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College of Basic Education Research Journal, Vol.20/2.1 January 2024
Green Synthesis ZnO NPs and Their Effects on
Plant Growing
Himdad H. Azeez
Physics Department, College of Education/Shaqlawa, Salahaddin University-Erbil, Iraq.
Hiwa H. Hasan
Biology Department, College of Education/Shaqlawa, Salahaddin University-Erbil, Iraq.
Dr. Hawbash H. Karim
Department of Physics, Faculty of Science and Health, Koya University, Koya KOY45,
Kurdistan Region - F.R. Iraq
Article Information
Abstract
Article history:
Received: January 25,2024
Reviewer: March 17,2024
Accepted: March 27,2024
Available online
This study investigates the production of ZnO NPs by the green method
and the effects of these nanoparticles on plant growth. Artemisia
abrotanum L. leaf extract was used as a chelating and capping agent. It
was found that the Artemisia abrotanum L. leaf extract has the potential
to biosynthesize ZnO NPs. FT-IR, UV-Vis, XRD, FE-SEM, and EDX
techniques were utilized in this study. The energy band gap of ZnO-NPs
was calculated to be around 2.48 eV. The average size crystal structure
of NPs was 50.29 nm, as confirmed by XRD. However, the ZnO
nanoparticles have a nanorod shape with a size of 66.93 nm, as evaluated
by FE-SEM, and the EDX confirmed that the sample is pure ZnO
nanoparticles. In addition, The nanoparticles were used to grow
Lepidium sativum L. plants. The total growth of Lepidium sativum L.
dramatically increased with the use of ZnO NPs, and the content of
natural dyes like chlorophyll a and b increased twofold compared to the
control. It was explained that zinc oxide nanoparticles are a good
candidate for growing plants instead of chemical fertilizers.
Keywords:
Green synthesis; ZnO NPs; Plant
Growing; XRD and EDX techniques
Correspondence:
himdad.azeez@su.edu.krd
hiwa.h.hasan@su.edu.krd
hawbash.hamadamin@koyauniversity.org
ISSN: 1992 – 7452
CC BY
College of Basic Education Research Journal, Vol.20/2.1 January 2024
KOY45
Artemisia abrotanum L
Artemisia abrotanum LZnO NPsFT-IR
UV–VisXRDFE-SEMEDXZnO NPs
50.29XRD
FE-SEMEDX
Lepidium sativumLepidium sativum LZnO NPs
ab
:
College of Basic Education Research Journal, Vol.20/2.1 January 2024
I. INTRODUCTION
Using materials and machines at the nanoscale is known as nanotechnology (Vidya
et al., 2013b). Nanotechnology has numerous applications in various fields including
medicine, agriculture, pharmacy, and renewable energy. Nanoparticles are particles that
measure between 1 and 100 nm in size (Sadatzadeh et al., 2018). These nanoparticles can
have various shapes. Materials or particles at this scale exhibit different properties compared
to bulk materials, such as electrical, chemical, optical, and thermal properties (Bala et al.,
2015, Karim and Omar, 2020). Furthermore, nanoparticles have a large surface area in
relation to their volume. This occurs because when the number of surface atoms exceeds the
number of atoms within the particle's volume, the particle size decreases and its surface
energy increases (Sharma et al., Barman, 2021).
The zinc oxide nanoparticles have attracted researchers due to their incredible
applications in various fields, including dental care, cosmetics, agriculture, solar cells, and
water purification (Narayana et al., 2018, Mahdi et al., 2020). Furthermore, zinc oxide
exhibits excellent specifications such as conductivity and transparency. As a semiconductor,
zinc oxide has a crystal structure that classifies it as an n-type semiconductor with a wide
band gap (Sagar Raut and Thorat, 2015). This compound involves the elements zinc and
oxygen, which are located in the second and sixth groups of the periodic table respectively
(Narayana et al., 2018).
ZnO nanoparticles can be produced using various techniques, including chemical,
physical, and environmentally friendly methods (Hiremath et al., 2013, Hedayati, 2015).
Unlike chemical and physical synthesis, the use of plants for nanoparticle synthesis is unique
and offers an affordable and environmentally friendly option (Senthilkumar et al., 2017,
Olajire and Adesina, 2017). Moreover, the use of plants allows for easy scaling up of
production without the need for hazardous chemicals, high pressures, or high temperatures
(Devi and Gayathri, 2014, Behzadi et al., 2022).
In this study, ZnO nanoparticles were synthesized using green synthesis, specifically
by utilizing the leaf extract of Artemisia abrotanum L. and zinc nitrate hexahydrate. These
zinc oxide nanoparticles were then utilized as a fertilizer.
II. METHOD AND MATERIALS
College of Basic Education Research Journal, Vol.20/2.1 January 2024
A. Sample Preparation
In this study, fresh leaves of Artemisia abrotanum L. were collected in Lass Village,
Shaqlawa, Erbil Government, Iraqi Kurdistan region. The following equipment was used: a
hotplate stirrer, zinc nitrate hexahydrate, beakers, filter paper, an oven, a mortar, and a pestle.
First, the fresh leaves of Artemisia abrotanum L. were placed into the mortar and ground
into a paste using the pestle. Then, 6 g of the paste was mixed with 100 mL of distilled water
in a beaker. The mixture was heated and stirred at 70°C for one hour. Subsequently, the
product was filtered using filter paper. Next, 30 mL of the plant extract was heated at 70°C
on the hot plate and stirrer. Afterwards, 3 g of zinc nitrate hexahydrate was added to the
heated extract. The mixture was further heated and stirred until it formed a paste. The paste
was then burned at 350°C for 2 hours, resulting in the formation of ZnO NPs powder.
Figure 1.(a) Washing leaves of Artemisia abrotanum L., (b) The leaves being crushed by
mortar and pestle, (c) The paste mixed with distilled water was heated and stirred at 70°C,
(d) Filtration of the product and mixing it with zinc acetate dihydrate, (e) The mixture was
heated and stirred until it became a paste, (f) Burning the paste at 350°C, and (g) Obtained
ZnO NPs powder.
B. Application of Synthesized Nanoparticles in Plant Growth
College of Basic Education Research Journal, Vol.20/2.1 January 2024
Four soil samples were prepared for this study. Seeds of Lepidium sativum L. were
then used and planted in these samples. Meanwhile, 3 gm of ZnO NPs were mixed with 3 L
of tap water. Two of the samples were watered with the zinc oxide nanoparticles solution,
while the other two were watered with tap water. After a period of two weeks, the results
were observed.
C. Morphological Study
Plant height and stem diameter were measured at the end of the experiment. In
addition, petiole and leaf length were also measured.
D. Determination of The Dye Content in Plant Leaf
To create the extract, dimethyl sulfoxide (DMSO) and 0.1 g of plant material were
combined. After being left at room temperature for approximately an hour in the dark, the
samples were incubated at 65 °C (water bath) for 30 minutes. The amount of chlorophyll
dyes was measured using a UV/VIS8453 spectrophotometer at the following wavelengths:
663 nm for chlorophyll a, 645 nm for chlorophyll b, 470 nm for carotenoids, and 534 nm for
anthocyanins. The Arnon formula (1949), modified by Richardson et al. (Richardson et al.,
2002), was used to determine the amount of vegetable dyes present. The dosage of individual
dyes was measured in mg/g of fresh weight.
III. RESULTS AND DISCUSSION
A. Characterization of plant extract
Researchers are currently creating customized nanoparticles by synthesizing them
using specific phytochemicals derived from plants. These phytochemicals, found within the
plant extract, are responsible for transforming metal ions into metal nanoparticles. As a
result, the plant extract acts as both a stabilizing and reducing agent in this process. The
progress of this reaction is monitored using UV-visible spectroscopy.
UV-Vis analysis of Artemisia abrotanum L. leaf extract
Before using Artemisia abrotanum L. leaf extract, it must be characterized by UV-
Vis spectroscopy to determine the presence of phytochemicals within the plant. These
College of Basic Education Research Journal, Vol.20/2.1 January 2024
phytochemicals function as reducing and stabilizing agents for nanoparticle synthesis
(Reddy et al., 2021).
The UV-Vis spectra of Artemisia abrotanum L. leaf extract are shown in Figure 2.
The highest peaks at 262 nm and 326 nm represent phenolic components and flavonoids,
respectively, in Artemisia abrotanum L. leaf extract. Artemisia abrotanum L. leaf extract
contains flavonoids, polyphenols, carotenoids, lipids, polysaccharides, tannins, free organic
acids, and essential oils. This indicates that this plant is rich in OH groups, which are
believed to be potential bio-reducing and stabilizing agents for nanoparticle synthesis(Lee et
al., 2013).
Figure 2. UV-Vis spectra of Artemisia abrotanum L. leaf extract.
FT-IR analysis of Artemisia abrotanum L. leaf extract
The FT-IR spectra obtained from the leaf extract of Artemisia abrotanum L., as
shown in Figure 3, display a wide range of distinctive peaks covering the entire spectral
range. In the field of Fourier-transform infrared (FT-IR) spectroscopy, these spectra are
traditionally divided into two main regions: the functional group region, which spans the
College of Basic Education Research Journal, Vol.20/2.1 January 2024
wavenumber range from 1800 to 4000 cm-1, and the fingerprint region, which extends from
0 to 1500 cm-1. Within the observed FT-IR spectrum, there are several prominent peaks that
can be identified. Specifically, the bands occurring between 1600 cm-1 and 3200 cm-1
indicate the stretching vibrations of tertiary amide C=O bonds and the O-H stretching
vibrations of phenolic groups. Furthermore, the presence of bands at 1645 cm-1 is attributed
to the stretching vibrations of tertiary amide C=O bonds, which is a characteristic feature as
reported by Isaac et al. in 2013(Isaac et al., 2013).
Figure 3. FT-IR spectra of Artemisia abrotanum L. leaf extract.
B. Characterization of ZnO NPs
UV-Vis analysis of ZnO NPs
In response to electromagnetic waves, surface plasmon resonance, which collects
conduction band electron oscillations, showed a peak absorption in UV-Vis spectroscopy,
indicating the production of nanoparticles and the reduction of metal ions. (Evanoff Jr and
Chumanov, 2005).
The peak at 373 nm observed by UV-Vis spectroscopy (Figure 4) was typical of ZnO
NPs. The surface plasmon resonance (SPR) was responsible for this, and the sharper peak
proved the formation of mono dispersed ZnO NPs (Pai et al., 2019). The absorption peak
maxima for ZnO NPs are typically between 300 and 380 nm (Varadavenkatesan et al., 2019).
The calculated value is lower than 380 nm and exhibits a blue shift in excitonic absorption.
College of Basic Education Research Journal, Vol.20/2.1 January 2024
ZnO NPs are known for having a peak at 373 nm, which was observed
by UV-Vis spectroscopy (Figure 3). This was caused by the surface plasmon resonance
(SPR), and the sharper peak indicated the formation mono dispersed ZnO NPs (Pai et al.,
2019). ZnO NPs typically have absorbance peak maxima between 300 and 380 nm
(Varadavenkatesan et al., 2019). The computed value shows an excitonic absorption shift to
the blue and is less than 380 nm.
The straightforward band-gap energy E (g) for ZnO NPs is obtained by fitting the reflection
data to the straight transform formula 𝛼ℎ𝑣 = 𝐴(ℎ𝑣 − 𝐸𝑔)𝑛, where 𝛼 is the absorption
coefficient, hv is the photon energy, E (g) is the straight band-gap, A is the absorbance, and
the power n is an integer for the direct band gap n=1/2. Extending the rectilinear component
of (𝛼hv)2 vs hv to the x-axis provides information about the exact band gap magnitude.
Figure 4 shows that the straight band gap for ZnO NPs is 2.48 eV. Utilizing plant
extract is expected to result in a decrease in the band-gap, and for the greenly produced
nanoparticles, this result does not conflict with the spatial quantum confinement effects
because some plant extract components cover/modify the surface and reduce the band-gap
of the nanoparticles (Khan et al., 2019). Nanoparticles produced through biosynthesis are
typically more reactive than identical nanoparticles produced through other methods
(Pantidos and Horsfall, 2014).
College of Basic Education Research Journal, Vol.20/2.1 January 2024
Figure 4. UV-Vis spectroscopy of ZnO NPs.
FT-IR analysis of ZnO NPs
In Figure 5, we present the Fourier-transform infrared (FT-IR) spectrum. This
spectrum has been used to rigorously assess the purity and composition of biosynthesized
zinc oxide nanoparticles (ZnO NPs). The absence of any discernible peaks within the
monitoring range clearly demonstrates the remarkable purity achieved through our
environmentally friendly synthesis approach. This unequivocally shows that the "green"
method used to produce ZnO NPs has successfully yielded highly pure nanoparticles.
Upon closer examination of the FT-IR spectrum, we can observe specific
vibrational signatures that are associated with the zinc and oxygen bonds. These signatures
manifest as a distinctive peak at 435 cm⁻¹, which has been previously reported by
(Yuvakkumar et al., 2015, Bhuyan et al., 2015). Further analysis of the spectrum reveals
additional prominent peaks. One notable band is observed at 1384 cm⁻¹, and it is attributed
College of Basic Education Research Journal, Vol.20/2.1 January 2024
to the C–O stretching of the carboxylic acid group. This finding is consistent with the
existing literature on these chemical bonds. Additionally, the spectrum exhibits a discernible
peak at 1647 cm⁻¹, which indicates the –C=C– stretching vibrations commonly found in
aromatic compounds, as reported by (Hu et al., 2019). We also observe a striking and
relatively wide band at 3498 cm⁻¹, corresponding to the O-H stretching vibrations associated
with phenolic compounds, as reported by (Ganesh et al., 2019). This observation provides
valuable insight into the presence of these chemical constituents in our synthesized ZnO
NPs.
Figure 5. FT-IR spectrum of ZnO NPs.
XRD analysis of ZnO NPs
X-ray diffraction spectra (Figure 6) show eleven peaks at 31.77°, 34.42°, 36.25°,
47.53°, 56.56°, 62.85°, 66.36°, 67.91°, 69.04°, 72.59°, and 76.96°, which correspond to the
(100), (002), (101), (102), (110), (103), (200), (112), (201), (004), and (202) planes. This
spectrum corresponds to JCPDS card No. 89-0510, which represents the hexagonal Wurtzite
crystal structure of ZnO NPs. The crystal size of Zinc Oxide nanoparticles was determined
using the (Debye-Scherrer) equation: D=kλ/β cosθ, where D represents the crystalline size,
k is the shape factor equal to 0.9, λ is the X-ray wavelength (λ=0.154 nm) for Cu K_α, θ is
the diffraction angle, and β is the full width at half maximum (FWHM) (Vijayalakshmi and
College of Basic Education Research Journal, Vol.20/2.1 January 2024
Rajendran, 2012). The average crystal size was found to be 50.29 nm. Similar results were
obtained for the green-synthesized Zinc Oxide nanoparticles using Peltophorum
pterocarpum leaf extract [(Pai et al., 2019).
Figure 6. XRD spectra of Zinc Oxide nanoparticles.
FE-SEM analysis of ZnO NPs
FE-SEM analysis was used to evaluate the surface morphology of green-
synthesized ZnO NPs, and the resulting picture is shown in Figure 7. The majority of the
ZnO NPs are in nanometer size, with an average particle size of 66.93 nm and nanorod
shapes. More of the particles have the same size. However, the nanoparticles are
agglomerated, which is normal for the green synthesis method. Due to the large surface area
of the green-synthesized nanoparticles, their strong affinities produce aggregation or
agglomeration (Agarwal et al., 2017). It may be concluded that ecological considerations
have a significant impact on NP stability and agglomeration. As a result, during the
nanoparticle production process, NPs adhere to one another and spontaneously form
asymmetrical clusters (Vidya et al., 2013a).
College of Basic Education Research Journal, Vol.20/2.1 January 2024
Figure 7. XRD spectra of Zinc Oxide nanoparticles.
Also, EDX analysis was used to determine the elemental analysis of the
nanoparticles. The EDX spectra (Figure 8) of the ZnO NPs show that the sample contains
Zinc and Oxygen elements, with weight percentages of 89.49 and 32.44, respectively. The
EDX spectra exhibit two distinct zinc peaks at 1.1 keV and 8.7 keV, as well as one sharp
oxygen peak at 0.5 keV, which are characteristic of ZnO NPs (Shim et al., 2019). The
presence of gold is attributed to the sample being coated with a thin layer of gold during FE-
SEM analysis.
College of Basic Education Research Journal, Vol.20/2.1 January 2024
Figure 8. XRD spectra of Zinc Oxide nanoparticles.
C. Application of Synthesized Nanoparticles in Plant Growth
Morphological study:
Our results indicate significant differences in plant length between the control group
and the plants treated with nanoparticle Zinc Oxide Lepidium sativum. The control group
had a plant length of 103 mm, while the treated plants had a length of 66 mm. Furthermore,
the application of Zinc Oxide NPs also affected the stem, petiole, and leaf length and width
of the treated plants, as can be seen in Table 1 and Figure 9, when compared to the control
group.
Table 1. Plant length, stem, and leaf dimensions of Lepidium sativum controls and those
treated with ZnO NPs, in millimeters*
Plant
Plant
lengt
h
Stem
Petiole
Leaf
length
width
A L
W
length
width
A
L
W
length
width
A L
W
Contro
l
66
(37-42) 39.5
(0.8-1.2) 1.0
39.5
(14-15)
14.5
(0.5-0.6)
0.55
26.
3
(11-13)
12
(3-5) 4
3
Treate
d ZnO
NPs
103
(56-60) 58
(1.4-1.5) 1.45
40
(20-27)
23.5
(0.8-0.9)
0.85
27.
6
(13-15)
14
(6-7) 6.5
2.1
* The measurements were taken from (10-15) specimens, with the numbers inside the bracts denoting the minimum and maximum,
and the numbers outside the bracts denoting the average. A L
W = Average Length/Width.
College of Basic Education Research Journal, Vol.20/2.1 January 2024
The content of vegetable dyes:
Carotenoids are a class of 750 naturally occurring dyes and non-enzyme antioxidants
produced by plants in response to unfavorable environmental conditions. These compounds
play a vital role in protecting cells from damage caused by free radicals. Anthocyanins, on
the other hand, are water-soluble phenolic compounds that contribute to the vibrant colors
seen in flowers and fruits. Chlorophyll, a natural substance found in plants, is responsible
for their green color. It facilitates the absorption of solar energy and its conversion into
chemical energy through the process of photosynthesis. Chlorophylls also play a role in
converting magnesium ions to divalent metal ions like iron (resulting in a grey color), copper
(giving a green color), or zinc (also resulting in a green shade) (MacFarlane and Burchett,
2001).
According to the results shown in Figure 10, Lepidium sativum treated with ZnO
nanoparticles displayed a two-fold increase in both chlorophyll a and chlorophyll b
compared to the control group. The plant cells were subjected to zinc compounds, and the
levels of carotenoid and anthocyanin were measured using spectrophotometry. The studies
indicated that, regardless of the form or concentration of the Zn ions, the content of
carotenoids and anthocyanins increased when exposed to them, in comparison to the control
sample. This finding demonstrates the activation of the plant's antioxidant system, as
depicted in Figure 11.
According to tests conducted by Wang et al., a decrease in chlorophyll a, chlorophyll
b, and carotenoids was observed regardless of the dose of nano-Zn used (400–32000 mg
nano-Zn/kg soil). Additionally, Tiecher et al.'s research on maize showed that chlorophyll a
and b levels were lower compared to control plants at all investigated dosages (30, 90, 180,
or 270 mg Zn / kg soil). In contrast, Radha et al. found that sugarcane (Saccharum spp.)
grown in soil with an excess amount of zinc exhibited an increase in the levels of carotenes
and chlorophyll.
College of Basic Education Research Journal, Vol.20/2.1 January 2024
Figure 10. Concentration of carotenoids and anthocyanins for various variants of the experiment.
V. CONCLUSION
The biosynthesis of ZnO nanoparticles (NPs) using plant extract has greatly improved
due to their diverse needs. This study focuses on the rapid and cost-effective production of
zinc oxide (ZnO) NPs using zinc nitrate and Artemisia abrotanum L. aqueous leaf extract.
Various techniques such as FT-IR analysis, UV-Vis spectroscopy, XRD, FE-SEM, and EDX
were employed to analyze the plant components, structure, morphology, chemical
composition, and optical properties of the ZnO nanoparticles. The band-gap of the ZnO NPs
was calculated to be approximately 2.48 eV. However, FE-SEM revealed that the ZnO NPs
have a nanorod shape with a size of 66.93 nm, and the EDX confirmed that the sample
consists of pure ZnO nanoparticles. Additionally, XRD confirmed that the average crystal
structure size of the ZnO NPs was 50.29 nm.
Our objective is to utilize biosynthesized ZnO NPs as an alternative to chemical
fertilizers, which have adverse effects on human beings, for the growth of plants (Lepidium
sativum L.). It has been suggested that zinc oxide nanoparticles can be used for plant
cultivation instead of chemical fertilizers.
ACKNOWLEDGMENTS:
We would like to thank Salahaddin University-Erbil, as well as the Soran Research
Center and Koya University, for helping us to characterize the samples during the research.
College of Basic Education Research Journal, Vol.20/2.1 January 2024
Disclosure statement:
Regarding their involvement in the research and writing of this study, the authors have
not disclosed any potential conflicts of interest.
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