Plant Nano Biology 4 (2023) 100031

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Plant Nano Biology

journal homepage: www.journals.elsevier.com/plant-nano-biology

Bio-genic synthesis of calcium coated zinc oxide nanoparticles from

beetroot extract and their photo-degradation study on methylene blue and ]]
]]]]]]
]]

rhodamine B
Darshan Singha, , Anuradhaa, Divya Mathura, Surendra Kumarb, Balaram Panic, Amar Kumard,
⁎

Rajni Kanojiae, Ravi Guptaf, , Laxman Singhg,

⁎ ⁎

a
Department of Chemistry, Daulat Ram College, University of Delhi 110007, India
b
Department of Chemistry, Hansraj College, University of Delhi, 110007, India
c
Department of Chemistry, Bhaskaracharya College of Applied Sciences, University of Delhi, 110075, India
d
Department of Chemistry, Ram Krishna Dwarika College, Patna, Patliputra University Patna, 800020 Bihar, India
e
Department of Chemistry, Shivaji College, University of Delhi 110027, India
f
College of General Education, Kookmin University, Seoul 02707, South Korea
g
Department of Chemistry, Siddhartha University, Kapilvastu, Siddharth Nagar, 272202, India

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

Keywords: The advent of nanotechnology has helped in several invention in science & technology. Contamination of surface
Beet root extract water, ground and soil by various industrial dyes causes several ecological problems. Zinc oxide nanoparticles as
Zinc oxide photocatalysts and semiconductor materials show unique physical properties at the nanoscale and can be used to
Photocatalyst solve these problems to some extent. In this paper, we synthesized ZnO NPs and calcium-coated ZnO nano-
Rhodamine B
particles using extract of beetroot and then, for industrial point of view, we studied the photocatalytic de-
Methylene blue
gradation of methylene blue and rhodamine B using sun as a natural light source (sunlight). We have synthesized
Degradation
calcium-coated ZnO nanocomposites with 20–50 nm particle size. Synthesized nanomaterials were characterized
by using the different physio-chemical techniques such as - FT-IR, XRD, TEM, SEM, EDX and UV-spectro-
photometer do their photocatalytic degradation.

1. Introduction Photocatalyst” is derived from two words “photo,” which means

The latest advancements in the nano-science field have gathered
significant focus from different sectors since they could be used for
diagnosing, detecting, purifying industrial waste (Sharma et al.,
2017), catalysis (Gangadoo et al., 2017), therapeutics (Venkatasubbu
et al., 2016), food technology (Menon et al., 2018)and role in energy
utilization or production. The contamination of water resources,
including groundwater and reservoirs, as a consequence of increased
industrialization and population growth, has made water treatment
one of today’s most concerning challenges. As a result, it’s critical to
clean and recycle industrial and municipal wastewater. Due to their
outstanding benefit as adsorbents and for use in filtering, nano-
particles have attracted much attention in the last decades to treat
wastewater (Iravani, 2011). Photocatalysts play a very important
role in pollutant treatment (Nayak and Sahu, 2022). The keyword
“photocatalysis” first appeared in the scientific literature in 1911
(Bruner and Kozak, 1911).
light, & “catalyst” which is chemical agent that fastens the chemical
reactions by absorbing light or photons. Since they have a potential in
reducing environmental pollution, improvements have been made to
the photocatalytic activity or reaction rates (Abdeltif et al., 2020a). This
field has a lengthy history, and several useful catalysts have already
found a market. Because pollutants should be adsorbable on catalyst
surfaces, materials with a wide surface area and strong interactions
have been considered for the general removal of pollutants. Under
Visible Light illumination, photogenerated holes and electrons play
crucial roles, as depicted in Fig. 1 In the CB, the electrons travel to the
catalyst’s surface and react with oxygen to form active: O2- radicals, and
holes in VB react to the OH/H2O-absorbed species to form OH free
radicals. The h+ , O2-, and OH free radical species consequently have
an vital role in the degradation of pollutants. Over 16,000 scientific
papers, including the terms “photocatalyst” or “photocatalysis”, have
been published in the previous 10 years, and the number of publica-
tions is rising each year (Rueda-Marquez et al., 2020).

⁎
Corresponding authors.
E-mail addresses: darshnachem2004@gmail.com (D. Singh), ravigupta@kookmin.ac.kr (R. Gupta), laxmanresearcher84@gmail.com (L. Singh).

https://doi.org/10.1016/j.plana.2023.100031
Received 11 January 2023; Received in revised form 8 May 2023; Accepted 12 May 2023
2773-1111/© 2023 The Author(s). Published by Elsevier B.V. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
D. Singh, D.Anuradha, D. Mathur et al. Plant Nano Biology 4 (2023) 100031

Fig. 1. Photogenerated holes and electrons using Under Visible Light illumi- Fig. 2. Schematic representation of formation of ZnO nanoparticles.
nation.

Methylene blue dyes (AR grade) were procured from Sigma-Aldrich

Different photocatalysts like the CdS, Fe-TiO2, SrO2, WO3, ZnS, and
(USA). All of the chemicals used were not purified any further. The
ZnO are used. The most commonly used of these catalysts is the ZnO
experiments were carried out at optimum pressure and temperature
(Zinc Oxide), having high stability, economical and environmental
conditions. The reaction water used was distilled doubly.
friendliness. The photocatalyst’s performance is affected by several
factors that include surface area, size, pore structure, etc. In general,
nano-photocatalysts have large surface areas and high surface densities. 3. Method
Zinc oxide nanoparticles have been employed as photocatalysts for air
filtration and primarily for degradation. Compared to bare nano- 3.1. Preparation of beetroot extract
particles, when combined with other doped metals have shown better
efficiency (Schulte et al., 2019). To treat harmful gaseous effluent, Beetroots were purchased from the local market and washed with
biological and photocatalysis treatment should be combined using distilled water(DW) to remove dust particles. 20 g were grinded in a
nano-photocatalysts (Abdeltif et al., 2020b). beaker and boiled with distilled water (100 ml) for 15 min. Then we
The development of photocatalysts has stimulated much research on kept the beaker for cooling. After cooling, the solution was filtered to
using light-absorbing materials to remove pollutants. Photocatalysts remove the insoluble part. The wine-colored solution was then stored in
produce scavenger radicals like Superoxide (O2) and Hydroxyl Radicals glass bottles and kept in the fridge for further use.
(OH), which are produced from the water when it comes close to
photocatalyst. The organic pollutants are eventually attacked by these
3.2. Synthesis of zinc oxide nanoparticles
scavenging radicals and get broken down into less hazardous & simple
molecules. Organic pollutant mineralization would form only oxygen
To synthesise ZnO nanoparticles, 40 ml of beetroot extract was
and water molecules (Houas et al., 2001). It is reported that calcium
transferred in a 100 CC beaker having a magnetic bead. The solution
coating can enhance the properties of ZnO nanoparticles (Srivastava
was heated to 60 °C followed by the addition of 4.5 g of zinc acetate.
et al., 2014). In this work, ZnO NPs with or without calcium coating
The pH of the solution was maintained between 8.0. It was stirred at
were synthesized using beetroot extract, where beetroot acts as a re-
this temperature and pH for 40 min. till the formation of pale-yellow;
ducing agent. The nanoparticles hence resulted, were grouped ac-
the whole nanoparticles formation process is demonstrated in Fig. 2 The
cording to their surface functionality, size, crystallinity, and optical
resulting solution was centrifuged and washed using double distilled
behavior. Their dye degradation capacity was then studied using Rho-
water at 80 °C in the oven were dried. After that, the dried powder was
damine B (RhB) and methylene blue (MB).
calcinated at 450 °C in a muffle furnace for 15 min. The synthesized
In this paper, we propose the synthesis of blank (without calcium)
sample was named BRA and stored for further analysis.
and Ca-ZnO nanoparticles in the weight ratios (1:1 &1:2) for study of
their effective photocatalytic activity. For this we have chosen two
different dyes i.e., methylene blue and rhodamine. Our aim is to study 3.3. Synthesis of calcium coated zinc oxide nanoparticles
the effect of calcium coating on synthesized NPs for their photocatalytic
properties. Also, as we all know that calcium is a biodegradable ma- Ca-coated ZnO nanoparticles were synthesized by the coprecipita-
terial having no side effects, this further add advantage to these na- tion method. Zn (CH3COO)2. 2 H2O and calcium chloride were taken in
noparticles to be used for drug delivery purposes. Enhancing or de- 1:1 and 1:2 wt ratios (Singh et al., 2022). To synthesise Ca-ZnO nano-
creasing the efficiency will be helpful for us as we will use the particle particles, weighed Zn(CH3COO)2. 2 H2O was dissolved in 40 ml of
for sustainable release if coating decreases the efficiency. Beetroot extract at temperature 60 °C and pH app. 8, followed by
adding a weight amount of calcium chloride. It was stirred for 1 h and
then centrifuged to separate the nanocomposite. It was washed with
2. Materials doubly distilled water to remove impurities and dried. It was calcinated
at 450 °C in a muffle furnace for 15 min. The Ca-ZnO nanoparticles
Zn (CH3COO)2. 2 H2O (AR grade), calcium chloride (AR grade) and prepared in 1:1 and 1:2 ratio was named BRA1:1 and BRA 1:2, re-
NaOH (AR grade) were purchased from SRL (India). Rhodamine B & spectively. The samples were stored for further analysis.

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D. Singh, D.Anuradha, D. Mathur et al.
Fig. 3. The TEM, SEM and Elemental composition of the synthesized zinc oxide nanoparticles (a,c,e) and (b,d,f) for calcium-coated zinc oxide nanoparticles.
3.4. Characterization of (ZnO) and (Ca-ZnO) nanoparticles
The surface morphology of dried Zinc oxide NPs with or without
coating were determined using “Scanning Electron Microscopy” (SEM),
(JEOL JSM-6610LV) and “Transmission Electron Microscopy” (TEM).
For TEM, Aqueous dispersion of nanoparticles was drop-coated, soni-
cated and dried into the Formvar Coated 200Mesh copper grids (Ted
Pella, USA), and after this, the imaging was done with a TECNAI
G2–30 U TWIN TEM (FEI, Eindhoven, The Netherlands), with a 300 kV
acceleration voltage. This tool framework was used to probe
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Plant Nano Biology 4 (2023) 100031
nanoparticle’s elemental compositions (using EDX, Energy Dispersive X-
ray spectroscopy). High-Resolution powder XRD (X-ray diffraction) is
used for analysing the nano particles phase composition using the
Brukar D8 Discover X-ray spectrometer, with a 2ϴ range of 10–60º,
1.5 s/step scan rate, a 0.02 step-size, 40KV voltage, and 40milli-amp
current, using λ = 1.5406 A˚ Cu-Kα radiation. The FT-IR Spectra
(Fourier Transform Infrared Spectroscopy) were recorded for the na-
noparticles formed using the Perkin Elmer RX1 Tool. The optical
properties of the nanoparticles were estimated using a Shimadzu UV-
1601 spectrophotometer (Shimadzu, Kyoto, Japan). The photocatalytic
D. Singh, D.Anuradha, D. Mathur et al.
properties of zinc oxide (ZnO) and calcium-coated zinc oxide (Ca-ZnO)
nanocomposites with Methylene Blue (MB) and Rhodamine B (RhB)
were studied using a dye degradation experiment. Here, in an aqueous
medium, Zinc Oxide (ZnO) and calcium-coated zinc oxide (Ca-ZnO)
were treated with MB and RhB. First, the stock solution was formed by
mixing 10 mg of the respective dye in 1 L deionised water. Dried ZnO
and calcium-coated zinc oxide (1:1,1:2) were dispersed in a 100 ml
solution in a 250 ml conical flask on magnetic stir for 15 min. Control of
both dyes was also kept without the samples. Then, it was exposed to
sunlight until maximum, and 2 ml was taken every 10 min. to de-
termine the photocatalytic degradation activity. The Dye degradation
was measured with the absorbance spectrum of different supernatant
solutions using a UV-Visible spectrophotometer. The concentration of
the dye was calculated by observing the absorbance of methylene blue
at 660 nm and rhodamine B at app. 554 nm at different time intervals.
The photocatalytic properties of the synthesized samples were analysed
by photocatalytic degradation of RhB and MB. Photocatalytic experi-
ments were conducted under sunlight irradiation. MB solution was
photocatalytic decomposed using photocatalyst ZnO NPs (0.01 g) in
100 ml conical Flask with continuous stirring and under sunlight irra-
diation for achieving solution homogeneity. A UV-VI
Fig. 4. XRD-Spectra of reported ZnO nanoparticles (a) Synthesized zinc oxide nanoparticles (b) and (c, d) for calcium-coated zinc oxide nanoparticles (1:1,1:2).
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Plant Nano Biology 4 (2023) 100031
spectrophotometer was used to measure the absorbance.
4. Results and discussion
The uniform distribution of ZnO NPs can be estimated with SEM.
The SEM, TEM images and elemental composition of the synthesized
zinc oxide and calcium-coated zinc oxide nanoparticles are depicted in
Fig. 3(a, b, c, d, e and f, respectively). TEM imaging depicted that the
nanoparticle diameter varied in the 20–100 nano-meters range, with a
regular ‘Hexagonal shape.
Calcium coating into ZnO does not change the morphology of the ZnO-
Ca nanoparticles (Pham et al., 2020) EDX analysis results of nanoparticles
confirmed the elemental composition of the nanoparticles with oxygen
and zinc in the case of zinc oxide (ZnO) nanoparticles and calcium, zinc
and oxygen in the case of calcium-coated zinc oxide nanocomposites (Ca-
ZnO). The EDX data demonstrated the chlorine present because of con-
tamination from calcium salt used to coat ZnO nanoparticles. Powder XRD
patterns of Zinc oxide nanoparticles (ZnO) and calcium-coated zinc oxide
nanoparticles with 1:1 and 1:2 is shown in Fig. 4.
The XRD pattern of synthesized ZnO nano-powder is in the hex-
agonal phase. The stronger and sharper diffraction peaks were noted.
D. Singh, D.Anuradha, D. Mathur et al.
Fig. 5. FT-IR spectrum of reported zinc oxide nanoparticles (a) Synthesized zinc oxide nanoparticles (b) and (c,d) for calcium coated zinc oxide nanoparticles
(1:1,1:2).
Diffraction peaks on 31.75°, 34.40°, 36.23°, 47.53°, 56.59°, 62.85°,
66.39°, 67.94° and 69.07° 2θ are linked to planes at (100), (002), (101),
(102), (110), (103), (200), (112) and (201), respectively. All diffraction
peaks are indexed to the reported data of pure ZnO nanoparticles
(JCPDS file 79–2205). In the XRD pattern, there was no other impurity
detected (Manyasree et al., 2018). To detect several functional groups
connected to the generated nanoparticle, synthesized zinc oxide nano-
particles were subjected to FT-IR spectrum analysis. FT-IR spectrum of
zinc oxide nanoparticles synthesized was demonstrated in Fig. 5. Peaks
on 450 cm−1 (Gnanasangeetha and Sarala, 2013) and 603 cm−1 are
assigned to Zn-O stretching vibration (Ravichandrika et al., 2012).
Peaks on 1638 cm−1 and 3405 cm−1 are assigned to a carbonyl group
and hydroxyl groups in the beet root extract (Pham et al., 2020) that
impart stability to the nanoparticles.
For removing dye from the sewage water, Photocatalytic technology
acts as an important tool. Photocatalytic degradation characteristics of
synthesized ZnO nanoparticles with or without calcium coating with
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Plant Nano Biology 4 (2023) 100031
MB and RhB Dye solutions were estimated under natural sunlight. The
electrons produced under sunlight excited to CB (Conduction Band)
from VB (Valence Band) by creating holes in the VB.
Photocatalytic reactions involve the generation of electron-hole
pairs by semiconductor NPs. On irradiation to sunlight, photons can
excite the electrons to CB (Conduction Band) from VB (Valence Band)
to produce electrons and holes. These species then generate oxidizing
agents such H2O2, O2-, and OH free radicals after interaction with
oxygen and water on the surface of the ZnO. These strong oxidizing
agents can decompose organic dyes into carbon dioxide and water
(AlHamedi. et al., 2009; Muruganandham. and Swaminathan., 2006).
Generally, electrons and holes produced on the photocatalyst’s surface
react with O2 and H2O molecules to form reactive O2 species and hy-
droxyl free radicals. Hence, the produced oxidizing species led to the
oxidation of organic pollutants. Usually, the photocatalytic activity of
ZnO nanomaterials is related to the total number electron and hole pair
produced (Liang. et al., 2020; Ngoepe et al., 2018; Meena et al. 2016).
D. Singh, D.Anuradha, D. Mathur et al.
Fig. 6. (a), (b) and (c) show absorption spectra of blank ZnO and calcium coated ZnO NPs (1:1,1:2) with methylene blue, (d) Summary data by irradiation with
natural Sunlight.
Figs. 6(a), 6(b) & 6(c) represent a decrease in optical density (OD) at
625 nm (absorption maximum for Methylene Blue) as a function of the
time of exposure to sunlight for blank ZnO nanoparticles, Ca+ZnO (1:1)
and Ca+ZnO (1:2) NPs, respectively. Data has been compiled in Fig. 6(d)
for better understanding. The plots for ZnO and Ca+ZnO (1:1 and 1:2)
NPs show a considerable reduction in optical density (owing to photo-
bleaching of MB by singlet oxygen with the time of sunlight exposure,
indicating the effective generation of singlet oxygen. Free ZnO nano-
particles show appreciable results i.e., 100 % degradation was obtained
after 40 min. while in the case of calcium-coated nanoparticles nearly 90
% degradation was observed after 60 min. This experiment shows the
sustainable release of photocatalysts, indicating that we can alter the result
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Plant Nano Biology 4 (2023) 100031
when ZnO particles are coated with calcium, as shown in Fig. 6(b, c).
The same type of analysis on MB dye photocatalytic degradation
with ZnO-Camellia sinensis NPs also resulted in nearly 80 % degrada-
tion in 120 min. (Nava et al., 2017; Alamdari. et al., 2020). The same
experiment was done with Rhodamine that B showed a decrease in OD
(Optical Density) at 554 nm (absorption maximum for Rhodamine B) as
a function of the time of sunlight exposure for blank ZnO nanoparticles,
Ca+ZnO (1:1) and Ca+ZnO (1:2) nanoparticles, respectively shown in
Fig. 7. The degradation efficiency of the rhodamine B dye by synthe-
sized ZnO NPs increased with time, and its calcium-coated nano-
particles showed almost no release. Combined data for all the 3 graphs
are shown in Fig. 7(d).
D. Singh, D.Anuradha, D. Mathur et al. Plant Nano Biology 4 (2023) 100031

Fig. 7. (a), (b) and (c) show absorption spectra of blank ZnO and calcium coated ZnO nanoparticles (1:1,1:2) with Rhodamine B, (d) Summary data by irradiation
with natural Sunlight.

5. Conclusion CRediT authorship contribution statement

The present research work showed that the synthesized blank ZnO Darshan Singh-Proposed, designed and performed the experiments,
NPs and calcium coated ZnO NPs (1:1 &1:2) act as good photocatalysts. Anuradha-Collected Data, Divya Mathur-Contributed data analysis,
Degradation efficiency for ZnO NPs with methylene blue is 100 % after Surendra Kumar- Contributed data analysis, Balaram Pani-Performed
40 min and for Ca-ZnO (1:1, 1:2) in the range of 80–90 % after 60 min. the analysis, Amar Kumar- contributed in data analysis, and Rajni
Coating of calcium on the surface of ZnO NPs decreases the degradation Kanojia-wrote and Ravi Gupta & Laxman Singh-Investigate and revised.
of methylene blue. While in case of rhodamine B dye, blank ZnO na-
noparticles show 55.2 %. While coated with calcium shows 5.4 % in
Data availability
case of 1:1 ratio and 3.4 % in 1:2 ratio. Blank ZnO nanoparticles show
good degradation with methylene blue, while coated one shows a sus-
Data will be made available on request.
tainable reaction. Rhodamine B dye degradation with blank ZnO na-
noparticles show good activity; while coatedones shows almost no re-
lease. These results indicated the potential application of ZnO Declaration of Competing Interest
nanoparticles with or without the calcium coating in degrading toxic
organic compounds from wastewater. The authors declare that they have no conflict of interest.

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D. Singh, D.Anuradha, D. Mathur et al. Plant Nano Biology 4 (2023) 100031

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Mota-González, M.L., et al., 2017. Influence of Camellia sinensis extract on zinc oxide
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