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Please cite this article as: Bourakadi KE, Merghoub N, Fardioui M, Mekhzoum MEM, Kadmiri IM,
Essassi EM, Qaiss AEK, Bouhfid R, Chitosan/polyvinyl alcohol/thiabendazoluim-montmorillonite bio-
nanocomposite films: Mechanical, morphological and antimicrobial properties, Composites Part B
(2019), doi: https://doi.org/10.1016/j.compositesb.2019.05.042.
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Chitosan/Polyvinyl alcohol/Thiabendazoluim-Montmorillonite Bio-
nanocomposite Films: Mechanical, Morphological and Antimicrobial
Properties
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Rachid Bouhfida,*
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a
Moroccan Foundation for Advanced Science, Innovation and Research (MAScIR), Institute of
Nanomaterial and Nanotechnology (NANOTECH), Rabat Design Center, Rue Mohamed El Jazouli,
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Madinat El Irfane, 10100 Rabat, Morocco.
b
Mohammed V-Rabat University, Faculty of Science, Laboratory of Organique and Heterocyclic
Chemistry, Rabat, Morocco.
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c
Moroccan Foundation Advanced Science, Innovation and Research (MAScIR), Biotechnologie verte.
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Rabat Design Center, Rue Mohamed Jazzouli, Madinat El Irfane 10100, Rabat, Morocco.
d
Laboratoire de Matériaux, électrochimie et environnement, Université Ibn Tofail, Faculté des
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Abstract
Recently, the development of biopolymer/organoclay nanocomposites for food packaging,
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where at least one of the components is derived from nature or even biomass, has attracted
much attention since the properties of polymers can be enhanced and controlled by
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properties, Antibacterial activity.
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1. Introduction
Over the past few decades, food packaging industry has paid growing attention to renewable
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raw materials that are environmentally friendly and sustainable. These materials are
frequently formulated with natural biopolymer sources, such as starch, chitin, chitosan,
cellulose or biodegradable synthetic polymers namely, polyvinyl alcohol (PVA) and
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polylactic acid (PLA) which with effective reinforcement can generate “green” materials with
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interesting properties and functionalities.
Generally, Biopolymers are biodegradable polymers which contain monomeric units that are
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covalently bonded, forming chain like macromolecules. They have various advantages over
other synthetic polymers, including biodegradability, bioactivity, abundant availability, and
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therefore proves to be one of the most promising materials to be used as a eco-friendly food
packaging [1]. However, low mechanical, thermal, barrier and biological properties of
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biopolymers restrict their use in a wide-range of applications. For this reason, a considerable
amount of work has been devoted towards improvement of biopolymer properties. They are
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satisfied physical properties for use as packaging materials of non-food products, such as
water-treatment products [4]. In the same context, Fan-Long Jin et al. have been reviewed and
discussed the related mechanisms of biopolymer's modification methods such as nucleating
agent addition, fiber reinforcement, compounding, blending, stereoisomer complexation, and
chemical modification in order to enhance the poor heat resistance of poly(lactic acid)[5].
Among all the synthetic biopolymers, polyvinyl alcohol (PVA) provides a diverse range of
benefits, due to its easy preparation, good biodegradability, excellent chemical resistance,
completely water-soluble synthetic polymer, non-toxic and excellent film forming ability [6].
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PVA was commonly blended with different synthetic and natural polymers. It is useful in
many applications such as controlled drug delivery systems, recycling of polymers, and
packaging [7]. Besides, chitosan is a modified, natural biopolymer derived by deacetylation of
chitin, a major component of the shells of crustacean which is the second most abundant
natural polymer after cellulose [8]. Due to its non-toxicity, biocompatibility, biodegradability,
several distinctive biological activities, functional properties, film-forming ability [9], and
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weak permeability to oxygen, chitosan macromolecule was found to be one of the most
interesting polysaccharide polymers widely used for packaging applications in recent years
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[10].
Blends of synthetic and natural polymers have gained a considerable attention especially as
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biomaterial and in electrochemical devices. The combination of the properties of PVA
including its physically cross-linking ability and its bio-compatibility with antibacterial
properties of chitosan represents a promising way to make new class of hybrid materials with
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distinct characteristics and several biological activities [11,12]. Many studies have been
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carried out blending chitosan with PVA [10,14,15]. Literature survey [16] shows that
plasticized CS/PVA blend doped with NH4NO3 salt have been used as a film for electrolyte
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system and application in proton batteries. The conductivity of CS/PVA blend film can be
significantly enhanced to the order of 10−3 S.cm−1 by the addition of NH4NO3 and ethylene
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formaldehyde and glycerol as a plasticizer improved the thermal stability and mechanical
properties of the polymer blends. Tripathi et al. [18] developed a novel chitosan-based
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antimicrobial films consisting of chitosan and PVA blend with glutaraldehyde as a cross-
linker by solution casting method. The microbiological screening has demonstrated the
antimicrobial activity of the film against food pathogenic bacteria viz Escherichia coli,
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Staphylococcus aureus, and Bacillus subtilis. The obtained film may be used in food
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packaging applications. In another example, Jahan et al. [19] studied the effect of ionic cross-
linked with KNO3 on CS/PVA blend film to enhance the mechanical strength and percentage
elongation. The result shows that there was 298% improvement in tensile strength of the
CS/PVA (50/50) blend film and percentage of elongation from 7% to 67%.
Recently, more attention has been paid to nanocomposites-based silicates layers, in particular
montmorillonite (Ms) owing to its biological and pharmaceutical properties [16-17].
Interestingly, montmorillonite is among the most researched clay minerals because of its
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natural abundance and low cost. It is one of the most employed clay in nanocomposite
materials due to its unique properties such as high cation exchange capacity (CEC), swelling
capacity, good adsorption ability and large specific surface area [22]. However, it performs
badly in the organic environment. Thus in order to ensure a compatibility between polymer
and sodium montmorillonite clay in the preparation of nanocomposites materials, chemical
modification with hydrophobic agents becomes necessary by introducing organic molecules
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(cationic surfactants) such as quaternary ammonium compounds into the interlayer space
through ion exchange [19-20]. Several efforts have been made in the field of nanocomposites
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to modified montmorillonite with organic surfactants [25,26]. In light of these facts, the
functionalization of sodium montmorillonites using geminal hexadecyl hydrophobic buttress
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via a cationic exchange process has been carried out by Ennajih et al. [27]. The resulting
benzimidazolium-modified montmorillonite exhibits a large d-spacing of 3 nm between
silicate layers and showed a high thermal stability compared to the commonly used clay
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modified alkyl ammonium salts. Another study comparing sodium montmorillonite, Cloisite
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30B and Cloisite 20A was carried out by Rhim et al. [28] this time with polylactic acid
(PLA). They investigated the tensile, moisture barrier and antimicrobial properties of the
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PLA/clay nanocomposite films. The obtained results indicated that the concentration of clay
plays an important role in enhancing the properties of biofilms. Relatedly, sodium-
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The originality of the present work lies in the manufacture of new biodegradable blend
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composed of natural polymer chitosan and synthetic polymer PVA reinforced by organo-
modified-Montmorillonite in order to generate novel bioactive composite films. In this, new
cationic surfactants salts were synthesized and characterized through the 1H, 13C NMR, FTIR
and antibacterial activity. These cationic surfactants were used to produce the organically-
modified montmorillonites, the thiabendazole derivatives compounds have received
considerable interest in biology and medicine areas [31–33]. Whereas the organo-clays
produced were investigated through different techniques, using X-ray diffraction (XRD),
Fourier transform infrared (FTIR) and SEM analysis. Presently, to the best of our knowledge,
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this is the first report demonstrating the combination of chitosan, PVA and montmorillonite-
thiabendazolium (Mt) in composite preparation. The objective of this research was to evaluate
and compare the mechanical, morphological and antibacterial properties of reinforced films
prepared with the neat ones for potential application in packaging industry.
2. Materials and Methods
2.1. Materials
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High viscosity Chitosan (CS) with a degree of deacetylation about 85% and polyvinyl alcohol
(Mw=61000g/mol) were supplied by Sigma-aldrich. The commercial sodium montmorillonite
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(Cloisite-Na, Ms) used in the present work was provided by Southern Clay Product Inc.
(Gonzales, Texas). Ms was a fine particle powder with a cation exchange capacity of 96
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meq/100g and a basal spacing of the air-dried product of spacing d001=1.17 nm. All reagents
used for the synthesis of both thiabendazolium derivatives and organoclyas were provided by
Aldrich, solvents such as dimethylformamide and methanol were also supplied by Aldrich and
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used without further purification, Mueller Hinton broth purchased from Biokar Diagnostic
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and agar purchased from Sigma life science. The water used during synthesis and treatment of
organoclays was ultra-pure.
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cm−1. The vibrational transition frequencies are reported in wavenumbers (cm−1). 1H and 13C
NMR spectra were recorded in CDCl3 using an Advance 300 (Bruker) instrument, chemical
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shifts are given in parts per million. The following abbreviations are used for the proton
spectra multiplicities: s: singulet, d: doublet, t: triplet, q: quartet, qt: quintuplet, m: multiplet.
Coupling constants (J) are reported in Hertz (Hz). The melting points (mp) of the synthesized
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compounds were determined in open capillary tube on melting point apparatus Stuart SMP30.
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The tensile tests have been used to study the effect of chemical modification of
montmorillonite on the mechanical properties of nanocomposite films. The tests were
performed on a universal testing machine model (Tinius Olsen Horizon H10KT with a cross
header speed of 3mm/min at room temperature, the tensile properties have evaluated on terms
of Young’s modulus, tensile strength and elongation at break of neat CS/PVA and CS/PVA
nanocomposite films. Scanning electron microscopy was used to characterize the
nanocomposites surface morphology. The images were taken on a FEI Quattro S
Environmental Scanning Electron Microscope (ThermoFischer) operated at 20 kV. The Ms
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and Mt samples were prepared by fixing the powder particles on a microscope holder. The
surfaces of bio-nanocomposite films were first frozen in liquid nitrogen before being cryo-
fractured. The cross-section surfaces were coated with carbon in an ionization chamber to
avoid charging. The X-ray diffraction pattern of the MtCn are recorded on a Bruker D2-Phaser
diffractometer (Bruker Corp), using Cu Kα radiation, 1.5406 Å (30 kV, 10 mA).
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a. Antibacterial effect of synthesized Thiabendazoluim compounds
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The antibacterial activity of the synthesized compounds was evaluated against three
microorganisms, including two Gram-negative bacteria: Escherichia coli (ATCC-8739),
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Pseudomonas aeruginosa (ATCC-9027), and Gram-positive bacteria Staphylococcus aureus
(ATCC-6538). The minimum inhibitory concentrations (MICs) of the compounds were
determined using the broth microdilution method. Bacterial strains were cultivated with
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Mueller Hinton broth as growth media to obtain inocula at 105 CFU/mL. The samples were
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dissolved in DMSO, and tested at a final concentration ranging from 25µg/ml to 200µg/ml.
The final DMSO concentration was less than 1%. The volume of 180 µL of bacterial culture
and 20 µL of sample at various concentrations were dispensed in 96-well micro-titration
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plates. The plates were incubated at 37°C, 24 h. The Chloramphenicol was used as positive
control. Assays were carried out in triplicate, and repeated twice.
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Antibacterial activity of the films was assessed against the same bacteria strains used
previously by evaluating the inhibition zone of developed films. Bacterial strains were grown
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in nutrient broth liquid medium at 37 °C. After overnight incubation, bacterial suspensions
were diluted to obtain a concentration of 108 CFU/mL. The Chitosan/ Polyvinyl alcohol/
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To a large excess of alkyl bromide, one equivalent of thiabendazole was added, in the
presence of N,N-dimethylformamid (DMF), potassium carbonate, and an amount of
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tetrabutylammonium bromide as catalyst, the mixture was stirred at room temperature for 48
hours, then the solution was filtered by suction filtration. The solvent was removed under
reduced pressure, this step resulted in the formation of N-alkyl products t8, t12, t14 and t16 as
intermediates. The synthesized N-alkyle thiabendazole compounds were subjected to the
action of iodomethane in DMF at 100°C for 24 hours. This reaction leads to the formation of
N-alkylsubstituted quaternary thiabendazoluim salts tC8, tC12, tC14 and tC16. These organic
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surfactants have been obtained with a good yield and fully characterized by FTIR, NMR and
mass spectrometry.
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3-methyl-1-octyl-2-(thiazol-4-yl)benzimidazol-3-ium iodide (tC8):Yellow solid; Yield: 85%;
mp 86 °C; FTIR (KBr): 3494 (-CHaromatic), 2923, 2861 (-CHaliphatic), 1566 (-C=N), 1519, 1450
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(-C=Caromatic) cm-1.1H NMR (300 MHz, CDCl3) δ (ppm): 0.82 (t, J = 6.8 Hz, 3H, CCH3); 1.85-
1.04 (m, 12H, CCH2); 4.08 (s, 3H,NCH3); 4.60 (t, J = 7.4 Hz, 2H, NCH2); 8.28-7.67 (m, 4H,
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HAr); 8.96 (d, J= 1.7 Hz, 1H, NCHthiazolic); 9.61 (d, J = 1.7 Hz, 1H, SCHthiazolic). 13C NMR (151
MHz, CDCl3) δ (ppm): 14.4 (CCH3); 33.99 (NCH3); 44.6 (NCH2);46.5,31.6,29.1,28.9, 28.7,
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26.0, 22.5 (CH2), 144.4, 136.2, 132.4, 131.4 (Cq), 158.3, 131.7, 127.6, 127.5,114.3, 114.1
(CH). HRMS (ESI) analysis of compound tC8 gave a molecular ion [M]+ at m/z 328.1842
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73%; mp 70 °C; FTIR (KBr): 3016 (-CHaromatic), 2923, 2846 (-CHaliphatic), 1681 (-C=N), 1519,
1465 (-C=Caromatic) cm-1. 1H NMR (300 MHz, CDCl3) δ (ppm): 0.83 (t, J = 6.7 Hz, 3H,
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CCH3); 1.80-1.04 (m, 20H, CCH2); 4.05 (s, 3H, NCH3); 4.52 (t, J = 28.3 Hz, 2H, NCH2);
8.28-7.64 (m, 4H, HAr); 8.91 (d, J= 1.7 Hz, 1H, NCHthiazolic); 9.57 (d, J = 1.5 Hz, 1H,
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SCHthiazolic). C NMR (151 MHz, CDCl3) δ (ppm): 14.4 (CCH3); 33.89 (NCH3); 46.51
(NCH2);31.75,29.4,29.3, 29.2,29.1,28.7,28.7,26.0,26.0,22.6 (CH2), 144.4,136.2,132.4,131.4
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(Cq), 158.3, 131.6, 127.6, 127.5, 114.3, 114.1 (CH). HRMS (ESI) analysis of compound tC12
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gave a molecular ion [M]+ at m/z 384.2468 (calcd 384.2422 for C23H34N3S+).
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1.80-1.23 (m, 28H, CCH2); 4.25 (s, 3H,NCH3); 4.52 (t, 2H, NCH2); 7.84-7.69 (m, 4H, HAr);
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9.14 (d, J= 1.8 Hz, 1H, NCHthiazolic); 9.80 (d, J = 1.8 Hz, 1H, SCHthiazolic). C NMR (151
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MHz, CDCl3) δ (ppm): 14.11 (CCH3); 31.90, 29.67, 29.65, 29.64, 29.57, 29.57, 29.47, 29.34,
29.34, 29.30, 29.30, 28.86, 26.57, 22.67 (CH2); 34.6 (NCH3); 46.51 (NCH2);127.74, 127.77,
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113.67, 113.06 (CH), 131.23, 132.24, 135.66 ,144.12 (Cq). HRMS (ESI) analysis of
compound tC16 gave a molecular ion [M]+ at m/z 440.3080 (calcd 440.3094 for C27H42N3S+).
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2.5. Thiabendazolium –Montmorillonite preparation (Mt)
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1g of sodium-montmorillonite were dispersed in 100 mL of water: methanol (1:1) and
energetically stirred for 4 hours. After 5 min of sonification the suspension was heated at
80°C after which a solution of thiabendazolium (2CEC) in methanol (30 mL) was added
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dropwise. The stirring was continued for 24 hours at 80°C. The Mt was isolated by
centrifugation, washed by water: methanol (1:1) solution two times and then by deionized
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evaporation process. The matrix was constituted by 40% chitosan, 56 % PVA and 4 %
glycerol. Initially, chitosan solution was prepared by dissolving 0.5g of chitosan in 40 ml
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acetic acid solution (1%) and stirred for 3h for homogenization. Afterward, the solution was
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filtered with cheese cloth by vacuum aspiration to remove foam and any undissolved
impurities. On the other hand, PVA solution was prepared by dissolving 4g of PVA in water
at 90 °C. Both solutions were then mixed together for 1 hour using magnetic stirring after the
addition of an appropriate amount of glycerol as plasticized. Further, the suspensions of Mt
with varying surfactant lengths (C8, C12, C14 and C16) at a fixed weight content (5wt.%) were
prepared by dispersing appropriate amounts of MtCn into 10 ml of ethanol under vigorous
stirring. After 4 hours, the suspension was treated with an ultrasonic homogenizer for 1 hour.
Next, the dispersion of organo-montmorillonite nanoplatelets was added at the CS/PVA
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matrix solution and homogenized for 12 hours at room temperature first by magnetic stirrer
and then by ultrasonic dispersion. The solution was degassed to eliminate the air bulb, caste
on a Petri dish and left to evaporate at room temperature.
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The synthesis of a series of N-alkylsubtituted-3-methylthiabendazolium iodide has been
carried out in two steps (scheme 1), the first one concerns the N-alkylation of thiabendazole
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using four alkylating agents namely 1-bromooctane, 1-bromododecane, 1-bromotetradecane
and 1-bromohexadecane under phase transfer catalysis conditions. The second step in
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synthesis consists of the formation of thiabendazolium salts by quaternization using
iodomethane under reflux at 80°C for 24 hours. The synthesized products were crystallized in
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absolute ethanol affording the crude and targeted salts with a yield between 73-80%. The
compounds were characterized through 1H, 13
C NMR, IR and mass spectrometry. In the 1H
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NMR spectra of compounds tC8, tC12, tC14 and tC16, indicates a series of characteristic
signals of thiabendazolium ring bearing different long alkyl chains and methyl and methylene
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group at 1- and 3-position, respectively. As an example, signals at 4.08, 4.05, 4.5 and 4.25
ppm assigned to the N-CH3 group while the signals at 4.60, 4.52, 4.58 and 4.52 is
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C NMR spectrum, the signals of the NCH3 group were observed at 33.99, 33.89, 47.67, 34.6
ppm of the desired product tC12-tC16. Moreover, Mass spectrometry data confirmed the
structures of the synthesized salts.
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Fourier transform infrared (FTIR) was used to characterize the progress of the reaction, and
confirm the presence of surfactant agents in the clay gallery. Figure 1 shows the FTIR
spectrum of unmodified Ms and organically modified MtCn. Before modification, Ms showed
strong infrared adsorptions at 436-512 cm-1, 914-995 cm-1 and 3625 cm-1 assigned respectively
to (Al-O stretching), (Si-O-Si Stretching) and (O-H stretching in hydrogen bond water and
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free water) [34]. When the thiabendazolium was inserted into the Ms layered, the FT-IR
spectra of the hybrid material show the bands of the Ms as well as surfactant with slight shift.
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Two bands were observed in the MtCn at 1460, 1510 cm-1 can be attributed to the stretching’s
vibrations of C=C and C=N aromatic rings in the structure of thiabendazolium surfactant.
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Besides, two absorption bands corresponding to asymmetrical and symmetrical C–H
stretching modes of methyl (CH3) at 2916 cm-1 and methylene (CH2) groups at 2847 cm-1.
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Therefore, these results confirm the presence of the organic guest within the galleries of the
montmorillonite.
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The intercalation of thiabendazolium salts into the interlayer space of Ms has been confirmed
by X-ray diffraction where the value of the basic distance within lamellar Ms layers can be
determined. XRD patterns of the unmodified Ms and MtCn are presented in figure 2.
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According to the bragg’s equation, the exchange of the Na+ ions by thiabendazolium salts
increased the interlayer distance of Ms from 1.17 nm to 1.74 nm. The highest interlayer
distance of MtC16 is 1.74 nm, this increase in the interlayer space was obtained with surfactant
tC16, which has a relatively long alkyl chain length than other three surfactants. All d-spacing
of organoclays were larger than the raw Ms, which clearly confirmed that the organic salts
had been effectively intercalated into Ms.
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3.3.Chitosan-PVA/M-thiabendazoluim nanocomposites
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Figure 3. FTIR spectra of CS-PVA blends and its nanocomoposite films (CS/PVA-MtCn).
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FTIR spectroscopy was used to examine the interactions of the materials. The infrared spectra
of CS/PVA blend and the CS/PVA-MtCn films are presented in Figure 3. For CS/PVA blend,
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the band at about 3288 cm-1 is assigned to -OH and -NH2 stretching, the bands appearing
between 2750 and 3000 cm-1 are due to stretching vibrations of C-H bond in methylene (2927
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cm-1) and methyl (2860 cm-1) groups, the bands of the amide groups at 1649 and 1557 cm-1
are assigned respectively to the C=O and N-H stretching, the band at about 1033 cm-1
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corresponds to C-O stretching[14,30]. From the FTIR spectra of the nanocomposite films, it
can be observed the appearance of new bands at 920 cm−1 corresponding to Si-O-Si stretching
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and at 626 cm−1 assigned to Al-O stretching which is related to the presence of
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Young’s modulus, tensile strength and elongation at break of neat CS/PVA and CS/PVA-
MtCn nanocomposites films were shown in table 1. These results reflect the effects of the
chain length of the molecules used for the Mt modification on the final tensile properties of
the present nanocomposites. In addition, the enhancement in tensile strength of the elaborated
bio-composites compared with the neat film can be explain by the formation of hydrogen
bonds between the two polymers which make them more compatible with reinforcement
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agents at the interface filler-matrix. These results are in good agreement with those found in
SEM images.
These date shown how some clay features such as the clay organo-modification may affect
the polymer behavior as well as the properties of elaborated composite [36]. It is clearly
observed that for a fixed value of organoclay content (5 wt.%), the mechanical properties of
the nanocomposites improved with increasing the length chain of the organoclay[37], the
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comparison between the CS/PVA-MtC8 and CS/PVA-MtC16 films tensile properties and
those of pure Cs/PVA film allows to indicate this positive effect, while the Young’s modulus
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increased by 0.3 and 114% and the tensile strength was increased by 0.88% and 38.8% for
CS/PVA-MtC8 and CS/PVA-MtC16 nanocomposites compared to CS/PVA films. The
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ductility behavior of the elaborated composites was evaluated through the strain at yield or
elongation at break. The elongation at break decreases significantly for the reinforced films
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having an alkyl chain length varying between C8-C14 as compared with neat film, which
explain that the ductility of the new composite was affected by the reinforcement.
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For the elongation at break, there is almost no difference between the results of pure CS/PVA
and nanocomposites films.
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It is widely known that a high aspect ratio and good dispersion/distribution of nanofiller into
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the polymer matrix are the important factors needed to achieve polymer nanocomposites with
enhanced final properties. In the case of organoclay, these factors are controlled by the ability
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space with increasing alkyl chain length. These data results, which were discussed previously,
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the SEM image of the neat film (CS/PVA), The micrographs (b, c) show that the Mt are
uniformly dispersed in the CS/PVA matrix and the fabricated bio-nanocomposite films can be
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considered as homogeneous, with small amount of agglomerates are observed in some parts.
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Young’s Modulus Tensile strength Elongation at Break
(MPa) (MPa) (%)
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CS/PVA 66.9±3.7 24,9±1.6 36±5.5
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CS/PVA-MtC8 67.2±2.1 25,2±3.5 27±3.0
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Figure 4. SEM images of Na-montmorillonite (a) and montmorillonite modified with tC8 (b),
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tC16 (c).
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Figure 5. SEM images of bio-nanocomposite films (a) CsS/PVA, (b) CS/PVA-MtC8, (c)
CS/PVA-MtC16
Results of the in-vitro antibacterial activity of new thiabendazoluim derivatives against three
bacteria strains are listed in table 2, together with those of chloromphenicol used as a positive
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control. The highest antibacterial activity was found in the synthesized compounds tC12 and
tC14, which exhibited an inhibitory effect against three bacteria with a MIC values varying
between 25 to 50µg/mL. The product tC16 showed moderate inhibitory effect against E. coli
and P. aeruginosa with a MIC of 100 µg/mL and higher inhibitory activity against S. aureus
with a MIC of 25µg/mL. However, the product tC8 showed no effect under the applied
experimental conditions. In general, it is observed that the molecules tC12 and tC14 which
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have a chain length between twelve and fourteen carbons substituent exhibited excellent
activity when compared to the standard drug chloromphenicol. Nevertheless, the compound
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tC16 with long chain length of 16 carbons substituent showed equipotent to moderate activity
when compared with the standard drug chloromphenicol, while the compound tC8 having a
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short chain length displayed no activity against all the tested bacterial strains. According to
these results the major factor for the antibacterial activity of the synthesized compounds could
be due to the carbon chain length as well the positively charged amino groups at N3 in the
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thiabendazoluim molecules. These results are in agreement with those reported by Alessandro
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Busetti et al [40],that the quinolinium bromide with an alkyl in amino group at N1 position,
have a good antibacterial activity against a panel of bacterial strains. In order to confirm the
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antibacterial effect of the elaborated bio-nanocomposites films, we tested the activity of the
CS/PVA neat and CS/PVA-Mt nanocomposites films against the same bacteria strains.
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Table 2. Antimicrobial activities of synthesized compounds (tC8, tC12, tC14 and tC16).
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Chloramphenicol was used as positive control. The experiment was carried out in triplicate,
and repeated twice.
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tC8 NA NA NA
50 50 25
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tC12
tC14 50 50 50
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aeruginosa, S. aureus and E. coli bacteria (Table 3). From the above observations, it can be
concluded that all of the reinforced bionanocomposites films CS/PVA-MtC8, CS/PVA-MtC12,
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CS/PVA-MtC14 and CS/PVA-MtC16 displayed good antibacterial activity against all the tested
bacteria when compared with the neat CS/PVA blend. These results are in agreement with
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those found in thiabendazoluim salts previously evaluated in this study (Table 2), which
suggests that the antibacterial action of bio-nanocomposites films can be attributed to the
thiabendazoluim molecules. On the other hand, the enhancement of antibacterial activity of
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all the assessed bio-nanocomposite films as compared to the neat CS/PVA films, could be
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related to the high specific area of the montmorillonite nanoclay layers which can absorb the
bacteria from the solution and immobilize them on their surface, as previously reported in
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literature [41]. Aris Giannakas et al. [37], reported that the addition of organo-modified
montmorillonite to the CS/PVA blend leading into the enhancement of antimicrobial
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S. aureus - + + + +
E.coli - + + + +
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Bio-nanocomposite films based on CS/PVA and MtCn were prepared through the simple
casting-evaporation process. The effect of chain length of thiabendazolium surfactant on the
structure of montmorillonite as well as the structural, mechanical, morphological and
antibacterial properties of CS/PVA-MtCn bio-nanocomposites films was investigated. The
results obtained in this approach showed that the mechanical properties in term of tensile
ACCEPTED MANUSCRIPT
strength and young’s modulus of bio-composites films increases with the presence of
reinforcement agent (MtCn) when compared with the neat film. Also, the antimicrobial
activity of new elaborate films have been affected by the effect of thiabendazolium
compounds. Accordingly, the whole results presented here suggested that in the future
perspectives, the bio-composite films reinforced by thiabendazolium-Montmorillonite have an
excellent potential as antibacterial food packaging items.
PT
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Acknowledgement
This work was supported by MAScIR; Moroccan Foundation for Advanced Science,
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Innovation and Research, MESRSFC and CNRST, Morocco Grant no. 1970/15.
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