Journal of the Indian Chemical Society 102 (2025) 101772

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Journal of the Indian Chemical Society

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Bio-engineered copper oxide nanoparticle for high biocompatible improved

wound healing: Comprehensive characterization and in vivo & in
vitro assessment
Valikala Viswanath a,1 , Jayaprakash Meena b,1 , N. Kumaraguru c, N. Sami veerappa d,
Paik-kyun Shin e, Jiro Tatsugi f, Kannappan Santhakumar b,*
a
Annamacharya College of Pharmacy, Rajampeta, Andhra Pradesh, India
b
Department of Chemistry, School of Advanced Sciences, Vellore Institute of Technology, Vellore, 632 014, India
c
Department of Chemistry, Thanthai Periyar Government Arts and Science College, Tiruchirappalli, 620 023, India
d
Department of Education, Government College of Education for Women, Coimbatore, 641 001, India
e
School of Electrical Engineering, Inha University, South Korea
f
Department of Applied Chemistry, Aichi Institute of Technology, Toyota, Japan

A R T I C L E I N F O A B S T R A C T

Keywords: Copper oxide nanoparticles (CuO NPs) were synthesized using a green technique with Ceropegia fimbriifera floral
Antibacterial extract. UV–visible spectroscopy showed an absorption band at 302 nm with a 2.4 eV band gap, indicating an
Biocompatibility optimized size. FTIR analysis confirmed no incompatibilities, with characteristic peaks at 588 cm− 1 (M − O),
Ceropegia fimbriifera
1052 cm− 1 (aliphatic amines), 2340 cm− 1, 2912 cm− 1, and 3396 cm− 1 (N–H and O–H), 1532 cm− 1 and 1376
Copper oxide nanoparticles
Wound healing
cm− 1 (C–H vibrations), 1638 cm− 1 (N–H bending), 3323 cm− 1 (O–H), 1600 cm− 1 (C– –O), and 1719 cm− 1 (C–H).
Distinct peaks at 540 cm− 1, 596 cm− 1, and 1038 cm− 1 confirmed the formation of CuO NPs. Raman spectroscopy
displayed characteristic peaks at 296 cm− 1 (Ag), 617 cm− 1 (Ag), and 342 cm− 1 (Bg), along with three acoustic
modes (2Au + Bu) and nine optical phonon modes (4Au + 5Bu + 2Ag + Bg). Photoluminescence showed a 434
nm peak, indicating semiconductor behavior, while zeta potential (− 16.2 mV) suggested moderate stability and
XRD revealed an average size of 49.23 nm. Wound healing showed 85.54 ± 0.52 % contraction, while BET
reported a pore volume of 0.062 cm3/g and surface area of 12.45 m2/g, suggesting catalytic potential. TEM and
SEM analyses revealed spherical, cubic, and irregular particles with an average size of 40 nm, and weight and
atomic compositions of 14.58 %, 85.42 %, 9.25 %, and 93.16 %, respectively. SAED confirmed a polycrystalline
structure, while XPS showed peaks at 970 eV (Cu 2p), 965 and 945 eV (Cu2p1/2, Cu2p3/2), and 956 and 934 eV
(Cu2p1/2, Cu2p3/2). Antibacterial studies showed activity against gram-positive and gram-negative bacteria, while
cytotoxicity on A549 and K562 cells indicated reduced viability.

1. Introduction antimicrobials in order to provide the necessary antimicrobial activity

while minimizing toxicity [3]. The importance of copper in biological
The most common metal in the crust of the planet is copper, which sciences has increased as a result of nanotechnology advancements, and
has special physicochemical qualities [1]. It is a necessary component this has encouraged researchers to look into the material’s potential for
with a wide range of therapeutic applications that is only preferred in targeted medication administration [4,5]. According to definitions,
the development of bones, the healing of wounds, angiogenesis, and nanoparticles are discrete nanomaterials with sizes between one and one
enzymatic and antibacterial activity. Extreme amounts of copper will hundred nm. They have special physicochemical properties like
provide antibacterial action even though it is one of the metals that biocompatibility, tunable size, and singlet oxygen formation, which
microbes need most to customize their physiological function [2]. As a allow them to get more attention in nanoscience [6,7]. Cupric oxide and
result, it is advised to use copper at optimal concentrations in a variety of cuprous oxide are highly recommended antibacterial agents in the

* Corresponding author.
E-mail addresses: [email protected], [email protected] (K. Santhakumar).
1
Author for equal contribution.

https://doi.org/10.1016/j.jics.2025.101772
Received 21 February 2025; Received in revised form 18 April 2025; Accepted 13 May 2025
Available online 13 May 2025
0019-4522/© 2025 Indian Chemical Society. Published by Elsevier B.V. All rights are reserved, including those for text and data mining, AI training, and similar
technologies.
V. Viswanath et al. Journal of the Indian Chemical Society 102 (2025) 101772

biological sciences due to their exceptional physicochemical properties indicates colloidal stability, while the nanoparticles exhibit strong
and extended shelf life. Because ambient oxygen causes copper to wound healing potential (85.54 % contraction) and significant cyto­
oxidize into cuprous oxide, it is advisable to protect the nanoparticles toxicity against A549 and K562 cancer cells. These integrated physico­
with a suitable polymer and surfactant to maintain their stability [3,8]. chemical and biomedical traits underscore the functional superiority
It is possible to encapsulate the copper oxide nanoparticles in various and translational potential of the synthesized CuO NPs, marking this
supportive media while maintaining their antibacterial efficacy and work as a significant advancement in green nanoscience with implica­
investigating their potential therapeutic application [9–11]. Copper tions in catalysis, medicine, and materials engineering”. Despite the
primarily acts against bacteria through two mechanisms: cofactor remarkable advantages of Ceropegia firmbriifera-derived CuONPs, several
conjugation in metalloproteins or the generation of reactive oxygen species. critical research gaps remain unaddressed. The precise molecular
The copper oxide nanoparticles have greater antibacterial activity than mechanisms through which Ceropegia firmbriifera phytochemicals
their original form, yet the exact mechanism is still unknown. According contribute to nanoparticle bioactivity, stability, and therapeutic efficacy
to theories on dissolving, the nanoparticles’ enhanced surface-to-volume are yet to be fully understood. Furthermore, while these CuONPs show
ratio makes it easier for them to dissolve, interact with different bio­ promise, their potential in advanced biomedical applications such as
logical components of bacteria, and have an antibiotic effect [3,6,12, targeted drug delivery, tissue engineering, and bioimaging remains
13]. Using the "Trojan Horse Mechanism," the copper oxide nanoparticles largely unexplored due to insufficient pharmacokinetic and bio­
infiltrate cells, producing singlet oxygen that starts the structural distribution studies. Although preliminary findings suggest reduced
degradation of DNA and has antimicrobial properties, changes the po­ cytotoxicity, comprehensive in vivo and long-term safety evaluations
tentiality of the mitochondrial membrane, and produces reactive oxygen across diverse biological systems are crucial to validate their biocom­
species. The surfaces of gram-positive and gram-negative bacteria are patibility. Additionally, a lack of comparative efficacy studies against
negatively charged, whereas the copper oxide nanoparticles and the CuONPs synthesized via other botanical extracts or conventional
surrounding metal ions are strongly positively charged and interact methods limits the understanding of their true therapeutic superiority.
electrostatically. Adherent metal ions or nanoparticles induce fast de­ Beyond biomedical applications, the long-term ecological impact of
polarization of the cell membrane, internal component leakage, and these nanoparticles demands urgent attention to ensure sustainable and
ultimately lysis [14,15]. With 200 species, Ceropegia fimbriifera is one of responsible development.
the largest genera of flowering plants and is only sporadically found in Kumar et al. [18] synthesized bimetallic CuO/Cu2O–ZnO nano­
tropical and subtropical regions. Thirty-five of the fifty species found in composites using Ipomoea carnea leaf extract under microwave condi­
India are restricted to the Western Ghats and are used extensively in tions with varying Cu:Zn ratios (90:10 to 60:40). Characterization
ayurvedic medicine, food, and decorative plants [16,17]. Massive confirmed particle sizes ranging from 7 to 50 nm, band gaps of 1.47–2.9
diversification has resulted in an amazing spectrum of floral morphol­ eV, and high photocatalytic (up to 98.96 %) and antioxidant (up to
ogies, including reduction of the floral whorls and syn-organization 99.77 %) activities. Similarly, Sharma et al. [19] developed ZnO nano­
[16]. The endemic flower species Ceropegia fimbriifera is only found in particles using aloe vera extract at pH 11.5, which demonstrated diverse
a small area of India due to habitat destruction and heavy exploitation. morphologies, band gaps of 3.36–3.43 eV, and effective photocatalytic
We’ve started looking into its antibacterial activity because of the pre­ and antibacterial performance. In another study, Sharma et al. [20]
cise theoretical understanding of its therapeutic relevance. To provide synthesized CuO nanoparticles using Ocimum tenuiflorum, yielding par­
scientific proof of its antibacterial effect, we have created copper oxide ticles sized 6–18 nm that exhibited strong antibacterial activity against
nanoparticles encapsulated with Ceropegia fimbriifera flower extract. As Bacillus subtilis, Staphylococcus aureus, and Escherichia coli, along with
a result, the study seeks to create environmentally friendly CuONPs 96.4 % methyl orange degradation in 24 min. Thakur et al. [21] reported
using the floral extract of Ceropegia fimbriifera to minimize the impact on (Ag, Co) co-doped CuO nanoparticles synthesized via a
the environment. The nanoparticles will be subjected to multiple spec­ microwave-assisted method, showing morphology transitions from
troscopic, microscopic, and thermal degradation analyses to determine nanorods to nanospheres, increased band gap (1.42–1.58 eV), and
their structural and morphological features, as well as their thermal enhanced antibacterial activity against both Gram-positive and
resistance. The antibacterial efficacy of the nanoparticles will be eval­ Gram-negative bacteria. In a related study, Thakur et al. [22] also
uated against various common pathogens to gauge their antibacterial investigated (Ag, Fe) co-doped CuO nanoparticles with crystallite sizes
properties. The research also demonstrates the animal experiments that between 13.10 and 24.98 nm, which showed promising antibacterial
support the effectiveness of CuONPs in fighting bacterial infections. and photocatalytic activity for dye degradation. Furthermore, pure and
Hence, the study aims to promote a sustainable approach to nanoparticle (Ag, Ni) co-doped CuO nanoparticles synthesized by Thakur et al. [23]
production with significant medical potential. “The green synthesis of demonstrated particle sizes below 30 nm and strong antibacterial effects
copper oxide nanoparticles (CuO NPs) using Ceropegia fimbriifera floral against multidrug-resistant strains. The reports of Thakur et al. [24] on
extract presents a novel and eco-conscious approach in nanotechnology. (Ag, Zn) co-doped CuO nanoparticles revealed enhanced crystallinity,
Unlike conventional green synthesis methods that employ widely stud­ optical properties, and antibacterial efficiency. Additionally, Sharma
ied plants such as Azadirachta indica or Simmondsia chinensis, the use of et al. [25]synthesized CuO nanoparticles using aloe vera extract at pH
C. fimbriifera introduces unique phytochemicals that modulate nano­ 11.5 and 70 ◦ C, producing 5–20 nm particles that showed variable
particle morphology, surface chemistry, and biological performance. morphology, effective antibacterial activity, and 96 % methyl orange
Existing green syntheses often suffer from poor control over nano­ degradation under UV light in the presence of H2O2. This exclusive
particle size distribution, crystallinity, and colloidal stability due to feature enhanced its applicability in tissue engineering, wound healing
uncharacterized phytochemical variability. Moreover, limited use of materials, or bioactive coatings. Kumar and associates [26] synthesized
advanced characterization techniques such as Raman spectroscopy, eco-friendly Co/Ni doped hematite nanoparticles using both green and
photoluminescence (PL), and Brunauer–Emmett–Teller (BET) analysis chemical methods. In the chemical synthesis, polyvinyl pyrrolidone was
restricts the understanding of structural and functional dynamics in employed, while Azadirachta indica leaf extract was used as a stabilizer
many reports. In contrast, the current study overcomes these constraints in the green synthesis. The nanoparticles had sizes ranging from 15.98 to
by achieving well-defined CuO NPs with a narrow size range (40–59 25.91 nm, exhibiting a quasi-spherical shape with crystalline sizes be­
nm), a distinct band gap of 2.4 eV (UV–Vis), and high crystallinity (XRD, tween 14 and 21 nm. XPS spectra confirmed the presence of dopants,
SAED). The presence of a defect-rich multilayered CuO shell, confirmed and Vibrating Sample Magnetometry (VSM) studies evaluated the
through XPS and PL quenching, promotes efficient charge carrier sepa­ magnetic properties, including residual magnetism, magnetization, and
ration and extends carrier lifetimes, thereby enhancing photocatalytic coercivity. The nanoparticles demonstrated enhanced photocatalytic
and antimicrobial efficacy. A moderate zeta potential (− 16.2 mV) activity, with 93–95 % degradation of methyl blue and methyl orange

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V. Viswanath et al. Journal of the Indian Chemical Society 102 (2025) 101772

dyes within 90 min. Antioxidant activity, measured using DPPH and extraordinary, achieving a staggering 96.42 % degradation of Congo
nitric oxide assays, showed a scavenger activity of 83–88 %, which Red and an even higher 98.36 % degradation of Eosin Yellow.
increased with nanoparticle concentration. The IC50 values for the green Furthermore, antioxidant assays demonstrated exceptional free radical
synthesized and metal-doped nanoparticles were 18.33 μg/ml and scavenging capabilities, with DPPH inhibition ranging from 71.13 % to
16.09 μg/ml, respectively, suggesting their suitability for biomedical 90.35 %, and explored the immense potential of green synthesis in
applications. Sharma and colleagues [27] synthesized eco-friendly developing bioactive nanomaterials for environmental and biomedical
nickel-doped zinc oxide and neodymium-doped zinc oxide nano­ applications. The current investigation successfully achieved the green
particles, loaded with Vitex negundo, and assessed their antioxidant, synthesis of CuONPs using an aqueous extract of Ceropegia firmbriifera.
photocatalytic, and antibacterial properties. X-ray diffraction confirmed The structural, morphological, and optical properties of the synthesized
their hexagonal structures, and Transmittance Electron Microscopy CuONPs were comprehensively analyzed using advanced techniques,
(TEM), Scanning Electron Microscopy (SEM), and Energy dispersive including UV–visible spectroscopy, X-ray diffraction (XRD),
X-ray spectroscopy (EDX) revealed nanoparticle sizes ranging from 8 to Fourier-transform infrared spectroscopy (FTIR), scanning electron mi­
15 nm. Photocatalytic degradation using methyl green dye showed croscopy (SEM), transmission electron microscopy (TEM), and X-ray
promising results for both nanoparticles. Antibacterial studies indicated photoelectron spectroscopy (XPS). Furthermore, the study demonstrated
their effectiveness against E. coli and Bacillus subtilis, while antioxidant the antibacterial efficacy of the green-synthesized CuONPs against
activity, measured by the DPPH assay, demonstrated enhanced perfor­ various gram-positive and gram-negative bacteria and evaluated their
mance. Kumar et al. [28] successfully synthesized innovative cytotoxic effects on specific cell lines. These findings highlight the suc­
copper/zinc-doped hematite nanoparticles through a co-precipitation cessful development of CuONPs with enhanced biomedical applicability,
method, using polyvinylpyrrolidone and Azadirachta indica extract as sustainability, and biocompatibility. The green synthesis of CuONPs
stabilizing agents. These nanoparticles, measuring between 13.33 and using Ceropegia firmbriifera offers a novel, eco-friendly approach,
19.90 nm, not only exhibited remarkable ferrimagnetic behavior at 300 yielding stable colloidal nanoparticles with remarkable therapeutic po­
K but also demonstrated a significant enhancement in the photocatalytic tential. Acting as both a reducing and stabilizing agent, the floral extract
degradation of Rhodamine Blue and Congo Red dyes. Furthermore, their streamlines synthesis, eliminates hazardous chemicals, and reduces
antioxidant activity was rigorously evaluated using advanced assays, cytotoxicity while enhancing cost-effectiveness. These CuONPs exhibit
including 2,2′-Azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) and 2, potent anticancer, antibacterial, antiviral, antifungal,
2-Diphenyl-1-picrylhydrazyl, enumerating their potential in nano­ anti-inflammatory, and wound-healing properties, amplified by the
science. Verma and his research team [29] pioneered a green synthesis bioactive compounds of Ceropegia firmbriifera. Their well-defined
strategy, introducing a dual-doping technique that infused zinc oxide nanoscale size and morphology enhance cellular permeability,
structures with aluminum and iron elements. This innovative approach enabling superior antimicrobial action at minimal concentrations. With
aimed to engineer nanoparticles with exceptional structural integrity. unique phytochemical advantages boosting bioavailability and thera­
X-ray diffraction (XRD) analysis unveiled crystalline sizes ranging from peutic efficacy, these nanoparticles represent a promising frontier in
17 nm to 31 nm, dynamically influenced by aluminum doping. nanomedicine and biomedical research. Verma et al. [32] pioneered the
High-resolution imaging through TEM and SEM confirmed the forma­ eco-friendly synthesis of nanoparticles doped with zinc oxide, copper,
tion of nearly perfect spherical nanoparticles, averaging 20.56 nm in and cerium using Colocasia esculenta plant extract via advanced micro­
diameter. FTIR spectroscopy unveiled a complex and intricate nano­ wave technology. Their structural and functional properties were pre­
particulate composition, while optical studies revealed a remarkable cisely analyzed, revealing crystalline nanoparticles with a size range of
narrowing of the band gap energy from 2.99 eV to 2.88 eV. Beyond their 16–19 nm, as confirmed by XRD studies. The intricate surface
structural and optical finesse, these nanoparticles exhibited extraordi­ morphology and internal architecture were exposed through TEM and
nary functional capabilities demonstrating potent antibacterial and SEM-EDS imaging, while UV spectroscopy illustrated their remarkable
antioxidant properties, coupled with an astonishing 97 % degradation energy band characteristics, featuring their potentiality towards
efficiency of rhodamine dye, marking a significant advancement in UV-shielding applications. Beyond structural excellence, these nano­
nanomaterial applications. Devi et al. [30] unveiled the remarkable particles exhibited strong antibacterial and antioxidant properties,
biological potential of gadolinium-doped and zinc-doped manganese unlocking transformative possibilities for innovative applications in
oxide nanoparticles, shedding light on their powerful antibacterial, biosciences and nanomedicine. Thakur and colleagues [33] pioneered
antifungal, and antioxidant properties. These eco-friendly nanoparticles the synthesis of α-Fe2O3 nanoparticles infused with cobalt, nickel, and
were synthesized using Pinus roxburghii, harnessing nature’s chemistry zinc, comprising the natural essence of Azadirachta indica alongside
to create a sustainable and effective solution. Advanced characterization polyvinyl pyrrolidone coatings. Their XRD analysis unraveled a crys­
techniques confirmed their unique structural attributes, such as XRD talline architecture spanning 14–21 nm, while SEM and TEM disclosed
analysis revealed a well-defined tetragonal crystalline framework, while the presence of uniformly spherical nanoparticles within the 16–26 nm
SEM-DES, in conjunction with TEM, verified their precisely spherical range. Investigating into their magnetic essence, these nanoparticles
morphology, with sizes ranging between 8 and 12 nm. The antifungal exhibited antiferromagnetic behavior, at an irreversible temperature of
studies demonstrated their formidable ability to suppress fungal growth, 300 K. These remarkable nanostructures, with their striking structural
while their antibacterial efficacy showcased their potency in curbing and magnetic properties, emerged as potent catalysts, and revealed an
bacterial proliferation. Furthermore, their remarkable antioxidant po­ impressive degradation of Rhodamine Blue and Malachite Green dyes
tential was evident in their ability to combat oxidative stress, high­ when exposed to UV light. This transformation was driven by the gen­
lighting their immense promise in biomedical and environmental eration of hydroxyl radicals from dissolved oxygen, exhibiting their
applications. Kumar and his research team [31] explored the synthesis of strenuous oxidative capability. These nanoparticles demonstrated
cobalt and zinc-doped hematite nanoparticles, using polyvinyl pyrroli­ exceptional antioxidant potential, attributed to their rich phenol content
done and an eco-friendly approach using Azadirachta indica aqueous leaf which serves as a promising avenue for environmental and biomedical
extract. Their meticulous characterization produced nanoparticles with applications. Thakur et al. [33] developed eco-friendly zinc-cobalt and
sizes ranging from 8.64 to 15.32 nm, adopting a well-defined rhombo­ titanium dioxide nanoparticles using Tinospora cordifolia and poly­
hedral crystallite structure measuring 23.25 nm for chemically synthe­ vinylpyrrolidone via chemical encapsulation. The nanoparticles were
sized variants and 12.52 nm for their green-synthesized counterparts. spherical in shape with sizes ranging from 61 to 82 nm. Structural
The nanoparticles exhibited impressive magnetic properties, with analysis confirmed their stability and uniformity. Photocatalytic studies
magnetization values spanning 36–45 emu/g and remarkable super­ showed efficient degradation of methyl orange and methylene blue
paramagnetic behavior at 300 K. Their catalytic efficiency was equally under UV light. DPPH and hydrogen peroxide assays revealed excellent

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V. Viswanath et al. Journal of the Indian Chemical Society 102 (2025) 101772

antioxidant activity, with 96 % and 95 % scavenging efficiency, of copper nanoparticles (Scheme 1). Furthermore, the nanoparticles
respectively. These nanoparticles, with their dual potential in environ­ were rinsed with deionized water, chloroform, and ethanol and air-dried
mental cleanup and biomedical applications, represent a sustainable for 72 h. The plausible bioactive compounds which are present in the
advancement in green nanotechnology for water treatment. Balkrishna Ceropegia fimbriifera extract are vanillin, oleic acid, ferulic acid, lupeol,
et al. [20]synthesized eco-friendly silver nanoparticles (AgNPs) using Lup-1S-en-3beta-ol, 2-Geimatenol, cerpegin, gallic acid, and catechol
leaf extracts of Ocimum tenuiflorum and Catharanthus roseus. XRD anal­ are the reasons for the Bioreduction process (Scheme 2). The phyto­
ysis indicated particle sizes of 21.42 nm for O. tenuiflorum and 19.96 nm constituents can act as reducing agents, capping agents, and stabilizing
for C. roseus, while TEM revealed broader size ranges of 17–55 nm and agents to form stabilized Nps [34].
10–48 nm, respectively. SEM confirmed their predominantly spherical Chemical Grade & Purity Company
morphology. These green-synthesized AgNPs showed strong antibacte­ Copper acetate AR (Monohydrate) with 99 % purity CDH Fine Chemicals,
rial activity against E. coli and B. subtilis. Remarkably, O. tenuiflorum India
AgNPs demonstrated a lower Minimum Inhibitory Concentration of 3.90 Ethanol LR grade with 99 % purity Midaskem, India
± 0 μg/ml. Their enhanced antimicrobial effect is attributed to phyto­ Chloroform LR grade with 99.9 % purity JSK Chem zone
chemical surface functionalization facilitating bacterial membrane
disruption. These findings reinforce the potential of
phytochemical-assisted nanomaterials as promising candidates for the
2.4. Stoichiometric estimation of percentage yield analysis
development of effective antimicrobial agent. Despite of the numerous
advantages offered by the green synthesis of nanoparticles, there still
To initiate the synthesis, 100 ml of a 1 mM copper acetate solution
remains a gap in exploring the synthesis of copper oxide nanoparticles
was precisely prepared, corresponding to 1 × 10− 4 mol of copper ace­
using the rare plant species like ceropegia firmbriifera is underutilized.
tate. Under optimal reaction conditions, each mole of copper acetate
The ceropegia firmbriifera plant species comprise of miscellaneous plant
theoretically yields 1 mol of copper oxide (CuO) nanoparticles. Based on
extracts such as alkaloids, flavonoids, and phenols which play a signif­
stoichiometric calculations, the theoretical yield of CuO was determined
icant role in stabilizing the CuONPs and enhancing their therapeutic
to be 7.96 mg. Following the green synthesis process and purification
activity. However, the well documented evidence broad-spectrum
steps, the actual mass of CuO nanoparticles collected after drying was
antimicrobial activity, high redox potential, photocatalytic effi­
7.5 mg. This outcome reflects a highly efficient synthesis, achieving a
ciency, and cost-effectiveness, of copper oxide makes it a promising
percentage yield of 94.22 %, a strong indicator of reaction completeness
candidate for diverse applications in biomedicine, wastewater treat­
and minimal material loss during post-synthesis handling. The per­
ment, solar energy conversion, and environmental remediation.
centage yield was calculated using the formula:
The investigation not only serves as a paradigm to the green chemistry,
but also leverages the untapped potential of endemic flora for applica­ Actual mass
% Yield = X 100
tions in biomedicine, water purification, and environmental Theoretical mass
remediation. Hence, the research in this point of view is truly and
scientifically justified. 3. Evaluation techniques

2. Methodology 3.1. UV–visible spectroscopy

2.1. Collection of plant material Ceropegia fimbriifera embedded CuONPs optical properties were
examined using a double beam UV visible NIR spectrophotometer
The fresh matured flowers of hibiscus platanifolius (Ceropegia fim­ (PerkinElmer Lambda 950 at 10 mm path length) that was calibrated in
briifera floral extract) were collected from Jawathu hills (latitude 12.34◦ the 200–900 nm wavelength range. The spectral interpretation was done
60′ N and longitude 78◦ 60′E) are an extension of the Eastern Ghats for the structural characterization of the nanostructures, and the spec­
spread across parts of Tirupattur and Tiruvannamalai districts in the troscopic characterization was recorded using distilled water as the
northern part of the state of Tamil Nadu in south-eastern India. The standard solvent.
authentication of the plant species was done by Prof. N. Samiveerapa,
Department of Biotechnology, Bharathidasan University, Tamil Nadu, 3.2. Fourier-transform infrared spectroscopy
India. The voucher specimen (BDUA2024N0074) was also maintained in
the Department of Biotechnology, Government arts and Science College, The Brukner Alpha ATR-IR spectrophotometer was used to examine
Bharathidasan University, Trichy, Tamil Nadu, India. the FTIR spectroscopy of CuONPs throughout a spectrum range of
4000–370 cm− 1. The instrument was run at 37 ± 0.5 ◦ C with a resolu­
2.2. Extraction of Ceropegia fimbriifera floral extract tion of 4 cm− 1 and 64 scans on average per sample. A graph was created
based on the analysis results of the nanoparticles, and the functional
The flowers underwent cleaning, drying, and were exposed to heat at groups within the sample were identified by analyzing the peak values.
60 ± 0.5 ◦ C using Milli-Q water for 30 min. After filtration and centri­
fugation for 10 min at 8000 rpm to extract the aggregates, the final 3.3. Raman spectroscopy
extract was stored at 10 ◦ C for formulation.
The Raman spectroscopy was produced at 532 nm excitation and in
2.3. Synthesis of copper oxide nanoparticles using floral extract the 400 –50 cm− 1 range with a resolution of 0.8–4 cm− 1 using the
Brukner RFS 27 MultiRAM FT-Raman Spectrophotometer. Raman
The Ceropegia fimbriifera flowers were collected from the Javadhu spectroscopy is frequently employed to obtain information on the
hills, Eastern Ghats, Vellore, India. The flowers were cleaned, dried, and microstructural arrangement and structural bonds of nanomaterials. The
warmed with Milli-Q water at 60 ± 0.5 ◦ C for 30 min. The extract was use of nanomaterials is necessary to accurately identify their unantici­
subjected to filtration followed by centrifugation at 6000 RPM for 10 pated phase and crystallinity.
min to extirpate the aggregates, and the ultimate extract was preserved
at 10 ◦ C for formulation. The CuONPs were prepared by admixing 10 ml 3.4. Photoluminescence spectroscopy
of the extract with 100 ml of 1 mM Cu(OAc)2 solution and heating at 85
± 0.5 ◦ C for 1 h, which results in dark brown and signifies the formation The photoluminescence spectroscopy was measured with a LabRAM

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V. Viswanath et al. Journal of the Indian Chemical Society 102 (2025) 101772

Scheme 1. Schematic diagram of the preparation of CuO NPs.

(HR 800) spectrophotometer at 37 ± 0.5 ◦ C with a 150 W xenon lamp. 3.7. BET analysis
The photoluminescence was triggered and the amount of electron en­
ergy in the nanomaterials was counted at 325 nm. Carefully examining The Brunauer-Emmett-Teller (BET) plot was used to investigate the
the emission spectrum allows one to decipher the emission bandwidth surface area of the CuO nanoparticles and the Micromeritic ASAP 2460
and identify the distinctive peaks that reveal the structural surface area and porosity analyzer was used to establish the pore size
functionalities. distributions. The BET surface area and micropore surface area were
determined by using the t-plot and BET equation, respectively.
3.5. Zeta potential
3.8. SEM and EDX analysis
Using the dynamic light scattering approach, the surface charge of
the nanomaterials is determined by distributing the produced CuONPs The JEOL JSM-7600F equipped with Energy Dispersive X-ray (EDX)
according to their volume and density. Using the Zetasizer nanoseries- at an operating voltage of 25 kV was used to perform Scanning Electron
ZEN 3690 (Malvern Instruments) and the recorded spectrum, the zeta Microscopy (SEM) on the produced nanoparticles in order to examine
potential of the nanoparticles was examined by taking intensity (a.u.) their morphological and structural makeup. The elemental composition
into account on the ordinate axis and zeta potential on the abscissa axis. and impurity profile of the nanomaterials were utilized in the EDX study.
Analyzing the particle size distribution also involves plotting frequency
(%nm) on the y-axis and zeta potential (mV) on the x-axis. A higher 3.9. TEM and particle size distribution
surface charge suggests great colloidal stability, while a lower surface
charge suggests accumulation of the nanomaterials. The stability of the The JEOL JEM-F-200 operating at 200 kV was used to determine the
nanocomposites is directly related to their surface charge. transmission electron microscopy, and the sample for analysis was
prepared by dispersing the nanoparticles in aqueous media and then
3.6. X-ray diffraction studies (XRD) ultrasonification for 15 min to investigate the structural characteristics
of nanomaterials. Using the Zetasizer nanoseries-ZEN 3690 (Malvern
Using a Bruker Kappa APEXII diffractometer, XRD experiments were Instruments) to report the hydrodynamic diameter of the nanomaterials,
conducted to examine the crystalline structure of CuONPs. The radiation the particle size distribution of nanocomposites was ascertained. After
measured was CuKα radiation (1.54 Å) operated at 40 kV and 30 mA. the CuONPs were added to quartz cell cuvettes, diluted with distilled
Using a 0.029◦ step and a speed of 2.5◦ /min, the XRD pattern was water, and incubated at 200 rpm in an orbital shaker, the particle size
captured in continuous scanning mode for the 2θ range of 30◦ to 75◦ . distribution was determined.
Bragg’s law was used to calculate the basal spacing in the crystalline
architecture, and the findings were interpreted appropriately.

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V. Viswanath et al. Journal of the Indian Chemical Society 102 (2025) 101772

Scheme 2. A plausible mechanism of Bioreduction process for the formation of CuONPs.

3.10. X-ray photoelectron spectroscopy (XPS) 3.12. Phytochemical screening

An examination of nanomaterials chemical and elemental composi­ A phytochemical analysis was performed on the Ceropegia fimbriifera
tion using XPS is more accurate, repeatable, and advantageous from a flower extract in order to identify different primary and secondary
scientific standpoint. Peak fitting techniques utilizing synthetic elements metabolites.
with background correction from inelastic electron scattering are Wagners test: 100 mL of distilled water is diluted with 1.2 g of po­
necessary for the spectrum interpretation. The PHI 5000 Versaprobe III tassium iodide and 2 g of iodine that have been dissolved in 5 mL of
spectrophotometer, equipped with an Al Kα monochromator that oper­ distilled water. After adding 1 mL of Ceropegia fimbriifera flower extract,
ates at 350 W and produces an X-ray beam smaller than 10 μm, was used the completed concoction was set aside. The presence of alkaloids is
to perform the XPS. The test chamber pressure was maintained below 1 indicated by the emergence of reddish-brown colour.
× 10− 9 Torr during the acquisition process in order to provide high General test: Two drops of 1 % lead acetate were added to 1 ml of
resolution images and count the structural features of the Ceropegia fimbriifera flower extract, and the resultant white precipitate
nanocomposites. indicated the presence of tannins in the extract.
Salkowski reaction test: 1 ml of Ceropegia fimbriifera flower extract
3.11. TGA-DTA analysis was treated with chloroform and added with sulphuric acid slowly at the
walls of the test tube. The appearance of red colour indicates the pres­
Thermogravimetric analysis is a popular method for studying the ence of steroids in the extract.
kinetics of thermal deterioration in a variety of settings, such as dynamic Shinodas test: 1 ml of Ceropegia fimbriifera flower extract was dis­
and isothermal environments with nitrogen and oxygen atmospheres solved in 95 % ethanol and added with magnesium metal and 4 drops of
present. CuONPs underwent DTA and TGA analysis using PerkinElmer’s concentrated hydrochloric acid. The appearance of red colour indicates
DTA7 and TGA7, with a temperature range of 25–800 ◦ C and a rate of the presence of flavonoids.
10 ◦ C/min in an environment of nitrogen and oxygen (60 ml/min). The NaOH test: 1 ml of Ceropegia fimbriifera flower extract was added to
temperature range specified above is used to hold a crucible containing 1 ml of water and 2 ml of sodium hydroxide solution and kept aside. The
10 mg of the sample under investigation. The loss that happened during appearance of yellow colour indicates the presence of glycosides.
heating is automatically measured and plotted against temperature. Hydroxy anthroquinone test: The Ceropegia fimbriifera flowersa re
dried and boiled with dilute sulphuric acid. The filtrate is cooled and
extracted with chloroform and added with dilute ammonia and kept
aside. The appearance of pinkish red colour indicates to anthraquinone

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V. Viswanath et al. Journal of the Indian Chemical Society 102 (2025) 101772

glycosides. and 45 ± 5 % RH for the duration of 12 h of day and night cycles. The
Fehling’s test: Equal volumes of Ceropegia fimbriifera flower extract institutional ethics committee and the CPCSEA guidelines (reference
and the Fehling’s solution are mixed with each other kept aside. The number IAEC No. 439/PO/01/a/CPCSEA) imposed stringent guidelines
appearance of red precipitate indicates the presence of reducing sugars. on the study on animals. The rats were handled carefully and split up
Molisch’s test: 1 ml of Ceropegia fimbriifera flower extract was taken into six groups, each consisting of six rodents. The animals were divided
in a test tube and 2–3 drops of Molisch’s reagent was added. The into two groups: group II contained 0.5 % v/v of Ceropegia fimbriifera,
resultant was added with concentrated sulphuric acid dropwise at the group III contained bare CuO NPs, and group IV contained Ceropegia
side walls of the test tube. The formation of purple layer indicates the fimbriifera loaded CuO NPs. Group I served as the reference group in this
presence of carbohydrates. arrangement. The rats’ dorsal hair was removed and they were entirely
shaved before the trials began, leaving an exposed area that was six
3.13. In-vivo: antibacterial studies square centimeters. Following a shave, the region was cleaned with 0.9
% NaCl and alcohol to prevent infection. To induce sleepiness, rats
The well diffusion assay technique was employed to examine the received intraperitoneal injections of Xylazine hydrochloride (50 mg/
antibacterial properties of the synthesized copper oxide nanoparticles. kg) and Ketamine hydrochloride (10 mg/kg). A 500 mm2 section of each
The Institute of Microbial Technology in Chandigarh, India’s Microbial rat’s injury was excised using a surgical knife, and it was then placed in a
Type Culture Collection and Gene Bank (MTCC) provided the gram- separate housing for post-wound protection. After applying 1 g of
positive bacteria S. aureus (MTCC96), B. cereus (MTCC430), E. coli medication to the wound twice daily for 14 days, the region around the
(MTCC6133), and 1-g negative bacteria, K. Oxytoca (MTCC2275). Yeast wound was observed for changes and any indications of skin irritation
extract (2.0 g m), peptone (5.0 g m), meat extract (1.0 g m), agar (15.0 g [3–5]. The following equation (1) was used to estimate the circumfer­
m), and sodium chloride (2.0 g m) of distilled water were the ingredients ence of the wound closure:
used to cultivate the subcultures of bacteria to create fresh bacterial
Ad
cultures. Using a sterile cork-borer, the Muller Hilton agar plates were % wound contraction = 1- x 100 (1)
A0
sliced to a diameter of 5 mm after being streaked with a varied strain of
bacteria. The agar plates were completely dried, and then each well was In the above equation, Ad stands for area of wound closure on successive
filled with 15 μl, 20 μl, and 25 μl of the synthesized nanoparticles. These days and A0 stands for area of wound at the initial day.
were then incubated at 37 ◦ C for a period of 24 h. The resultant showed a Fourteen days later, skin tissue samples were submitted to histo­
zone of inhibition, which was found to surround the wells; this indicated logical investigations to determine the extent of wound healing. After
that the experiment was successful. Moreover, the minimum inhibitory being preserved in a 10 % Formalin solution for a full day, the specimens
concentration was determined by means of the broth micro dilution were stained with eosin and hematoxylin for pathological examination.
technique, in which the synthesized copper oxide nanoparticles were The histo-photographs were made using an OLYMOUS BX 51 micro­
cultured in Muller Hilton agar media at 37 ◦ C, and their density was scope that was 10X magnified.
adjusted to 108 CFU/ml. Furthermore, a 96-well plate was filled with 90
μl of Muller Hilton broth, 10 μl of bacterial inoculum, and 10 μl of CuO
3.16. Photocatalytic studies
NPs in various concentrations. After that, the plate was incubated for a
full day at 37 ◦ C. After the incubation period, the samples’ microbial
The study involved an examination of the ability of CuONPs, derived
growth was visually assessed, and the sample with no growth was found
from Ceropegia Firmbriifera floral extract, to catalyze the degradation of
to have the lowest inhibitory concentration.
Remazol brilliant blue and malachite green dyes under UV light. For the
preparation of the initial solution, 10 mg of each dye was combined with
3.14. Cytotoxicity studies
1 L of distilled water. Subsequently, 20 mg of CuONPs were mixed with
100 ml of each dye solution. The experimental group was composed of
The ability of the active ingredients to specifically target and produce
the solution with nanoparticles, while the control group was devoid of
harm in healthy cells is assessed using cell cytotoxicity. Human cancer
nanoparticles. The suspension containing nanoparticles was then sub­
cell line research, such as MCF 7, Caco-2, HT-29, and A549, acquired
jected to UV light for the purposes of photocatalytic degradation. Sam­
from the National Center for Cell Science, Pune, India, are used for the
ples were collected at specific time points (0, 30, 60, 90, and 120 min),
cytotoxicity analyses. Under 37 ± 0.5 ◦ C and 5 % CO2, the cells were
where 5 ml of the mixture was extracted, centrifuged, and then sub­
cultured in Dulbecco’s Modified Eagle’s Medium, which contained 10 %
jected to UV visible spectroscopy.
v/v fetal bovine serum, 100 μg/ml streptomycin, and 100 μg/ml of
penicillin. Cell density was optimized at 2 × 104 cells/cm2 and the cells
4. Discussion
were cultivated in 96-well plates that were treated for 24 h at 37 ±
0.5 ◦ C and 5 % CO2 with CuO NPs (5–500 μg/cm2). The cells were then
4.1. UV–visible spectroscopy
suspended in culture media (250μl/well) to achieve a concentration
range of 10–100 μg/ml. Subsequently, each well was filled with 25 μl of
A UV–visible spectrum of copper oxide nanoparticles synthesized
Dulbecco’s Modified Eagle’s Medium containing MTT (5 % mg ml− 1)
from Ceropegia Firmbriifera extract supernatant is illustrated in Fig. 1A.
and incubated for 5 h at 37 ± 0.5 ◦ C with 5 % CO2. 150 μl of DMSO was
The surface plasmon resonance of the copper oxide semiconductor
used in place of the medium, and its optical density was calculated to be
excitation is responsible for the absorption band that the copper oxide
302 nm. The process was run in triplicate, and the statistical values are
nanoparticles at 302 nm showed to be rather broad. The creation of pure
reported as mean ± SD. Non-regression analysis was used to determine
copper oxide nanoparticles was verified by the UV absorption spectra,
the procedure’s IC50 values.
which also showed the absence of impurity peaks. Using statistical
equation (2), the Tauc plot was created to ascertain the band gap energy
3.15. Wound healing activity
of copper oxide nanoparticles.
( )
The capacity of copper oxide nanoparticles to promote wound αhυ(1/n) = A hυ − Eg (2)
healing was evaluated using the excision wound healing model. The
research employed 300–400-g Wistar albino rats, which were kept in Here h, α, and υ stand for planks constant, coefficient of absorption, and
propylene cages with sterile husk bedding and access to food and water. frequency of light. Eg represents band gap energy; "n" stands for tran­
The ambient temperature and humidity were maintained at 25◦ ±2 ◦ C sition; ½ represents indirect transition, and 2 represents direct

7
V. Viswanath et al. Journal of the Indian Chemical Society 102 (2025) 101772

Fig. 1. (A)UV–Visible spectroscopy of Ceropegia Firmbriifera extract and copper oxide nanoparticles and the insert for Tauc Plot of Copper oxide nanoparticles and
(B) FTIR spectra of Copper oxide nanoparticles and Ceropegia Firmbriifera extract.

transition. The copper oxide nanoparticles’ band gap energy was found Bg are present in copper oxide. With characteristic peaks at 296 cm− 1
to be 2.4 eV, which grew gradually with the ordinate. This indicates that (Ag), 617 cm− 1 (Ag), and 342 cm− 1 (Bg) (Fig. 2A), three asymmetric
the nanoparticles’ size has decreased, ensuring that the formulation has modes (2Ag + Bg) are active, while the remaining modes are in radio­
improved pharmacokinetics. The absence of distinctive absorption frequency (IR). The shift of the oxygen atom is seen in the Ag and Bg
peaks in the Ceropegia Firmbriifera extract’s UV–visible spectrum sug­ modes, with the Bg mode shift being perpendicular to the b-axis. With
gested that the active ingredients were converting copper ions into the induced dipole moment parallel to Au modes and perpendicular to Bu
copper oxide nanoparticles [35]. modes, respectively, Cu and O exhibit high levels of activity in the
infrared mode [37].
4.2. Fourier-transform infrared spectroscopy
4.4. Photoluminescence spectroscopy
Utilizing extract from Ceropegia Firmbriifera, we assessed the func­
tional groups of copper oxide nanoparticles by means of FTIR spec­
Photoluminescence (PL) spectroscopy is a pivotal technique for
troscopy. For copper oxide nanoparticles, the spectrum with an
investigating electron-hole recombination dynamics in copper oxide
inscribed range of 500 cm− 1 to 4000 cm− 1 produced distinct peaks at
(CuO) nanoparticles, offering critical insights into their electronic
588 cm− 1, 1052 cm− 1, 1376 cm− 1, 1532 cm− 1, 1638 cm− 1, 2340 cm− 1,
structure, defect states, and photoactive performance. In this study, PL
2912 cm− 1, and 3396 cm− 1 (Fig. 1B). A distinct absorption peak at 1052
spectra were obtained using a LabRAM HR 800 spectrophotometer at an
cm− 1 for aliphatic amines and 588 cm-1 for the M − O stretch of copper
excitation wavelength of 325 nm, enabling precise evaluation of photon-
oxide were identified. The amides and amines’ NH- and hydrogen-
induced transitions and carrier dynamics. CuO nanoparticles, charac­
bonded –OH stretch is represented by the infrared broad peaks at
terized by p-type semiconducting behavior, exhibited a distinct emission
2340 cm− 1, 2912 cm− 1, and 3396 cm− 1. An IR band at 1532 cm− 1 for
peak at 434 nm, along with a broad luminescence profile. This spectral
C–H vibration and C–H bending of aliphatic methyl groups were
broadening stems from radiative and non-radiative recombination
discovered at 1376 cm− 1, while the NH-bending of the amine group was
pathways, influenced by surface defects, oxygen vacancies, interstitial
represented by the IR band at 1638 cm− 1. The distinctive peaks at 3323
Cu2+ ions, and band tail states, which introduce localized trap levels
cm− 1, 1600 cm− 1, and 1719 cm− 1 in the FTIR spectra of the Ceropegia
within the bandgap. The trap states influence the recombination dy­
Firmbriifera extract indicate the existence of O–H, C– –O, and C–H
namics based on their energy levels and spatial distribution, while the
stretching. These experimental findings list the electrostatic interactions
observed PL quenching reflects a notable suppression of radiative
that exist between the copper ions that are produced by copper oxide
recombination, an indicator of enhanced charge carrier separation ef­
nanoparticles and the ketonic (C– –O), alcoholic (-OH), and amino
ficiency. This phenomenon is chiefly ascribed to the presence of a
groups in the Ceropegia Firmbriifera extract. After comparing the previ­
multilayered CuO shell that functions as an electron sink, effectively
ously described characteristic peaks with the investigation findings of
capturing photogenerated electrons and prolonging their recombination
Ibrahim et al. [36] it was determined that copper oxide nanoparticles
with holes. The abundant defects within the shell enable effective
were present at three significant distinctive peaks: 540 cm− 1, 596 cm− 1,
trapping of charge carriers at various energy depths, thereby prolonging
and 1038 cm− 1.
their lifetimes, a key contributor to the improved photocatalytic per­
formance and antimicrobial efficacy of the nanoparticles. Upon photo­
4.3. Raman spectroscopy excitation, electrons are excited from the valence band to the conduction
band, leaving behind holes. The Trapped electrons in the oxide shell
Copper oxide features optical photon modes focused on 2Ag and Bg, participate in secondary redox reactions, contributing to the formation
with a monolithic structural symmetry of C2 H6. Copper oxide crystal’s of reactive oxygen species such as hydroxyl and superoxide anion rad­
well-resolved characteristic bands at 296 cm− 1, 617 cm− 1, and 342 cm− 1 icals which are instrumental in microbial inhibition and organic
are associated with the Ag, Ag (1), and Bg modes. A statistical exami­ pollutant degradation. Thus, PL spectroscopy not only elucidates the
nation of the vibrational properties of copper oxide indicates the pres­ optical and structural integrity of CuO nanoparticles but also reflects
ence of four atoms, resulting in twelve phonon branching units. The their charge separation efficiency, making it an essential tool for eval­
quantum confinement effect is responsible for the blue shift and Raman uating their suitability in biomedical applications [38]. Due to the
active peak broadening. Three (2Au + Bu) acoustic modes and nine zone- artifact and band edge emission, the copper oxide nanoparticles dis­
centered optical phonon modes with symmetries of 4Au + 5Bu + 2Ag + played a characteristic emission peak at 434 nm (Fig. 2B) and a broad

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V. Viswanath et al. Journal of the Indian Chemical Society 102 (2025) 101772

Fig. 2. (A) Raman spectra (B) photoluminescence spectra and (C) Zeta potential of Ceropegia Firmbriifera mediated CuO nanoparticles.

range of luminescence. found to be 49.23 nm.

4.5. Zeta potential 4.7. BET analysis

A considerable degree of repulsion between identical and oppositely The BET surface area of the samples was investigated using the ni­
charged particles was reflected in the zeta potential of Ceropegia Firm­ trogen adsorption/desorption isotherms displayed in Fig. 3B. The cor­
briifera assisted copper oxide nanoparticle dispersion and the magnitude responding isotherms revealed that the pore volume and surface area
of this repulsion was found to be − 16.2 mV (Fig. 2C). According to the were, respectively, 0.062 cm− 3g− 1 and 12.45 m2g. The results of the
investigation, nanoparticles were unstable at room temperature. On the experiment demonstrated that the CuO NPs that were produced had a
other hand, when the zeta potential was low, the formulation caused the low surface area and resembled type IV isotherms (IUPAC). The iso­
nanoparticles to become attracted to one another and flocculated. In therms also showed that the pore size distribution of the CuO NPs varied
contrast, the High zeta potential prevented the nanoparticles from from 10 to 50 nm, indicating that the nanoparticles were mesoporous in
coagulating. More stable suspensions were produced by nanoparticles nature. The experiment results demonstrated that the increased surface
closer to the tolerable threshold of 30 mV, which separated stable from area and porosity of nanoparticles had meritoriously boosted the pace of
unstable suspensions. catalytic processes, which in turn enhanced the active sites and enabled
charge transfer while suppressing charge carriers. The data from the BET
4.6. X-ray diffraction studies isotherms would indicate that CuO NPs would be more obvious in
eliciting the catalytic performance [39,40].
The structural properties and crystalline phase information of Cer­
opegia Firmbriifera-assisted copper oxide nanoparticles were listed using 4.8. SEM and EDX characterization
XRD analyses. The study utilized 100 mg of sample and recommended
applying Bragg’s law, which states that nλ = 2dsinθ, for determining the The SEM analysis utilized 100 mg of the sample and captured images
interplanar spacing (d).The XRD absorption peaks of the copper oxide at a high resolution of 100 nm in Fig. 4A(a), showed clusters of CuO NPs
nanoparticles at 2θ = 32.35◦ , 35.92◦ , 38.86◦ , 49.54◦ , 54.63◦ , 58.28◦ , due to the adhesive properties of the plant extract. The average particle
62.34◦ , 65.38◦ , 68.64◦ were assigned the following values: (110), (002), size was found to be 40 nm which indicated a well-established synthesis.
(111), (202), (020), (202), (113), (311), and (113) (Fig. 3A). The peaks Copper dioxide or oxide was formed, as evidenced by the oxygen-copper
matched the copper oxide nanoparticle monolithic phase JCPDS card interaction and the Energy Dispersive Spectroscopy (EDS) Fig. 4A(b)
number 01-080-1268 [38,39]. The average particle size of copper oxide that showed the purity of CuO NPs. According to the EDS spectra, the
nanoparticles was calculated using Debye-Scherrer’s equation and weight compositions of copper and oxygen were found to be 14.58 %

Fig. 3. (A) XRD spectra and (B) Nitrogen adsorption-desorption isotherms of CuO NPS.

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V. Viswanath et al. Journal of the Indian Chemical Society 102 (2025) 101772

Fig. 4. (A) Scanning Electron Microscopy (b) Energy Dispersive Spectroscopy of CuO NPs. (B) (a) TEM image (b) Particle Size analysis (c) Selected Area Electron
Diffraction image of CuO NPs.

and 85.42 %, respectively, whereas their expected atomic compositions chemical bonding, oxidation status, elemental composition and surface
were 9.25 % and 93.16 %. The spectral aberrations indicated the chemistry of nanomaterials. It is commonly used to clarify the chemical
interference of traceable ions with extract during synthesis. and peripheral properties of NPs. The high-resolution spectrum was
constructed using Gaussian fitting and Shirley background deduction,
4.9. TEM and particle size distribution and the survey scan spectra (Fig. 5a) were generated from 100 mg of
sample. This revealed a distinct peak for Cu 2p at 970eV, along with two
Transmission Electron Microscopy (TEM) was conducted using 100 well-defined peaks for Cu 2p1/2 and Cu 2p3/2 states exhibiting different
mg of CuO NPs, resulting in the production of homogeneous clusters binding energies at 965 and 945eV. Additionally, there were two sat­
with an average size of 100 nm in the images. The image displayed a thin ellite peaks for Cu 2p1/2 and Cu 2p3/2 states at 956eV and 934eV,
layer of organic material acting as a capping agent on particle clusters. A respectively (Fig. 5b). The NPs produced a distinctive characteristic
strong aggregation was seen in the image, along with a few segregated peak a characteristic peak at 285 eV corresponding to the C–C bond at
particles that displayed their morphological traits. The TEM investiga­ the C 1s region (Fig. 5c) and the potent O 1s area with an improved
tion revealed that the produced nanoparticles were spherical and hex­ degree of resolution at 528 eV (Fig. 5d). The presence of a spin-orbit
agonal in shape, with significant variations in diameter as a result of doublet at the corresponding binding energies suggested that the NPs
cluster formation. More precisely, as seen in Fig. 4B(a), the TEM ex­ were in a +2 oxidation state [41,42]. The emergence of a distinct
amination supported the particle morphology as spherical, cubic, and spin-orbit doublet at specific binding energies in the XPS spectrum
irregularly shaped. Nonetheless, the XRD analyses validated the crys­ confirmed that the copper oxide nanoparticles existed predominantly in
talline character of the nanoparticles and supported the aforementioned the +2 oxidation state, characteristic of CuO. This spectral signature also
treatments. A histogram, which is a bar graph that shows particle size indicated the development of multilayered CuO films encasing the
versus frequency, is a common way to display the particle size distri­ nanoparticle cores, an architectural feature that plays a pivotal role in
bution. The histogram for particle size analysis is shown in Fig. 4B(b) on modulating both electronic behavior and photocatalytic performance.
a linear scale. From the peak position, the average diameter of the CuO These CuO-rich surface layers act as strategic electron traps, effectively
NPs was found to be 30.7 nm, with the particle size distribution ranging suppressing the rapid recombination of photogenerated electron-hole
from 15 to 45 nm. If sufficient numbers of particles are counted and their pairs. The mechanism lies the oxidation of terminal copper ions,
size interval is at least ten, a smooth size frequency curve created across which enhances the growth of a chemically active CuO outer shell. When
the histogram becomes legitimate. Since the bulk of the samples were exposed to light, this shell absorbs photons, resulting in the generation
plotted on the linear scale, the histogram showed the size distribution of electron-hole pairs. The electrons become localized within the oxide
skewed towards the larger terminal point of the scale. The findings film, while the holes remain mobile, leading to enhanced charge sepa­
(Fig. 4B(c)) obtained from subjecting the nanoparticles to Selected Area ration and a prolonged lifetime of charge carriers. This phenomenon is
Electron Diffraction (SAED) demonstrated the presence of tiny, sporadic crucial for driving more efficient photocatalytic reactions. Furthermore,
bright spots on the concentric ring structure, which validated the the CuO shell also imparts unique surface chemistry, enhancing terminal
polycrystalline structure of CuO NPs. This could explain why the bio- reactivity and molecular interaction which plays an influential role in
reduced aqueous region has a macroscopic output of nanoparticle electronic structure modulation, thereby governing charge transfer dy­
dispersion. At high magnification, the fringed lattice areas of CuO NPs namics and interfacial reactivity. These characteristics are not only vital
were clearly apparent despite their severe aggregation. for boosting photocatalytic activity but also contribute significantly to
the antimicrobial potency of the nanoparticles, enabling them to disrupt
4.10. X-ray photoelectron spectroscopy (XPS) microbial systems through sustained oxidative stress and surface in­
teractions. Altogether, the structural and electronic sophistication
The XPS is a potent analytical tool in furnishing the knowledge on imparted by the CuO layer makes these nanoparticles multifunctional

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V. Viswanath et al. Journal of the Indian Chemical Society 102 (2025) 101772

Fig. 5. (a) Broad scan X-ray Photoelectron Spectrum (XPS) of CuO NPs; (b) High-resolution survey scans spectrum of Cu 2p NPs; (c) XPS of the spectrum of C 1s; (d)
XPS of the spectrum of O 1s.

and highly efficient in both environmental and biomedical applications.

4.11. TGA-DTA analysis

Utilizing thermal gravimetric analysis (TGA) and differential thermal

analysis (DTA), the thermal characteristics of the nanoparticles were
examined. The study utilized a 100 mg sample for conducting the
experiment, and the degradation commenced at distinct temperature
intervals: 25 to 415 ◦ C, 415 to 520 ◦ C, and 520 to 630 ◦ C, as demon­
strated by the TGA plot. This revealed a three-stage decomposition
process for the formation of copper oxide. The CuONP calcinations were
conducted within a temperature range of 25–800 ◦ C, with a nitrogen
temperature thermal range of 10 ◦ C/min. A main weight loss (̴ 10 %)
below 100 ◦ C is shown by 2-min endothermic peaks at 96 ◦ C on the DTA
curve. This weight loss could be caused by coordinated bond cleavage,
dehydration, or the system loss of organic solvent. At a temperature
between 415 and 520 ◦ C, the second fragment of decomposition in the
Fig. 6. DTA/TGA analysis of CuONPs.
TGA curve was observed. This was ascribed to the combustion and py­
rolysis of organic molecules, which were expelled from the system as
Table 1. The phytochemical screening of flower extract revealed that the
COx and organic gases (X denotes 1, 2). The DTA curve shows a large
phytochemical constituents acted as a capping stabilizer in the forma­
exothermic peak at 677 ◦ C (Fig. 6), indicating that the nanoparticles
tion of CuO NPs. The capping substances are used for designing the
were calcined at 700 ◦ C for 2 h and that the remaining functional
morphological characteristics of the NPs to impart stability and impede
moieties were oxidized. A broad endothermic peak at 390 ◦ C indicates
their agglomeration. The phytochemical investigations revealed the
the loss of chemisorbed functional groups (̴ 15 %) But there was no
presence of crucial components such as alkaloids, tannins, steroids,
further loss after 677 ◦ C, which resulted in the creation of stable
flavonoids, glycosides, anthraquinone glycosides, reducing sugars, and
nanoparticles.
carbohydrates that resulted in the formation of NPs. Furthermore, GC-
MS for Ceropegia fimbriifera flower ethanol extract was studied to
4.12. Phytochemical screening confirm phytochemicals used for bio-capping agents, and stabilizing
agents which are able for the bioreduction process to form CuONPs
The synthesized CuO NPs were subjected to various tests to investi­ (Fig. 7, Table 2).
gate the presence of various phytochemical constituents enclosed in

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V. Viswanath et al. Journal of the Indian Chemical Society 102 (2025) 101772

Table 1 resistant strains. Control samples of S. aureus, B. cereus, and E. coli

Phytochemical analysis of Ceropegia fimbriifera flower extract. exhibited a zone of inhibition of 19 ± 0.2 mm, while K. oxytoca showed
Test Name Test Method Flower Extract 20 ± 0.2 mm (Table 3, Fig. 8). Notably, E. coli displayed the highest
antibacterial activity among the gram-positive bacteria, indicating its
Alkaloid Wagner’s Test +
Tannin General Test + effectiveness at the control concentrations. In experiments conducted
Steroid Salkowski reaction Test + with E. Coli, S. aureus, B.cereus, and K. oxytoca bacteria, different con­
Triterpenes Salkowski reaction Test – centrations of CuO nanoparticles showed varied zones of inhibition. For
Flavonoids Shinoda Test + E. Coli, the zones of inhibition at 5.0, 7.0, and 10 mg/mol were 12 ± 0.3,
Saponin Shake or Foam Test
14 ± 0.1, and 18 ± 0.1 mm, respectively. Similarly, for S. aureus, the
–
Resin General Test –
Glycoside NaOH Test + zones of inhibition were 11 ± 0.3, 13 ± 0.2, and 15 ± 0.1 mm at the
Cardiac Glycoside Keller-Killiani Test – same nanoparticle concentrations. For B.cereus, the inhibition zones
Anthraquinone glycoside Hydroxy anthraquinone Test + were 12 ± 0.1, 13 ± 0.3, and 16 ± 0.2 mm. Notably, K. oxytoca, a gram-
Phenol Ferric chloride Test –
negative bacterium, demonstrated a concentration-dependent increase
Reducing sugar Fehling’s Test +
Carbohydrate Molisch’s test + in antibacterial activity, with zones of inhibition of 13 ± 0.4, 15 ± 0.1,
Protein Biuret Test – and 18 ± 0.1 mm, respectively (Table 3, Fig. 9). Additionally, it was
Fats and Fixed Oils General Test – observed that the smaller-sized nanoparticles exhibited higher antibac­
terial activity compared to bigger ones, suggesting a size-dependent
antimicrobial activity of the CuO NPs. The nanoparticles were likely
able to interact with the cell membrane more effectively due to the
increased surface-to-volume ratio, leading to degradation and ulti­
mately cell death. Furthermore, with p-values of less than 0.001, the
results were found to be statistically significant [2].

4.14. Cytotoxicity studies

According to cytotoxicity experiments on several cell lines, CuSO4

and CuONPs produced increased toxicity on the HT-29 cell line in
comparison to the other cell line studies. In all four cell lines, the IC50
values for CuONPs showed a substantial reduction (p < 0.005) in
comparison to CuSO4, suggesting that CuONP effectiveness induced
copper cytotoxicity. The physical and chemical properties of nano­
particles, including their size, shape, and surface area, influence their
toxicological consequences. Following a 72-h treatment with serially
diluted concentrations of CuONPs (10 μg/ml), A549 cells (Fig. 10)
showed reduced cell viability due to the nano composites. Fahmy et al.

Table 3
Fig. 7. GC-MS chromatograms of Ceropoegia fimbriifera flower extracted with
Antibacterial study of Ceropegia fimbriifera flower mediated CuO NPs on Gram-
ethanol solvent.
positive and Gram-negative bacteria.
Concentration (mg/mol) Gram-positive Gram-negative
4.13. In vitro: antibacterial studies
S. aureus B. cereus E. coli K. oxytoca
The study investigated the antibacterial activity of CuO NPs against a Control 19 ± 0.2 19 ± 0.2 19 ± 0.2 20 ± 0.2
variety of gram-positive bacteria including S. aureus, B. cereus, and E. 5.0 11 ± 0.3 12 ± 0.1 12 ± 0.3 13 ± 0.4
7.5 13 ± 0.2 13 ± 0.3 14 ± 0.1 15 ± 0.1
coli, as well as gram-negative bacteria such as K. oxytoca using the well
10 15 ± 0.1 16 ± 0.2 18 ± 0.1 18 ± 0.1
diffusion assay technique. The results revealed significant zones of in­ P value <0.001 <0.001 <0.001 <0.001
hibition for both types of bacteria, with smaller zones observed for the

Table 2
Phytochemical compounds of Ceropoegia fimbriifera flower extracted with ethanol solvent.
Tables Identified Name Tables Molecular formula Tables M.Wt Tables Rt(min) Tables Peak Area (%)

Gallic acid C6H2(OH)3COOH 170.12 9.43 0.42

Vanillin C8H8O3 152.15 12.01 0.56
Dodecanal C12H24O 184 14.06 0.72
Diethyl Phthalate C12H14O4 222 17.34 0.80
Myristoleic acid C14H26O2 226 22.64 1.02
n-Hexadecanoic acid C16H32O2 256 24.47 1.21
Oxiraneundecanoic acid C11H20O3 200.28 26.68 2.27
Palmitic acid C16H32O2 256.43 28.43 35.5
9,12- Octadecadienoic acid C18H32O2 280.45 30.02 4.31
Oleic acid C18H34O2 282.47 31.75 5.12
N-Normethadol C12H17NO 191.27 36.21 2.32
Oleic anhydride C36H66O3 550.91 38.02 2.28
Friedelan-3-one С30Н50O 426.73 40.42 1.83
Strychane 1-acetyl- 20-hydroxy-16- methylene C23H28N2O3 380.48 44.48 7.04
Trilinolein C57H98O6 879.37 45.23 4.51
Lupeol С30Н50O 426.73 45.67 3.14

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V. Viswanath et al. Journal of the Indian Chemical Society 102 (2025) 101772

Fig. 8. Studies on the zone of inhibition for control samples involving different types of gram-positive and gram-negative bacteria.

[43]research on the cytotoxic and genotoxic effects of CuONPs on cancer reducing agent. Photo degradation experiments were conducted using
lung cell lines showed comparable outcomes. However, the results of malachite green dye at different pH levels, revealing a peak degradation
Cronholm experiments [35,44] and those of Midander and colleagues of 94.15 % observed at pH 6.4. In the same way, CuONPs were syn­
[35] showed that the CuONPs were more harmful than free copper thesized by Muthukumaran and colleagues [48] through the sono­
fragments because of the Trojan Horse Effect, which makes the CuONPs chemical approach, and their efficiency in degrading malachite green
capable of acting as ion carriers during cell transport. One possible dye was studied. The analysis found that the CuONPs displayed a band
explanation for the toxicity could be that CuONPs dissolve and release gap of 2.26eV and achieved a maximum degradation rate of 97 % when
copper ions, which oxidize cytochrome c, an essential component in the exposed to visible light. Prakalathan and others [37] synthesized Copper
electron transfer across the mitochondrial cell wall. A number of studies oxide-based nanomaterials containing extracts from Curcuma amada
were carried out to document the toxicity of CuONPs and copper ions in seeds and Strychnos potatorum leaves and analyzed them to determine
both malignant and normal cells. According to Shafagh and colleagues’ their structural and morphological properties. The degradation of Congo
research findings [45], CuONPs only showed cytotoxicity on K562 cells red dye was studied catalytically under UV light irradiation, and the
and not on the healthier cells. Chemotherapeutic molecules incompe­ results showed a degradation efficacy of 91 %. CuONPs containing citrus
tence to discriminate between cancerous and healthy cells is a major limon extract were created by Kumar and colleagues [49], and they
problem when it comes to chemotherapy. Nevertheless, their research found that these nanoparticles could degrade Remazol brilliant blue dye
results demonstrated CuONPs ability to eradicate cancer cells on their with a 90 % efficiency within 180 min (Fig. 10) when exposed to UV
own and highlighted their therapeutic importance. irradiation. Based on the results of the experiments and the existing
literature review, it can be concluded that the CuONPs exhibit strong
photolytic activity, making them suitable for a range of applications in
4.15. Photocatalytic degradation studies nanoscience.

Investigations were carried out to study the photo catalytic degra­

dation of CuONPs synthesized using green methods. The degradation 4.16. In vivo: wound healing activity
process of malachite green, and Congo red dye was studied under the
influence of UV radiation. The CuONPs, which were obtained using The wound healing mechanism is associated with the synchronized
Ceropegia Firmbriifera floral extract, demonstrated approximately 74.6 % phases of tissue remodeling, inflammation, proliferation, and hemosta­
degradation efficacy in malachite green dye, 50 % in Remazol brilliant sis. The body’s natural wound-healing mechanism takes over as soon as
blue (Fig. 11). In the absence of UV light, the dyes showed minimal there is an injury, encouraging the synthesis of collagen and the
degradation even after 1 h, with Congo red dye exhibiting the highest regeneration of epithelial tissue. The differences between the treatment
degradation of 5 % after 1 h. Tai and his team [46] created CuONPs and control groups were documented, and an examination of the rate of
using environmentally friendly methods and assessed their ability to wound healing was initiated immediately after the damage had
degrade malachite green dye under visible light at 1.64eV, showing an occurred. Metal nanoparticles have unique physiochemical properties
impressive effectiveness of 95.39 %. Murshed and colleagues [47] pro­ that make them useful for targeted drug delivery and effective against a
duced copper oxide nanoparticles using bee propolis as a capping and range of bacterial infections. These properties include their particle size

13
V. Viswanath et al. Journal of the Indian Chemical Society 102 (2025) 101772

Fig. 9. Studies were conducted on the zone of inhibition for different gram-positive and gram-negative bacteria using different concentrations, including (A) 5 mg/
mol, (B) 7.5 mg/mol, and (C) 10 mg/mol.

and application. According to CPCSEA guidelines suggested by the granulation, collagen loading, and fibrosis, which indicate increased
Institutional Animal Ethical Committee (IAEC), the wound-healing po­ wound contraction, were listed in the histopathological analyses for
tency of CuONPs was investigated on albino Wistar rats. The animal group IV. Groups II and III showed signs of epidermal healing, including
studies were conducted with ethical approval number minute inflammation and well-organized granulation, which suggested
BDUAS2024B0132 at the Department of Biotechnology, Government a light closure of the lesion (Fig. 11). On the other hand, group I showed
Arts and Science College, Bharathidasan University, Trichy, Tamil Nadu, signs of severe edema, poor vascularization, and excessive cell infiltra­
India. The investigative results revealed the absence of skin irritation tion, all of which suggested insufficient wound healing during the
and edema in the first two weeks of the study and a minute redness in the research (see Table 5).
test group which can be ignored. The fundamental hypothesis for the
redness was due to the bacteria on the skin’s surface that might have 5. Conclusion and future directions
penetrated the injured tissues and interfered with the healing process.
Consequently, the anti-inflammatory properties of the copper oxide The present study demonstrates the successful green synthesis of
nanoparticles were elicited by reducing cell infiltration and cytokine copper oxide nanoparticles (CuONPs) using Ceropegia fimbriifera flower
modulation, as well as initiating collagen synthesis for re-epithelization extract, offering a cost-effective, eco-friendly, and sustainable alterna­
and NFKB (Nuclear factor kappa B) signaling pathway for enhancing the tive to traditional chemical methods. The synthesized CuONPs exhibited
wound healing mechanism. Fig. 12 shows the optical images of the full a negative surface charge and optimal particle size, contributing to their
wound healing investigations that were taken at predefined intervals of stability, biocompatibility, and enhanced therapeutic efficacy. Notably,
time (1, 5, 10, and 14 days). The experimental results showed that the the nanoparticles showed significant potential in promoting rapid
rate of wound healing was 71.13 ± 0.32 % in group I (control) and wound healing with minimal toxicity. Phytochemical analysis
85–93 % in groups II to IV. Group III (87.46 ± 0.67 %) and Group IV confirmed the presence of various natural bioactive compounds in the
(92.89 ± 1.04 %) showed improved wound healing compared to Group plant extract, which are likely responsible for the enhanced therapeutic
II (85.54 ± 0.52 %) (Table 4). The combined effects of Ceropegia fim­ effects and stability of the nanoparticles. Furthermore, the CuONPs
briifera and copper oxide nanoparticles resulted in increased wound displayed strong antibacterial activity against both gram-positive and
contraction in group IV. Compared to bare nanoparticulate formulations gram-negative bacteria, including drug-resistant strains, highlighting
(group III) and standard formulations (group II), formulation IV showed their broad-spectrum antimicrobial potential. This study is innovative in
considerable wound contraction. The formulations demonstrated strong utilizing the underexplored Ceropegia fimbriifera in nanoparticle syn­
bioavailability, minimal toxicity, and rapid wound contraction, all of thesis, bridging the fields of nanotechnology and ethnobotany. This
which were indicative of the CuO-NPs’ potential for therapeutic use. study represents a compelling fusion of traditional plant-based knowl­
Several indicators of wound healing, including keratinization, edge and modern nanotechnology, unveiling a sustainable route to

14
V. Viswanath et al. Journal of the Indian Chemical Society 102 (2025) 101772

Fig. 10. Effect of CuO NPS on the viability of (a) A549 (b) MCF 7 (c) HT-29 and (d) CaCo-2 cell lines determined by MTT assay.

Fig. 11. Photocatalytic dye degradation studies of CuONPs.

therapeutic innovation. The green synthesis of copper oxide nano­ precision, stability, and therapeutic depth. Their engineered surface
particles (CuONPs) using Ceropegia fimbriifera extract was validated chemistry and biocompatibility make them ideal for integration into
through detailed spectral, morphological, and structural characteriza­ antimicrobial scaffolds, wound healing matrices, and targeted drug
tions, affirming the precision and efficiency of the process. These carriers. This green synthesis approach elevates functional material
nanoparticles displayed outstanding optical and catalytic properties, design through sustainable innovation. Altogether, the findings position
coupled with notable stability and purity. Furthermore, biological CuONPs as powerful, multifunctional candidates for future biomedical
evaluations revealed low cytotoxicity, enhanced bioavailability, and and environmental applications. “Future research should emphasize
accelerated wound healing and highlighted their effective cellular advanced surface functionalization through biopolymer conjugation or
interaction. The study unveils a new horizon in bioinspired nano­ ligand engineering to reinforce colloidal stability and facilitate
material development, where phytochemical-mediated CuONPs exhibit precision-targeted therapeutic delivery, while strategic dopant

15
V. Viswanath et al. Journal of the Indian Chemical Society 102 (2025) 101772

Fig. 12. (A) Photographic images of wounds treated with various nanoparticles and (B) Histopathological images of wounds undergoing treatment with Ceropegia
fimbriifera loaded CuO-NPs on the 14th day of the investigation.

Table 4
Wound healing activity of control and treatment groups of Ceropegia fimbriifera loaded CuO-NPs.
Treatment groups % Wound contraction

3rd Day 5th Day 10th Day 12h Day 15th Day

Control (I) 12.18 ± 0.16 23.34 ± 0.26 48.42 ± 0.46 61.24 ± 0.26 71.13 ± 0.32
Ceropegia fimbriifera (II) 23.48 ± 0.17 32.59 ± 0.32 62.37 ± 0.29 74.57 ± 0.28 85.54 ± 0.52
CuO-Nps (III) 26.56 ± 0.21 35.18 ± 0.31 65.18 ± 0.27 79.26 ± 0.19 87.46 ± 0.67
Ceropegia fimbriifera loaded CuO-NPs (IV) 29.18 ± 0.32 39.35 ± 0.23 69.31 ± 0.15 84.23 ± 0.23 92.89 ± 1.04

incorporation or nanocomposite fabrication could significantly amplify comprehensive evaluation of long-term physicochemical stability, in
antimicrobial efficacy and modulate the electronic band structure for vivo toxicodynamics, and pharmacokinetic behavior remains essential
enhanced biomedical performance. Mechanistic elucidation via inte­ for successful clinical translation. Optimizing green synthesis via
grative omics and molecular docking studies can inform the rational phytochemical profiling will ensure reproducibility and scalability of
design of nanostructures with enhanced bioactivity, while CuO NPs.”

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V. Viswanath et al. Journal of the Indian Chemical Society 102 (2025) 101772

Table 5
The current work is compared with the other relevant works.
Plant extract used Particle size Crystallinity Characterization techniques Zeta Biological actives Reference
potential

Saponin (from Bare CuO: 244.1 ± Confirmed (XRD) UV–Vis, FTIR, XRD, SEM Not Photocatalytic degradation of [50]
Sapindus 36.3 nm Reported methylene blue (86.32 % vs. 13.14
mukorossi) Green CuO: 147.1 ± % for bare CuO)
41.6 nm (aggregates of
<30 nm NPs)
Morinda citrifolia 26 nm Confirmed (XRD) DLS, Zeta Potential, UV–Vis, − 70 mV (at Antimicrobial activity (stronger [51]
(Noni-leaf tea by- FTIR, XRD, SEM, TEM basic pH) against E. coli and B. cereus);
product after Gompertz model for microbial
nanofiltration) growth kinetics
Mangifera indica 211 nm (DLS) Confirmed via XRD DLS, FTIR, UV–Vis, SEM, Not Antioxidant (DPPH assay, IC50 = [52]
kernel extract TEM, EDX, XRD reported 453.72 μg/ml), Antibacterial (E. coli
& S. aureus), Anticancer (MTT assay
on SH-SY5Y cells, IC50 = 24.14 μg/
ml)
Eucalyptus globulus 30–300 nm (SEM) Confirmed via FTIR UV–Vis, FTIR, SEM, EDS Not Bactericidal activity [53]
(aqueous extract) reported
Solanum melongena 47.97 nm Monoclinic CuO SEM-EDS, XRD, FTIR, TGA, Not Dye degradation: Rose Bengal (60 [54]
Seed Extract BET, UV–Vis DRS Reported min) - Antibacterial - Antioxidant
activity
Artemisia judaica Not explicitly Confirmed as GC-MS, XRD UV–Vis, FTIR, Not Antibacterial activity, Cytotoxicity [55]
mentioned polycrystalline by SEM, TEM, DLS mentioned studies, Photocatalytic studies and
XRD larvicidal studies
Tecoma stans Not explicitly Monoclinic phase XRD, UV–Visible, FTIR, SEM Not Photocatalytic activity, [56]
mentioned; described (confirmed by XRD) with EDS, HRTEM, XPS, BET mentioned Antibacterial activity, Cytotoxicity
as spheroidal (via and Langmuir surface area
HRTEM) analysis
Peganum harmala 38.62 nm (via SEM) Amorphous (via FTIR, UV–Vis, SEM, PXRD Not Antibacterial, Antidiabetic, [57]
PXRD) mentioned
Rosmarinus officinalis 18–26 nm (XRD) Monoclinic (XRD) UV–Vis, XRD, FTIR, SEM Not Antimicrobial, [58]
mentioned
Luffa acutangula (peel) 26 nm (XRD) Monoclinic (XRD) XRD, SEM, FTIR, DLS − 34 mV Antibacterial [59]
Carica papaya leaf Not mentioned Crystalline (XRD XRD, FTIR, SEM Not Antimicrobial activity [60]
extract confirmed) mentioned
Aegle marmelos leaf Not mentioned Crystalline UV–Vis spectroscopy, XRD, Not Antibacterial activity, plasmid DNA [61]
extract (confirmed by XRD SEM, EDS, In silico docking mentioned fragmentation, potential drug
(AutoDock Vina, PyMOL) candidacy based on Lipinski’s rule
Abelmoschus 20–25 nm Monoclinic crystalline TGA/DTA, UV–Vis, DRS, Not Anticancer activity, low cytotoxicity [62]
esculentus (Okra) structure FTIR, XRD, BET, TEM, reported to normal fibroblast cells
fruit extract FESEM/EDX, PSA
Spinacia oleracea leaf 134.8 nm Monoclinic CuO UV/Vis Spectroscopy, FT-IR Not Antibacterial, Antifunga, Larvicidal [63]
extract (confirmed by XRD at Spectroscopy, X-Ray Mentioned
various 2θ values) Diffraction (XRD), SEM
Analysis
Moringa oleifera leaf Nanometer range Monoclinic structure XRD, SEM with EDAX, FTIR, Not Antimicrobial [64]
extract (exact size not (confirmed by XRD) UV–Vis spectroscopy mentioned
specified)
Tragopogon collinus Average size: 50 nm Crystalline UV–vis spectroscopy, FTIR, − 33.91 mV cytotoxicity [65]
leaf extract Hydrodynamic radius: XRD, XPS, TEM, DLS
88.31 nm
PDI: 0.193
Eucalyptus globulus Mean particle size: 88 Crystalline; SEM, DLS, XRD − 16.9 mV Photocatalytic/adsorption activity [66]
leaf extract nm Monoclinic phase
Crystalline size: 85.80 (XRD confirmed)
Pimenta dioica leaf 20–50 nm Confirmed via XRD; UV–Visible spectroscopy, Not Antibacterial, Cytotoxicity/ [67]
extract crystalline structure FTIR, XRD, SEM-EDS, TEM, specified Anticancer, Antioxidant,
present TGA-DT Antidiabetic
Calotropis procera 20–80 nm (based on High crystalline XRD, FTIR, UV–Visible Not Antibacterial & Antifungal, [68]
aqueous extract SEM and TEM); nature confirmed by spectroscopy, SEM, TEM specified Antioxidant
40–100 nm (based on XRD
XRD
Azadirachta indica 15–16 nm (based on Monoclinic crystal XRD, FTIR, UV–Visible Not Anticancer, Comparative modeling [69]
(Neem) leaf extract crystallite size from structure confirmed spectroscopy specified
XRD) by XRD
Mussaenda frondosa – 2–10 nm (grain size Monoclinic crystalline XRD, UV–Vis spectroscopy, Not Antioxidant, Anticancer, [70]
leaf, stem, and leaf- from XRD) nature confirmed by SEM, TEM, EDS, FTIR, DLS, specified Photocatalytic
derived callus XRD BET analysis
Ceropegia fimbriifera 40 nm (From XRD, Cubic, crystalline XRD, UV–Vis, IR,Raman, − 16.2 Dye degradation, antibacterial, This work
floral extract TEM, SEM) nature confirmed by TEM, SEM, zeta potential, Wound healing, cytotoxicity
XRD BET, PL, TGA, HRMS.

17
V. Viswanath et al. Journal of the Indian Chemical Society 102 (2025) 101772

CRediT authorship contribution statement [16] K. Sivakumar, S. Eswaramurthy, K. Subramanian, S. Natarajan, Structure of
cerpegin, a new alkaloid, Acta Crystallogr. C 46 (1990) 839–841, https://doi.org/
10.1107/S0108270189009595.
Valikala Viswanath: Writing – original draft, Methodology, Inves­ [17] H. Sauquet, M. von Balthazar, S. Magallón, J.A. Doyle, P.K. Endress, E.J. Bailes,
tigation, Formal analysis. Jayaprakash Meena: Writing – original draft, E. Barroso de Morais, K. Bull-Hereñu, L. Carrive, M. Chartier, G. Chomicki,
Validation, Resources, Methodology, Investigation, Formal analysis. N. M. Coiro, R. Cornette, J.H.L. El Ottra, C. Epicoco, C.S.P. Foster, F. Jabbour,
A. Haevermans, T. Haevermans, R. Hernández, S.A. Little, S. Löfstrand, J.A. Luna,
Kumaraguru: Writing – review & editing, Visualization, Supervision. N. J. Massoni, S. Nadot, S. Pamperl, C. Prieu, E. Reyes, P. dos Santos, K.
Sami veerappa: Writing – original draft, Visualization, Supervision. M. Schoonderwoerd, S. Sontag, A. Soulebeau, Y. Staedler, G.F. Tschan, A. Wing-Sze
Paik-kyun Shin: Writing – review & editing, Visualization, Supervision. Leung, J. Schönenberger, The ancestral flower of angiosperms and its early
diversification, Nat. Commun. 8 (2017) 16047, https://doi.org/10.1038/
Jiro Tatsugi: Writing – review & editing, Validation, Supervision. ncomms16047.
Kannappan Santhakumar: Writing – review & editing, Visualization, [18] R. Kumar, K. Kumar, S. Sharma, N. Thakur, N. Thakur, Multifunctional properties
Validation, Supervision, Funding acquisition. of microwave assisted CuO/Cu2O-ZnO mixed metal oxide nanocomposites,
J. Mater. Sci. Mater. Electron. 34 (2023) 1255, https://doi.org/10.1007/s10854-
023-10693-3.
Declaration of competing interest [19] S. Sharma, K, Kumar, Aloe-vera leaf extract as a green agent for the synthesis of
CuO nanoparticles inactivating bacterial pathogens and dye, J. Dispersion Sci.
Technol. 42 (2021) 1950–1962, https://doi.org/10.1080/
The authors declare that they have no known competing financial 01932691.2020.1791719.
interests or personal relationships that could have appeared to influence [20] S. Sharma, K. Kumar, N. Thakur, S. Chauhan, M.S. Chauhan, Eco-friendly Ocimum
tenuiflorum green route synthesis of CuO nanoparticles: characterizations on
the work reported in this paper. photocatalytic and antibacterial activities, J. Environ. Chem. Eng. 9 (2021)
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