Accepted Manuscript

Novel composite films based on cellulose reinforced with chitosan and polyvinyl
alcohol: Effect on mechanical properties and water vapour permeability

Patricia Cazón, Manuel Vázquez, Gonzalo Velazquez

PII: S0142-9418(18)30470-7
DOI: 10.1016/j.polymertesting.2018.06.016
Reference: POTE 5513

To appear in: Polymer Testing

Received Date: 22 March 2018

Revised Date: 30 April 2018
Accepted Date: 9 June 2018

Please cite this article as: P. Cazón, M. Vázquez, G. Velazquez, Novel composite films based on
cellulose reinforced with chitosan and polyvinyl alcohol: Effect on mechanical properties and water
vapour permeability, Polymer Testing (2018), doi: 10.1016/j.polymertesting.2018.06.016.

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 ACCEPTED MANUSCRIPT
1 Novel composite films based on cellulose reinforced with chitosan and polyvinyl

2 alcohol: effect on mechanical properties and water vapour permeability

4 Patricia Cazón1,2, Manuel Vázquez2*, Gonzalo Velazquez1

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1
6 Instituto Politécnico Nacional. CICATA unidad Querétaro. Cerro Blanco No. 141.

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7 Colinas del Cimatario, Querétaro, 76090, México.
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8 Departament of Analytical Chemistry, Faculty of Veterinary, University of Santiago de

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9 Compostela, 27002-Lugo, Spain

10 *Corresponding author: manuel.vazquez@usc.es

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12 ABSTRACT
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13 Novel composite films were prepared by dissolving microcrystalline cellulose (3-5%

14 w/w) in NaOH/urea solution, followed by coagulation in acetic acid solution. The

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15 regenerated cellulose films were immersed in chitosan-polyvinyl alcohol solutions at

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16 concentrations of 0-1% w/w and 0-4% w/w, respectively. Tensile strength, percentage

17 of elongation at break, Young’s modulus and water vapour permeability were measured
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18 to assess the effect of each compound on the mechanical and barrier properties.

Polynomial models were obtained to evaluate the effect of the formulation on the
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19
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20 measured properties. The microstructure was analysed by scanning electron

21 microscopy. Results showed tensile strength values in the range 27.75-78.48 MPa,

22 similar to usual synthetic polymer films. Percentage of elongation at break ranged from

23 0.98 to 12.82 %, increasing when polyvinyl alcohol and chitosan increased. Young’s

24 modulus ranged from 2727.04 to 4217.25 MPa, showing values higher than pure

25 chitosan and polyvinyl alcohol films. The highest value was reached combining

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26 cellulose and polyvinyl alcohol without chitosan. The water vapour permeability

27 (1.78·10-11-4.24·10-11 g/m·s·Pa) showed 2 orders of magnitude higher than that of

28 synthetic polymers, but lower than pure chitosan and polyvinyl alcohol films. Results

29 showed that it is feasible to obtain cellulose-chitosan-polyvinyl alcohol composite films

30 with improved mechanical properties and water vapour permeability.

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31

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32 Keywords: Regenerated cellulose; tensile strength; elongation at break; Young’s

33 modulus; water vapour permeability.

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34

35 1. Introduction

36
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Recently, many studies have been conducted to develop biopolymers made from
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37 agricultural commodities and/or food waste products to obtain new food packaging
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38 materials. These bio-based materials can be degraded by bacteria, yeast, or fungi [1].

39 The interest in biopolymers has increased due to the depletion of the fossil fuel reserve
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40 and the environmental impact caused by the accumulation of plastic-based packaging

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41 materials. Synthetic polymers are available in large quantities at low cost, and possess

42 good properties such as heat sealability, tensile, tear strength, barrier to gas and aroma
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43 compounds. However, these synthetic plastics are no-biodegradable, leading to serious

environment problems by accumulation. Research efforts have focused in the

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44
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45 development of biodegradable films and coatings as a potential solution to reduce

46 environmental problems associated with food packaging.

47 In addition, it is necessary to study the mechanical and permeability properties to

48 assess their performance as food packaging material [2,3]. Mechanical properties of

49 films are important to determine if they could have an adequate mechanical strength and

50 integrity during transportation, handling and storage of packaged foods [4]. Barrier

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51 properties of a polymeric film play a key role in maintaining food product quality.

52 Moisture may transfer through the polymer package, resulting in product quality change

53 that decreases shelf-life [5].

54 The raw material used to develop biopolymeric films must proceed from renewable

55 sources such as proteins, lipids, polysaccharides and all possible combinations among

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56 them. Among the polysaccharides used to develop biodegradable films, cellulose and

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57 chitosan are the most abundant biopolymers on earth. As raw material to elaborate

58 films, chitosan and cellulose are readily available, ease to modify and with several

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59 potential applications [3].

60 Cellulose represents yearly about 1.5·1015 kg of the total biomass production. It is

61
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considered an almost inexhaustible source of raw material for the increasing demand for
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62 environmentally friendly and biocompatible products. Wood pulp is still the most
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63 important raw material source for the processing of cellulose. Approximately, 2 % of

64 cellulose is used for production of regenerated cellulose fibers and films, as well as for
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65 the synthesis of a large number of cellulose esters and ethers [6].

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66 Cellulose is a linear chain with two anhydroglucose rings ((C6H10O5)n), covalently

67 linked through an oxygen in a β-1-4 glycosidic bond. The number of repeated units per
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68 chain depends on the source [7]. Cellulose applications can be roughly classified into

three aspects: direct use, degradation into small molecules and chemical modifications.
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69
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70 Direct use without any chemical derivation has received considerable attention recently,

71 although cellulose is not soluble in usual solvents, due to its structure [8]. Some solvent

72 systems for cellulose dissolution have been developed, including N-methylmorpholine

73 N-oxide, ionic liquids, LiCl/N,N-dimethylacetamide, NaOH aqueous solution,

74 NaOH/urea or NaOH/thiourea aqueous solution (Lindman, Karlström, & Stigsson,

75 2010; S. Wang et al., 2016).

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76 Cellulose-based films possess some drawbacks including low mechanical and

77 barrier properties [11]. A feasible strategy to improve the film properties is the

78 combination of cellulose with other biopolymers. Chitosan, poly(β-(1–4)-2-acetamido-

79 D-glucose), is structurally identical to cellulose, except that a secondary hydroxyl on the

80 second carbon atom of the hexose repeated unit is replaced by an acetamide group.

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81 Cellulose and chitosan show some different properties and behaviors. For example,

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82 chitosan is soluble in acid conditions and chitosan films have a selective permeability

83 to gasses (CO2 and O2) and good mechanical properties [12]. However, the high

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84 permeability to water vapour of chitosan films limits their use since an effective control

85 of moisture transfer is a desirable property for most foods, especially in moist

86
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environments. Therefore, strategies have been followed to improve the physical
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87 properties of chitosan-based films [13,14].
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88 Other polymer that could be combined with cellulose and chitosan is polyvinyl

89 alcohol (PVA), a semi-crystalline polymer extensively studied [15]. It is biodegradable,

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90 soluble in water, slightly soluble in ethanol, but insoluble in other organic solvents.
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91 PVA is suitable to combine with other polymer to increase the physical, thermal and

92 barrier properties due to its high polar character, good mechanical properties and easy
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93 processability [16,17].

Usually, a combination of materials allows obtaining better mechanical and barrier

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94
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95 properties than those of pure polymers. Thus, combining chitosan and PVA with

96 cellulose could produce composite films with novel properties. Studies on cellulose

97 films combined with chitosan or PVA has been published but the combination of both

98 together with cellulose has not been published yet. Many studies on combined cellulose

99 and chitosan have been focused in chitosan films reinforced with microfibrillated,

100 nanofibrillated cellulose or cellulose fiber [16,18–20], cellulose nanowhiskers,

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101 nanocrystalline cellulose [21–25] or cellulose particles added directly to a chitosan

102 dissolution [26].

103 Therefore, the aim of this study was to develop biodegradable copolymer films

104 based on cellulose, chitosan and PVA. Polynomial models were obtained to evaluate the

105 effect of the composition of the blend on mechanical properties (tensile strength,

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106 percentage of elongation at break, Young’s modulus) and water vapour permeability

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107 (WVP).

108

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109 2. Materials and methods

110 Microcrystalline cellulose, extra pure (average particle size 90 µm), urea (99.5 %)

111
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for analysis, sodium hydroxide (98 %), sodium bromide (99 %) extra pure anhydrous
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112 and acetic acid (99.5 %) were purchased from Acros organics (Geel, Belgium).
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113 Polyvinyl alcohol, degree of hydrolysis (calculated on dried substance) ≥98 % and

114 molecular weight (MW) approx. 30000, was obtained from Merck KGaA (Darmstadt,
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115 Germany). Chitosan (Mw 100000-300000) was purchased from Acros organics (Geel,
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116 Belgium).

117
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118 2.1 Preparation of films

The NaOH/urea method was used for dissolving cellulose. Films were elaborated
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119
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120 according to a modified method [27]. Briefly, the aqueous solution containing

121 NaOH/urea/H2O at 7:14:79 (w/w) ratio was used as solvent. The desired amount of

122 cellulose was dispersed in the solvent blend at room temperature, and the mixture was

123 vigorously stirred for 1 h. The cellulose suspension was frozen at -20 ºC for 24 h. Then,

124 the mixture was thawed at room temperature stirring vigorously during 1 h to obtain a

125 transparent solution.

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126 A defined volume was cast in Petri dish to reach a 0.5 mm thick layer, and then

127 immersed in a coagulation bath of 4% (v/v) acetic acid to regenerate. The resulting wet

128 films were washed with running distillate water until pH 7 and then immersed in a final

129 bath with a mixture of chitosan and PVA. Finally, the films were dried on the Petri dish

130 at room temperature for 2 days. The dried films were peeled from the plates and

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131 samples were cut to specific sizes for each test. The samples were stored in desiccators

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132 for 5 days at 51 % or 0 % relative humidity (RH) as required for each test.

133

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134 2.2. Film thickness measurement and mechanical properties

135 Thickness (mm) was measured at 5 locations of each film using a Thickness Meter

136
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ET115S (Etari GmbH, Stuttgart, Germany). Measurements were carried out prior to the
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137 characterization tests and mean values were reported.
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138 Tensile tests of the cellulose films were performed at room temperature using a

139 texturometer with load cell of 30 kg and software Exponent version 6.1.12.0 (TA-
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140 XTplus, Stable Micro System, UK), according to the ASTM D882 method (ASTM),
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141 with some modifications. Tensile strength (TS), percentage of elongation at break point

142 (%E) and Young’s modulus (YM) were measured. An initial grip separation of 40 mm
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143 and cross head speed of 4 mm/s were set up. Ten samples of 15 × 100 mm were cut

from each film formulation and stored in desiccators at 51 % RH for 5 days. The films
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144
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145 were fixing at both ends using mechanical grips and the strength was measured.

146 Curves of stress (MPa) as a function of strain (%) were automatically recorded by

147 the texturometer TA-XTplus software. Tensile strength (maximum force divided by

148 film cross-section), deformation at break (extension at the rupture point, mm), percent

149 of elongation at break (deformation divided by initial sample length and multiplying by

150 100, %) and Young’s modulus (yield stress divided by yield strain in the viscoelastic

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151 region, MPa) were calculated from the stress–strain curves according to the ASTM

152 D882 method.

153

154 2.3. Water vapour permeability

155 Water vapour permeability values were measuared gravimetrically following the

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156 ASTM E96 method, known as the “cup method”, using an especially designed

permeability cell. A wide-mouth cup with a cap allowing 5·10-3 m2 area was filled with

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157

158 distilled water. The film was sealed on the mouth of the cup and placed at 20 °C inside a

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159 desiccator with silica gel. Weight was measured each 24 h for 5 days. The slope of del

160 plot of weight vs time was calculated by linear regression (r2 > 0.99). Water vapour

161
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transmission rate (WVTR) was calculated as the ratio slope (g/s) / test area (m2).
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162 Water vapour permeability (WVP) was calculated according to the combined Fick
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163 and Henry laws for gas diffusion through films, using Equation 1:

164 = ·∆ (Eq. 1)
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165 Where x (m) is the film thickness and ∆P (Pa) is the differential of water vapour
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166 pressure through the film [28]. A driving force of 2339 Pa was used as differential

167 vapour pressure of water. Each test was performed by triplicate.

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168
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169 2.4. Scanning electron microscopy (SEM)

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170 The films were coated on slides with adhesive carbon tape, metalized with Au

171 and observed using a high-vacuum microscope (JEOL JSM-6360LV, Jeol Ltd, Tokyo,

172 Japan) at an accelerating voltage of 20kV.

173

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174 2.5. Statistical design and analysis

175 The experiments followed a second-order, incomplete, factorial design. Cellulose,

176 chitosan and polyvinyl alcohol concentration were considered as operational variables

177 (denoted A, B and C, respectively) and the effects on the select dependent variable (TS,

178 %E, YM and WVP) were evaluated. The Box–Behnken design [29] was used. Table 1

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179 summarizes the experiment design. The statistical analysis was performed by Design

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180 Expert® 10.0.6 software (Stat-Ease, Inc., Minneapolis, MN, USA). Cook´s distance was

181 applied the detect outliers [30].

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182

183 3. Results and discussion

184
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Biodegradable films were prepared using cellulose, chitosan and polyvinyl alcohol
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185 (PVA) at several concentrations following the experimental design showed in Table 1.
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186 Cellulose ranged from 3 to 5% (w/w), to obtain a complete dissolution of cellulose

187 using NaOH/urea method. Mixing baths of chitosan ranged from 0 to 1% (w/w) and
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188 PVA ranged from 0 to 4% (w/w). At higher concentrations of chitosan and PVA, the
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189 bath was very viscous resulting in too thick films.

190 The results of the dependent variables (TS, %E, YM and WVP) are also showed in
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191 Table 1. The relationship between the independent (cellulose, chitosan and polyvinyl

alcohol concentration) and dependent variables (TS, %E, YM and WVP) of the films
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192
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193 was modelled using a second-order polynomial equation or partial polynomial

194 equations.

195

196 3.1. Effect on Tensile strength (TS)

197 Films obtained showed values of TS in the range from 27.75 to 78.48 MPa.

198 Experimental data fitted well to a linear model. Experiment 12 was ignored because it

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199 was detected as an outliner. The multifactor analysis of variance (ANOVA) is shown in

200 Table 2. F-value was 9.48 (p-value = 0.0029) implying that the model was significant.

201 The value of r2 was 0.74, and the adequate precision obtained was 9.38 indicating an

202 adequate signal.

203 The evaluation of F-values of the three variables confirmed that TS was highly

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204 affected by the linear effect of cellulose and chitosan concentration. PVA concentration

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205 slightly affected TS properties. Equation 2 predicts the response of TS for real variables.

206 = −5.97 + 10.15 % + 20.17 ℎ"# $ % +

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207 2.59 % % (Eq. 2)

208 The response surface describing the effect of the concentration of cellulose,

209
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chitosan and polyvinyl alcohol on TS is shown in Figure 1. In general, TS values
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210 increase by increasing cellulose and chitosan concentration. PVA concentration also
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211 affected this value, although with a lower effect, reaching higher TS values at higher

212 cellulose, chitosan and PVA concentration. Keeping constant the concentration of
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213 chitosan and PVA, by increasing the cellulose concentration, TS value increased from
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214 45.58 to 78.48 MPa. For other hand, keeping constant the cellulose and chitosan

215 concentration, TS values increased from 55.44 to 78.48 MPa when the PVA
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216 concentration increased. Similar effect was observed keeping constant cellulose and
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217 PVA concentration as and increasing chitosan concentration TS values changed from
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218 47.16 to 66.53 MPa.

219 Comparing with literature, a TS value of 25.64 MPa for PVA films prepared from a

220 dissolution of 2.3% (w/v) is reported [31]. Films with PVA at 5% (w/w) showed a TS

221 ranging from 40 to 50 MPa [25]. Chitosan films prepared with 1-2% (w/v) showed TS

222 values ranging from 22.2 to 79 MPa [18,22,31–33]. Variations in TS values in chitosan

223 films could be due to several factors, including chitosan composition, chitosan source,

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224 film preparation and storage [33]. Cellulose-based films from cotton linter at 4% (w/w)

225 using NaOH/thiourea as solvent and H2SO4 as coagulation bath, showed TS values of

226 68.10 MPa [34]. Cellulose-based films from a 5% (w/w) solution, using NaOH/urea as

227 solvent and acetic acid as coagulation bath, had TS values from 60 to 90 MPa as a

228 function of the molecular weight (2.2·104- 8.2·104 g/mol) [35].

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229 TS values obtained in this work are in the same range of those reported in

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230 literature. The model obtained suggests that tensile strength of the blend films was

231 dominated by cellulose. Interactions between cellulose and chitosan at higher

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232 concentrations resulted in higher TS values. This behavior can be explained by the

233 formation of hydrogen bonds between both polymeric chains. Blend films of cellulose

234
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and chitosan, prepared using N-methylmorpholine-N-oxide as solvent also showed
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235 increased TS values when the concentration of chitosan was increased [12]. The same
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236 authors reported lower TS values for cellulose films using N-methylmorpholine-N-

237 oxide as cellulose solvent compared to those values obtained in this study. Mechanical
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238 properties depend on the solvent employed and the molecular weight of cellulose used
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239 as raw material [36].

240
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241 3.2. Effect on elongation at break (%E)

Films showed values of %E in the range from 0.98% to 12.82%. Experimental data
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242
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243 fitted well to a two-factor interaction equation. Experiment 11 was ignored because it

244 was detected as an outliner. The multifactor analysis of variance (ANOVA) is shown in

245 Table 2. The F-value of the quadratic equation was 45.41 (p-value = 0.0365) implying

246 that the model was significant. The value of r2 was 0.79. The adequate precision ratio

247 obtained (7.51) was higher than 4 indicating an adequate signal.

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248 The F-values of the three variables confirmed that %E was highly affected by the

249 linear effect of the chitosan concentration, and the effect of the interaction chitosan-

250 PVA, followed by the linear effect of PVA concentration. Small effect of the cellulose

251 concentration was observed. The interaction cellulose-PVA was negligible. Equation 3

252 predicts the response on %E.

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253 %& = 2.73 − 0.025 · % − 6.79 · ℎ"# $ % − 1.29 · % % +

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254 1.19 · % · ℎ"# $ % + 0.014 · % · % % + 3.60 ·

255 ℎ"# $ % · % % (Eq. 3)

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256 The response surface of the model is shown in Figure 2. Overall increasing PVA

257 and chitosan concentration, %E values increases. The maximum value of %E (12.82%)

258
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was reached at the highest concentration of chitosan and PVA, reflecting the effect of
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259 the interaction between these two polymers. Increasing cellulose concentration at lower
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260 concentration of chitosan and PVA, the value of %E increased slightly from 3.04 to

261 6.09 %.
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262 In previous research, cellulose-based films at 5 % (w/w) solution, using NaOH/urea

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263 as solvent and acetic acid as coagulation bath, reported %E values from 2 to 5%, as a

264 function of the molecular weight (2.2·104- 8.2·104 g/mol) [35]. Cellulose-based films
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265 from a 4% (w/w) solution of cotton linter using NaOH/thiourea as solvent and H2SO4 as
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266 coagulation bath, showed a %E value of 6.98 % [34]. Cellulose-based films at 4%

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267 (w/w) solution showed values ranging from 1.9 to 2.9 % as function of the molecular

268 weight using NaOH/urea, LiCl/DMAc system and NMMO and ionic liquids as solvent

269 [36].

270 On the other hand, a %E value of 105.47 % was reported for films prepared from a

271 2.3 % (w/v) solution of PVA, similar to the values between 100 and 115% reported for

272 films from PVA at 5% (w/w) [25,31]. Comparing %E of chitosan films in literature,

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273 several studies reported %E values for pure chitosan films in the range from 13 to

274 53.85%, most of them in the lower values [18,33,37,38]. The variability in the values

275 can depend on chitosan composition, chitosan source, film preparation and storage

276 conditions.

277 PVA had a plasticizing effect, consequently intermolecular hydrogen bonds were

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278 formed between PVA and cellulose molecules, weakening the hydrogen bonds between

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279 cellulose chains. The interaction with PVA increased the mobility of cellulose chains.

280 This plasticizing effect was also observed in regenerated cellulose films bathed with

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281 glycerol [34,39].

282 In our study, PVA and chitosan improved elastic properties of cellulose-based films

283
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reaching %E values higher than those of cellophane, a commercial cellulose derivate.
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284 However, %E values were far from those of synthetic polymers (633-859 %) [3].
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285

286 3.3. Effect on Young’s Modulus (YM)

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287 The films showed YM values in the range from 2727.04 to 4217.25 MPa.
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288 Experiment 11 was ignored because it was detected as an outliner. Experimental data

289 fitted well to a quadratic model. The multifactor analysis of variance (ANOVA) is
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290 shown in Table 2. The F-value of the two-factors interaction equation was 6.27 (p-

value = 0.0146) implying that the model was significant. The value of r2 was 0.84. The
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291
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292 adequate precision ratio obtained (10.52) was greater than 4 indicating an adequate

293 signal.

294 F-values of the three variables showed that YM was highly affected by the linear

295 effect of the chitosan concentration, followed by the interaction chitosan-PVA.

296 Cellulose concentration and interaction cellulose-chitosan also affected the Young’s

297 modulus values, but the effect was smaller than that observed by the chitosan

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298 concentration and the interaction chitosan-PVA. Finally, the effect of PVA

299 concentration and the interaction cellulose-PVA concentrations were negligible.

300 Equation 4 predicts the response of YM.

301 * = 2562.98 + 212.65 · % + 1362.86 · ℎ"# $ % + 116.49 ·

302 % % − 307.62 · % · ℎ"# $ % + 35.60 · % ·

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303 % % − 457.65 · ℎ"# $ % · % % (Eq. 4)

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304 The response surface of the model is shown in Figure 3. Young’s modulus values

305 of the developed films depend mainly on the interaction between chitosan and PVA.

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306 The maximum values were obtained when PVA or chitosan was used in the bath

307 formulation. It was observed a negative interaction between chitosan and PVA was

308
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observed decreasing the Young’s modulus values. Therefore, increasing chitosan and
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309 PVA concentration, decreased Young’s modulus values from 4217.25 MPa (without
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310 chitosan) and 3750 MPa (without PVA) to 3150.48 MPa when both polymers were

311 added into the formulation. Formulation with PVA had higher Young’s modulus values
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312 than formulation with chitosan. Also, in formulations containing PVA, increasing the
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313 cellulose concentration increased Young’s modulus values. The opposite effect was

314 observed combining chitosan with cellulose resulting in a decreasing of Young’s

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315 modulus values. The maximum values of Young’s modulus were observed in films
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316 from formulation containing maximum concentration of PVA and cellulose.

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317 It was reported In literature, Young’s modulus values of chitosan films from 1-2 %

318 (w/w) solutions ranged from 1874 to 3000 MPa [18]. Variations in Young’s modulus

319 values of chitosan films depend on concentration, molecular weight, degree of

320 polymerization, storage conditions and preparation method. Also, decreasing the degree

321 of polymerization negatively affected Young’s modulus of chitosan films [18].

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322 Films from PVA at 5% (w/w) showed Young’s modulus values in the 1000-1500

323 MPa range [23,25]. Regenerated cellulose, using ionic liquid 1-butyl-3-

324 methylimidazolium chloride as solvent, showed a Young’s modulus value of 1800 MPa

325 [40]. Regenerated-cellulose films from 6% (w/w) cellulose solution mixed with NaOH,

326 urea, ZnO and distilled water, showed a Young’s modulus value of 6100 MPa, and 3700

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327 MPa for cellulose films from 7% (w/w) cellulose solution with NaOH and urea [41],

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328 indicating that Young’s modulus values of regenerated cellulose films depend on the

329 cellulose concentration, solvent method and preparation conditions. Regenerated-

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330 cellulose films showed higher Young’s modulus values than those of chitosan and PVA

331 films suggesting that mechanical properties in the obtained films are driven by the

332
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cellulose content. Increasing PVA or chitosan concentration allowed obtaining higher
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333 Young’s modulus values. This behavior could be explained due to the intermolecular
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334 hydrogen bonds formed between PVA and cellulose, or chitosan and cellulose. Higher

335 Young’s modulus values were obtained blending PVA-cellulose than chitosan-cellulose.
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336 However, using both PVA and chitosan in the formulation resulted in lower Young’s
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337 modulus values. Probably, the interaction between PVA and chitosan was not as strong

338 as the interaction of each polymers alone with cellulose.

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339

3.4. Effect on water vapour permeability (WVP)

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340

Obtained films showed WVP values in the range from 1.78·10-11 to 4.24·10-11
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341

342 g/m·s·Pa. Experimental data were well fitted using a quadratic equation. The

343 multifactor analysis of variance (ANOVA) is shown in Table 2. The F-value of the

344 two-factors interaction in the mathematical model was 13.75 (p-value = 0.0008)

345 implying that the model was significant. The value of r2 was 0.91. The adequate

346 precision ratio obtained (11.35) was greater than 4 indicating an adequate signal.

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347 The F-values of the three variables showed that water vapour permeability was

348 highly affected by the cellulose-chitosan and cellulose-PVA interactions. The linear

349 effect of cellulose, chitosan, PVA concentration and the interaction chitosan-PVA

350 showed a smaller effect on the water vapour permeability. Equation 5 predicts the

351 response on water vapour permeability.

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352 = −4.66 · 10,-- + 1.88 · 10,-- · % + 6.85 · 10,-- ·

$ % + 1.42 · 10,-- · % % − 1.74 · 10,-- ·

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353 ℎ"# % ·

354 ℎ"# $ % −3.67 · 10,-- % · % % + 3.24 · 10,-. · ℎ"# $ % ·

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355 % % (Eq. 5)

356 The response surface of the model is shown in Figure 4. The water vapor

357
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permeability of the films depends mainly on the cellulose-chitosan and cellulose-PVA
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358 interaction. In films obtained from a combination of cellulose with chitosan or PVA, at
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359 low chitosan or PVA concentrations, increasing cellulose concentration resulted in

360 higher values of WVP (from 1.86·10-11 to 3.95·10-11 g/m·s·Pa). On the contrary, at high
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361 concentrations of chitosan and PVA, increasing cellulose concentration caused a

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362 decreased the WVP values. Keeping cellulose concentration constant, the increase of

363 chitosan and PVA resulted in slight increases of WVP values. This increase was less
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364 significant than the effect due to the increase of cellulose concentration. The effect of
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365 cellulose on WVP was smaller when chitosan or PVA were added into the film
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366 formulation. When adding both the chitosan and the PVA into the formulation, the

367 VWP values were increased, especially at lower concentration of cellulose.

368 The obtained results agree with values reported in literature. Films from 1% (w/v)

369 chitosan solution, showed WVP in the 1.5·10-10-6.9·10-10 g/m·s·Pa range, depending on

370 the molecular weight and the solvent used [15,42,43]. PVA films showed WVP values

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371 between 1.2·10-11 and 1.7·10-11 g/m·s·Pa [15,44]. Fish gelatin films also showed similar

372 WVP values (0.38 × 10−10 g/m·s·Pa) [45].

373 WVP of biodegradable films depend on several factors including structure, ratio of

374 crystalline and amorphous zones, hydrophilic–hydrophobic ratio and interaction

375 between the main polymeric chains and presence of plasticizers or other additives [13].

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376 WVP values obtained in the composite films were driven by cellulose permeability

RI
377 properties. WVP values increased when chitosan and PVA concentration increased due

378 to the high permeability of these component which are more hydrophilic than cellulose.

SC
379 Cellulose permeability depends on its crystallinity as high-crystalline regions are

380 inaccessible to water molecules. Cellulose also contains a para-crystalline component

381
U
(amorphous area) which is accessible to water molecules, thus making them hydrophilic
AN
382 [28]. Water molecules preferably diffuse through amorphous areas of the polymer
M

383 matrix, so the degree of crystallinity is also important in the permeability behavior of

384 films [22]. Increasing cellulose concentration, resulted in higher WVP values, probably
D

385 due to the increasing of amorphous areas promoting water molecules diffusion.
TE

386 WVP values in the 1.0–1.5·10-13 and 3–4·10-14 g/m·s·Pa range have been reported

387 for low density polyethylene (LDPE) and high-density polyethylene (HDPE) films,
EP

388 respectively, at 90% RH and 38 ºC. The water vapor permeability values of transparent

cellulose films are up to 2 orders of magnitude higher than the those of polyethylene
C

389
AC

390 films [28].

391 For other hand, WVP values of developed films in this work were in the same

392 range as those of cellophane (8.4.10-11 g/m·s·Pa), due to the similar chemical structure

393 of both polymers [46].

394

395 3.5. Microstructure

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396 In previous studies, films with chitosan has a slight yellowish colour [46]. Figure

397 5 shows that the films obtained were translucent. Perhaps the colour of the cited study

398 can be due to impurities in the chitosan used. The surfaces of both sides are different as

399 showed by scanning electron microscopy (SEM) employed to analyse the morphology

400 of the film surfaces. Figure 6 shows the top and bottom surface of the films obtained in

PT
401 experiments 1, 5 and 6. The selected experiments allowed analysing the effect of the

RI
402 cellulose (3% or 5%) and chitosan (0% or 1%) on the surface at constant PVA

403 concentration (2%). Bottom side showed a smoother and continuous surface. It can be

SC
404 due to this is the side in contact with the Petri dish when the films were formed. Both

405 surfaces depended on the composition of the blending. Films showed a continuous

406
U
matrix, no pores or cracks were detected, indicating a dense structure. Surface of films
AN
407 with lower cellulose concentration looked more homogeneous than films with higher
M

408 cellulose content that have a rougher surface. The rough surface of composite films with

409 higher cellulose content were reduced adding chitosan. Previous studies using different
D

410 coagulation media and conditions showed also a decrease in the presence of voids and
TE

411 cracks as increasing PVA and chitosan [27].

412
EP

413 4. Conclusions

The polynomial models allowed selecting the formulation of cellulose-chitosan-

C

414
AC

415 PVA to prepare films with similar TS values of synthetic polymer films. The

416 combination of cellulose-based films with chitosan and PVA, improved %E values

417 compared to pure cellulose films. Chitosan and PVA, showed a plasticizing effect on

418 the cellulose-based films, due to interactions among the polymers. However, %E values

419 of the developed films are far from those of synthetic polymers, as LDPE and HDPE,

420 but they have better elongation properties than commercial cellophane. Interaction

17
 ACCEPTED MANUSCRIPT
421 between cellulose-chitosan or cellulose-PVA showed an important effect on Young’s

422 modulus values, increasing YM values when PVA or chitosan concentration increased.

423 Blending PVA-cellulose resulted in higher YM values than those of chitosan-cellulose

424 films; although using PVA and chitosan resulted in lower YM values due to a stronger

425 interaction between PVA and chitosan than the interaction of both polymers with

PT
426 cellulose. WVP values were 2 orders of magnitude higher than those of synthetic

RI
427 polymers. Composite films had lower WVP values than those of pure PVA and chitosan

428 films. Therefore, the combination of cellulose with PVA or chitosan improved WVP

SC
429 properties. Results showed that it is feasible to obtain cellulose-chitosan-polyvinyl

430 alcohol composite films with improved mechanical properties and water vapour

431
U
permeability. Results of this study could provide valuable information on fabricating
AN
432 cellulosic materials, further expanding cellulose applications.
M

433

434
D

435 Acknowledgements
TE

436 A grant from CONACYT (Mexico) to author Patricia Cazón is gratefully

437 acknowledged. The financial support for this project was provided by Consellería de
EP

438 Cultura, Educación e Ordenación Universitaria, Xunta de Galicia (ES) (Project #

ED431B 2016/009) and where applicable co-financed by the FEDER, within the
C

439
AC

440 PT2020 Partnership Agreement.

441

442 References

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588
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589
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590

591

592

593

594

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Table 1

Formulations assayed for the study of cellulose based films reinforced with chitosan and

polyvinyl alcohol and experimental results for the properties of the films achieved. TS is

Tensile strength, %E is elongation at break, YM is Young’s modulus and WVP is water

vapour permeability.

PT
RI
Cellulose Chitosan PVA TS %E YM WVP
Exp.
% (w/w) % (w/w) % (w/w) MPa % MPa g/s m Pa

SC
1 5.00 0.00 2.00 47.16 2.23 4217.25 4.00·10-11

2 4.00 1.00 0.00 44.63 2.08 3262.49 2.57·10-11

U
3 5.00 0.50 4.00 78.48 3.61 3752.91 2.76·10-11

3.84·10-11
AN
4 4.00 1.00 4.00 67.16 12.82 2727.04

5 5.00 1.00 2.00 66.53 6.09 3150.48 2.98·10-11

6 3.00 0.00 2.00 27.75 1.57 3681.91 1.78·10-11

M

7 3.00 0.50 0.00 44.45 1.90 3599.97 1.86·10-11

2.89·10-11
D

8 4.00 0.50 2.00 57.27 3.48 3718.31

9 5.00 0.50 0.00 55.44 2.59 3750 3.95·10-11

TE

10 3.00 1.00 2.00 48.66 3.04 3230.38 4.24·10-11

11 4.00 0.00 4.00 33.10 2.94 2833.9 2.82·10-11

EP

12 4.00 0.50 2.00 29.76 0.98 3531.45 2.59·10-11

13 3.00 0.50 4.00 45.58 2.82 3318.12 3.60·10-11

C

14 4.00 0.00 0.00 38.30 1.55 3153.93 2.85·10-11

AC

15 4.00 0.50 2.00 43.99 1.64 3554.6 2.63·10-11

595

596

597

598

599

25
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600

601

PT
RI
U SC
AN
M
D
TE
EP
C
AC

26
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602 Table 2

603 Analysis of variance (ANOVA) for each of the study dependent variables (tensile

Tensile Elongation at Water vapour

Young’s Modulus
strength break permeability

PT
F
Source p-value F value p-value F value p-value F value p-value

RI
value

Model 9.48 0.0029 4.41 0.037 6.271 0.0146 13.75 0.0008

SC
A-
12.64 0.0052 0.95 0.3610
Cellulose 3.49 0.1039 6.97 0.0297

U
AN
M
D
TE
EP

604 strength, elongation at break, Young’s Modulus and water vapour permeability).
C
AC

27
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B-Chitosan 12.48 0.0054 11.89 0.0107 24.55 0.0016 6.73 0.0319

C-PVA 3.31 0.0991 2.23 0.1794 0.58 0.4717 4.64 0.0634

AB 0.40 0.5451 2.44 0.1621 34.67 0.0004

AC 8.35·10-4 0.9778 0.52 0.4930 24.69 0.0011

BC 9.33 0.0185 13.71 0.0076 4.83 0.0593

PT
Lack of Fit 0.71 0.7341 2.55 0.3050 4.83 0.1804 4.19 0.2055

RI
605

606

SC
607

U
608 AN
609

610
M

611

612
D

613
TE

614 Figure legends

EP

615 Fig. 1. Prediction of the model for the effect of cellulose, chitosan and polyvinyl alcohol

616 (PVA) concentration on tensile strength (TS). A) Films with 0% (w/w) PVA; B)
C

617 Films with 2% (w/w) PVA; C) Films with 4% (w/w) PVA.

AC

618 Fig. 2. Prediction of the model for the effect of cellulose, chitosan and polyvinyl alcohol

619 (PVA) on elongation at break (%E). A) Films with 3% (w/w) cellulose; B) Films

620 with 4% (w/w) cellulose; C) Films with 5% (w/w) cellulose.

621 Fig. 3. Prediction of the model for the effect of cellulose, chitosan and polyvinyl alcohol

622 (PVA) on Young’s modulus (YM). A) Films with 3% (w/w) cellulose; B) Films

623 with 4% (w/w) cellulose; C) Films with 5% (w/w) cellulose.

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624 Fig. 4. Prediction of the model for the effect of the cellulose, chitosan and polyvinyl

625 alcohol (PVA) concentration on water vapour permeability. A) Films with 0%

626 (w/w) PVA; B) Films with 2% (w/w) PVA; C) Films with 4% (w/w) PVA.

627 Fig. 5. Visual appearance of the films obtained.

628 Fig. 6. Scanning electron microscopy images of the surface of the films of experiment 1,

PT
629 5, and 6.

RI
630

631

SC
632

633

634
U
AN
635
M

636

637
D
TE
C EP
AC

29
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A.
65

54.75

TS (MPa)
44.5

34.25

24

PT
1.00 5
0.75 4.5

RI
0.50 4
0.25 3.5
% Chitosan (w/w) 0.00 3
% Cellulose (w/w)

SC
B.
71

U
60
TS (MPa)

49
AN
38

27
M

1.00 5
0.75 4.5
D

0.50 4
0.25 3.5
% Chitosan (w/w) % Cellulose (w/w)
0.00 3
TE

C.

79
EP

67.5
TS (MPa)

56
C

44.5
AC

33

1.00 5
0.75 4.5
0.50 4
0.25 3.5
% Chitosan (w/w) 0.00 3 % Cellulose (w/w)
638

639

640 Figure 1

30
 ACCEPTED MANUSCRIPT
A.
10

6.75

3.5

%E
0.25

PT
-3

4.00 1.00
3.00 0.75

RI
2.00 0.50
1.00 0.25
% PVA (w/w) 0.00 0.00 % Chitosan (w/w)

SC
B.
13

U
9
AN
5
%E

1
M

-3

4.00 1.00
D

3.00 0.75
2.00 0.50
TE

1.00 0.25
% PVA (w/w) 0.00 0.00 % Chitosan (w/w)

C.
EP

12

8.25
C

4.5
AC
%E

0.75

-3

4.00 1.00
3.00 0.75
2.00 0.50
1.00 0.25
0.00 0.00
% PVA (w/w) % Chitosan (w/w)
641

642 Figure 2

31
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A. 4100

Young's Modulus (MPa)

3750

3400

3050

2700

PT
1.00

0.75

0.50
% Chitosan (w/w)

RI
0.25
0.00
1.00
3.00 2.00
0.00 4.00

% PVA(w/w)

SC
B. 4500
Young's Modulus (MPa)

U
4050 AN
3600

3150

2700
M

1.00

0.75
D

0.50
% Chitosan (w/w)
0.25
TE

0.00
2.00 1.00
3.00
0.00 4.00

% PVA(w/w)
EP

C. 4900
Young's Modulus (MPa)

4350
C

3800
AC

3250

2700

1.00

0.75

0.50
% Chitosan (w/w)
0.25
0.00
2.00 1.00
3.00
0.00 4.00

% PVA(w/w)
643

644 Figure 3

32
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A.

4.8·10-11

3.825·10-11

WVP (g/m·s·Pa)
2.85·10-11

1.875·10-11

PT
9·10-12
5.00
4.50

RI
1.00 4.00
0.75 % Cellulose (w/w)
0.50 3.50
0.25
0.00 3.00
% Chitosan (w/w)

SC
B.

4.3·10-11

U
3.625·10-11
WVP (g/m·s·Pa)

AN
2.95·10-11

2.275·10-11
M

1.6·10-11
5.00

4.50
D

1.00 4.00
0.75
0.50 3.50 % Cellulose (w/w)
0.25
TE

0.00 3.00
% Chitosan (w/w)

5.2·10-11
C.
EP

4.45·10-11

3.7·10-11
WVP (g/m·s·Pa)
C

2.95·10-11
AC

2.2·10-11

1.00

0.75
% Chitosan (w/w) 0.50

0.25

0.00 3.00 3.50 4.00 4.50 5.00

% Cellulose (w/w)
645

646 Figure 4

647
33
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PT
RI
U SC
AN
M
D
TE

648
EP

649
C

650
AC

651

652

653 Figure 5

654

34
 ACCEPTED MANUSCRIPT

659

658

657

656

655
Figure 6
Exp 6. C 3%, Ch 0%, Exp 5. C 5%, Ch 1%, Exp 1. C 5%, Ch 0%,
PVA 2% PVA 2% PVA 2%

PT
RI
SC

Top
U
AN
M
D
TE
EP

Bottom
C
AC
35
 ACCEPTED MANUSCRIPT
Highlights

- Cellulose reinforced films could be an alternative to synthetic packaging films

- Composite films based on cellulose, chitosan and polyvinyl alcohol were
developed.
- Tensile strength is similar to that of synthetic polymers
- Young’s modulus was higher than that of pure chitosan and polyvinyl alcohol
- Water vapour permeability is higher than that of synthetic polymers

PT
RI
U SC
AN
M
D
TE
C EP
AC