Received: 21 August 2024 Revised: 9 January 2025 Accepted: 24 February 2025

DOI: 10.1002/ep.14583

ORIGINAL RESEARCH

Evaluation of the preservative efficacy of green-synthesized

ZnO Nanoparticles using Cucumis sativus in cream formulations

Sanaya Yousaf 1 | Anam Ashraf 1 | Shaista Ali 1 | Akasha Rafiq 1 |

Abid Mahmood 2 | Farrukh Bashir 3 | Muhammad Akhyar Farrukh 4 |
Fizza Naseem 5 | Durr-e-Shahwar 6

1
Department of Chemistry, Government
College University Lahore, Lahore, Pakistan Abstract
2
Department of Environmental Sciences, Due to concerns about the safety of preservatives in cosmetic formulations, there
Government College University, Faisalabad,
Faisalabad, Pakistan is an urgent need to explore eco-friendly alternatives that do not pose a risk to con-
3
Department of Chemistry, Sardar Bahadur sumers. Zinc oxide nanoparticles (ZnO NPs) are promising preservatives for phar-
Khan Women's University, Quetta, Pakistan
maceutical and cosmetic products. In this study, ZnO NPs were synthesized using
4
Department of Basic and Applied Chemistry,
Cucumis sativus (cucumber) extract as a stabilizing agent and zinc acetate dihydrate
University of Central Punjab, Lahore, Pakistan
5
Department of Chemistry, Superior as a precursor. Polyphenols, terpenoids, and flavonoids in cucumber extract act as
University, Lahore, Pakistan capping agents and facilitate the reduction of metal ions to form ZnO NPs. Synthe-
6
Kauser Abdulla Malik School of Life Sciences,
sized ZnO NPs were characterized using various techniques, including powder
Forman Christian College (A Chartered
University), Lahore, Pakistan x-ray diffraction, x-ray fluorescence, scanning electron microscopy, Fourier trans-
form infrared spectroscopy, photoluminescence and spectroscopy UV–Vis. The
Correspondence
Shaista Ali effectiveness of ZnO NPs as a preservative in cream formulation was evaluated
Email: [email protected];
along with their antioxidant and antibacterial properties against Gram-positive and
[email protected]
Gram-negative bacteria. This study introduces a new green synthesis method to
produce ZnO NPs using C. sativus extract. This eco-friendly and cost-effective
approach is a sustainable alternative for nanoparticle synthesis. It is worth noting
that this is the first time that zinc oxide nanoparticles are used as a natural preser-
vative in a moisturizing cream formulation. The resulting cosmetic formulation rep-
resents a unique advancement in cosmetic science that meets consumer demands
for safe and eco-friendly skin care products while utilizing the preservative proper-
ties of ZnO NPs. This research has significant potential to change the cosmetics
industry by providing natural and sustainable alternatives to traditional
preservatives.

KEYWORDS
anti-bacterial activity, green synthesis, nano-cosmetics, preservative cream, ZnO NPs

1 | I N T RO DU CT I O N technology enables the production of incredibly tiny particles that,

when used on the skin or hair, can have distinctive qualities and
Nano-cosmetics are cosmetic products that incorporate nanotechnol- effects.1 It has several potential benefits including enhanced penetra-
ogy to manipulate and process materials at the nanoscale, or one- tion, improved stability, targeted delivery, texture, and secondary
billionth of a meter, for enhanced formulation and design. This experience in sunscreen formulation.2 However, the safety of nano-

Environ Prog Sustainable Energy. 2025;e14583. wileyonlinelibrary.com/journal/ep © 2025 American Institute of Chemical Engineers. 1 of 14
https://doi.org/10.1002/ep.14583
2 of 14 YOUSAF ET AL.

cosmetics remains in question due to their small size, ability to pene- Various plants were reported for the synthesis of ZnO nanoparti-
trate the skin deeply, and enter the bloodstream, potentially causing cles: Agathosma betulina, Solanum nigrum, Calotropis gigantea, Hibiscus
unforeseen health effects.3 sabdariffa, Brassica oleracea, orange fruit peel, onion, and so forth.9
In response to these concerns, organizations like the US Food ZnO NPs are considered potential preservative agents for cosmetic
and Drug Administration (FDA) and the European Commission formulations.10 Additionally, the inclusion of Cucumis sativus extracts
have provided recommendations and regulations for the use of not only complements the green synthesis approach but also
4
nanoparticles in different industries. The growing demand for cos- enhances the beneficial properties such as antioxidant activity and
metic products such as shampoos, toothpaste, mouthwash, antimicrobial efficacy, thus potentially increasing the preservative
makeup, and other creams to improve physical appearance and capacity of the cream.11
beauty is continually expanding. To address the potential risks of This study seeks to determine the effectiveness of green syn-
hazardous materials and microbial contamination, this expanding thesized ZnO NPs using Cucumis sativus extract as preservatives in
industry must concentrate on multifaceted control. Microbial con- cream formulation. This study aims to confirm the potential of
tamination is a common issue in cosmetic formulations, as patho- these eco-friendly NPs as preservative agents in cream formula-
genic organisms can impact product stability and pose health risks tions through investigation of microbiological growth suppression.
5
to consumers. Furthermore, understanding the processes underlying the antibac-
Cream-based formulations are particularly common in many terial activity of ZnO NPs will contribute to the development of
pharmaceutical and cosmetic items, from topical drugs to skincare safer and more sustainable preservative approaches in cosmetic
creams. The organic components and water content present in and pharmaceutical formulations. The green synthesis method used
creams create a favorable environment for microbial growth, posing to synthesize ZnO NPs involved other ingredients like phytochemi-
a significant challenge for product stability and safety. To address cals, which may have specific interactions with NPs that can
consumer demand for products with extended shelf lives, the cos- enhance their antimicrobial properties and is a sustainable approach
metics industry employs various compounds to extend the shelf life for synthesis. The nanoparticles were confirmed by ultraviolet visi-
6
of cosmetic products. Preservative agents are thus essential; these ble (UV–Vis) spectroscopy, photo luminescence (PL), Fourier trans-
compounds prevent microbial growth, inhibit the production of form infrared (FTIR), scanning electron microscope (SEM), and x-ray
reactive oxygen species, and slow the oxidation processes. How- fluorescence (XRF).
ever, conventional preservatives such as triclosan, parabens, formal-
dehyde, benzalkonium chloride, and phenoxyethanol are linked to
harmful effects in humans, including allergic reactions, environmen- 2 | M A T E R I A L S A N D M ET H O D S
tal and animal toxicity, cytotoxicity, and genotoxic effects on
human lymphocytes, as well as DNA damage and antiandrogenic 2.1 | Materials
activity. Consequently, the cosmetics industry is under pressure to
find new, safe, and eco-friendly substitutes.7 Zinc oxide nanoparti- Zinc acetate dihydrate, sodium hydroxide, deionized (DI) water,
cles (ZnO NPs) are increasingly valued as a safe preservative in distilled water, agar, peptone, sodium chloride, nutritional yeast,
cream formulations due to their antimicrobial, antioxidant, antifun- 2,2-di phenyl picryl hydrazyl (DPPH), penicillin, DMSO, beeswax,
gal properties, and lower toxicity compared to conventional chemi- liquid paraffin, and borax were obtained from the department
cal preservatives. Unlike some synthetic preservatives linked to chemical store, and fresh Cucumber fruit was purchased from the
adverse health effects, ZnO NPs are non-irritating at lower concen- nearby market.
trations, making them suitable for applications in skincare. Addition-
ally, ZnO offers UV-protective benefits, which further stabilize the
formulation and add value to cream products as a multifunctional 2.2 | Preparation of cucumber extract
component.
Various chemical methods are present for the synthesis of To make cucumber extract (Cucumis sativus), a fresh cucumber
ZnO NPs, but the green synthesis techniques provide an eco- weighing about 50 g was taken and cleaned using DI water to
friendly approach to synthesize nanoparticles by eliminating remove impurities. The weighed cucumber was turned into fine
the use of hazardous chemicals as well as decreasing the environ- pieces and crushed in a grinder. A transparent soup of extract was
mental impact associated with nanoparticle synthesis.8 Natural formed by filtering it after 5 min of centrifugation at 5000 rpm
reducing agents that come from bacteria, plant extracts, or other (Figure 1).
sustainable sources can be used to produce ZnO NPs that have
a lower environmental effect and improved biocompatibility.8
Natural ingredients with therapeutic qualities could be added 2.3 | Green synthesis of ZnO NPs
to nano preparations to greatly increase their efficacy and
enhance the quality of life without the negative effects of syn- Two solutions of 25 mL were made separately containing 0.2 M
thetic formulations. Zn (CH3COO)2.2H2O and 0.2 M NaOH with DI water,
YOUSAF ET AL. 3 of 14

FIGURE 1 Preparation of cucumber extract.

respectively. The solutions were mixed carefully as shown in 2.4 | Mechanism for green synthesis of ZnO NPs
Figure 2, and 1 mL of pure cucumber extract was added to the
mixture. When everything had dissolved, the solution was kept The synthesis of metal oxide nanoparticles using the green syn-
under continuous stirring for 2 h. The off-white precipitate thesis method occurs only if the reduction potential of the metal
obtained was centrifuged for 10 min at 10,000 rpm to obtain a ions is greater than the reduction potential of phenolic com-
pellet. The pellet was washed with DI water and dried overnight pounds present in the plant extract. The reduction potential of
in an oven at 60
C; after drying, an off-white powder formed most of the phenolic compounds is approximately in between 0.3
(Figure 2). and 0.8 V. the plant extract used in this research work, Cucumis
4 of 14 YOUSAF ET AL.

FIGURE 2 Green synthesis of ZnO nanoparticles (NPs).

sativus has a number of phenolic compounds like gallic acid, vanil- in a beaker and heated at 50
C. Mix both the prepared oil phase and
12
lic acid, kaempferol, and protocatechuic acid. The reduction aqueous phase along with 1 mg of synthesized ZnO NPs. Continu-
potential of these phenolic compounds is in between 0.2 and ously stir the mixture until a homogeneous semi-solid mixture forms,
0.8 V.13 The reduction potential for Zn2+/Zn0 is 0.76 V, which is as shown in Figure 4.
very low compared to the reduction potential of the phytocom-
pounds present in the cucumber extract, so Zn2+ cannot be
reducible. That is why alkalinity must be induced in the reaction 2.6 | Antibacterial activity
mixture through the addition of NaOH to deprotonate the OH
and COOH groups present in phytocompounds, which decrease To study the antibacterial activity of ZnO NPs against gram +ve and
the reduction potential.14 gram –ve bacteria, the well diffusion method was performed using the
The synthesis mechanism of ZnO NPs using the plant extract nutrient broth solution. The nutrient broth was prepared by taking
involves forming metal ion complexes with the phytochemicals present 2.5 g peptone, 1.25 g yeast, and 2.5 g NaCl in 50 mL DI water; pH 7
in the extract and the conversion of these complexes into metal oxide was maintained by adding acid or base, and then 200 mL DI water
nanoparticles. The first step involves the oxidation of phytochemicals and 3.75 g agar were added. Two gram +ve bacterial strains, Bacillus
present in cucumber extract. In the next step, the anionic forms of phy- subtilis and Staphylococcus aureus, and 2 g ve strains, Escherichia coli
tochemicals make complexes with the Zn2+ ions through the chelation and Salmonella Typhimurium, were used to estimate the antibacterial
15
process. In the third step, the complex is hydrolyzed to form zinc potential of ZnO NPs. The solution was carefully mixed, and then the
hydroxide, and the phytochemicals are also recovered in this step. In solution and other glass equipment were autoclaved. After this,
the last step, the thermal treatment during drying converts the zinc the agar plates were prepared by putting the solution in the petri
hydroxide to ZnO NPs with the capping of phytochemicals Figure 3.16 dishes in a laminar flow hood to avoid any contamination. When the
agar solution solidified, wells were made in the agar plates by punch-
ing the agar plates with a microtip. Agar plates were inoculated with
2.5 | Preparation of cream bacteria and evenly spread on the plates using the glass spreader. A
solution of synthesized NPs was made using DMSO (1 mg/mL) and
To prepare the oil phase, 10 g of beeswax and 30 mL of liquid paraffin was put down in the wells. After 24 h of incubation at 30
C, the


were mixed in a beaker and heated in a sand bath at 70 C. To prepare radius of the inhibition zone surrounding the wells was measured to
the aqueous phase, 0.1 g of borax is added to 2 mL of deionized water estimate the antibacterial potential of ZnO NPs.
YOUSAF ET AL. 5 of 14

F I G U R E 3 Details of
phytochemicals involved in green
synthesis of ZnO nanoparticles (NPs).

2.7 | Antioxidant activity concentration of synthesized NPs by making them up to a 1 mL vol-

ume, and add 1 mL of DPPH in each concentration. After adding the
Antioxidant activity of ZnO NPs has been observed using the DPPH DPPH, vials had been kept in the dark at room temperature for approxi-
assay. Take an aliquot of 25–100 μL of synthesized NPs dissolved in DI mately 30 min; later, at 517 nm, absorbance had been calculated. The
water (1 mg/mL). Add methanol in the range of (900–975 μL) in each formula to measure the remaining DPPH is given in Equation (1).
6 of 14 YOUSAF ET AL.

Absorbance of sample obtained using ED-XRF by Rigaku NEX CG-I with the Fundamental
Percentage of DPPH remaining ¼  100, ð1Þ
absorbance control parameters x-ray analysis (Secondary targets RX9-kα, Cu-kα, Mo-kα,
Al-kα), Diaphragm (20 mm), Atmosphere (Air), Scan rate (0–50 eV),
• The absorbance of the sample around = 517 nm Tube Voltage (50 V), Current (1.00 mA) and Fine powder sample
• Absorbance of sample after 30 min = 1.2094 nm (≤75 μm) and powder x-ray diffraction (pXRD) measurements were
carried out using an analytical X'pert PRO diffractometer operating
with Cu Kα radiation (λ = 0.15406 nm).
2.8 | Preservative efficacy of ZnO NPs

ZnO NPs had been tested as preservatives in moisturizing creams by 3 | RESULTS AND DISCUSSION
comparing the anti-bacterial potential of cream that was preserved
with NPs to unpreserved cream. In a nutshell, 0.9% of a sterile NaCl 3.1 | Powder x-ray diffraction spectroscopy
solution was used to dilute 15 g of both preserved and unpreserved
cream. Salmonella Typhimurium bacteria were injected into both Figure 5 clearly indicates the pXRD spectrum of green synthesized ZnO
creams and kept at room temperature for 30 days. One gram of each NPs. The average particle size and structure of the synthesized nanocom-
contaminated cream was taken and diluted. After dilution, the solu- posite had been determined with XRD data. Each peak was sharp and
tions were dispersed in separate agar plates to measure the colony corresponded to ZnO, with specific 2θ values of 21.50
, 27.087
, 28.34

forming units (CFUs)s of bacteria after 2,7,14, and 28 days. The for- 32.89
, 33.34
, 34.14
, 39.91
, and 47.60
with diffraction planes (hkl)
mula to calculate CFU is given in Equation (2): values of (114), (400), (022), (217), (009), (224), (325) and (332) respec-
tively that were well congruent with orthorhombic (JCPDS#00-020-
No: of colonies counted on plate  diluation factor 1435). The remaining peaks in Figure 5 of 2θ values 23.20
, 29.40
, and
Colony forming unit ðCFUÞðgÞ ¼ :
volume of cultured plate
58.99
with diffraction planes (hkl) values of (220), (301), and (350) show
ð2Þ
the pXRD pattern of the orthorhombic structure of the synthesized NPs
2.9 | Characterization and were well congruent with (JCPDS #:01-071-2215). From the pXRD
data, the crystallite size of the synthesized ZnO NPs obtained was calcu-
The synthesized nanoparticles were characterized using Fourier lated to be 2.46 nm using the Scherer equation (Equation 3)
1
Transform infrared spectroscopy (FTIR) in the range 400–4000 cm
with a Bruker IFS 66v/S spectrometer. UV–Vis spectra were recorded Kλ
Crystallite size ¼ ð3Þ
β Cosθ
using Cary-60 UV–Vis spectrophotometer in the range 190–1100 nm,
PL studies were done using the Florence spectrometer Agilent Cary
Eclipse with the Data mode (Fluorescence), Fast Scan rate (1200 nm/ and 11.91 nm using Williamson-Hall equation (Equation 4)
min), Data rate interval (2 nm), Excitation filter (Auto) and emission fil-
ter (Open) & distilled water as solvent. Scanning Electron Microscopy βε ¼ 4ε tanθ: ð4Þ
was done by the Nova Nano SEM 450 instrument. XRF graph was

F I G U R E 5 Powder x-ray diffraction pattern of ZnO

FIGURE 4 Preparation of cream. nanoparticles (NPs).
YOUSAF ET AL. 7 of 14

3.2 | X-ray fluorescence spectrometry analysis, it was frequently employed.17 The elements in the sample
were represented by the peaks in the spectrum. Each peak's x-ray
In x-ray fluorescence (XRF) examination, a material was exposed to concentration was correlated with its atom density.18 Characteristic
high-energy X-rays, which caused the molecule since the sample peaks of Zn (KA) were at 8.34 keV, and Zn (KB) at about 9.55 keV had
to absorb energy as indicated in Figure 6. For chemical and elemental been depicted in the spectrum. The XRF analysis shows possible

FIGURE 6 X-ray fluorescence (XRF) spectrum of ZnO nanoparticles (NPs).

FIGURE 7 Scanning electron microscopy image of ZnO nanoparticles (NPs) with size distribution curve.
8 of 14 YOUSAF ET AL.

FIGURE 8 Fourier transform infrared spectroscopy spectrum of ZnO nanoparticles (NPs).

F I G U R E 9 Photoluminescence (PL) spectrum of ZnO

FIGURE 10 UV–Vis spectrum of ZnO nanoparticles (NPs).
nanoparticles (NPs).

confirmation of the formation of ZnO, and the concentration of Zn

was 74.2%. In the literature, the peaks of Zn (Kα) and Zn (Kβ) were at
8.2 and 9.4 keV respectively, and the concentration of Zn had been
described at 79.35%.19

3.3 | Scanning electron microscopy

In SEM imaging, a high magnification of 7000 at 10 kV results in

extremely exact images of a wide range of materials.20 Using the
SEM, the surface morphology of the synthesized NPs has been esti-
mated, which shows the presence of tightly packed clusters of ZnO
nanoflakes, as shown in Figure 7 (a) at 1 μm, (b) at 500 nm, and (c) at
2 μm. The image clarity was due to the instrumental parameters:
accelerating voltage, SEM magnification, and working distance. The
size distribution of the particles is homogeneous with non-uniform FIGURE 11 Band gap of ZnO nanoparticles (NPs).
YOUSAF ET AL. 9 of 14

sizes. By fitting the particle size distribution histogram to the log- absorption pattern in the range of 500–1000 cm1 was responsible
normal distribution function in Equation (5), the average particle size for different forms of collision of carboxylic groups. The peaks
for each sample has been determined, which is represented as, observed at 604 and 665 cm1 were due to the stretching and
secondary vibration of ZnO NPs.17,23 The cucumber fruit extract

 h i
f ðDÞ ¼ 1=√2πσ D exp ln2 ðD=D0 Þ=2σ 2 , ð5Þ contains bioactive functional compounds such as phenols, amines,
flavonoids, terpenoids, carotenoids, and others that act as capping
where σD is the standard deviation and D is the average particle size. agents for ZnO NPs and stabilize the colloidal particles during
An example of estimating the particle size distribution histogram for interaction22; comparison with reported peaks is shown in table in
ZnO NPs using a log-normal distribution function is illustrated in Figure 8.
Figure 7d. The estimated average particle size for ZnO NPs is 106 nm
with a standard deviation of 22.12.
3.5 | PL spectroscopy

3.4 | FTIR spectroscopy The crystal quality of nanomaterials was studied using photolumi-
nescence spectroscopy.26 The recombination of surface states was
FTIR is a significant and easy method for providing molecular-level where the photoluminescence comes from. As noted, the quantum
information in a frequency range of 4000–400 cm1.21 FTIR yields of band edges will fall exponentially with an increase in the
studies of ZnO NPs prepared using extract, as shown in Figure 8 depth of surface state energy levels; the surface states stay rela-
exhibited absorption peaks at 3363, 1542, 1383, 1327, 1020, tively shallow, usually exhibited by strong PL.27 As shown in
1 1
917, 665, and 604 cm . The broad peak observed at 3363 cm Figure 9 the PL spectra show a strong emission peak at 412 nm and
shows the presence of the OH group.24 At 1542 cm1 exhibits a green band at 536 nm.28 The ZnO lattice contains the zinc vacan-
because of the stretching vibration of amide. 1383 cm1 indicates cies that were responsible for the visible violet emission peaks at
24 1
an aromatic C C bond. The peak around 1327 cm was because 412 nm.29 The green emissions at 536 nm are because of the transi-
of C N stretching vibration of amines. 19,25
The absorption peak tion of e from the deep single-ionized oxygen vacancy to the
1 18
at 1020 cm was due to the stretching vibration of C O. The valence band.30,31

FIGURE 12 Measurement of zone of

inhibition.

T A B L E 1 Calculation of
Zone of inhibition (mm)
inhibition zone.
Sr. No. Bacteria ZnO NPs Penicillin (+ve) control DMSO (ve) control
1. Bacillus subtilis 0.9 3.5 No activity
Gram +ve
2. Staphylococcus aureus 1.2 4.25 No activity
Gram +ve
3. Escherichia coli 0.5 1.5 No activity
Gram ve
4. Salmonella typhimurium 1.4 9.5 No activity
Gram ve
10 of 14 YOUSAF ET AL.

3.6 | UV–Vis spectra

The study of the optical characteristics of nanosized particles is

frequently done using the UV–Vis absorption spectroscopy tech-
nique.32,33 Figure 10 shows the UV–Vis absorption spectra of the
synthesized NPs generated from a cucumber extract, which con-
tains organic biomolecules that were thought to be useful reduc-
ing and capping agents for nanoparticle formation.34 ZnO NPs
had a distinctive absorption peak at 362 nm, as shown in
Figure 10.20
The optical band gap (Eg) was calculated with the help of the Tauc
Equation (Equation 6). The curve generated between (hv) and (αhν)2
was extrapolated to get the direct band gap of the ZnO NPs.21

2
½αhv  =n ¼ K ðhv  EgÞ, ð6Þ

F I G U R E 1 3 Graph showing the zone of inhibition of ZnO where K is a constant, hv represents the energy of the photon, and n
nanoparticles (NPs).
is the nature of the transition that could have discrete values like ½

FIGURE 14 Mechanisms involved in the antibacterial activity of ZnO nanoparticles (NPs).

F I G U R E 1 5 Mechanism for
2,2-diphenyl picryl hydrazyl
(DPPH) free radical scavenging
activity of ZnO
nanoparticles (NPs).
YOUSAF ET AL. 11 of 14

and 2. The optical band gap (Eg) could be attained by the extrapola- 3.8 | Antioxidant activity
tion of the linear portion of the plot (αhv)2/n versus hv (Equation 7).34
For a direct transition n = 1, the equation becomes, Antioxidant activity of ZnO was observed with the help of the
DPPH assay. The dark purple color of the freshly prepared DPPH
½αhv2 ¼ K ðhv  EgÞ: ð7Þ solution disappears in the presence of antioxidant agents. The color
of the DPPH solution disappears due to the capturing of DPPH free
The extrapolated curve yields a band gap energy of about 2.9 eV, radicals, due to which it transforms into a colorless product that
as shown in Figure 11.35 decreases the absorbance at 517 nm, corresponding to the reduced
form of DPPH, which is 2,2-diphenyl-1-picrylhydrazine (DPPH-H)
illustrated in Figure 15.38 Absorbance for each concentration of
3.7 | Anti-bacterial activity synthesized NPs was determined. The percentage of DPPH radical
scavenging capability had been determined by using the formula
The antibacterial activity of ZnO NPs has been observed after 24 h given in Equation (1) indicated in Figure 16. The results of antioxi-
(Figure 12). The formation of inhibition zones in each agar plate dant activity were summarized below in Table 2. The maximum free
depicts that NPs have performed activity against bacterial species. For radical scavenging activity was obtained at a 100 μL concentration
each sample, the inhibition zone is calculated, and the NPs depict a of ZnO NPs.
different inhibition zone for each bacterium. The calculated inhibition
zone for each bacterium is shown in Table 1. The inhibition zone of
ZnO NPs acting as antibacterial agents against different types of bac- 3.9 | Preservative efficacy of ZnO NPs
teria was shown in Figure 13.
The mechanism involved in the antimicrobial activity of ZnO Due to frequent exposure, the danger associated with the use
NPs includes specific interactions like adsorption, Zn2+ ion release, of various preservation agents in cosmetics has increased
production of reactive oxidative species (ROS), and intercellular microorganism resistance and caused a variety of negative side
responses of microbial cells like lipid peroxidation, cell membrane effects.5 These issues call for the deployment of cutting-edge
damage, inhibition of energy metabolism, and disruption of DNA eco-friendly solutions that permit cosmetic products to be prop-
replication Figure 14.36 The interaction between positively charged erly preserved without compromising human health. ZnO NPs are
ZnO NPs and negatively charged bacterial cell walls occurs, and after a safe and skin-friendly component for cosmetics and surfaces
adsorption, the ZnO NPs are internalized into the bacterial cell, that come in contact with human skin.10 ZnO NPs show bacteri-
resulting in membrane rupture, loss of cell integrity, and increased cidal effects on both Gram-positive and Gram-negative bacteria,
oxidative stress from lipid peroxidation. This led to the production of
reactive oxygen species (ROS), damage to DNA, and inhibition of
bacterial growth.37 T A B L E 2 Percentage of 2,2-diphenyl picryl hydrazyl (DPPH)
radicals remaining.

Sr. Concentration % DPPH radicals

No. (μL) Absorbance scavenging activity (%)
1. 25 1.0916 90.25
2. 50 0.9646 79.75
3. 75 1.2677 82.22
4. 100 0.9444 100

T A B L E 3 Measurement of colony forming unit (CFU) (mL) in

preserved and unpreserved cream.

CFU (mL)

Sr. Number Preserved Unpreserved

No. of days cream cream
1. 2 53.51 46.67
2. 7 61.44 16.67
3. 14 67.68 0
F I G U R E 1 6 Graph showing the % of 2,2-diphenyl picryl hydrazyl
4. 28 71.57 0
(DPPH) radical scavenging activity.
12 of 14 YOUSAF ET AL.

as well as spores resistant to high temperatures and high

pressure. The increase in the surface area of ZnO NPs over
its microparticles was linked to their enhanced antibacterial anti-
oxidant activities, which make ZnO NPs a good preservative
agent for cosmetic formulations.
The preservative efficacy of ZnO NPs in the formulated mois-
turizing cream was observed by measuring the bacterial growth
suppression, and it can be concluded from Table 3 given below
that the number of CFU reduces with the passage of days and
reaches zero in cream preserved with ZnO NPs, indicating the
excellent preservative efficacy of green synthesized ZnO NPs.
While the unpreserved cream did not show any preservative effi-
ciency against test bacteria39 (Figure 17). Thus, the anticipated
cream formulation confirmed that the green synthesized ZnO NPs
are effective preservatives for cosmetics. Similar results have been
reported with green synthesized silver nanoparticles using Iris

F I G U R E 1 7 Preservative efficacy of preserved and unpreserved tuberosa and thymoquinone loaded chitosan nanoparticles in the
cream formulations. literature (Table 4).25,40

TABLE 4 Reported methodologies of the green synthesis of ZnO nanoparticles.

Volume of Reaction time and Average

Zinc source Plant extract (mL) temperature size (nm) Shape Reference
Zinc acetate Cucumis sativus 5 Stirring for 2 h at room 20 Spherical [41]
(20 g) temp
Zinc sulfate Cucumis sativus 1 Stirring for 24 h at 70
C 60 Clusters [42]
(0.1 M)
Zinc acetate Curcuma zedoaria 100 Stirring for 2 h at 60
C 19.65 Round [43]
(6 g)
Zinc acetate Syzygium Cumini 100 Stirring for 4 h at 80
C 10–12 Agglomerated [44]
(10 g)
Zinc acetate Orange peels 42.5 Stirring for 1 h at room 12 Spherical [45]
(2 g) temp
Zinc nitrate (2 g) Euphorbia hirta 5 Stirring till the temp – Spherical [46]
reached 90
C
Zinc sulphate Tecoma castanifolia 10 Incubated at room temp 70–75 Spherical [47]
(1 mM) for 4 days
Zinc nitrate Pontederia crassipes 10 Stirring for 6 h at 70
C 39 Spherical and [48]
(2.975 g) polyhedral
Zinc nitrate (6 g) Phoenix roebelenii 100 Stirring for 20 h at room 15.6 Spherical [49]
temp
Zinc nitrate Deverra tortuosa 25 Stirring for 1 h at 60
C 9–31 Hexagonal [50]
(2.5 g)
Zinc nitrate (2 g) Cinnamomum verum 25 Stirring till ppt formed at 48 Agglomerated [51]
60
C
Zinc acetate Pelargonium 20 Stirring till ppt formed at 14 Spherical and [52]
(1.095 g) odoratissimum (L.) 50
C hexagonal
Zinc nitrate Coffea arabica 20 Stirring till ppt formed at 40 Cubic [53]
(1 M) 80
C
Zinc nitrate (5 g) Pomegranate fruit 20 Stirring for 1.5 h at room 18–30 Aggregated [54]
temp nanoplates
Zinc acetate Cucumis sativus 1 Stirring for 2 h, room temp 2.46 Clustered Present
(0.2 M) nanoflakes study
YOUSAF ET AL. 13 of 14

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The ZnO NPs were successfully synthesized using a green synthesis
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method with Cucumis sativus extract. pXRD analysis confirms the syn- vacrol and eugenol chitosan nanoparticles as eco-friendly preserva-
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Res. 2010;27:1746-1749.
tions and was found to be 2.46 and 11.91 nm, respectively. SEM images
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AUTHOR CONTRIBUTIONS odology for the extraction of polyphenol contents and hplc profiling
Sanaya Yousaf: Writing – original draft; formal analysis. Anam Ashraf: of cucumis sativus peels. J Jilin Univ. 2022;41:41-52.
Writing – original draft; validation; data curation. Shaista Ali: 13. Eslami S, Ebrahimzadeh MA, Biparva P. Green synthesis of safe
zero valent iron nanoparticles by Myrtus communis leaf extract
Writing – review and editing; supervision; methodology; investigation;
as an effective agent for reducing excessive iron in iron-
funding acquisition. Akasha Rafiq: Formal analysis; validation. Abid overloaded mice, a thalassemia model. RSC Adv. 2018;8(46):
Mahmood: Resources; visualization. Farrukh Bashir: Software; valida- 26144-26155.
tion; resources. Muhammad Akhyar Farrukh: Investigation; conceptu- 14. Mubeen I, Farrukh MA. Mechanisms of green synthesis of iron nano-
particles using Trifolium alexandrinum extract and degradation of
alization; project administration. Fizza Naseem: Resources; data
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ACKNOWLEDGMENTS 5120.
16. Ahmed AA, Rizvi ZR, Shahzad H, Farrukh MA. Neodymium oxide
We would like to thank CASP, Government College University for
nanoparticles synthesis using phytochemicals of leaf extracts of dif-
providing the FTIR facility and the Material Chemistry Lab, Depart- ferent plants as reducing and capping agents: growth mechanism,
ment of Chemistry, Government College University Lahore for the optical, structural and catalytic properties. J Chin Chem Soc. 2022;
UV–Visible facility. 69(3):462-475.
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