Journal of Dispersion Science and Technology

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Aloe-vera leaf extract as a green agent for the

synthesis of CuO nanoparticles inactivating
bacterial pathogens and dye

Saurabh Sharma & Kuldeep Kumar

To cite this article: Saurabh Sharma & Kuldeep Kumar (2020): Aloe-vera leaf extract as a green
agent for the synthesis of CuO nanoparticles inactivating bacterial pathogens and dye, Journal of
Dispersion Science and Technology, DOI: 10.1080/01932691.2020.1791719

To link to this article: https://doi.org/10.1080/01932691.2020.1791719

Published online: 20 Jul 2020.

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JOURNAL OF DISPERSION SCIENCE AND TECHNOLOGY
https://doi.org/10.1080/01932691.2020.1791719

Aloe-vera leaf extract as a green agent for the synthesis of CuO nanoparticles
inactivating bacterial pathogens and dye
Saurabh Sharma and Kuldeep Kumar
Department of Chemistry, Career Point University, Hamirpur, H.P., India

ABSTRACT ARTICLE HISTORY

The present work describes the aloe-vera, as a green agent for the facile synthesis of copper oxide Received 5 February 2020
NPs and their deliberation for antibacterial and photocatalytic activities. The synthesis has been Accepted 13 June 2020
carried out at pH 11.5 and 70
C with detailed characterization investigation through UV-spectros-
KEYWORDS
copy, XRD (X-ray Diffraction), SEM (Scanning electron microscope), EDX (Energy-dispersive X-ray
Aloe-vera; antibacterial
spectroscopy) and TEM (Transmission electron microscope) techniques. The study reveals the activity; copper oxide
altered morphology of CuO NPs with a change in concentration of precursor. After characteriza- nanoparticle; photo-catalyst;
tion, CuO NPs with size 5–20 nm have been examined against pathogenic bacteria like Bacillus transmission electron
subtilis, Staphylococcus aureus, and Escherichia coli to calculate the zone of inhibition, MIC (min- microscope
imum inhibitory concentration) and MBC (minimum bactericidal concentration). Photo-catalytic
degradation has been performed in the presence of H2O2, which results in the degradation of MO
(methyl orange) dye up to 96% in 24 min under exposure of UV light.

GRAPHICAL ABSTRACT

Introduction by many researchers in the past two decades due to special

features including optical/electrical properties. From centu-
The efficiency of nanoparticles (NPs) over bulk material
ries, copper and its compounds have been used as disinfec-
focuses, the recent research toward nanoscience and nano-
tants due to their antibacterial as well as antiviral properties.
technology. The various novel metal oxide NPs like Co, Ni,
From the ancient time, the copper utensils have been used
Cu, Zn, Ag, Au, etc are the for-front in the research. These to store water for drinking purposes.[3]
metal NPs are capable to stand under harsh conditions and For the treatment of dye-bearing wastewater, various
are comparatively safe to humankind. In this framework, the physical and chemical methods, such as oxidation, adsorp-
copper oxide NPs are known for their various biomedical tion, ion exchange, UV radiation, ozonization, and so on,
and industrial applications. The CuO NPs are particularly have been used in the past few decades.[4] However, these
attractive because of copper’s high natural abundance, low processes face certain limitations including secondary pollu-
cost, and straight forward multiple ways of preparing Cu- tion problems, cost effectiveness, and low adsorption cap-
based nanomaterials.[1] The low-cost CuO NPs are known acity. Ultimately, researchers focused on advanced oxidation
for effective antifungal and antimicrobial properties related processes like Fenton process, biodegradation, and heteroge-
to the high-cost silver and gold NPs. Yoon et al.[2] have neous photo-catalysis as an efficient tool for degradation of
demonstrated the highest susceptibility for the copper NPs organic dyes.[4] Fenton reaction catalyzed by Fe2þ has been
(0.0734 mL/lg) in comparison to silver NPs (0.0236 mL/lg) used by Youssef et al.[5] for the oxidation of 97.8% methyl
with the pathogen E. coli. Copper NPs have been considered orange (MO) dye. However, in heterogeneous photo-

CONTACT Kuldeep Kumar kuldeep.sharma.753@gmail.com; kuldeep.che@cpuh.edu.in Department of Chemistry, Career Point University, Hamirpur, H.P.
176041, India
ß 2020 Taylor & Francis Group, LLC
2 S. SHARMA AND K. KUMAR

catalysis, which involves a semiconductor photo-catalyst, the for N-arylation of nitrogen-containing heterocycles with aryl
CuO NPs can be employed as photo-catalyst for aqueous- halides. In another study on green synthesis, stable Pd/CuO
phase photo degradation of organic dyes due to their envir- NPs have been synthesized by using Theobroma cacao L.
onmental-friendly benefits.[6] In this direction, Sinha and seed extract without adding any outside capping agent.[30]
Ahmaruzzaman,[7] have reported the effective degradation of These particles were found as a separable and reusable cata-
MO dye in the presence Cu NPs as the photo-catalyst in lyst for the cyanation of a range of aryl halides.
the sunlight. Consequently, the green methods being environment
Nanomaterials fabricated from earth-abundant and inexpen- friendly, cost-effective, and novel route for the synthesis of NPs,
sive metals have attracted considerable attention because of may be a prospective commercial alternate for large-scale pro-
their potential as viable alternatives to the rare and expensive duction of the nanomaterial.[31] Furthermore, the aloe-vera, a
novel-metals, catalysts used in many conventional commercial rich source of a large number of phytochemicals, has been
chemical processes.[8,9] Recently, Nasrollahzadeh et al.[10] have known to hold a large number of biological applications like
presented a review study on biosynthesis and its future chal- UV protective, immunomodulatory, anti-inflammatory, and
lenges in case of Ag-based nanocatalysts. Further, the catalytic wound- and burn-healing promoting properties. The phyto- or
activities of these nanocatalysts in numerous chemical reactions bio-chemicals present in the aloe-vera leaves are found to be
like cyanation, oxidation, cycloaddition, reduction, and coupling effective in the reduction of metal ions and, therefore, used suc-
reactions have also been discussed. A more efficient, econom- cessfully to synthesize the transition metal (gold, silver, zinc,
ical, and reusable Cu NPs,[11] magnetic Cu/Fe3O4 NPs,[12] Cu/ titanium, copper, etc.) NPs.[32–37] Therefore, in continuation of
reduced graphene oxide/Fe3O4 nanocomposite,[13] CuO NPs,[14] our previous study on green synthesis of ZnO nanoparticles,[38]
Cu/ZnO NPs,[15] and CuO/ZnO nanocomposite[16] have been at present, an attempt has been made to use aloe-vera leaf
reported for the catalytic reduction of the variety of chemicals extract as a green mediator for the synthesis of CuO nanopar-
like nitroarenes, 4-nitrophenol, rhodamine B, etc. ticles. The so-obtained CuO nanoparticles have been character-
The various synthesis strategies and reaction parameters ized and employed for inactivating bacterial pathogens and dye
significantly affect the chemical and physical properties of degradation.
the NPs. The use of mixing, doping, and alloy capping/stabi-
lizing ligand techniques also significantly contribute to alter- Experimental details
ing the properties of NPs.[1] In order to synthesize the metal
NPs, numerous methods have been developed, dominantly Materials
divided into three classes, physical, chemical, and biological
The deionized distilled water with a conductivity of 2–3 106
methods.[17] The chemical methods involve the use of envir-
scm1 and a pH of 6.8–7.0 (at 298.15 K) was used for all experi-
onmentally toxic and biologically hazardous substances as
ments. Copper acetate mono-hydrate (Cu(CH3COO)2H2O),
well as high temperature to synthesize the stable NPs.[18]
potassium hydroxide (KOH), and methyl orange (MO) dye
This demands the eco-friendly/cost-effective methods and,
were obtained from Merck Pvt. Ltd. and are used as such with-
hence, augmented the interest of the researchers to minimize
out any further purification. Microbes, Bacillus subtilis (MTCC
the use of chemicals with the insinuation of the fundamental
441), Staphylococcus aureus (MTCC 737), and Escherichia coli
principles of green chemistry. In this context, the last two
(MTCC 739) were obtained from the CSIR-IMTECH,
decades witnessed a lot of work on the green synthesis of Chandigarh in the freeze-dried ampoule. Aloe-vera plant leaves
NPs, especially, on silver based nano catalysts.[10] In the were collected from the nearby area of Career Point University
green methods, the biological material from different parts Hamirpur (HP).
of the plant like stem, leaf, flower, seeds, fruit, etc. act as the
reducing, oxidizing, and stabilizing agents in the absence of
hazardous chemicals and capping agents.[19,20] As synthe- Methods
sized metal oxide NPs are found to be more useful in the
Preparation of the aloe-vera leaf extract
field of catalysis industries, particularly for the degradation
Fresh aloe-vera leaves were collected, weighed (25 gm), and
of hazardous environmental pollutants.[21–24] However, in
washed with tap water as well as with distilled water to
some cases, a blend of green and conventional methods like
remove the surface impurities. These leaves were then
the combustion, sol-gel, and precipitation are used to syn-
crushed and boiled for about 15 min in distilled water
thesize novel metal oxide nanostructures.[25–28]
(100 ml). After that extract was filtered through Whatman
In the literature, the leaves of Thymus vulgaris L. have
No. 1 filter paper and stored in the refrigerator for further
been successfully used as reducing and stabilizing agents in
use in the synthesis of CuO NPs.[38]
the green synthesis of CuO NPs of size less than 30 nm.[29]
The so synthesized CuO NPs was then used as an excellent
heterogeneous catalyst for N-arylation of indoles and amines Preparation of CuO NPs
with good yield. This catalyst was reported to be recycled The 5, 10, and 50 mmolkg1 aqueous solutions of copper
and reused up to five times with almost no loss in the activ- acetate monohydrate were prepared in the distilled water.
ity. Further, Nasrollahzadeh et al.[14] have first time reported The aloe-vera leaf extract was added dropwise to these solu-
the Tamarix gallica leaf extract mediated synthesis of CuO tions (5 ml/min) in a round bottom flask under constant
NPs. The particles were then examined as recyclable catalyst stirring at 70
C. About 15 min after this addition, freshly
 JOURNAL OF DISPERSION SCIENCE AND TECHNOLOGY 3

Figure 1. Biosynthesis, characterization and applications of CuO NPs using aloe-vera leaf extract.

prepared 0.2 molkg1 KOH solution was added dropwise to

the reaction mixture to maintain the pH ¼ 11.5 (monitored
with the digital pH meter). During the reaction time, the
formation of CuO NPs was confirmed by the appearance of
brownish to black color precipitates in the solution mixture.
After stirring for 4 h, the CuO NPs were collected and Figure 2. Chemical structure of methyl orange (MO) dye.
washed with distilled water/ethanol. These NPs were dried
in a hot air oven at 55–60
C before further use.
Photocatalytic degradation of dye
To investigate the photocatalytic activity of the synthesized
Characterization of CuO NPs CuO NPs, 100 ml of 10 ppm MO (Figure 2) solution has
been treated with 5, 15, and 25 mg of CuO NPs catalytic
The synthesized CuO NPs have been confirmed from the load. The reaction was monitored in the presence of the UV
UV–visible spectra, which show a characteristic peak of light with stirring up to the dye de-coloration with the add-
absorbance. X-ray powder diffraction (XRD) has been used ition of 1 ml H2O2 solution (30%). Before exposing to the
to study the crystallinity, phase identification, and purity of UV light the reaction mixture was stirred in the dark for
the NPs. A field emission-scanning electron microscope 30 min to obtain adsorption-desorption equilibrium between
(FE-SEM) was used for the morphological study with the organic molecules and the catalyst. The decomposition of
attached SEM-EDX for the elemental detection analysis of MO was examined by evaluating the change in absorbance
the synthesized NPs. Transmission electron microscopy intensity in the UV–Visible spectrophotometer Cary 100 Bio
(TEM) is used for the determination of the shape and size at k ¼ 465 nm.[40]
of the CuO NPs. Figure 1 illustrates the biosynthesis, char-
acterization and applications of CuO NPs. Results and discussion
UV–visible analysis

Antimicrobial activity The optical properties of obtained CuO NPs have been
investigated by carrying out UV–visible spectroscopic studies
The inactivation effect of obtained CuO NPs over three in the range of 200–800 nm. The NPs were dissolved in
pathogens, Bacillus subtilis, Staphylococcus aureus, and methanol with the help of a sonicator to prepare a clear sus-
Escherichia coli has been investigated by using well diffusion pension. The kmax values of UV–visible spectra presented in
assay (NCCLS).[39] The zones of inhibition, minimum Figure 3 at different concentrations of Cu(CH3COO)2H2O
inhibitory concentration (MIC) and minimum bactericidal (5, 10, and 50 mmolkg1) are in contest with the values
concentration (MBC) of so synthesized CuO nanoparticles reported in the literature.[41] The kmax values given in table
were measured as per the detailed procedure given in our 1 are used to calculate the direct optical energy band gap
previous study on ZnO NPs.[38] (Eg) with the following relation (1).
4 S. SHARMA AND K. KUMAR

Figure 4. XRD pattern of CuO NPs at different concentrations of

Cu(CH3COO)2H2O. (a) 5, (b) 10 and (c) 50 mmolkg1.

XRD analysis
X-ray diffraction analysis has been carried out to validate
Figure 3. UV–visible spectra of CuO NPs at different concentrations of
the crystalline nature, phase purity, and formation of the
Cu(CH3COO)2H2O. (a) 5, (b) 10 and (c) 50 mmolkg1. monoclinic crystal system of the synthesized CuO NPs. XRD
patterns (Figure 4) show peaks at angle 2h  32.5, 35.5, 38.7,
Table 1. The kmax and Eg values for the synthesized CuO NPs at different con- 48.7, 53.4, 58, 61.5, 66.2 and 67.9 degree. These peaks cor-
centrations of Cu(CH3COO)2H2O. respond to the ð
110Þ, ð002Þ, ð200Þ, ð
202Þ, ð020Þ, ð202Þ,
a
m (mmolkg1) kmax (nm) Eg (eV) ð
113Þ, ð
311Þ, and ð113Þ crystal planes, respectively, of CuO
5 368 3.4 NPs matched with JCPDS file no. 45-0937. Similar planes
10 372 3.3
50 384 3.2
reported by Dhineshbabu et al.[43] for the CuO NPs. In all
the spectra, no extra peaks have been seen, which confirm
a
m is the molality of Cu(CH3COO)2H2O in water.
the phase purity for all the synthesized CuO NPs. Further,
hc the XRD technique also applied to investigate the structural
Eg ¼ [1] arrangement of the particles and the size of the unit cell of
k
the compound. Clear and sharp peaks at the definite posi-
tions explain the pure monoclinic crystal phase, and broad-
where, h is Plank’s constant ¼ 6.626  1034 Js, C is velocity
ening of the peaks indicates the smaller particle size of the
of light ¼ 3  108 ms1 and k corresponds to the wavelength
synthesized CuO NPs. The crystallite size (D) is determined
of the peak with maximum intensity.[42] The Eg values are by taking the full-width half maxima (FWHM) for the two
tabulated in Table 1. adjacent most intense peaks in the spectra (Figure 4) using
The band gap values are found to fall in the range of the Debye–Scherrer Equation (2).[46] Inter-planer spacing
3.2–3.4 eV, which are almost similar to the values reported (d), lattice parameters (a, b, c), and volume for the mono-
by Udayabhanu et al.[41] and Dhineshbabu et al.[43] clinic unit cell (V) have been calculated by using the Bragg’s
However, the slight difference between the values may be Equation (3), (4), and (5), respectively.[47,48] All these
due to the change in the shape or size of the CuO NPs syn- parameters are summarized in Table 2.
thesized at different concentrations of metal salt used as a
0:94k
precursor. These large band gap values as compared to the D¼ [2]
b cos h
bulk material may be attributed to a decrease in the size of
the particles.[38,44] Further, the Eg values decrease with the nk ¼ 2d sin h [3]
concentration of the precursor, which again can be attrib-  
1 1 h2 k2 l2 2hl cos b
uted to the small size of NPs synthesized at low concentra- ¼ þ þ sin 2
b þ  [4]
tion of the precursor.[38,45]
2
dhkl sin 2 b a2 b2 c2 ac
 JOURNAL OF DISPERSION SCIENCE AND TECHNOLOGY 5

Table 2. Crystallite size (D), inter-planer spacing (d), lattice parameters (a ¼ b V ¼ abc sin b [5]
and c) and volume (V) calculated from XRD measurements at different concen-
trations of Cu(CH3COO)2H2O. where, k ¼ 1.5406 Å, b (in D calculation) ¼ full width at
a
m (mmolkg1) D (nm) d (nm) a (nm) b (nm) c (nm) V (nm3) half maximum (FWHM), h ¼ Bragg’s angle, (h, k, l) ¼
5 4.64 0.242 0.470 0.341 0.512 8.206 Miller indices, dhkl ¼ inter-planar spacing, bo ¼ 98.54
10 6.91 0.242 0.471 0.340 0.509 8.147 degree[49], V ¼ volume of the unit cell, and (a, b, c) ¼ lat-
50 7.28 0.242 0.473 0.341 0.510 8.192
a tice parameters for the monoclinic system of CuO NPs.
m is the molality of copper acetate mono-hydrate in water.

Figure 5. SEM/EDX spectra of CuO NPs at concentrations of Cu(CH3COO)2H2O. (a) 5,(b) 10 and (c) 50 mmolkg1.
6 S. SHARMA AND K. KUMAR

10 mmolkg1 of Cu(CH3COO)2H2O, the lattice parameters

‘b’ and ‘c’ are smaller, which further results in the V value
for monoclinic CuO. The lattice parameter ‘a’ subsequently
increases with the increase in the concentration of
Cu(CH3COO)2H2O.

SEM/EDX analysis
As illustrated in Figure 5, the SEM images show that the
shape of synthesized CuO NPs is in capsule and spherical
fluffy like arrangement.
EDX spectroscopy is applied to enumerate the elemental
composition of the synthesized NPs. The peaks in the EDX
spectra illustrated in Figure 5 confirm the presence of Cu,
O, and C in all the synthesized CuO NPs. However, some
other peaks corresponding to Au have been obtained due to
the gold coating used for SEM/EDX micrograph analysis.
The peaks corresponding to carbon may be attributed to the
carbon tape used in SEM/EDX analysis. The EDX spectral
images can be computed with the help of peaks assigned by
Khan et al.[50] have explained Cu and O elemental peak’s
position in EDX spectra for the cubic shaped CuO NPs.
Rajesh et al.,[51] have decorated the spherical shaped par-
ticles using the sodium borohydride and poly vinyl alcohol,
which are also reported in the present results of SEM/
EDX spectra.

TEM analysis
The TEM microscope used to study the shape and size of the
nanometer size particles. Figure 6 represents the TEM images
of CuO NPs synthesized at different concentrations of
Cu(CH3COO)2H2O. The particles were analyzed by image
processing software tool, Image J for the size calculation. From
the analysis of TEM images, it can be concluded that the shape
of CuO NPs varies with Cu(CH3COO)2H2O concentration. At
5 mmolkg1 Cu(CH3COO)2H2O the synthesized NPs are
spherical in shape. At 10 mmolkg1 Cu(CH3COO)2H2O, the
NPs are small rod/capsule in shapes that are arranged/aggre-
gated in the wheat grain like morphological arrangement. At
50 mmolkg1 Cu(CH3COO)2H2O particles are mostly spher-
ical in shapes that are found to aggregate in an irregular man-
ner. The examination of TEM images shows that a layer has
capped the CuO NPs. From TEM image analysis, the size of
approximately 5–9 nm has been obtained. It further substanti-
ates the particle size obtained from the calculations of the
Debye–Scherrer equation using XRD patterns. The size of the
particles arranged in the wheat grain or elliptical shapes at
10 mmolkg1 of Cu(CH3COO)2H2O are with the width/length
Figure 6. TEM images of CuO NPs at different concentrations of of 45/95 nm.
Cu(CH3COO)2H2O. (a) 5, (b) 10 and (c) 50 mmolkg1.
Gunalan et al.[36] have reported the spherical shape CuO
NPs prepared using varying amounts of aloe-vera extract
and a fixed concentration of copper sulfate with particle size
The values of crystallite size, volume, and lattice parame- ranging from 15 to 30 nm. It concludes that the concentra-
ters (Table 2) are in close agreement with the values tion of aloe-vera extract did not change shape, but affected
reported by Singh and Bedi in case of CuO NPs.[48] The the size of NPs. Kumar et al.[52] have also synthesized the
crystallite size (D) increases with [Cu(CH3COO)2H2O] at spherical shaped CuO NPs using aloe-vera and copper
constant inter-planer spacing (d) values. Moreover, at nitrate with an average size of 20 nm. Yugandhar et al.[53]
 JOURNAL OF DISPERSION SCIENCE AND TECHNOLOGY 7

Figure 7. Antibacterial activity of CuO NPs at different concentrations of Cu(CH3COO)2H2O. (i) 5, (ii) 10 and (iii) 50 mmolkg1 against (a–c) Bacillus subtilis, (d–f)
Staphylococcus aureus and (g–i) Escherichia coli, respectively.

reported the synthesis of CuO NPs from the Syzygium alter- Antimicrobial activity
nifolium plant with size 8.8 nm and spherical shape. In
In order to examine the antibacterial efficiency of the syn-
another work, Zhu et al.,[54] have carried out by the synthe-
thesized NPs, agar well diffusion method is applied.[39] The
sis of CuO NPs by precipitation method at 100
C and
activity of the CuO NPs against the Bacillus subtilis,
obtained the spherical-shaped particles.
8 S. SHARMA AND K. KUMAR

Table 3. The zones of inhibition of CuO NPs at different concentrations of Cu(CH3COO)2H2O.

Bacteria a
m (mmolkg1) NPs dosage (mg/ml) Zone of inhibition (mm)
Bacillus subtilis 5 10 15.96 ± 0.82b
25 22.20.75
50 26.0 ± 0.35
10 10 18.33 ± 0.33
25 20.73 ± 0.47
50 25.23 ± 0.39
50 10 18.83 ± 0.17
25 23.23 ± 0.39
50 24.83 ± 0.44
Cipro-floxacin Cipro-floxacin 0.1 mg/ml 35 ± 1
Staphylococcus aureus 5 10 6.10 ± 0.72
25 7 ± 0.25
50 13 ± 0.58
10 10 6.82 ± 0.21
25 8.67 ± 0.33
50 11.5 ± 0.29
50 10 7.12 ± 0.1
25 10 ± 0.76
50 13.88 ± 0.48
Cipro-floxacin Cipro-floxacin 0.1 mg/ml 30 ± 1.2
Escherichia coli 5 10 5.89 ± 0.89
25 6.43 ± 0.72
50 11.76 ± 0.46
10 10 7±0
25 11 ± 0.58
50 12.33 ± 0.67
50 10 10.27 ± 0.39
25 11.3 ± 0.67
50 14.7 ± 0.91
Cipro-floxacin Cipro-floxacin 0.1 mg/ml 33 ± 0.7
a
m is the molality of zinc acetate di-hydrate in water.
b
Standard error in measurement of zone of inhibition.

Table 4. Minimum inhibition concentration (MIC) and minimum bactericidal

concentration (MBC) values of CuO NPs synthesized at different concentrations
of Cu(CH3COO)2H2O.
Bacteria a
m (mmolkg1) MIC (mg/mL) MBC (mg/mL)
Bacillus subtilis 5 98 1562
10 391 1562
50 391 1562
Cipro-floxacin <1 <0.5
Staphylococcus aureus 5 391 3125
10 781 1562
50 391 781
Cipro-floxacin 16.65 1
Escherichia coli 5 391 1562
10 391 1562
50 391 781
Cipro-floxacin <1 0.5
a
m is the molality of copper acetate mono-hydrate in water.

Staphylococcus aureus, and Escherichia coli has been investi-

gated on the Mueller Hinton agar (MHA). The zones of Figure 8. A representative plot of UV–visible absorption spectra of methyl
inhibition of CuO NPs, as illustrated in Figure 7, have been orange dye in the presence of 25 mg CuO NPs synthesized at 10 mmolkg1
[Cu(CH3COO)2H2O].
measured for each of the bacteria and summarized in Table
3. From this table, it can be seen that the largest zone of
inhibition for all the synthesized CuO NPs has been synthesized at 50 mmolkg1 CuSO45H2O. The possible
obtained generally for Bacillus subtilis, which is followed by cause for this may be the variable shape/size of these NPs,
Escherichia coli and Staphylococcus aureus. which are aggregated in an irregular manner. Whereas at
Biological activity with respect to the microbes has been other concentrations of CuSO45H2O, the obtained particles
evaluated for 10, 25, and 50 mg/ml CuO NPs in methanol are of uniform shape/size. The values of MIC and MBC cal-
solvent. The 30 mL of the solution of CuO NPs was loaded culated for the CuO NPs by using the 96th well microtiter
into the wells by taking the methanol and Ciprofloxacin plate are tabulated in Table 4. Moreover, the MBC values of
(100 mg/mL w/v) as a negative and positive control, respect- the CuO NPs synthesized at 50 mmolkg1 CuSO45H2O for
ively. The best results can be seen for 10 mg/ml of NPs Escherichia coli and Staphylococcus aureus, are at the lowest.
 JOURNAL OF DISPERSION SCIENCE AND TECHNOLOGY 9

Table 5. Percentage of photocatalytic degradation of 100 ml of 10 ppm MO

dye solution at different dosages of CuO NPs.
Percentage of photo-catalytic
a
m (mmolkg1) NPs dosage (mg) degradation of MO Dye (%)
5 5 89.5
15 91.3
25 93.0
10 5 86.9
15 93.4
25 96.1
50 5 86.7
15 90.1
25 91.3
a
m is the molality of copper acetate mono-hydrate in water.

method. In a work reported by Ananth et al.[56] it has been

observed that plate-like CuO NPs are more efficient for anti-
bacterial activities in comparison to grain or needle-shaped
NPs against the Streptococcus iniae, Streptococcus parauberis,
Escherichia coli, and Vibrio anguillarum. Similarly, in the
present study, we have observed the shape-dependent anti-
bacterial efficiency of CuO NPs.

Photocatalytic degradation of methyl orange dye

In order to examine the photocatalytic efficiency of the syn-
thesized CuO NPs have been used as photo-catalyst for deg-
radation of 10 ppm MO dye solution in the presence of
H2O2 and UV light.[57] The decrease in the intensity of the
absorbance peak at 465 nm (Figure 8) was measured in a
UV spectrophotometer to examine the degradation of dye
with the passage of time.
The UV light plays an efficient role in the degradation of
the dye and the degradation can easily be examined by the
decolorization of the dye solution in the presence of NPs.
The degradation follows the hydroxyl ion mechanism.[58] In
the presence of the H2O2, the degradation increases at an
alarming rate, and approximately 90% of the dye degrad-
ation has been observed in only 24 min. Possibly, the add-
ition of H2O2 results in a large number of OH free radicals
that further result in the electron-hole pairs generation on
the surface of the CuO nanoparticle photo-catalysts. Similar
approximations are reported by Zhang et al.[59] on the effect
of H2O2 in the case of graphene decorated CuO photo-cata-
lyst for the methyl orange dye degradation. The percentage
degradation of dye was calculated from the following
relation.
Ao  At
% Degradation ¼  100 [6]
Ao
where, Ao ¼ Absorbance of pure dye solution and At ¼
Absorbance of the reaction mixture at a time t.
The photocatalytic activities have been investigated at 5,
15, and 25 mg/100 ml of CuO NPs. The catalytic efficiency
Figure 9. Plot for extent of decomposition of 10 ppm MO dye with the passage
of time in the presence of CuO NPs (5, 10, and 25 mg) at different concentra- of CuO NPs has been evaluated by plotting the extent of
tions of Cu(CH3COO)2H2O. (a) 5, (b) 10 and (c) 50 mmolkg1. decomposition (At/Ao) versus reaction time (Figure 9). From
Table 5 and Figure 10, it can be concluded that the percent-
age degradation rate increases slightly with the dosage of
The MIC and MBC values vary between 98 to 781 mg/mL NPs, and the shape and size arrangement of NPs affected
and 781 to 3125 mg/mL, respectively. Similar results have the results specifically. Comparatively, the rod-shaped par-
been reported by Kronvall[55] using the disk diffusion ticles having the spheroidal and parabolic arrangement
10 S. SHARMA AND K. KUMAR

Figure 11. Representative plot of photodegradation kinetics of 10 ppm MO dye

with the passage of time in the presence of 25 mg CuO NPs synthesized at dif-
ferent concentrations of Cu(CH3COO)2H2O (5, 10 and 50 mmolkg1).

et al.[60] have pointed out the same effect of concentration

of nano-catalyst in the degradation of the methylene blue
(MB) dye. Liang et al.[57] reported the 95% degradation of
MB dye in 65 min in the presence of Cu/CuO NPs. They
have also decorated the reusability of the CuO photo-catalyst
for a series of reactions, with nearly the same degradation
rate.[57] In another study, Wang et al.[58] discussed the deg-
radation of rhodamine B in the presence of CuO NPs and
the sun-light. They obtained only 7.4% rhodamine B degrad-
ation in the presence of H2O2, whereas with the addition of
the CuO NPs catalyst 93.4% rhodamine B degradation was
obtained. Sharma et al.[61] have reported the ZnO catalyzed
degradation of the MO in 180 min with maximum degrad-
ation of 87.9% at 50 mg dosage of catalyst. Youssef et al.[5]
have reported 77.01, 86.25, and 90.12% removal of the MO
dye with the use of 1.0  104, 1.3  104, and 1.9  104 M
Fe2þ catalyst in a Fenton reaction.

Kinetics study of the photocatalytic dye degradation

The kinetics of the dye degradation reaction has been
studied by considering the pseudo-first-order reaction
kinetics:

ln ðAo =AÞ ¼ ln ðCo =CÞ ¼ kt (7)

where Co is the initial concentration of the dye having

absorbance Ao, C is the dye concentration at time t, having
Figure 10. Percentage photo degradation of 10 ppm MO dye with the passage absorbance A and k is the pseudo-first-order rate constant.
of time in the presence of CuO NPs (5, 10, and 25 mg) at different concentra-
tions of Cu(CH3COO)2H2O. (a) 5, (b) 10 and (c) 50 mmolkg1.
Figure 11 depicts a linear relationship between ln Ao/A
and time t. This clearly demonstrates the pseudo-first-order
reaction kinetics.[60] The calculated value of pseudo-first-
order rate constant k was found to be minimum
synthesized at the 10 mmolkg1 Cu(CH3COO)2H2O pro- (0.0827 min1) and maximum (0.1313 min1) for CuO NPs
vide better results. This may be due to the reason that a synthesized at 50 and 10 mmolkg1 Cu(CH3COO)2H2O
high interstitial space has been provided which is believed to with 5 and 25 mg catalytic load, respectively. Further, the
be responsible for higher degradation of MO dye. Farrouji correlation constant for the fitted lines was found to be
 JOURNAL OF DISPERSION SCIENCE AND TECHNOLOGY 11

R2 ¼ 0.94539 and 0.90886, respectively, for the above men- Green Synthesis of Ag-Based Nanoparticles for Environmental
tioned two cases.[62] and Catalytic Applications. Chem. Rec. 2019, 19, 2436–2479.
DOI: 10.1002/tcr.201800202.
[11] Nasrollahzadeh, M.; Sajadi, S. M.; Hatamifard, A. Anthemis
xylopoda Flowers Aqueous Extract Assisted in Situ Green
Conclusions
Synthesis of Cu Nanoparticles Supported on Natural Natrolite
The present attempt of eco-friendly synthesis reports the Zeolite for N-Formylation of Amines at Room Temperature
under Environmentally Benign Reaction Conditions. J. Colloid
small rod and spherical shaped CuO NPs with size ranges
Interface Sci. 2015, 460, 146–153. DOI: 10.1016/j.jcis.2015.08.
from 5 to 9 nm at a varying concentration of the precursor 040.
metal salt. This also shows the concentration effect on the [12] Sajadi, S. M.; Nasrollahzadeh, M.; Maham, M. Aqueous Extract
shape and size of as-synthesized CuO NPs. The TEM and from Seeds of Silybum marianum L. as a Green Material for
XRD results provide a correlation in the particle size calcu- Preparation of the Cu/Fe3O4 Nanoparticles: A Magnetically
Recoverable and Reusable Catalyst for the Reduction of
lation. As compare to rod-shaped CuO NPs, the spherical-
Nitroarenes. J. Colloid Interface Sci. 2016, 469, 93–98. DOI: 10.
shaped NPs have shown high antibacterial activity. Whereas 1016/j.jcis.2016.02.009.
the rod-shaped CuO NPs are found to be better photo-cata- [13] Atarod, M.; Nasrollahzadeh, M.; Sajadi, S. M. Green Synthesis
lyst for the degradation of MO dye. In the dye degradation of a Cu/Reduced Graphene Oxide/Fe3O4 Nanocomposite Using
reaction, the maximum value of the rate constant was Euphorbia wallichii Leaf Extract and Its Application as a
observed to be 0.13131 min1. Further, the shape and size of Recyclable and Heterogeneous Catalyst for the Reduction of 4-
Nitrophenol and Rhodamine B. RSC Adv. 2015, 5,
NPs have affected both types of properties. The results 91532–91543. DOI: 10.1039/C5RA17269A.
reflect that NPs synthesized by using plant extract are more [14] Nasrollahzadeh, M.; Sajadi, S. M.; Maham, M. Tamarix gallica
efficient photo-catalysts and antibacterial agents as compare Leaf Extract Mediated Novel Route for Green Synthesis of CuO
to those NPs reported via chemical methods. Nanoparticles and Their Application for N-Arylation of
Nitrogen-Containing Heterocycles under Ligand-Free
Conditions. RSC Adv. 2015, 5, 40628–40635. DOI: 10.1039/
Disclosure statement C5RA04012D.
[15] Momeni, S. S.; Nasrollahzadeh, M.; Rustaiyan, A. Green
There are no conflicts of interest to declare. Synthesis of the Cu/ZnO Nanoparticles Mediated by Euphorbia
prolifera Leaf Extract and Investigation of Their Catalytic
Activity. J. Colloid Interface Sci. 2016, 472, 173–179. DOI: 10.
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