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*Correspondence: rasha.ha@yahoo.com
(Received: 18 August 2019; accepted: 12 November 2019)
Citaon: Rasha Saam Hameed, Raghad J. Fayyad, Rasha Saad Nuaman, Noor T. Hamdan and Sara A.J. Maliki, Synthesis and
Characterizaon of a Novel Titanium Nanoparcals using Banana Peel Extract and Invesgate its Anbacterial and Inseccidal
Acvity, J Pure Appl Microbiol., 2019; 13(4):2241-2249. hps://doi.org/10.22207/JPAM.13.4.38
© The Author(s) 2019. Open Access. This arcle is distributed under the terms of the Creave Commons Aribuon 4.0 Internaonal License which
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Hameed et al. J Pure Appl Microbiol, 13(4), 2241-2249 | December 2019
Arcle 5601 | hps://doi.org/10.22207/JPAM.13.4.38
Print ISSN: 0973-7510; E-ISSN: 2581-690X
RESEARCH ARTICLE OPEN ACCESS
www.microbiologyjournal.org2241Journal of Pure and Applied Microbiology
Synthesis and Characterizaon of a Novel Titanium
Nanoparcals using Banana Peel Extract and
Invesgate its Anbacterial and Inseccidal Acvity
Rasha Saam Hameed*, Raghad J. Fayyad, Rasha Saad Nuaman, Noor T.
Hamdan and Sara A.J. Maliki
Biology Department, Collage of Science, Mustansiriyah University, Iraq.
Abstract
Titanium nanoparcles (TiNPs) have been synthesized due to its certain characteriscs that are expected
like non-toxic, eco-friendly, and bioacvity. In this study, the researchers used Banana Peels Extract (BPE)
with tanium dioxide to prepare new nanoparcles which are never carried before. These nanoparcles
were biologically synthesized using an aqueous soluon of banana peel extract as a bioreductant.
The novel TiNPs were successfully prepared and characterized using Ultraviolet–Visible Spectroscopy
(UV-VIS), Atomic Force Microscopy (AFM), X-Ray Diractometer (XRD), and examined its anmicrobial
acvity against several pathogenic bacteria as well as inseccidal agent against Musca domesca. The
instrumental analysis conrms the presence of TiNPs with average diameter: 88.45 nm and volume
31.5 nm as resulted by AFM and XRD respecvely, while the bioacvity exam to TiNPs shows inhibitory
eect against several pathogenic bacteria, as well as it cause a high mortality percentage against three
larval stages of house y.
Keywords: Banana Peel Extract (BPE), eco-friendly, tanium nanoparcals, Musca domesca.
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Journal of Pure and Applied Microbiology
INTRODUCTION
Green synthesis of nanoparcles seeks to
minimize generated wastes and apply prospecve
progression1,2. In latest years, materials with
nano-sized aspect have paying an aenon to the
researchers all the way through the world. In up to
date nano science and technology, the interface
between inorganic nanoparcles and biological
structures are one of the majority excing area
of research. In addition to that another thing
is concern in this field of study like health,
environmental (eco-friendly), non-toxic materials
in synthesis procedures3,4.
Besides that, biosynthesis of nanoparcles
has other properes like opcal, catalyc, and
magnetic properties which allow them to be
applied in biosensing, catalysis, imaging, drug
delivery, and in medicine and since it has this wide
applicaons the producon of nanoparcles is a
signicant aspect of nanotechnology5,6.
Plants, enzymes, and microorganisms
were suggested to be used as probable natural
alternaves products to the lethal chemicals that
are non-biodegradable, hazardous to all living
creatures on the earth, as well as its high cost7.
Plants extracts serve as capping and reducing
agents in the preparation of nanoparticles
which are more benecial comparing with other
biological procedures8 these techniques of
nanoparcles synthesis which based on plants are
favored due to its properes as, ecofriendly, cost-
eecve, a single-step biosynthesis process and
non-toxic to workers and researchers9. Classically,
dierent parts of the plant can be used as the main
source to obtain the extract such as, fruit, fruit
peels, bark, callus, and root. These parts have been
examined in the synthesis of gold, silver, tanium
nanoparcles in various shapes and sizes10.
Banana (Musa paradaisica), belongs to
the Musaceae family, and it is a standout amongst
the most vital tropical fruits in the world market
and usually after pulp consumption, banana
peels are usually discarded, the peels represent
approximately 18-33% of the whole fruit, and
presently peels are not used for any other purposes
and somemes it is used as animals food in a very
limited extent11. It is therefore a signicant and
vital to discover applicaons for these peels in
ecological topics. It was found that banana peels
are rich in pecn, lignin and hemicelluloses which
encourage the researchers in current study to
use them in green synthesis of nanoparticles,
furthermore, banana peels contains large amounts
of phenolic compounds that can act as a ligand
and coordinate with the metal ion and form the
metallic nanoparcles12.
Titanium dioxide (TiO2) is naturally
formed and usually used as a white pigment
and paints, food colorants, papers, plascs, inks,
and toothpastes because it is nontoxic, it is also
considered as low-cost metal due to its wide
existence in nature. Due to its ability to absorb UV
light and high refracve index, TiO2 nanoparcles
have been used to synthesize nanoparcles with
many plants like Nyctan, arbortriss extract, leaf
extract of Catharanthus roseus, Eclipta prostrate
aqueous leaf extract, and peel extract of Annona
squamosa L.13,14.
In the current paper, tanium dioxide
has been used for the rst me with the water
extract obtained from banana peels to synthesize
the titanium nanoparticles TiNPs and was
characterized using several instrumental analysis
(UV–visible spectroscopy, XRD and AFM). Acvity
of nove TiNPs against bacteria and inseccide
agents as well as against several pathogenic
bacteria and (Muscadomesca) respecvely has
been examined and discussed.
MATERIALS AND METHODS
Preparaon of Banana Peel Extract (BPE)
Fresh banana was gained from local
markets; banana peels were cut it into small
pieces, washed three mes with dislled water
to remove external dirt layers and contaminants
from it then, the peels pieces were dried on paper
toweling. A 75 g of peels were place in a beaker
containing 150 ml double dislled water and then
boiled at 100°C for 20 min, and ltered through
Whatman No. 1 lter paper for two mes. The
extract was stored in fridge at 4°C15.
Preparaon of tanium dioxide nanoparcles
(TiO2NPs)
Titanium dioxide was obtained from
Sigma Aldrich, China. Molar mass was 79.87g/
mol and density was 4.2 g/cm3. Their average size
of the tanium oxide bulk parcles (TiO2BPs) was
(550 nm). Deionized dislled water (DW) was used
as a solvent to prepare a soluon of (100 mg/ml)
concentraon, and then only 5 ml of this soluon
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Hameed et al. J Pure Appl Microbiol, 13(4), 2241-2249 | December 2019 | hps://doi.org/10.22207/JPAM.13.4.38
was added in dropwise to 50 ml of Banana Peel
Extract (BPE) soluon with a connuance srring
(300 rpm) for one hour, this mixing process was a
achieved on a hot plate using magnec srrer and
at ambient temperature.
Characterizaon of TiO2 nanoparcles
UV–visible Spectroscopy
The UV–VIS absorpon spectra of novel
TiNPs soluon was achieved using Schimadzu 1601
spectrophotometer and 200–800 nm range16.
Atomic Force Microscopy (AFM)
This analysis was used to characterize
many properties of biosynthesized NPs such
as, NPs size, NPs surface, NPs topography, and
granularity volume distribuon of NPs. A thin lm
of the NPs sample was prepared on a glass slide
using 100µl of the sample on the slide, allowed
to dry for 5 min, then the slide was scanned using
AFM, (AA-3000, USA)17.
X-Ray Diracon
X-Ray Diffractometer (XRD) was used
to determine and idenfy the formaon of the
synthesized NPs, the XRD apparatus (Shimadzu
6000, Japan) operate at a voltage of (40 kv) and
current of (30 mM) with Cu Kα radiaon in a (θ-2θ)
conguraon.
Microorganisms
Microorganisms were collected from
laboratory of post graduate in biology department
/ College of Science / AL-Mustansireyah University.
These micro-organisms are: Gram-positive
bacteria (Bacillus sp., Staphylococcus aureus,
Staphylococcus epidermidis, Streptococcus sp.)
and Gram-negative bacteria (Pseudomonas
aeruginosa, Escherichia coli, Kleipseilla sp.,) the
strains of bacteria were maintained on nutrient
agar slants at 4°C.
Antibacterial activity of synthesized NPs and
banana peel extract (BPE)
Three concentraons of TiO2NPs were
prepared in (100, 50, and 25 mg/ml)m there
bioacvity were evaluated using well diusion
method according to (CLSI)18.
Insects rearing and Larvicidal bioassay
The Insects colony was collected from
house y (Musca domisca) which is free from
inseccides and pathogenic organisms, this animal
house belong to Biology Department, College of
Science, Al-Mustansiriyah University. The colony
was maintained under 28 ± 2 °C (starng from 1st
larval stage reaching to 3rd instar stage). The 1st,
2nd, and 3rd instar larvae were completed in healthy
environment; each test was replicated for three
mes. The collected Musca domisca larvae were
placed in a cup contains 10 g of nutrional media
(this media consist of 10 g of yeast extract, 200 g of
sh food, and 100 ml of dislled water). 2 mL from
each concentraon of TiO2NPs were added to each
cup; all cups were covered with muslin cloth and
kept in room temperature for 24 hrs. Larvae were
considered dead if there is no signicant molity19,
mortality percentage was calculated by applying
Abbo’s formula20:
X - Y
XX 100 = Percent of corrected control
X: Percent of alive in the check
Y: Percent of alive in the treatment
RESULTS AND DISCUSSION
Idencaon of TiO2 nanoparcals
Titanium nanoparcles were idened
via dierent consideraons, visually the reacon
mixture of TiNPs turned to white grayish color
aer 30 min. comparing with the white color of
tanium oxide indicang the formaon of tanium
nanoparcles as shown in Fig. 1.
Table 1. Dimensions of synthesized TiO2 NPs
Roughness average 1.53 nm
Root mean square 1.77 nm
Average diameter 88.45 nm
Fig. 1. Shows the comparison between three soluons:
A: Titanium Dioxide bulk soluon
B: Water extract of banana peel soluon
C: Titanium Dioxide nanoparcles soluon
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UV–Visible Spectral
Fig. 2 represent the TiNPs UV-Visible
spectra, the TiO2 absorbance was 4.2 at wave
length 208 nm, while Fig. 3 shows the value of
energy gap (Eg) of TiNPs which was 4.7(ev) these
results are agreed with Jawad 201721.
Atomic Force Microscopy (AFM)
Table 1 indicates the size of TiNPs ranged
between (65-115 nm) with average diameter
(88.45 nm), while the roughness average (RA)
and root mean square were 1.53 nm and 1.77 nm
respecvely. Fig. 4 shows AFM topographic images
of the TiNPs while Fig. 5 shows granularity volume
distribuon of TiNPs.
X-Ray Diracon (XRD)
Fig. 6 shows the X-Ray Diractometer
paern, the diagram indicates the presence of
three peaks, strong diffraction peaks (2-theta
28.2956°), (2-theta 40.4901°), and (2-theta
50.1821°), while the average crystallite size
of TiNPs was 31.5 nm according to Scherer’s
equaon.
Anmicrobial acvity of tanium nanoparcles
Bioacvity of the novel TiNPs showed that
these nanoparcals exhibited anmicrobial impact
against the pathogenic microorganisms, the
Fig. 2. Absorbance spectrum of synthesized (TiO2NPs)
Fig. 3. Energy gap diagram for (TiO2NPs) thin lm
Fig. 4. AFM topographic images
inhibion zone diameter was varies as depicted in
Table 2, while Banana Peels Extract (BPE) did not
shows any acvity against these bacteria.
All the prepared concentrations of
tanium nanoparcals shows bioacvity against
the gram positive bacteria (Stapylococcus
aureus), and (Steptococcus spp.) while the rst
concentraon (25%) doesn’t shows any acvity
against (Staphylococcus Epidermidis, Escherecia.
coli, Klebsiella spp., and Bacillu ssp.) as it is clearly
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Table 2. Bioacvity of TiNPs against certain bacteria
Compound Pathogenic Microorganism
Gram Posive Gram Negave
Stapylococcus Steptococcus Staphylococcus Bacillus Escherecia Klebsiella
aureus spp. Epidermidis Sp. coli spp.
Banana Peels
Extract (BPE)
TiNPs 25% conc. 6 18
TiNPs 50% conc. 27 16 18 15 21 13
TiNPs 100% conc. 16 12
5-10 week acvity against bacteria, 10-15 Moderate acvity, 15+ Over 15 consider to be strongly acve.
Fig. 6. X-Ray Diractometer paern of TiNPs
Fig. 5. Granularity volume distribuon of TNPs
appear from Table 2. Also the results show that
there was no anmicrobial acvity for banana peel
extract against all tested pathogens.
Table 2 indicates that best effective
concentration to the newly synthesized TiNPs
was on 50% conc., this concentration shows
strong bioacvity against gram posive bacteria
than negative bacteria, that’s because Gram
negave bacteria have cell walls with a thin layer
of pepdoglycan and an outer membrane with a
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lipopolysaccharide component which is not exist or
found in Gram posive bacteria and this property
enable the TiNPs to penetrate the cell membrane
of Gram posive bacteria and cause its fatality,
this results agreed with the results obtained by
Adam and his coworkers22 and Shrevastava and
his coworkers23.
The most logical explanaon proposed
that the possible mechanisms involving the
interacon between TiNPs with the biomolecules
which propose that microbes have negave charge
(represented by the membrane of the microbe)
and the posive charge (represented by TiNPs),
this “electromagnec” aracon between the
metal ion and the microbe membrane will leads
to microbial oxidaon and rapid death24.
Zhang and his coworkers suggested that
posive ion in nanoparcals form a coordinaon
bond with the thiol group (-SH) which is part
of protein molecule which is part of bacteria
membrane, this deactivation of protein will
increase the permeability of the bacteria
membrane leading to fast death25.
Larvicidal eect of TiNPs
Fig. 7 indicates the larvicidal analysis
by which the larvae death was scored aer 24
hrs, the results shows that there was a marked
mortality recorded during the development of
(Musca domisca) larvae depending upon the
concentraon of each dose. The highest mortality
was observed for 1st stage followed by 2nd and
3rd larval stage respecvely. While insignicant
mortality observed in 1st larval stage vials treated
with water and BPE (6.6%, 6.6%) as shown in Fig.
8.
The mortality percentage is proporonal
to the concentraon of the synthesized TiNPs which
indicates that the essenal role of concentraon
in larvicidal acvity. Each test included a control
group with three replicates for each concentraon.
Fig. 7. Mortality percentage of house y larvae treated with synthesized TiO2NPs
Fig. 8. Phenotypic variaon observed in A: 2nd larval stage, B: 3rd larval stage of Musca domisca treated with 100
mg/ml prepared TiNPs
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The results agreed with Sabat and his coworkers26.
After hatching from the egg, the larva starts
feeding with TiNPs treated food, once it pass in
larval gut, it interferes with the a certain process
that leads to unusual phenotypes which end with
neuronal defect, ending with larvae death.
Hassan and his coworkers27 reported
that the possible mechanism behind the death of
house y is the dispersion of metal nanoparcles
via oral route or through rupturing of the cucle
membrane, by this means the TiNPs enter into
the cavity of insect body to aect the surviving
the ability of generating the Reactive Oxygen
Species (ROS), these parcles are causave agent
for oxidave stress, which leads to eggs damage
due to its toxic tension, then the egg are not able
to proceed on its next developmental stages. This
analogous mechanism was recorded by magnete
NPs which shows bioacvity against Drosophilla
melanogaster28 and silver NPs bioacvity against
Culex pipienes29.
In producing TiNPs, there several factors
aecng its yield among them are temperature of
the synthesis process, pH of the reacon, purity
of the metal salt in addion to the extract itself,
these factors play an eecve role on the yield and
characteriscs of TiNPs, these factors was studied
by Njagi and his coworkers30.
In this study, the researchers used banana
peels extract in a new green synthesis method to
synthesized tanium nanoparcals, this method
has not been carried before and it was found that
TiNPs show bioactivity against bacteria which
increase its future applications against many
serious human pathogens. In addition, TiNPs
shows good larvicidal applicaons against Musca
domisca resulted. These results will open wide
and new biomedical applicaons.
CONCLUSION
The researcher concludes the following
points:
1. A novel, green, eco-friendly TiNPs was
synthesized successfully.
2. The synthesis procedure was rapid,
simple, and considered as a new approach method
to synthesize TiO2NPs.
3. The synthesis was based on using
agricultural waste material (banana peels).
4. Banana peels were chosen in this
research due to its composion (rich with pecn,
cellulose, hemicelluloses…) in addion to its low
cost.
5. Due to the bioacvity of the novel
TiNPs against bacteria and insects (house y),
the researchers expect a wide and various
applicaons of green synthesized nanoparcles
precisely against infecous bacteria, biomedical,
and pharmaceucal applicaons
ACKNOWLEDGEMENTS
The authors would like to thank all the
sta at Biology Department, College of Science,
Mustansiriyah University, Iraq for their support
and help to achieve this research.
CONFLICT OF INTEREST
The authors declares that there is no
conict of interest.
AUTHOR'S CONTRIBUTION
All authors listed have made a substanal,
direct and intellectual contribuon to the work,
and approved it for publicaon.
FUNDING
None.
DATA AVAILABILITY
All datasets generated or analyzed during
this study are included in the manuscript and/or
the Supplementary Files.
ETHICS STATEMENT
This arcle does not contain any studies
with human parcipants or animals performed by
any of the authors.
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