OpenNano 8 (2022) 100067

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Eco-friendly synthesis of zinc oxide nanoparticles as nanosensor,

nanocatalyst and antioxidant agent using leaf extract of
P. austroarabica
Faiza A.M. Alahdal a, **, Mohsen T.A. Qashqoosh a, b, Yahiya Kadaf Manea a, b,
Mansour A.S. Salem a, Amjad M.T. Khan a, Saeeda Naqvi a, *
a
Department of Chemistry, Aligarh Muslim University, Aligarh, 202002, Utter Pradesh, India
b
Department of Chemistry, University of Aden, Aden, Yemen

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

Keywords: To reduce the hazardous effects of toxic compounds employed in the synthesis of nanoparticles,
Green synthesis researchers are looking at biological agents of reduction. The current work describes a green
Zinc oxide nanoparticles method for the biosynthesis of zinc oxide nanoparticles (ZnONPs) using leaf extract of
Metal sensing
P. austroarabica plant as biological reduction agent. The conditions of green synthesis were
Catalytic activity
Antioxidant activity
optimized to produce ideal size range of ZnONPs. ZnONPs were observed to have spherical and
FCC crystals with particle size of 13.9 ± 4.3 nm, as confirmed by PXRD, SEM, and TEM results.
The fluorescence technique demonstrated that ZnONPs are efficient probes for Cr(VI) ion, where
LOD was 1.30 nM, that is one of the lowest LOD reported for sensing substances. The findings also
explored that ZnONPs are very effective nano-catalyst for degrading of organic dyes, with
decolorization of congo red and methylene blue dyes attaining 93.42 and 95.74 % in 10 and 15
min, respectively. Furthermore, the results indicated that ZnONPs have high antioxidant activity,
due to the existing bioactive compounds on their surfaces. This study concludes that green syn­
thesized ZnONPs can be used as environmental sensor for detection of hazardous substances,
nanocatalyst for toxic dyes degradation and as antioxidants. This synthesis is highly effective, eco-
friendly, low-cost, and non-toxic.

1. Introduction

Synthetic techniques that employ non-toxic solutions like water and plant extract are widely used in biosynthesis procedures to
synthesize metallic nanoparticles. The biosynthesis of metallic nanoparticles has gained significant importance in recent years
compared to chemical synthesis because of avoiding the use of toxic chemicals in the synthesis protocol, and it also has several other
merits, such as being a single-step method, non-toxic, eco-friendly, low in cost, ease of application, and is carried out under normal
conditions. Additionally, these nanoparticles show compatibility for bio-medical and pharmaceutical applications [1–4]. In recent
years, the plants have acquired more medicinal importance and significant dimension due to the presence of a broad spectrum of
metabolites (primary and secondary) having antioxidant activities in their extracts [5]. Furthermore, because metal nanoparticles have

* Corresponding author.
** Corresponding author.
E-mail addresses: fzfz83@yahoo.com (F.A.M. Alahdal), fzchem83@gmail.com (S. Naqvi).

https://doi.org/10.1016/j.onano.2022.100067
Received 6 June 2022; Received in revised form 13 August 2022; Accepted 15 August 2022
Available online 18 August 2022
2352-9520/© 2022 The Author(s). Published by Elsevier Inc. This is an open access article under the CC BY-NC-ND license
(http://creativecommons.org/licenses/by-nc-nd/4.0/).
F.A.M. Alahdal et al. OpenNano 8 (2022) 100067

various physicochemical features, such as sensing, catalytic, antibacterial, antioxidant, magnetic and electronic properties, scientists
have demonstrated an interest in their production using novel techniques [6–8]. Nanoparticles of a large number of metals, like Zn, Ag,
Au, Cu, Ti, and, Pt, etc. have been produced employing a range of methods using bio-organisms, such as plant, fungi, bacteria, and
algae [9,10]. Researchers have shown interest in ZnONPs in recent years because of their unusual chemical and optical properties.
ZnONPs are nanoparticles that have been used in a wide range of cutting-edge applications such as sensing, catalytic activity, anti­
oxidant activity, biology, environmental protection, cosmetics, electronics, communication, and the pharmaceutical fields [1,11,12].
Furthermore, ZnONPs are of great utility in bio- applications such as bio-sensing, drug delivery, gene delivery, and nano-medicine
[12–15], in addition to their antibacterial, antifungal, anti-diabetic, and larvicidal activities [16–18]. The use of plant extract is a
promising technique for the easy synthesis of metallic nanoparticles using green methods. So, the extracts of many types of plants have
been used in ZnONPs synthesis in different works [1–3,19–23]. In the present work, ZnONPs have been synthesized using Phrag­
manthera austroarabica plant (Yemeni Mistletoe) as a promising plant to prepare green metallic nanoparticles.
Yemeni Mistletoe is type of species Phragmanthera austroarabica that grows parasitically on the tree of Yemeni Sidr (Ziziphus spina-
christi) and belongs to Loranthaceae family. For many years, mistletoe extracts had been used in herbal medicine in many regions of
Africa and Asia to treat a variety of diseases due to its wide range of biological activities, such as anticancer, antimicrobial, antiviral
activities, etc. The physiological, biochemical and therapeutic properties of this plant have been attributed to its phytochemical
constituents, which are largely phenolic and flavonoid molecules [24–30].
In the present study, we report the easy synthesis of ZnONPs by an eco-friendly procedure involving the reduction of Zn2+ by
P. austroarabica extract which acted as a reducing, stabilizing, and capping agent. The metal sensing, catalytic, and antioxidant ac­
tivities of the biosynthesized ZnONPs were then investigated using multi-spectroscopic techniques.

2. Materials and methods

Zinc acetate dehydrate (Zn(CH3COO)2.2H2O, ≥ 99%), hydrogen peroxide, 2,2-diphenyl-1-picrylhydrazyl (DPPH,99%), ascorbic
acid (≥ 99 %), methylene blue, congo red, borohydride sodium (NaBH4) and methanol were purchased from Sigma-Aldrich. All the
chemicals were of high analytical quality, which meant they could be employed without additional purification.

2.1. Collection of plant

P. austroarabica was gathered from Jayshan district-Yemen (Fig. 1). Plant leaves were washed and dried at room temperature for 4
weeks till it reached a consistent weight ground into powder and then was saved in a container in the refrigerator for analysis.

2.2. Preparation of plant extract

P. austroarabica extract was prepared by adding 8 g of powdered plant leaves to 100 mL of double-distilled water in a 500 mL flask
and was boiled for 30 min on a magnetic stirrer. Subsequently, the extract was allowed to cool until it attained the normal temperature,
and then was filtered through filter paper Whatman No.1. The filtrated extract was centrifuged at 5000 rpm for 10 min. The extract was
collected and stored in the refrigerator at 4◦ C for further experiments.

2.3. Green synthesis of zinc oxide nanoparticles

ZnONPs were prepared utilizing zinc acetate dihydrate (Zn(CH3COO)2.2H2O) as explained by Gnanasangeetha and Thambavani
[31]. About 70 mL of 10 mM of zinc acetate solution was added drop wise to 30 mL of the hot aqueous P. austroarabica extract which
was heated for 10 min at 55◦ C. The mixture was left to react with stirring for 4 h at pH 11. The reduction of Zn2+ and formation of
ZnONPs were monitored by UV-Vis spectrophotometry by observing the emergence of the characteristic spectrum of ZnONPs and

Fig. 1. P. austroarabica plant (Yemeni Mistletoe) on Yemeni Sidr tree.

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F.A.M. Alahdal et al. OpenNano 8 (2022) 100067

measuring the absorbance of the reaction mixture at different time intervals (5 min – 24 h). The precipitate was collected by
centrifugation at 10000 rpm for 15 min and rinsed twice with deionized water, followed by ethanol in order to remove any organic
residues. Then, the precipitate was dried overnight in an oven at 50◦ C, and was preserved for further studies.

2.4. Assessment of experimental conditions of ZnONPs formation

In order to obtain optimum synthesis conditions for preparation of ZnONPs, the different experimental parameters such as in­
fluence of metal ion concentration (5 - 30 mM), incubation time (5 min - 24 h), and pH (5 - 13) were investigated for green synthesis
process of ZnONPs using P. austroarabica. The sample absorbance was measured at 362 nm by employing UV–Vis spectrophotometry.

2.5. Characterization of ZnONPs

The UV–Vis spectra were measured using a double beam spectrometer (PerkinElmer UV Win Lab Lambda 800) in the wavelength
range 200–800 nm to characterize the production and stability of ZnONPs. The functional groups in the biosynthesized ZnONPs were
characterized by Fourier transformed infrared spectroscopy (FTIR, Perkin Elmer) using KBr pellets in the range of 4000–400 cm− 1. The
crystalline nature of ZnONPs was characterized by X-ray diffraction (XRD, Shimadzu 6100) in the range of 10–80 (2θ) with Cu Kα
radiation (λ = 0.155 nm), operated at a voltage of 30 kV and current of 15 mA. Scanning electron microscopy (SEM, JSM-6510LV,
JEOL Japan), energy dispersive X-ray spectroscopy (EDS), transmission electron microscopy (TEM, EM-410 LS Philips) and selected
area electron diffraction (SAED) were used to analyze the morphology, elemental investigation, structure and particle size distribution
of ZnONPs.

2.6. Sensitivity and selectivity of Cr (VI)

The fluorescence assessment of Cr (VI) was carried out by adding 25μL of Cr (VI) to 1000 μL of ZnONPs, and the final concentration
of Cr (VI) was 1000 mg/L, was allowed it to stand at room temperature for 5 min. Then, the fluorescence intensity of all solutions was
analyzed by Hitachi 2700 (Tokya, Japan) spectrophotometer with a cuvette of path length 1 cm equipped with a Xenon flash lamp.
Emission spectra were recorded by exciting the solutions at 280 nm and fixing the emission and excitation slit widths to 10 nm each.
Scans were performed in the emission range of 300–700 nm with a scanning rate of 1500 nm.min− 1. A buffer blank spectrum was
subtracted from the measured spectra for the fluorescence background correction. Selectivity of Cr (VI) was also investigated using a
range of coexisting ions (anions: CO2− 2− 2− 2+ 3+ 2+ 2+
3 , SO4 , CH3COO , NO3 , HPO4 , F , Cl , Br , and I , and cations: Co , Fe , Cu , Cd , Mn ,
− − − − − − 2+
2+ 2+ 3+
Pb , Sn , and As ) while maintaining the concentrations of all metal salts constant.

2.7. Catalytic activity (dye degradation)

The degradation of congo red (CR) and methylene blue (MB) dyes in the presence of NaBH4 was used to examine the catalytic
activity of ZnONPs. Briefly, 1 mL of freshly prepared NaBH4 solution (10 mM) was added to 1 mL of dye (40 ppm) (CR and MB), and the
volume was made up to 10 mL with distilled water with mild mixing. Likewise, 1 mL of freshly prepared NaBH4 solution (10 mM) was
added to 1 mL of dye (40 ppm) (CR and MB), and the volume was made up to 10 mL with distilled water, and also 100 μL of ZnONPs (1
mg/mL) were added to the solutions with mild mixing. The catalytic reduction of dyes was monitored by measuring the time-
dependent absorbance of the reaction mixture using UV-Vis spectrophotometry.

2.8. In vitro antioxidant activity

2.8.1. Screening of DPPH free radical scavenging activity

According to the method described by Bhakya et al. with slight modifications, the free radical scavenging activity of ZnONPs and
ascorbic acid was investigated using DPPH as stable radical [32]. Briefly, 1 mL of ZnONPs at various concentrations (10, 20, 40, 60, 80,
and 100 μg/mL) were mixed with 1 mL of fresh DPPH solution (1 mM in methanol). The system was then incubated in the dark for 30
min at room temperature. The absorbance was measured at 517 nm using UV-Vis spectrophotometer. Methanol was used as a blank,
and all the reagents were used as control. The inhibition percentage was computed using the following equation to indicate the free
radical scavenging activity:
Ac − As
Scavenging activity (%) = × 100 (1)
Ac

where (Ac) represents the control absorbance, and (As) represents ZnONPs or ascorbic acid absorbance.

2.8.2. Assay of scavenging activity of hydrogen peroxide

According to the technique described by Keshari et al. [33], the scavenging activity of H2O2 was performed to test the activity of
ZnONPs. Briefly, 0.4 mL of ZnONPs at various concentrations (10, 20, 40, 60, 80, and 100 μg/mL) was mixed with 1.2 mL of H2O2 (2
mM in the phosphate buffer) and 0.8 mL of phosphate buffer (50 mM and pH = 7.4), with continuous shaking the solution. The mixture
was then incubated for 15 min. The solution absorbance was controlled by UV-Vis spectrophotometer at 230 nm. Ascorbic acid and

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F.A.M. Alahdal et al. OpenNano 8 (2022) 100067

phosphate buffer were utilized as standard and blank, respectively. The inhibition percentage of the scavenging activity of H2O2 was
computed utilizing the following equation:
Ac − As
Scavenging activity (%) = × 100 (2)
Ac

where (Ac) represents the control absorbance, and (As) represents the absorbance of ZnONPs or ascorbic acid.

3. Results and discussion

3.1. Phytochemical analysis

Although the essential mechanism for biosynthesis of ZnONPs has not been fully known, but from the phytochemical analysis of the
extract, it can be observed that flavonoids, phenolic and other molecules, which are found in the extract of the plant may cause the
reduction of zinc ions, and control the biosynthesized nanoparticles size, where the hydroxyl, carboxylic and amino groups of phenolic,
flavonoids, alkaloids, proteins, etc., which exist in the extract of plant may link to the zinc surface and trigger the formation of ZnONPs,
which was also corroborated by FTIR analysis. C=C, C=O-C and C=O groups of heterocyclic compounds, as well as amide from
proteins may behave as stabilizers [34–36]. Earlier studies confirmed that P. austroarabica extract is essentially made up of phenolic
and flavonoid compounds such as quercetin, chrysophanic acid, catechin, emodin, methyl gallate, lupeol, ursolic acid, etc., that they
are enriched in functional groups such as hydroxyl and carboxylic groups [30,37]. Therefore, on the basis of the results of earlier
studies and FTIR results of the current study, and the well-known reducing power of the phenolic and flavonoid compounds, the
mechanism of the synthesis of ZnONPs has been proposed based on one of the phenolic compounds (quercetin) because it is present in
high content in P. austroarabica extract [38–40]. Fig.S1 and S2 present the suggested scheme for the reduction of zinc ions using
quercetin, which is present in high content in the extract of P. austroarabica plant. Therefore, the ZnONPs are formed in the
co-existence of phenolic and flavonoid compounds which enhance the reduction of Zn2+ to ZnONPs via release of electrons [30,
37–40].

3.2. Formation of ZnONPs

The ZnONPs formation began after the addition of 70 mL of 10 mM of zinc acetate solution drop wise to 30 mL of the hot
P. austroarabica extract with stirring. The reaction mixture changed to yellowish color, then to cream coloured precipitate, because of
Surface Plasmon Resonance (SPR) in ZnONPs, indicating ZnONPs formation [22,41]. ZnONPs formation and Zn2+ reduction to
ZnONPs was monitored using UV-Vis spectrophotometry. The characteristic band of ZnONPs was at 362 nm as shown in Fig. 2, which
is a distinctive fingerprint for ZnONPs. According to Mei’s hypothesis, a synthesized nanoparticle’s form is spherical if a single ab­
sorption peak is found in the UV–Vis spectra [42]. After 4 h, there was slight increase in the absorbance of solution, and there was no
change in the color and the absorbance of the solution after 24 h, indicating that the reduction of Zn2+ in the solution was complete.

3.3. Optimum experimental conditions of ZnONPs formation

All the experimental conditions were optimized to get the best circumstances for the production of ZnONPs such as incubation time,
Zn2+ concentration, and pH, which are necessary for the biosynthesis of ZnONPs.

Fig. 2. UV–Vis spectra of biosynthesized ZnONPs using P. austroarabica extract at different time intervals at 362 nm.

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F.A.M. Alahdal et al. OpenNano 8 (2022) 100067

3.3.1. Effect of Zn2+ concentration

The biosynthesis of ZnONPs is known to be affected by changes in metal concentration and biological components [43]. When zinc
acetate concentrations were increased from 5 to 30 mM, the peak of SPR for ZnONPs became visible, with the greatest intensity of peak
observed at 10 mM of zinc acetate (Fig. 3A). However, increasing the concentration of zinc acetate to 30 mM caused the absorbance to
drop. As a result, it was also observed that increasing Zn2+ concentrations beyond a threshold value resulted in decrease in nano­
particle synthesis. So, 10 mM of zinc acetate was shown to be the ideal concentration for the green synthesis of ZnONPs. Previous
studies suggest that high concentrations of zinc acetate cause unstable particles with a higher particle size [23,44].

3.3.2. Effect of incubation time

The effect of the incubation period on ZnONPs synthesis when observed the absorbance was found to increase as the incubation
period increased, as shown in Fig. 3B. The color and low value of absorbance at 362 nm indicated that the Zn2+ reduction rate was
sluggish during the first 5 min. But, after 5 min, an increase in the absorbance values along with color intensity were observed, and this
trend continued during the next 4 h. After this time, there was a slight rise that lasted up to 24 h. Then the increase halted, indicating
that the Zn2+ reduction was completed and ZnONPs were synthesized fully. According to the previous studies, the time required for full
reduction of zinc ions was to range from 3 - 24 h [42,45]. The remarkable reducing capability of the active chemicals in P. austroarabica
extract as flavonoid and phenolic compounds may be responsible for the rapid synthesis of ZnONPs in this work [1,3,19,23,38].

3.3.3. Effect of pH
To investigate the effect of pH on the biosynthesis of ZnONPs, the green synthesis technique was carried out at various pH values
ranging from 5 to 13. As demonstrated in Fig. 3C, increasing the pH from 7 to 11 resulted in an increase in the absorbance of the
solution. The spectra at pH 5 and 6 exhibited a nearly straight absorption line with no peak, but a typical absorption peak was observed
at pH 10 and 11. So, the better pH value for ZnONPs synthesis was at pH 11. The slight decrease in the absorbance at pH 12 and 13 may
be due to the low ZnONPs monodispersity, indicating that ZnONPs form in basic and neutral medium, whereas they do not form in the
acidic medium. The influence of pH investigated in this study was consistent with the results of prior studies [23,46,47].

3.4. Characterization of green synthesized ZnONPs

3.4.1. Fourier transform infrared spectroscopy

The primary functional groups of potential biomolecules available in the extract of plant and responsible for the reduction of Zn2+
ions and stability of biosynthesized ZnONPs were identified using FTIR. The FTIR spectrum exhibited numerous absorption peaks, as
shown in Fig. 4A, suggesting certain biological material components that contributed to the biosynthesis of ZnONPs. The characteristic
peaks at 3420 and 3244 cm− 1 are assigned to -OH stretch of phenolic or alcoholic molecules and -NH stretch of amines, whereas the
band at 2050 cm− 1 may be assigned to C=C stretch. The peaks at 1619, 1560 and 1413 cm− 1 attributed to C=O stretch of primary
amines, symmetric stretch of -C-C=C and -C=O group from aromatic ring, respectively. The characteristic peaks at 1230, 1115, 1025,
815 and 625 cm− 1 are attributed to secondary alcoholic group, bending of alcoholic -C-OH, -C-N stretch of aliphatic amines or
phenolic/alcoholic molecules, amine deformation (-NH), or plane bending of C-H and -OH bend in alkyl or alkyne halide, like C-Cl,
respectively. The characteristic peak at 480 cm− 1 attributed to Zn-O stretching, that indicates the presence of ZnO and the formation of
ZnONPs [1,19,20,23,48]. These results suggest that biological components existing in the P. austroarabica extract, such as phenolic and
flavonoids compounds, have a major role in the reduction of Zn ions and act as capping agents for ZnONPs. It is known that the
biological components react with metal salts via these functional groups, resulting in the reduction of metallic ions to metallic
nanoparticles [1,19,23,30,37–39,49,50].

3.4.2. Powder X‑ray diffraction analysis

The pattern of PXRD in Fig. 4B shows the peaks at 31.93◦ , 34.62◦ , 36.49◦ , 47.75◦ , 56.61◦ , 62.51◦ , 66.44◦ , 67.85◦ , 69.01◦ , 72.21◦ ,
and 76.50◦ which corresponded to (100), (002), (101), (102), (110), (103), (200), (112), (201), (004) and (202) reflection lines of

Fig. 3. Effect of diverse parameters on biosynthesis of ZnONPs: (A) effect of concentrations of zinc ions; (B) influence of incubation period; (C)
influence of pH medium.

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F.A.M. Alahdal et al. OpenNano 8 (2022) 100067

Fig. 4. (A) FTIR spectra and (B) PXRD of ZnONPs.

ZnONPs respectively, indicating hexagonal wurtzite structure and high crystallinity of ZnONPs. JCPDS (36-1451) was used as a
reference according to the peaks obtained for assigning the lattice planes. The average size of particles was calculated from the
distinctive peak corresponding to the 101 plane utilizing equation of Debye – Scherrer [1,23,51]:

D = K.λ / β.cosθ (4)

where K assigned to the constant of Scherrer (0.9), λ assigned to the X-ray wave length (0.15406 nm), β represents the full width at half
maximum (FWHM) of the diffraction line in radian and θ assigned to half diffraction angle of the peak, which is located at 2θ = 36.49◦ .
According to the FWHM for the peak matching to 101 plane under our experimental circumstances, the average size of the
nanoparticles was 14.8 ± 3.6 nm.

3.4.3. Scanning electron microscopy

According to the SEM results, P. austroarabica has a high efficiency to contribute in the production of ZnONPs, and revealed that
ZnONPs had a spherical morphology but were non-smooth with an approximately fair and uniform distribution (Fig. 5A).
EDS analysis revealed the existence of zinc oxide in green synthesized ZnONPs, as shown in Fig. 5B, indicating that the needed
phase of Zn and O is existent in the sample. The existence of carbon indicates the participation of plant phytochemicals in reduction
and capping of the bio-synthesized ZnONPs [1,41].

3.4.4. Transmission electron microscopy

According to the TEM results (Fig. 6A), the biosynthesized ZnONPs were approximately spherical in shape. Moreover, ZnONPs
were extremely crystalline and well distributed, with an average particle size of 6.1 – 26.6 nm and a mean particle size of 13.9 ± 4.3
nm (Fig. 6B). The selected area electron diffraction (SAED) pattern revealed (as shown in Fig. 6C) that the diffraction rings of the
biosynthesized ZnONPs exhibited Debye–Scherrer rings, indicating that ZnONPs are nano-crystalline in nature. Therefore, we found
that the size of particles and SAED pattern determined from TEM results are in good agreement with those of the results of XRD analysis

Fig. 5. (A) image of SEM and (B) EDS spectrum for ZnONPs.

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F.A.M. Alahdal et al. OpenNano 8 (2022) 100067

Fig. 6. A,B,C. TEM images, size distributions and SAED pattern of ZnONPs.

[52].

3.5. Sensitivity and selectivity of Cr (VI)

The ZnONPs sensitivity to detect the hexavalent chromium ions was investigated by measuring the concentration change of Cr (VI)
ions. The fluorescence intensity of ZnONPs was reduced when Cr (VI) concentration is increased, as shown in Fig. 7A. The FL intensity
vs. logarithmic values of Cr (VI) concentration (Fig. 7.B and C) revealed a linear decrease in FL intensity when the concentration of Cr

Fig. 7. (A) Fluorescence intensity of ZnONPs in existence of Cr (VI) with different concentrations (B) log [Cr (VI)] vs. fluorescence intensity, (C)
Fluorescence intensity of ZnONPs in terms of the ratio of decreasing values to the initial values at 470 nm against the log values of Cr (VI)
concentration.

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F.A.M. Alahdal et al. OpenNano 8 (2022) 100067

(VI) increased from 0.1–1.0 mg/L, with LOD of 1.30 nM and correlation coefficient of 0.998. The good sensitivity of ZnONPs for ions of
Cr (VI) could be imputed to the existing functional groups on the ZnONPs surface such as hydroxyl and amine, etc [53]. As a result,
ZnONPs can be employed as good eco-nanosensor since the detection limit is lower than that of numerous other Cr (VI) ion nanosensors
reported earlier [54–56]. Furthermore, the effect of pH on the ZnONPs sensitivity to Cr (VI) was investigated, and it was found that the
FL intensity decreased in the basic, neutral and acidic mediums, but that the decline was similar in the basic and neutral mediums,
while being slightly lower in the acidic media, indicating that the ZnONPs sensitivity to ions of Cr (VI) were efficient in all pH me­
diums, with a minor feature in basic and neutral medium. The minor change in FL intensity in acidic media might be due to loss of
certain chromophoric groups on ZnONPs surface.
In the presence of a combination of ecologically relevant cations such as Co2+, Fe3+, Cu2+, Cd2+, Mn2+, Pb2+, Sn2+, and As3+, the
selectivity of ZnONPs for the ions of Cr (VI) were assessed. In the presence of this mixture without Cr (VI), the fluorescence intensity of
ZnONPs decreased slightly, whereas adding Cr (VI) to this mixture resulted in a large emission decrease (Fig. 8A), indicating that
ZnONPs have excellent selectivity for detecting Cr (VI) while other cations also exist, without using any exterior masking factors.
Furthermore, ZnONPs selectivity for the ions of Cr (VI) were investigated in the existence of anion combination such as CO2− 2−
3 , SO4 ,
CH3COO− , NO−3 , HPO2− 4 , F−
, Cl −
, Br−
, and I−
. With the presence of this mixture without Cr (VI), the fluorescence intensity of ZnONPs
decreased slightly (Fig. 8B). The minor decrease in fluorescence intensity might be attributable to excess anions present in the mixture,
which was probably linked with the functional groups on ZnONPs surface. Whereas adding Cr (VI) to the anionic mixture resulted in a
large emission decrease, indicating that ZnONPs have excellent selectivity for the ions of Cr (VI) with the existence of anionic mixture.

3.6. Degradation of dyes

The degradation of dyes, when present in the excess of NaBH4 has generally been used to investigate the catalytic effectiveness of
metallic nanoparticles, whether loaded on other materials such as polymers as supports, or used alone. In this study, the catalytic
capability of green synthesized ZnONPs was investigated for the CR reduction (anionic dye) and MB reduction (cationic dye). As the
plasmon band of dyes is in the visible wavelength range, which is wholly different from the zinc range, allowing an obvious expla­
nation of the catalytic ability of ZnONPs [57–59].
This study demonstrated the ability of synthesized ZnONPs as a catalyst in the degradation of dyes such as congo red (CR) and
methylene blue (MB) when present in excess of NaBH4. UV–Vis spectrophotometry was used to monitor the color change of the so­
lution to colorless and the process of dye degradation, which were due to the dye reduction and leuco dye formation. The bands of
absorption for CR and MB dyes were at 494 and 663 nm, respectively, that might be due to the transition of electrons from π or n to π*
[60,61]. The findings revealed that catalytic degradation of dyes (CR and MB) in the presence of NaBH4 as reducing factor was
extremely slow, and the absorbance of dyes remained unaltered even after 60 min without ZnONPs (Fig. 9A,B). After the addition of
ZnONPs, a high decrease in the dye absorbance (CR and MB) and a rapid change in the color of the system (dye and BH−4 ions) to
colorless were showed within 10 and 15 min, respectively (Fig. 10A,B), which indicates the high catalytic effectiveness of green
synthesized ZnONPs and full reduction of CR and MB. In comparison to CR and MB, a high NaBH4 concentration was employed in the
system, resulting in a rise in the pH of the whole system. As a result, the breakdown of the BH−4 ion is slowed down. The ions of
hydrogen (H+) generated create a purging environment above the system, effectively inhibiting aerial oxidation of CR and MB dyes.
The catalytic degradation helped by ZnONPs is carried out efficiently by transferring electrons to electron acceptor (CR and MB) from
electron donor (BH−4 ), which contributes to the stability of the system and lowers the activation energy [57,62]. The rate of reaction
may be regarded as independent of the NaBH4 concentration because the NaBH4 concentration in the mixture is substantially larger
than that of CR and MB. So, the rate of kinetic reaction of catalytic degradation of system may be evaluated utilizing the following
pseudo-first-order kinetics equation [63]:

Fig. 8. Fluorescence spectra of (A) Cationic mixture (Co2+, Fe3+, Cu2+, Cd2+, Mn2+, Pb2+, Sn2+, and As3+, 10 mg/L each one) without /with ions of
Cr (VI), and (B) Anionic mixture (CO2− 2− − 2−
3 , SO4 , CH3COO , NO3 , HPO4 , F , Cl , Br , and I , 10 mg/L each one) without/with ions of Cr (VI) (10
− − − − −

mg/L).

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Fig. 9. UV-Vis spectra of dye degradation: (A) MB, and (B) CR.

Fig. 10. UV-Vis spectra of dye degradation catalyzed by ZnONPs: (A) MB, and (B) CR.

( )
A0
ln = − kt (4)
At

where At represents absorbance at final time, A0 represents absorbance at initial time, t assigned to time of reduction, and k represents
the apparent constant of first-order rate (min− 1).
The following formula was used to calculate the percentage of dye degradation:
A0 − At
% of dye degradation = × 100 (5)
A0

where At assigned to absorbance at final time and A0 assigned to the initial absorbance.
The slope of the pseudo-first order reaction equation was used to compute the rate constant of dye degradation. As seen in Fig. 11A,
B, A linear curve of time vs. ln(A0 / At) for CR and MB was seen at room temperature, confirming that the system reaction is of pseudo-
first order. Table 1 shows the dye degradation percentage and rate constant. So, the findings of this work were superior to several
previously published studies and agree with some others [64–69].
It can be safely deduced from the above study that the green synthesized ZnONPs behaved as effective catalyst for the reduction
process of both dyes. Furthermore, the ZnONPs have the ability to operate as an electron relay system. The catalytic dye reduction is
commenced through ZnONPs by transferring electrons from the donor (BH−4 ) to the acceptor (dye). Dyes could readily transfer to
surface of ZnONPs in the early step due to their high reduction and adsorption abilities. Throughout this procedure, the H2 produced
can stimulate the nanoparticles and water convection, eliminating the alternate products and keeping the surface fresh while main­
taining the high ZnONPs reactivity.

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F.A.M. Alahdal et al. OpenNano 8 (2022) 100067

Fig. 11. Plot of ln (A0/At) vs. time for pseudo-first order reaction of catalytic reduction of (A) MB, and (B) CR with NaBH4 in the existing of ZnONPs.

Table 1
The dye degradation percentage and rate constant (min− 1).
Dyes R2 Rate constant (min− 1) Degradation percentage of dye (%)

CR 0.998 0.2671 93.42

MB 0.999 0.1948 95.74

3.7. Antioxidant activity of ZnONPs

At normal temperature, DPPH is a stable free radical that accepts an electron or hydrogen radical to generate stable diamagnetic
molecules [70–72]. Scavenging ability of DPPH has been commonly used to disclose antioxidants such as polyphenolic substances. The
scavenging activity of free radicals for ZnONPs was measured by comparing changes in color from purple to yellow, depending on a
control that did not display any color change. When compared to P. austroarabica extract and ascorbic acid, ZnONPs had a high DPPH
scavenging activity due to their phytochemical substances, like flavonoids and phenols (Fig. 12A and Table 2). It’s also been shown
that when the concentration of ZnONPs increases, so does their antioxidant activity and they show higher inhibition with DPPH
scavenging activity of 85.17 %. So, the findings of this work were superior to those of several previously published studies [70,73,74].
The H2O2 radicals are weak oxidizing agents biologically. When H2O2 radicals convert to hydroxyl radicals with the presence of
metallic ions, they will be regarded as toxicant to the cell that initiates the lipid peroxidation propagation. Hydroxyl radicals are highly
reactive free radicals when generated in the bio-systems, which are capable of damaging large molecules like proteins, lipids, and DNA
[75,76].
The scavenging ability of H2O2 for ZnONPs was assessed using UV-Vis spectrophotometry at 230 nm, as shown in Fig. 12B and
Table 2. The findings of this work demonstrated that ZnONPs have a high reducing ability when compared to P. austroarabica extract
and ascorbic acid. The high activity of ZnONPs for scavenging of free radicals may be due to their propensity to lose electrons, as well

Fig. 12. Antioxidant activity of green synthesized ZnONPs and extract of P. austroarabica at various concentrations: (A) scavenging activity of
DPPH, (B) scavenging activity of H2O2.

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F.A.M. Alahdal et al. OpenNano 8 (2022) 100067

Table 2
Inhibition percentage of DPPH and radical scavenging activity of H2O2 for green synthesized ZnONPs and extract of P. austroarabica
Concentration (μg/mL) Assay of DPPH Assay of H2O2

Inhibition percentage (%) Inhibition percentage (%)

ZnONPs±SD* Extract±SD Ascorbic acid±SD ZnONPs±SD* Extract±SD Ascorbic acid±SD

10 30.62±0.69 37.13±0.63 41.32±0.17 31.45±0.51 35.24±0.83 38.10±0.47

20 47.30±0.57 53.21±0.49 58.96±0.24 41.32±0.63 46.09±0.54 50.80±0.74
40 58.26±0.46 67.24±0.71 78.42±0.11 53.11±0.49 63.36±0.92 66.08±0.81
60 64.53±0.72 80.83± 0.08 86.18±0.33 59.84±0.88 74.51±0.48 79.22±0.71
80 76.72±0.78 88.45±0.51 90.42±0.22 70.36±0.74 80.23±0.76 84.52±0.57
100 85.17±0.84 94.17±0.60 96.84±0.09 78.13±0.86 85.13±0.59 89.12±0.68

SD*: Standard deviation (N = 3)

as the existence of excellent capping and oxidazing agents on their surfaces. The findings are in agreement with the previous works that
have been reported [77–80].
Based on these findings, we conclude that these biosynthesized ZnONPs using P. austroarabica extract can be utilized as an effective
natural antioxidant agent to protect health from oxidative processes.

4. Conclusion

In this study, ZnONPs were produced using extract of P. austroarabica (Yemeni mistletoe), and characterized utilizing multi-
techniques including FTIR, PXRD, UV-Vis, SEM, EDS, TEM, and FL spectroscopy. As per the results of the FTIR analysis, ZnONPs
were formed, and phytochemicals contained in P. austroarabica extract worked as agents to reduce Zn2+ to ZnONPs and as stabilizing
and capping agents. The surface plasmon resonance of ZnONPs was confirmed by UV–Vis analysis, and the XRD spectrum also revealed
that ZnONPs have a FCC crystal structure. SEM and TEM results clarified that ZnONPs have spherical shapes with size from 6.1–26.6
nm and an average size of 13.9 ± 4.3 nm. The assay results of sensitivity and selectivity for Cr (VI) showed that ZnONPs are very
efficient as a nanosensor for the detection of Cr (VI), where this study revealed that LOD was 1.30 nM for Cr (VI), which is one of the
lowest limits of detection that have been reported in previous works. The results of this work also showed that ZnONPs were an
effective nanocatalyst for the degradation of toxic organic dyes. In addition, they had high antioxidant activity compared to
P. austroarabica extract and ascorbic acid. This indicates that the green synthesized ZnONPs from P. austroarabica extract could be a
promising material as bio-environmental probe to detect and remove the hazardous pollutants in various fields, in addition to be used
as antioxidant agents, because they are highly effective, ecofriendly, nontoxic, and low in cost.

CRediT authorship contribution statement

Faiza A.M. Alahdal: Conceptualization, Methodology, Investigation, Writing – original draft. Mohsen T.A. Qashqoosh: Meth­
odology, Visualization, Writing – review & editing, Resources. Yahiya Kadaf Manea: Data curation. Mansour A.S. Salem: Software.
Amjad M.T. Khan: Writing – review & editing. Saeeda Naqvi: Supervision, Writing – review & editing.

Declaration of Competing Interest

The authors declare that they have no conflict of interest.

Acknowledgments

The authors are grateful to the Chemistry Department at Aligarh Muslim University for supporting this work. The university
administration should also be praised and appreciated for supplying us with the required facilities.

Supplementary materials

Supplementary material associated with this article can be found, in the online version, at doi:10.1016/j.onano.2022.100067.

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