ extract is then mixed with a salt solution to create
cles made using an aqueous fruit extract from the nanoparticles. Kumar and co-researchers syn­
Rhuscoriaria L. Semi-spherical forms with diameters thesised copper nanoparticles using seed extract
ranging from 22–27 nm were found via TEM investi­ from Nigella sativa. Different concentrations of the
gation. According to the XRD investigations, the seed extract have been used, i.e. 5%, 6%, 8%, and
average crystalline size was 18 nm [46]. Copper nano­ 10%, for the evaluation of its antibacterial and anti-
particles produced with the help of Capparis spinosa obesity effect, out of which 8% was found to be the
fruit extract displayed SPR at 414 nm in wavelength. optimum concentration for the synthesis of nano­
The particle size was determined by SEM examination sized copper nanoparticles (98.23 nm) [14]. Copper
to be between 17 and 41 nm [47]. UV-vis spectroscopy nanoparticles were created by Jasrotia and co-
performed for copper nanoparticles made from the researchers [49] utilising extracts of Vigna radiata,
extract of Duranta erecta resulted in a strong absorp­ Allium cepa, and Cicer arietinum. The ability of pro­
tion peak at 588 nm. The synthesised nanoparticles duced copper nanoparticles to scavenge DPPH rad­
were 70 nm in size and were monodisperse, as per icals was used to assess their antioxidant properties.
FESEM [12]. For the green production of copper nano­ The fungal strain Aspergillus sp. and the bacterial
particles, extract Ziziphus spina-christi fruits (L.) Willd. strains E. coli, S. aureus, and S. typhi were used to
Were utilised as a reductant. The UV-Vis spectra investigate the antibacterial activity. Copper nanopar­
revealed a peak at 551 nm. The copper nanoparticles’ ticles were made from Persea americana seed extract
cubic size range of 8–15 nm were identified using and exhibited an absorption peak at 357 nm wave­
XRD. According to the FESEM micrographs, the gener­ length. XRD tests estimated the average diameter of
ated copper nanoparticles ranged in size from 5–20 the copper nanoparticles to be between 42 and 90
nm [48]. nm. According to the SEM investigations, the nano­
particles have a spheroid form, polydisperse, and
45–100 nm size. The agar well diffusion test was per­
3.3. Using seed, root, flower, rhizome, and husk
formed to measure the antifungal activity against
extract
A. fumigatus, A. niger, and F. oxysporum [50]. Studies
The nanoparticles can be prepared using various have shown that green copper nanoparticles have
plant parts like seeds, flowers, roots, rhizomes, etc. been produced using the flower extracts of
In the beginning, the selected part of the plant is S. lavandulifolia, E. crassipes, Rhizome extracts of

Table 1. Synthesis of copper nanoparticles using plant entities.

Plants Plant Parts (extract) Salt Morphology Size Concentration of the substrate Reference
T. spicata Leaf extract CuSO4 Spherical 21 nm 1 mmol [51]
A. muricata Leaf extract CuSO4 Spherical 30-40 nm 0.25mM [52]
C. asiatica Leaf extract CuSO4 Spherical 20-30 nm 5 Mm [53]
C. papaya Leaf extract CuSO4 Square - 1,5,20 mM [54]
M. oleifera Leaf extract Cu (II) - 35-49 nm 0.04 M [55]
Phoenix dactylifera L. Fruit extract CuSO4 Spherical 78 nm - [45]
F. ananassa Fruit extract CuSO4 Rice-grain shaped 10-30 nm 0.04 M [44]
A. cepa Seed extract CuCl Cubic 20 nm 10 m M [49]
Z. officinale Rhizome CuSO4 Spherical 20-10 nm 0.5,1,5 m M [56]
Rhuscoriaria L. Fruit extract CuSO4 Semi spherical 22-27 nm 0.01 M [46]
P. ciliata Leaf extract Cu(NO3)2 Spherical 87-79 nm 1mM [57]
G. glauca Leaf extract CuSO4 Quasi-spherical 1-5 nm 1mM [58]
C. incanus Leaf extract CuNO3 Spherical 15-25 nm - [59]
T. belerica Fruit extract Cu(NO3)2 Spherical 9–14 nm 0.1 M [60]
A. racemosus Root Cu(NO3)2 Rod 3-29 nm 1M [61]
A. laxiflora Leaf extract CuSO4 Spherical 3-29 nm 1mM [62]
T. catappa Leaf extract CuSO4 Spherical 21-30 nm 5, 25, 125, 250 mM [63]
A. indicum Leaf extract Cu(NO3)2 Hexagonal 16-78 nm - [64]
A. indica Leaf extract CuCl2 Cubical 48 nm - [65]
100 A. BANGER ET AL.

Zingiber officinale, Curcuma longa, Asparagus racemo­ time can significantly influence nanoparticle size,
sus, husk extracts of Zea mays L. and many other shape, and crystallinity during synthesis. The
plants and their entities as shown in Table 1. average diameter of gold nanoparticles increased
Figure 5 represents the synthesis of copper nano­ from 33.08 nm after 10 min–37.12 nm after 70 min,
particles via the green synthesising routes using showing a 4 nm growth as they reached their final
leaves, husks, seeds, and roots, as discussed earlier stage [69]. In contrast, reaction time with ascorbic
in this section. acid up to 60 min produced smaller, well-defined
nanoparticles by promoting nucleation, while 90
min led to unclear resonance, suggesting an even
4. Parameters affecting the synthesis of
smaller particle size [70].
nanoparticles
Numerous parameters, like reaction time, tempera­
4.2. Temperature
ture, pH, and reactant concentration, may be altered
to synthesise nanoparticles and characterise the mor­ Around the globe, various levels of research are
phology, as shown in Figure 6. The consequences of being conducted to comprehend the effect of temp­
ecological factors on the fabricated nanoparticles erature on the nanoparticles. The most substantial
have been extensively recognised via the undefined factor that affects the dimensional hierarchy, form,
parameters, and the elements tend to serve an essen­ and level of fabrication of the nanoparticles is temp­
tial role in the optimisation of metal nanoparticles erature. It may be used to alter the size and shape
[66]. of nanoparticles in spherical, triangular, octahedral,
etc. The formation of nucleation centres increases
with the increase in temperature due to alteration
4.1. Reaction time
in the reaction rate [71]. The temperature varies
Reaction time plays a crucial role in the structural from 30°–40°C [14,19] to higher temperatures >60°
behaviour of the nanoparticles and other parameters C [35,36]. The reaction time is an essential factor
like temperature and pH. It is essential in fabricating that significantly affects the size, shape, and yield
magnetic nanoparticles [67] and varies from a few of nanoparticles synthesised via the green approach
minutes to hours [68]. Extending or reducing reaction [72].

Figure 5. Flow chart representing a synthesis of copper nanoparticles via the green method.
 ENVIRONMENTAL TECHNOLOGY REVIEWS 101

Figure 6. The figure depicts the role of parameters during the synthesis of nanoparticles.

4.3. pH collective oscillations of the conduction band elec­

trons in response to electromagnetic waves using
The morphology and structure of nanoparticles are
UV-visible spectroscopy. It gives in-depth information
greatly affected by the pH of the reaction. In particu­
regarding the aggregation of nanoparticles, their
lar, the nucleation process of nanoparticles is affected
stability, structure, and size. When an incident light
by temperature and pH. An increase in the pH conse­
with a specific energy is inclined, the conduction
quently results in more nucleation centres, which
band electrons form resonance on the surface of
plays a vital role in accelerating the synthesis of
nanoparticles. The general diagrammatic represen­
metal nanoparticles. The structural morphology and
tation of the structure of nanoparticle formation
size of nanoparticles are determined mainly by pH
analysis is shown in Figure 7. The absorption range
[4]. Generally, biomolecules like flavonoids and terpe­
of peaks for metallic nanoparticles changes depend­
noids behave as reductants and stabilisers in the syn­
ing on the size of the nanoparticles since metallic
thesis of nanoparticles. Hence, neutral pH (pH 7) is
nanoparticles have discrete absorbance bands in
maintained during the transformation of flavonoids
their characteristic spectra. Gold nanoparticles have
from enol to keto form to reduce Cu2+ ions, facilitated
a characteristic absorbance peak in the 500–550 nm
by the release of hydrogen atoms [13].
range, while silver nanoparticles have a characteristic
5. Characterisation techniques peak in the 400–450 nm range [73]. Copper nanopar­
ticles have an absorption band between 550 and 600
Before using copper nanoparticles in any application, nm [74].
it is essential to have a general understanding of the
characterisation approaches that need to be applied.
Therefore, selecting the appropriate methodologies 5.2. Extraction of nanoparticles
for the characterisation of nanoparticles is crucial. It
An analytical technique for cloud point extraction is
will help in choosing a proper characterisation
used to extract nanoparticles. Amplification is necess­
approach to investigate nanoparticles. A few charac­
ary before analytical analysis because of the low con­
terisation techniques are listed below.
centration of nanoparticles and the matrix effects
present in environmentally friendly samples. This
5.1. Nanoparticle formation analysis
may be attained by introducing a surfactant at a con­
The synthesis of nanoparticles is confirmed by centration more significant than the critical concen­
measuring Surface Plasmon Resonance and the tration to the specimen. Surfactants create micelles
102 A. BANGER ET AL.

Figure 7. General diagram representing the structure of nanoparticle formation analysis.

that contain non-polar compounds as the tempera­ particles have a more significant portion of the drug
ture rises over a particular cloud point. The micelles stacked on them to be presented to the target. As a
aggregate, dehydrate, settle at the bottom of the sol­ result, there is a balance between nanoparticle stab­
ution, and then centrifuged to remove the nonpolar ility and small size.
contents. Using this technique, metal nanoparticles
including gold [75], silver [75], and copper [76] have
been extracted. 5.5. Scanning tunnelling microscopy
This approach is utilised to gain comprehensive
5.3. Tracking of nanoparticles knowledge of the surface of nanoparticles. The first
method used to investigate the shape and size of
It is a method used to track particle sizes in the nm nanostructures was scanning tunnelling microscopy
range when their refractive indices constrain the (STM). Its key characteristic is that it works with
detection of nanoparticles. This technique allows for various materials, including semiconductors and
direct visualisation of the nanoparticles suspended metals. Additionally, growth, topography, and nuclea­
in a liquid. In this method, an optical metal-assisted tion of thin films in vacuum have all been studied
surface of an element is illuminated by a laser using this technique. It is based on quantum tunnel­
beam, and software is built based on an optical micro­ ling, which involves moving a tip across the
scope for detection. It is generally used to administer sample’s surface. On moving the tip across the
targeted medications that relate to nanoparticles pre­ sample’s surface, variations in the tunnelling current
cisely and accurately [77]. generate a picture. For Scanning tunnelling
microscopy, 0.1 nm is the ideal lateral and depth res­
olution [79]. However, only spotless surfaces and
5.4. Analysis of shape and size
sharp conducting tips may be used for STM. This
Shape and size are the two critical components for method was applied to research the composition
nanoparticle applications. For instance, nanoparticles and structure of several materials, including the
are extensively used in medication delivery and tar­ development of copper nanoparticles on electrodes
geting. Particle size has been found to affect medi­ that were either bare or coated with thiols and
cation release. Because of their compact size, placed on gold-plated decamethiol [80]. The STM
nanoparticles provide a vast surface area for drug col­ images of copper on a clean gold surface are shown
laboration and are more effective [78]. Smaller in Figure 8.
 ENVIRONMENTAL TECHNOLOGY REVIEWS 103

Figure 8. STM images of copper on a clean gold surface at (a) 50 mV, (b) 40 mV, (c) 10 mV, and (d) 0 mV with permission from ACS.
[80].

5.6. Atomic force microscopy the surface directly. The map comprises many peaks
in either grayscale or various colour gradients, creat­
Using this method, one may collect quantitative and
ing a multi-colour picture of the sample’s surface top­
qualitative data depending on many characteristics
ology, which is particularly useful for locating and
such as morphology surface roughness, size, and
quantifying the investigated features. Other materials,
texture. Compared to other approaches like
such as silicon [81], chitosan [82], and composites
dynamic light scattering (DLS), electron microscopes,
comprised of copper nanoparticles and ZnO [81],
and optical analysis techniques, it is employed for 3D
are also examined using this approach to determine
characterising nanoparticles. Additionally, it offers
their shape and topography. The analysis of nanopar­
details on the nanoparticle’s geometries, analyses of
ticles with a size of 23.5 nm shows that AFM may
their hydration, and physical characteristics, including
properly investigate copper nanoparticles [83]; one
magnetic activity. Regardless of how conductible and
such example is shown in Figure 9, depicting the
opaque the synthetic materials are, this is also used to
coating of copper nanoparticles with cellulosic film.
analyze soft and hard synthetic materials.
The laser beam bounce method is used to quantify
5.7. Transmission electron microscopy
the cantilever detection that the probe is linked to
because of interaction. As a result, cantilever detec­ This technique is the most efficient among those
tions are used to measure the topographical map of often employed to ascertain the morphology of
104 A. BANGER ET AL.

nanoparticles. It is a sort of microscopy in which an 5.8. Scanning electron microscopy

intense electron beam shoots through a fragile
SEM offers several benefits for analyzing size and mor­
sample and its interfaces, and the electrons that
phology and operates on the same principles as an
pass through the sample are used to produce an
electron microscope. The device comprises a
image. The transmitted electron beam in a TEM
vacuum framework, condenser lenses, and an elec­
exposes the crystallized specimen, and the trans­
tron gun. If the material is non-conductive, the col­
mitted electrons are diffracted across the atomic
loidal solution of nanoparticles is coated with a
sites to create an image. The sample’s planes’ orien­
conductive metal using a sputter coater, dehydrated
tation about the electron beam determines how
to form a powder, and deposited on a sample
intense the direction will be. At the same time,
holder. A beam of very energetic electrons is then
High-resolution transmission electron microscopy
directed at it to extract information from the
(HRTEM) makes it possible to image a specimen’s crys­
sample. Detectors gather the signals and convert
tallographic structure right down to the nucleus level.
them into forms to extract information regarding
Copper nanoparticles may be examined using TEM
the materials’ surface (surface morphology), crystalli­
and HRTEM for various applications. Copper nanopar­
nity, atom orientation, and chemical configuration.
ticles were examined using the TEM to determine
The nanoparticles must tolerate vacuum pressure to
their size and shape, indicating that they were 20–
conduct effective research, as electron beams can
80 nm and spherical and hexagonal [85]. This tech­
harm the nano polymers. Using this technique, the
nique may also be utilised to analyze the morphology
morphology of different metal nanoparticles may be
of other metallic nanoparticles, while that of copper is
analyzed. The morphology of copper and its size dis­
shown in Figure 10.
tribution is shown in Figure 11. SEM images of

Figure 9. Three-dimensional AFM images of cellulose films coated with copper nanoparticles with permission from ACS. [84].
 ENVIRONMENTAL TECHNOLOGY REVIEWS 105

Figure 10. TEM images of copper nanoparticles coated with cellulose film with permission from ACS. [84].

nanoparticles, which demonstrated a spherical form The key drawback is that an aggregate cannot be
with little aggregation and a size range of 5–30 nm, used to determine the size of a specific particle. The
were used to corroborate the shape of copper nano­ concentration of a single aggregation was similarly
particles [86]. The disadvantage of this method is that intractable by the multimodal distributions. When
it is costly, takes a long time, and needs extensive size DLS was used to examine the size distribution of
distribution data. copper and nanoparticles made of other materials, it
was found that the size of copper nanoparticles was
40 nm with a 2 nm covering of silver [88].
5.9. Dynamic light scattering

The size and aggregation of nanoparticles are studied

5.10. For optical properties
using DLS, also known as quasi-elastic light scattering
or photon correlation spectroscopy. It is a quicker, Characterising nanoparticles’ optical characteristics
more potent, and more sensitive tool to characterise before use is essential since they have a wide range
colloidal solutions and macromolecules. In this of applications in optoelectronic devices. Research
method, a monochromatic light beam illuminated on the optical characteristics of nanoparticles uses
the solution containing spherical particles that the following instruments.
exhibit Brownian motion. Large particles move
slowly, and vice versa; a particle’s speed depends on (1) UV-Visible Spectroscopy
size. Doppler shift happens because of a change in (2) Fourier Transform Infrared Spectroscopy
the wavelength of light when it strikes a particle. (3) Raman Spectroscopy
106 A. BANGER ET AL.

Figure 11. SEM image and particle size distribution of copper nanoparticles with permission from ACS. [87].

5.10.1. UV-visible spectroscopy and the spectra demonstrated the metal particles’
One of the earliest techniques in molecular spec­ absorption. The typical signals for metal nanoparti­
troscopy, UV-Vis, is based on the Bouguer–Lambert– cles, like copper nanoparticles, have been shown in
Beer equation. This method is employed to test the Figure 12 a [89,90].
absorption of various materials or fluids. A gadget is
constructed to compare the intensity of two beams 5.10.2. Fourier transform infrared spectroscopy
as they travel throughout the specified wavelength The various functional groups connected to nanoparti­
range. A beam of light breaks into two pieces in UV- cles are identified using an FTIR spectrophotometer at
visible spectroscopy: (i) to analyze the substance in long wavelengths or in the low-frequency range. For
the quartz cell, 50% of the beams pass through it. instance, the FTIR spectra of two compounds are invari­
(ii) In a cell with the same solvent or reference ably distinct because each substance has a distinctive
material, the remaining 50% of the beam passes atom arrangement or vibronic coupling. Therefore,
through it. The intensity of the specimen beam is using infrared spectroscopy, different substances may
less than that of the reference beam. The chemical be positively identified (by a qualitative examination).
concentration in the sample is also determined Solids, powders, pastes, and other bulk or thin
using this method. Copper/corundum nanoparticles materials may be analyzed using FTIR. It can evaluate
were examined through UV-visible spectroscopy, materials quantitatively and inspect specimens up to
 ENVIRONMENTAL TECHNOLOGY REVIEWS 107

Figure 12. (a) UV-Vis spectra of copper chloride ethanol solution under Ultraviolet irradiation a. decrease and b. increase with
irradiation time with permission from ACS [90]. (b) FTIR spectra of Copper and Copper- Poly ethylene glycol nanoparticles with
the authorization from ACS [87]. (c) Zeta potential and pH of nano-copper oxide in nano-pure water with the permission of ACS
[96]. (d) TG data of Copper and Copper- Poly ethylene glycol nanoparticles with authorization from ACS [87]. (e) X-ray diffractometer
pattern of copper nanoparticles coated with cellulose film with permission from ACS [84].

11 mm in diameter. Pure composites’ FTIR spectra have rises, and that of C14O lowers. We infer an alkyl
an area that resembles the ‘fingerprint’ region [91]. group there [92]. The method may be utilised to
Cu(I)–nanoparticles FTIR spectrum revealed the exist­ obtain distinctive spectra of nanoparticles made from
ence of C14O, C–N, and C–H. Due to the contributions various materials, such as gold and copper [87], as
of two resonance structures, the bond order of C–N shown in Figure 12 b, silver, and zinc oxide [93].
108 A. BANGER ET AL.

5.10.3. Raman spectroscopy Copper nanoparticles, which generally vary in size

It is a powerful technique used to observe rotational, from 40–100 nm, were evaluated for colloidal stability
vibrational systems. When the light is scattered from [96] and have 50 mV zeta potential. The zeta potential
the laser, it interacts with the rotational or vibrational of nano-copper oxide is presented in Figure 12 c.
molecular excitations in the system, and the energy of
the photons gets shifted in upward and downward
5.12. Thermal gravimetric analysis
directions. This shifting of the molecular photons pro­
vides information regarding the material’s chemical The change in the mass of the specimen concerning
composition and molecular structure. Hence, this temperature is calculated using a thermobalance.
technique is based on the inelastic scattering of This combines a furnace with a temperature regulator
monochromatic light. When the photons are elasti­ having a suitable electric microbalance. To monitor
cally scattered on the interaction of the light with the environment, a balance is installed within the
the molecule, this process is known as Rayleigh scat­ encircled system. The temperature is plotted on the
tering. The scattered and incident light frequency x-axis, while the mass loss as a percentage is plotted
difference is known as the Raman shift. Plotting the on the y-axis, as shown in Figure 12 d. When the
spectra of the change provides information about sample weight changes, the linear variable differential
the bonds and materials corresponding to the transformer calculates the propensity to rise and fall.
respective peak. It is a sensitive, versatile, and non- The magnetic core of the counterbalance is subjected
destructive technique. This method can be used to to force by a current flowing through its coil, which
observe different spectra of nanoparticles [94]. causes the balance pan to return to its neutral pos­
Recent advancements [19,95] in this technique have ition. The mass change of the sample is matched by
enhanced the reaction rate for real-time monitoring the current required to maintain this position. Accord­
of their progression in different areas like ing to studies on thermal gravimetric analysis (TGA),
nanoscience, environmental monitoring, and biome­ the chemical composition changed during the oxi­
dical diagnostics. dation process, which also impacted the mass of the
copper succinate nanoparticles. Copper nanoparticles
underwent some modifications because of thermal
5.11. For surface charge
deterioration, including weight loss, enthalpy fluctu­
Surface charge is an additional significant aspect for ation, and moisture. Additionally, this method was
characterising the properties and utility of nanoparti­ utilised to determine synthetic nanoparticles’ copper
cles. The primary variables that control the aggrega­ and copper oxide contents. For instance, research dis­
tion, stability, and affinity of nanoparticles toward covered that copper was present in copper nanopar­
biological species and functional groups are the ticles synthesised at pH 1–7, but copper hydroxide
nature and magnitude of the surface charge. This was present at pH 8–11 [97].
knowledge makes the choice of nanoparticle appli­
cation settings and materials more accessible.
5.13. Chemical composition analysis

5.11.1. Zeta potential 5.13.1. Energy dispersive X-ray

The surface charge of nanoparticles is calculated An electron beam is used in the EDX method to
using the zeta potential to assess their colloidal stab­ bombard the sample, which causes it to emit X-rays.
ility. High zeta potential levels must be attained to Qualitative and quantitative assessments are made
guarantee stability and avoid particle aggregation, once these X-rays have been collected and evaluated.
whether positive or negative. The magnitude of Quantitative analysis uses peak intensities to estimate
surface hydrophobicity may be inferred from the an element’s concentration in a sample. Qualitative
zeta potential data. The substance that is contained analysis is used to find various X-ray peaks at pre­
inside the nanocapsules or applied to the surface defined spectrum positions. All elements can readily
can also be revealed by the zeta potential. The produce detectable X-rays. However, substances
surface charge of nanoparticles is assessed using a with lower energies, such as lithium (0.052 keV),
Zeta sizer, which is also used to determine stability. cannot. The method is commonly utilised in electrical,
 ENVIRONMENTAL TECHNOLOGY REVIEWS 109

physical, and chemical sciences. The current analytical [99]. This is around the same size as gold nanoparti­
electron microscope (AEM) has an EDX system and cles [100] and silver nanoparticles [101]. As a result,
TEM attached to it. SEM and EDX can identify and cal­ XRD has become the primary technique for describing
culate the amount of trace components. It cannot the structural characteristics of materials, as pre­
detect X-rays from elements with atomic numbers sented in Figure 12 e.
lower than sodium [79] due to the EDX limitation,
and an improperly calibrated instrument shifts 5.14.2. X-ray photoelectron spectroscopy
peaks to the erroneous energy. In contrast to The X-ray photoelectron spectroscopy (XPS) method
copper nanoparticles, which contain an oxide greatly benefits surface evaluation, which is applied
coating on their surface, EDX analysis of silver nano­ to various materials. It also goes by the name ‘electron
particles is pure, according to different research data spectroscopy for chemical analysis,’ or ‘ESCA,’ offering
[98]. quantitative and chemical state data about the
surface being studied. It delivers surface information
down to a 5 nm scale as a surface analysis approach.
5.14. Characterisations for structure analysis
This technique has a wide variety of information on
Using the characterisation procedures outlined elemental identification, valence band structure,
below, the different crystalline structures and non- chemical states of elements, and relative composition
crystalline materials or nanoparticles are quantified of the components in the surface region. Similar con­
or qualitatively recognised and analyzed. nections may be made to the structure and species of
the constituent parts that make up the compound
5.14.1. X-ray diffractometer composition of magnetic nanoparticles. There is a
Diffraction is investigated when incoming X-rays have minor quantity of Cu2O on the outer surface, and
a wavelength corresponding to the d-spacing of the oxygen was present because copper nanoparticles
sample’s crystalline structure. Plotting the angular were reduced during characterisation [102]. The XPS
coordinates and diffracted X-ray intensity will reveal spectra of Multilayer graphene-copper nanoparticles
the sample’s distinctive patterns. The X-ray diffract­ have been shown in Figure 13.
ometer is primarily used to characterise any crystalline
material and analyze its structure, including the
5.15. For magnetic properties
arrangement of atoms inside the crystal structure
and the angle and interatomic distance. For instance, These methods measure the whole magnetisation
Debye-Scherer’s formulas indicated that Cu and Cu2O rather than being element-specific. Below are some
would have cubic and FCC structures, respectively, of the methods that are frequently used to measure
and the particle size would be around 45.07 nm net magnetisation:

Figure 13. XPS spectra of Multilayer graphene-copper nanoparticles (left) and Multilayer graphene with permission from ACS. [103].
110 A. BANGER ET AL.

Table 2. Characterisation methods and their results.

Analysis of properties Characterisation methods Inference
Analysis of nanoparticle formation UV visible spectroscopy (UV-Vis) Confirmation of synthesised nanoparticles
Analysis of nanoparticle tracking Particles correlation spectrometer (PCS) The investigation of nanoparticles and their size dispersion
Scanning tunnelling microscope (STM) Surface modifications, topology, and chemical analysis
Atomic force microscope (AFM) Size, morphology, texture, and surface roughness
Transmission electron microscope Topography, size, morphology, and crystallographic structure
(TEM) Topography, morphology, and crystallographic structure, size, and
Scanning electron microscope (SEM) composition
Dynamic light scattering (DLS) Size distribution of nanoparticles
Hydrodynamic size of nanoparticles via Brownian motion
Analysis of optical properties UV visible spectroscopy Transmission, absorption, composition, size, concentration, and shape
(UV-Vis) Identification of chemical bonds and functional groups
Fourier transmission infrared
spectroscopy (FTIR)
Analysis of surface area Zeta potential Surface charge and colloidal stability
Brunauer-Emmett and Teller (BET) Gas adsorption data and specific surface area
Analysis of elements Energy dispersive X-ray (EDX) Chemical configuration and purity of elements present
X-ray absorption spectroscopy (XAS) Geometry, elemental composition, and its electronic structure
X-ray fluorescence (XRF) Chemical composition, coating thickness, and concentration
Analysis of structural analysis X-ray diffraction (XRD) Crystalline behaviour, type of structure, and size of the crystallite
X-ray photon spectroscopy (XPS) Crystallography, binding energy, chemical surface analysis, and
composition homogeneity
Analysis of magnetic properties Vibrating sample magnetometry (VSM) Magnetisation and magnetic behaviour of nanoparticles
Analysis of hydrophobicity of the X-beam photon relationship Identification of specific chemical groups on nanoparticles’ surface and their
surface spectroscopy surface hydrophobicity

5.15.1. Vibrating sample magnetometry antibacterial, antitumoral, and antifungal properties,

The Vibrating sample magnetometry (VSM) curve is enhanced by their catalytic activity. This improves
used to assess the magnetic behaviour of magnetic reaction efficiency and selectivity, reducing waste
nanoparticles over a wide temperature range. Mag­ and contaminants for safer, more sustainable pro­
netic nanoparticles are super magnetic, as shown by ducts. Due to their high surface area, copper oxide
the hysteretic loop from zero magnetic remanence nanoparticles are effective antibacterial and catalytic
at room temperature. The saturation magnetisation agents. Factors such as particle shape, size, and
can also be measured utilising the VSM curve’s exposed crystal planes directly influence catalytic
plateau. The core and shell materials must have and biological effectiveness [106,107]. Additionally,
sufficient magnetic and superparamagnetic charac­ copper nanoparticles are widely used for their afford­
teristics for practical use. For example, Zhao et al. ability, toxicity toward pathogens, stability, and
examined the effect of a silica cell on the saturation potential as catalysts [108]. Figure 14 illustrates the
magnetisation of Fe3O4 nanoparticles using VSM green synthesis of these nanoparticles and their appli­
[104]. Copper doping of ZnO has been shown to cations across various fields.
boost its saturation magnetisation and coercivity,
which increases the ferromagnetic behaviour of ZnO
6.1. The antioxidant effect
nanoparticles [105]. Table 2 represents the character­
isation techniques of nanoparticles and the infor­ Copper nanoparticles made from plant extracts are
mation they infer about their structure, morphology, proven to have antioxidant and antibacterial effects,
composition, and other properties. as shown in Figure 15. The behaviour of metallic
salts like copper acetate, copper chloride, and
copper nitrate demonstrates how the antioxidant
6. Biological applications of copper
capacity varies with concentration. Copper sulfate,
nanoparticles
however, is higher than these levels [50]. This quantity
Copper nanoparticles have significant roles in biologi­ was determined based on all the information that was
cal and industrial applications due to their high made accessible. Koelreuteria paniculata seed extract
efficiency, selectivity, and yield in catalytic processes, has less antioxidant activity (14.54%) than Solanum
chemical synthesis, and pollutant elimination. In bio­ nigrum leaf extract (90%). Despite these results, Tinos­
logical applications, copper nanoparticles exhibit pora cordifolia extracts had a greater half maximum
 ENVIRONMENTAL TECHNOLOGY REVIEWS 111

Figure 14. Synthesis and application of green nanoparticles.

inhibitory concentration (IC50) than Solanum nigrum active [107]. Therefore, there is an urgent need
leaf extracts and Borreriahispida (60 and 0.6 g/mL, for a safe, affordable, natural, and non-toxic
respectively). The plant parts that are used to make choice for therapy and prevention. Figure 15
extracts, such as the stems, leaves, roots, fruits, demonstrates the synthesis procedure of copper
flowers, and seeds, are substantial due to the connec­ nanoparticles’ antibacterial and antioxidant activity,
tion between the antioxidant capacity and the phyto­ and Table 3 represents the antioxidant effect of
chemical properties required for the reduction to nanoparticles with different plant species and
generate copper nanoparticles. The decline in pheno­ precursors.
lic compounds reduces the capacity to scavenge free
radicals because phenolic compounds are thought to
6.2. The antitumoral effect
be the primary contributors to plant extracts’ overall
non-enzymatic antioxidant activity [109]. In some One of the biggest reasons for deaths globally is
instances, like the bark of the trees, a link between cancer. Cancer has been treated with a variety of
total polyphenolic concentration and antioxidant techniques, including surgery, chemotherapy, and
activity has been seen [110]. It is significant to high­ radiation. However, these techniques have draw­
light that the extract’s phytochemicals can function backs, such as high costs and unwanted side
as reductant and stabiliser during the synthesis effects. Because of this, non-toxic, affordable treat­
[111]. Plant-based antioxidants can act as stabilising ments with little side effects are required. Nanopar­
agents throughout manufacturing to stop nanoparti­ ticles may aid in treating and diagnosing cancer
cles from aggregating. The phytochemicals with anti­ because of their distinct form, stability, and nanosize
oxidant properties may reduce oxidative stress when compared to other, more giant biological molecules
synthesising. Reactive antioxidant species can be pro­ like enzymes and receptors. Different kinds of nano­
duced due to oxidative stress, which may harm the particles have been studied and reported to have
stability and production of nanoparticles [111]. anticancer properties [127]. Among other nanoparti­
The antioxidant effect, however, inhibits cles, bioinspired copper nanoparticles have potential
ongoing hydrogen abstraction, prevents peroxides anticancer properties against many cancer cell types.
from oxidising, binds transition radical scavengers, Radiotherapy, chemotherapy, and surgery have all
and prevents metal ion catalysts from becoming been available cancer treatment options in recent
112 A. BANGER ET AL.

Figure 15. Synthesis procedure of antibacterial and antioxidant activity of copper nanoparticles.

years. However, these standard techniques do have by aggressively interacting with DNA’s phosphate
costs and usage limitations. Therefore, there is an groups, nitrogen bases, and intracellular protein func­
urgent need for a safe, affordable, natural, and non- tional groups [129]. In vitro tests were performed on
toxic choice for therapy and prevention. As a result, various human cell lines, including lung epithelial
methods employing copper nanoparticles will be cells, kidney cells, brain cells, liver cells, heart micro­
very economical. Copper nanoparticles are potential vascular endothelial cells, and others, to show that
tools for assessing and diagnosing cancer [128]. oxidative stress increased the cytotoxicity of nanopar­
They can operate by various cytotoxic pathways, ticles. Therefore, it is necessary to ascertain if the pro­
notably reactive oxidative species generation, apop­ duced nanoparticles are biocompatible. The
tosis, autophagy, and DNA damage, based on the proliferation and expansion of aberrant or malignant
source of the nanoparticles and the kind of cell cells are accelerated by the multiple mutations that
lines. They cause cytotoxicity in tumorous cell lines free radicals’ high concentration in normal cells’

Table 3. The antioxidant effect of copper nanoparticles.

Salt Plants Salt Concentration (mM) Plant parts Conclusion References
Antioxidant Activity (%) IC50 (µg/mL)
Copper acetate C. vitiginea – Leaves – 45.2 [112]
Copper acetate A. indica 200 Leaves – – [113]
Copper acetate M. buxifolia 50 Leaves 53.40 – [114]
Copper acetate E. prostrata 3 Leaves 53 – [115]
Copper nitrate T. cordifolia – Leaves – 566 [107]
Copper nitrate E. caucasicum 10 Leaves 40.02 – [116]
Copper nitrate S. nigrum 100 Leaves 90 – [117]
Copper nitrate A. noeanum 0.1 Leaves – 160 [118]
Copper sulfate P. ostreatus 2 Seeds – – [119]
Copper sulfate O. basilicum 100 Extract – – [120]
Copper sulfate T. spicata 1 Leaves – – [51]
Copper sulfate T. aestivum 40 Herb – – [121]
Copper sulfate F. vulgaris 40 Leaves – 190 [122]
Copper sulfate S. Hispanic 100 Leaves – – [123]
Copper sulfate L. nobilis 1 Leaves – – [124]
Copper sulfate M. oleifera – Leaves 60–70 – [125]
Copper sulfate U. dioica 10 Leaves 14.54 14 [126]
 ENVIRONMENTAL TECHNOLOGY REVIEWS 113

Figure 16. Schematic representation of antitumoral activity of copper nanoparticles against cancer cells.

DNA and RNA produces. According to some findings, delivery system needs to be tiny enough to exit the
copper nanoparticles may cause malignant cells to circulation, enter the arteries, and get to the tumour.
undergo apoptosis by forming reactive oxidative Figure 16 represents copper nanoparticles’ antitu­
species by altering the uptake of P53 and Bax/Bcl-2. moral activity acting as an antioxidant against
According to earlier studies, the reactive oxidative cancer cells [134]. Table 4 describes the antitumoral
species produced during the development of cancer effect of nanoparticles with their specific defined
cells and transported as radicals or free radicals are plant parts of different plant species with their
destroyed as part of the mechanism of action [130]. different precursor salts along with their defined
Their anticancer effectiveness appears to be concentration.
influenced by their size, shape, increase in oxygen
vacancies, surface area, capacity to diffuse reactant
6.3. The antibacterial effect
molecules, and release of Cu2+. Particle surface area
drastically rises as particle size decreases. As a result, The creation of new pharmaceuticals and the diagno­
there may be more reactive oxidative species sis and treatment of illnesses have benefited from
groups on the particle surface, which might further nano-based therapeutics in recent years. When
exacerbate any biological adverse effects [131]. tested against several harmful bacterial strains, the
Additionally, in cancer cells, tiny nanoparticles antibacterial properties of nanoparticles produced
provide a greater surface area for producing reactive significant results [152]. The antimicrobial effect of
oxidative species such as hydrogen peroxide, nanoparticles results from their interaction with bac­
hydroxyl radicals, and superanionic radicals. The terial cell walls, which are mainly formed of polymeric
reduced size of nanoparticles may lead to more excel­ sugars, amino acids, and peptidoglycans. Because of
lent tissue dispersion, deeper tissue penetration, their porous nature, it is simpler for nanoparticles to
better uptake of cells, and increased cytotoxicity get through. It also depends on the type of bacterium
against cancer cells [132]. According to a study, because Gram-negative cell walls only have one layer
several cancer cell lines are more active with particles of peptidoglycan. In contrast, Gram-positive cell walls
with a diameter of less than 50 nm [118]. Nanoparti­ have numerous layers [117], as shown in Table 5. The
cles around 50 nm have been shown to have much amination process, which enhances membrane per­
higher permeability in tumours than nanoparticles meability, is formed when carboxyl groups and aryls
near 125 nm [133]. As a result, a successful drug in the walls of gram-positive microbes react in the
114 A. BANGER ET AL.

Table 4. The antitumoral effect of copper nanoparticles.

Salt Plants Plant extract Con. of metals Type Size (nm) IC50 (µg·mL−1) Ref.
Copper sulfate A. indica Leaf extract – Breast cancer 26–30 56.16 [135]
Copper sulfate P. nepalensis Fruit
extract 1 mM Breast cancer 35–50 158.5 [129]
Copper sulfate J. glauca Leaf extract 0.1 M Breast cancer 19.72 28.72 [136]
Copper sulfate D. regia Leaf extract 5 mM Breast cancer 69–108 3.77 [137]
Copper acetate A. indica Leaf extract 0.2 M Breast cancer 12 25.55 [112]
Copper acetate Cucurbita spp. Seed
extract 3 mM Breast cancer 20 20 [132]
Copper acetate M. oleifera Leaf extract 0.2 M Breast cancer 12 26.71 [112]
Copper acetate B. Brevis Biomass 100 ul Breast cancer 2–28 122.3 [138]
Copper nitrate S. reticulata Leaf extract 1 mM Breast cancer 42 0.42 [139]
Copper sulfate M. pruriens utilis Seed extract 2.5 gr Cervical Cancer 28 22.48 [140]
Copper sulfate Black beans Beans 10 mM Cervical Cancer 26 – [141]
Copper acetate A. indica Leaf extract 0.2 M Epithelioma 12 26.73 [112]
Copper acetate M. koenigii Leaf extract 0.2 M Epithelioma 12 25.59 [112]
Copper acetate M. oleifera Leaf extract 0.2 M Epithelioma 12 29.58 [112]
Copper sulfate M. cochinchinensis Leaf extract 0.01 M Hepatocellular carcinoma 56 75 [142]
Copper sulfate C. myrica Brown alga 1 mM Hepatocellular carcinoma 21 – [143]
Copper acetate E. prostata Leaf extract 3 mM Hepatocellular carcinoma 23–57 – [114]
Copper acetate H. rosa sinensis Leaf extract 0.2 M Lung carcinoma 12 20.15 [112]
Copper acetate A. paniculata Leaf extract 0.5 M Lung carcinoma 23 14.76 [144]
Copper sulfate F. religiosa Leaf extract 5 mM Lung carcinoma 577 200 [145]
Copper nitrate T. asperellum Cell-free extract 5 mM Lung carcinoma 110 40.625 [146]
Copper acetate C. maxima Seed 3 mM Colorectal cancer 20 25 [147]
Copper sulfate O. europaea Leaf extract 1.7 gr Ovarian cancer 6–15 2.27 [148]
Copper sulfate Cressa spp. Leaf extract 2 mM Ovarian cancer 20–50 20 [149]
Copper sulfate B. oleracea - 5 mM Prostate cancer 4 - [150]
Copper nitrate Vibrio spp. Bacteria 1 mM Esophageal Cancer 8 18.26 [151]

presence of copper [153]. Copper nanoparticles have attraction of nanoparticles to the membrane, which
a deterrent or bactericidal effect precisely because the promotes the oxidation of nanoparticles. These ions
cell membrane enzymes are inhibited by the penetrate via the layers of lipids and proceed into

Table 5. The antibacterial effect of copper nanoparticles.

Plant Size Conc. of
Source Precursor extract (nm) Shape nanoparticles (µg/mL) Micro-Organisms Ref.
A. carambola Copper Fruit 15 Spherical, square, and 20 S. aureus, Bacillius spp., Pseudomonas spp. [159]
acetate extract hexagonal-shaped
S. Swartz Copper Whole 32 Spherical-shaped 25 V. parahaemolyticus [160]
acetate plant
B. braunii Copper Biomass 10–70 Spherical and cubical- 250 P. aeruginosa, E. coli, K. pneumoniae, [161]
acetate shaped S. aureus, F. oxysporum
A. sativum Copper Roots 20–40 Circular-shaped 150 E. coli, S. aureus, B. subtilis, S. pyogenes, [162]
nitrate P. aeruginosa, K. pneumonia
O. ficus- Copper Leaf 3–10 Spherical-shaped 100 E. coli [94]
indica nitrate extract
A. galangal Copper Rhizome 20–60 Nearly spherical-shaped 10 S. mutans, B. cereus, P. vulgaris, [163]
nitrate S. marcences
S. nigrum Copper Leaf 25 Spherical-shaped 100 B. subtilis, S. saprohyticus, E. coli, [117]
nitrate extract P. aeruginosa
C. frutescens Copper Leaf 20–40 Spherical and 150 K. pneumoniae, B. anthracis, [164]
nitrate extract rectangular-shaped L. monocytogenes
F. vulgaris Copper Leaf 20–25 Spherical-shaped 2 S. pneumonia, B. subtilis, C. guilliermondii, [122]
sulfate extract 4 C. krusei
8 P. aeruginosa, S. aureus, C. albicans,
4 C. glabrata,
8 S. typhimurium, E. coli
16 S. pneumonia, B. subtilis,
C. guilliermondii, C. kruse,
P. aeruginosa, S. aureus, C. albicans,
C. glabrata,
S. Typhimurium, E. coli
J. Copper Leaf 50– Flower-shaped 75 E. coli, S. Aureus [165]
gendarussa sulfate extract 100
 ENVIRONMENTAL TECHNOLOGY REVIEWS 115

Figure 17. Schematic representation of the antibacterial activity of copper nanoparticles against bacteria.

the cytoplasm, producing oxygen species like O2 and 6.4. The antidiabetic effect
H2O2 in the process [154]. This process causes the oxi­
Diabetes is a significant risk indicator in the develop­
dation of proteins and lipids. The size and form of
ment of early atherosclerosis. Oxidative stress is
nanoparticles, particularly those that are tiny and
essential since diabetic monocytes create more super­
spherical, may have the ability to act as inhibitors.
oxide anions [166]. In the past few years, type II or
By expanding the surface area, nanoparticles
non-insulin-dependent diabetes mellitus (chronic
improve membrane function and antibacterial activity
disease) has tended to be the most common illness
[155]. The nanoparticles tend to generate radicals of
in the world as well as in India. Because of this,
the hydroxyl group after reacting with substances
there is a dire need to create and evaluate new anti­
like phosphorus and sulfur, which then appear to
diabetic medications that specifically target high
have oxidised the proteins and damaged the Ribonu­
postprandial hyperglycemia since they identify the
cleic acid and Deoxyribonucleic acid, altering the
presence of diabetes in living organisms. Specific
helical helix [156]. Terpenoids, alkaloids, flavonoids,
efforts have been made to find alternatives for identi­
polyphenols, and tannins can disrupt ion transport
fying antidiabetic compounds where copper, zinc,
and affect the transportation of ions. When mixed
manganese, chromium, tungsten, and some other
with plant extracts, copper nanoparticles can further
metal ions revealed both in vivo and in vitro antidia­
improve the antibacterial action. Researchers [157]
betic properties. One such attempt includes synthe­
indicated that copper nanoparticles build up in the
sising copper nanoparticles via medicinal plants to
respiratory and alimentary pathways, rupturing the
characterise post-prandial hyperglycemia and the
tissue layers and affecting Anopheles subpictus in an
effective inhibition of α-amylase assay [167].
anti-larvicidal way. Fungi have polysaccharides like
Disaccharides and oligosaccharides are produced
chitin (N-acetylglucosamine) and lipids in their cell
via degradation of α-amylase and the gastro-intestinal
walls, which provide them resistance and rigidity to
enzyme α-glucosidase [168]. The clinical conditions of
nanoparticles [158]. Figure 17 represents the antibac­
type 2 diabetes include hyperglycemia along with
terial activity of copper nanoparticles against bacterial
excessive urination. Typically, it gets triggered by
degradation of DNA and its membrane.
116 A. BANGER ET AL.

low blood sugar levels. The disorder is distinguished 6.5. The antioxidant effect and cytotoxic effect
by elevated insulin levels and hepatic glucose
2,2-diphenyl-1-picrylhydrazyl (DPPH) is formed of free
secretion [113]. Because of impaired insulin actions,
radicals, which are stable and deep violet because of a
people with diabetes have high blood sugar levels.
strong absorption peak at 520 nm. The crystalline
Therefore, inhibiting enzymes like α-amylase and α-
powder has two primary purposes: monitoring chemi­
glucosidase is imperative to control insulin levels in
cal reactions in antioxidant assay and positioning and
living beings.
analyzing the intensity of electro-paramagnetic res­
Ghosh and co-researchers [169] synthesised
onance signals [172]. The DPPH radicals act as a sca­
copper nanoparticles from the tuber extract of a med­
venger to evaluate the capacity of other radicals.
icinal plant, D. bulbifera, via bioreduction. The syn­
The absorbance decreases when DPPH is exposed to
thesised copper nanoparticles inhibited α-amylase
antioxidants. That is why the reduction in the reaction
and α-glucosidase assay, potentially crucial for treat­
rate in the presence of DPPH indicates the nature of
ing type II diabetes mellitus. For the first time, the
the radical present in that reaction. On neutralisation,
inhibition of these enzymes is performed via the use
the deep violet colour of DPPH turns to pale yellow or
of synthesised copper nanoparticles. Their strong
colourless, allowing visual analysis of the response
potential to inhibit the gastrointestinal enzyme α-glu­
corresponding to the evaluation of the number of
cosidase and the scavenging of free radicals intensify
primary radicals after the transformation of the
their use in treating and evaluating diabetes.
electro-paramagnetic resonance signal.
Kirubakaran and co-researchers [10] synthesised
Studies reveal that copper is an essential element
copper/iron nanoparticles from the leaf extract of
in some antioxidant enzymes like superoxide dismu­
the S. cordifolia plant. On exploring the antidiabetic
tase, which maintains antioxidant activity in ver­
effect, the nanoparticles showed strong antidiabetic
tebrates class Aves. The particle size ranging from
effects as demonstrated by their efficient inhibition
1–100 nm tends to enhance the antioxidant activity
of α-Amylase and α-Glucosidase enzymes. Jamdade
of poultry-based birds [173].
and co-researchers [58] synthesised copper nanopar­
Ghosh and co-researchers [169] synthesised
ticles via aqueous leaf, stem, and flower extract of
copper nanoparticles from the tuber extract of a med­
G. glauca and aqueous leaf extract of P. zeylanica to
icinal plant, D. bulbifera, via bioreduction. The syn­
reduce copper ions. The synthesised copper nanopar­
thesised copper nanoparticles proved to be good
ticles proved to be antidiabetic solid agents as they
antioxidant agents shielding against oxidative stress,
effectively inhibited α-glucosidase (70% – 88%) and
preventing oxidative damage caused by free radical
porcine pancreatic α-amylase (30% – 50%) enzymes.
scavengers, which further helped prevent diabetes.
The mechanism involved in the inhibition corre­
Kirubakaran and co-researchers [10] synthesised
sponds to a change in the conformation of the sec­
copper/iron nanoparticles from the leaf extract of
ondary structure due to the use of copper
the S. cordifolia plant. The nanoparticles showed a
nanoparticles.
strong antioxidant effect since they effectively inhib­
Ameena and co-researchers [170] synthesised
ited the 2,2-Diphenyl-1-picrylhydrazyl (DPPH) assay
copper nanoparticles via leaves of C. hirsutus plant.
and 2, 2-azino-bis-3-ethylbenzothiazoline-6-sulphonic
The inhibition of α-amylase and α-glucosidase was
acid (ABTS) assay. Tahvilian and co-researchers [174]
carried out by infusing the leaves of C. hirsutus
reported the synthesis of copper nanoparticles via
plant and CH-copper nanoparticles. The synthesised
aqueous leaf extract of A. saralicum. The synthesised
nanoparticles were found to show high antidiabetic
nanoparticles showed excellent antioxidant, antifun­
efficiency, around 64% in the case of the α-amylase
gal, antibacterial, cytotoxic, and cutaneous wound
inhibitory assay and around 68% in the case of the
healing applications in both vivo and in vitro environ­
α-glucosidase inhibitory assay. Vardhana and co-
ments. A DPPH free radical test was performed to
researchers [171] synthesised copper nanoparticles
analyze the nanoparticles’ antioxidant behaviour,
via aqueous seed extract of V. vinifera. The inhibition
and the results were excellent.
of amylase enzymes by synthesised nanoparticles
Hasanin and co-researchers [175] biologically syn­
confirmed the antidiabetic effect of the
thesised copper nanoparticles in the presence of
nanoparticles.
 ENVIRONMENTAL TECHNOLOGY REVIEWS 117

starch. Copper nanoparticles with polysaccharide A. marmelos. The nanoparticles showed vigorous
starch-based nanocomposites showed excellent anti­ anti-inflammatory activity, with 78% inhibition for
oxidant effects compared to ascorbic acid, with the membrane stability test and 74% for protein
potent antimicrobial, anticancer, and cytotoxic denaturation. Prakash and co-researchers [180] syn­
effects revealing the safe use of nanocomposite. thesised copper nanoparticles from the leaves of the
Ameena and co-researchers [170] synthesised T. cordiofolia plant. The nanoparticles showed anti-
copper nanoparticles via leaves of C. hirsutus plant. inflammatory solid, antifungal, antibacterial, and anti­
The synthesised nanoparticles were found to show oxidant activity. The nanoparticles suppressed the
vigorous antioxidant activity and enhanced scaven­ protein’s denaturation, confirming the copper nano­
ging activity of free radicals in the H2O2 assay and particle extract’s anti-inflammatory action.
phosphomolybdenum assay. Thiruvengadam and
co-researchers [176] synthesised copper nanoparti­
6.7. The antifungal effect
cles from M. pinnata flower extract. The synthesised
nanoparticles strongly inhibit nitric oxide scavenging Fertiliser, fungicide, and insecticide manufacturers
assay and DPPH radical. The nanoparticles showed have used copper as a raw ingredient for many
excellent antioxidant, antidiabetic, anti-inflammatory, years. According to recent technological advance­
and antibacterial activity. Liu and co-researchers [177] ments, copper nanoparticles may be an effective fun­
synthesised copper nanoparticles using leaves of gicide. Copper nanoparticles are effective fungicides
C. zelanicum medicinal plant. Since copper nanoparti­ against various plant fungi, including C. lunata,
cles possess redox characteristics, they potentially R. solani, P. italicum, P. digitatum, F. oxysporum, and
deactivate the free radicals. DPPH test was conducted other plant fungi species [181]. For instance, copper
to evaluate the antioxidant properties of the nanopar­ nanoparticles produced using a green technique
ticles. The concentration of nanoparticles is directly that combines ascorbic acid and the chemical
proportional to antioxidant behaviour, i.e. the activity reduction of copper ions show antifungal efficacy
increases when the concentration of the nanoparti­ against F. oxysporum, F. solani, and Neofusicoccum
cles is increased. They successfully inhibited the sp. The fungal cell membranes were damaged, and
DPPH assay. reactive oxygen species were produced inside the
cell as part of the antifungal activity. So, using a con­
venient and cost-effective method, copper nanoparti­
6.6. The anti-inflammatory effect
cles can quickly be made with potent antifungal
The denaturation of protein may lead to inflam­ activity. Thus, it is possible to utilise copper to
mation. The anti-inflammatory effect of biologically inhibit and maintain fungi that threaten forest and
synthesised copper nanoparticles can be analyzed crop species.
by inhibition of protein denaturation. The release of Biosynthesized copper nanoparticles showed a
lysosomal neutrophils enhances the inflammation. high antifungal effect against numerous plant patho­
Thiruvengadam and co-researchers [176] syn­ genic fungi. Devipriya and co-researchers [182] syn­
thesised copper nanoparticles from M. pinnata thesised copper nanoparticles using
flower extract. In the inhibitory protein denaturation C. quadrangularis. The nanoparticles showed fungici­
percentage calculation, the copper nanoparticles dal activity against A. niger (86% inhibition) and
show great anti-inflammatory activity since they A. flavus (85% inhibition) by applying the Clinical
efficiently prevent the denaturation of albumin and Laboratory Standards Institute method. Rajesh
protein via the membrane stability method. Ananthi and co-researchers [183] synthesised copper nano­
and co-researchers [178] biologically synthesised particles using S. aromaticum. They showed inhibition
copper nanoparticles via an extract of T. rotundifolia of Penicillium species with a 6 mm zone of inhibition
flower, which showed a robust anti-inflammatory using the Kirby-Bauer disc diffusion method. Benassai
effect based on the concentration of the and co-researchers [184] synthesised copper nano­
nanoparticles. particles and estimated antifungal effects on
Angajala and co-researchers [179] biologically syn­ S. cerevisiae and C. albicans. Pariona and co-research­
thesised copper nanoparticles from leaves of ers [181] synthesised copper nanoparticles via green
118 A. BANGER ET AL.

and evaluated antifungal activity against F. solani, and animals, suggest various transition mechanisms
F. oxysporum, and neofusicoccum species. Tahir and are at play. The combined action of dissolved
co-researchers [185] synthesised copper nanoparti­ copper ions and intracellular particles is hypothesised
cles via leaves of G. asiatica. The synthesised nanopar­ to contribute to this toxicity.
ticles were tested against various bacteria and fungi. Several studies have investigated oxidative stress
The antifungal efficacy of nanoparticles was evaluated reactions induced by copper nanoparticles and
against A. niger (20 mm inhibition zone) and A. oryzae explored alternative cytotoxic pathways. These nano­
(23 mm inhibition zone). particles show promise as drug-delivery agents in
cancer therapy and exhibit antibacterial properties,
making them valuable in biomedicine. Given these
7. Conclusion
characteristics, it’s imperative to analyze and compre­
This review has provided a comprehensive overview hend copper nanoparticles’ harmful environmental
of the green synthesis and characterisation of and human health impacts.
copper nanoparticles using plant extracts, focusing The industrial application of green synthesised
on their biological applications. In recent years, nanoparticles could be challenging since it would
numerous endeavours have been made to synthesise be difficult to maintain their uniformity and reprodu­
nanoparticles through green methods, highlighting cibility. Obtaining high-quality plant extracts in large
the disadvantages of traditional physical and chemi­ quantities would not be economically feasible.
cal approaches. Green synthesis is cost-effective, Hence, their large-scale production could serve as a
environmentally friendly, and scalable for large-scale limiting factor. Proper evaluation of the disposal of
production. by-products generated is needed. The shelf life of
The shift towards green synthesis aligns with the the nanoparticles synthesised via the green
growing interest in eco-friendly technologies, driven approach can be a limiting factor since they are
by low toxicity, cost-effectiveness, and benign reac­ prone to degradation and oxidation, which could
tion conditions. While various organisms can be further affect their application and challenge their
employed for nanoparticle synthesis, challenges storage.
exist, particularly regarding scalability and residue for­ Developing analytical tools is crucial for assessing
mation. More research needs to be done in this area risks and regulating the toxic release of copper nano­
since green-synthesised nanoparticles get easily oxi­ materials and their byproducts into the environment.
dised, affecting their stability and storage. The full Understanding the mechanisms underlying copper
potential of copper nanoparticles awaits exploration, nanoparticle formation will further expand their appli­
with opportunities for innovation in diagnostic, thera­ cations in drug delivery, antimicrobial activities, and
peutic, and material applications. Integration with beyond.
cutting-edge technologies like nanomedicine and
nanoelectronics promises further advancements,
paving the way for impactful contributions across Acknowledgements
various industries. Sapana Jadoun is grateful for the support of the National
Research and Development Agency of Chile (ANID) for the
project FONDAP/1523A0006, Solar Energy Research Center
8. Challenges and future perspectives (SERC), Chile.
Copper plays a vital role in the metabolic functions of
various organisms, but elevated concentrations can
lead to hazardous effects. Using copper in its nano­ Disclosure statement
form extends its applications, amplifying its inter­ No potential conflict of interest was reported by the author(s).
actions with the environment. Genetic anomalies
can disrupt copper metabolism, typically regulated
by homeostatic mechanisms. Observations of cyto­ Funding
toxicity and biological accumulation of released This work was supported by Fondo Nacional de Desarrollo
nanomaterials in the environment, affecting plants Científico y Tecnológico: [Grant Number 3200850].
 ENVIRONMENTAL TECHNOLOGY REVIEWS 119

ORCID viability. Biocatal Agric Biotechnol. 2021;32:101932.

doi:10.1016/j.bcab.2021.101932
Sapana Jadoun http://orcid.org/0000-0002-3572-7934
[12] Ismail M, Gul S, Khan MI, et al. Green synthesis of zer­
Manish Srivastava http://orcid.org/0000-0002-6924-1609
ovalent copper nanoparticles for efficient reduction of
toxic azo dyes congo red and methyl orange. Green
Process Synth. 2019;8(1):135–143. doi:10.1515/gps-
2018-0038
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