Accepted Manuscript

Characterization of edible packaging films based on semi-refined kappa-

carrageenan plasticized with glycerol and sorbitol

Abdulaal Farhan, Norziah Mohd Hani

PII: S0268-005X(16)30639-7

DOI: 10.1016/j.foodhyd.2016.10.034

Reference: FOOHYD 3652

To appear in: Food Hydrocolloids

Received Date: 05 September 2016

Revised Date: 18 October 2016

Accepted Date: 22 October 2016

Please cite this article as: Abdulaal Farhan, Norziah Mohd Hani, Characterization of edible
packaging films based on semi-refined kappa-carrageenan plasticized with glycerol and sorbitol,
Food Hydrocolloids (2016), doi: 10.1016/j.foodhyd.2016.10.034

This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to
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Highlights
 Edible packaging films were produced from semi-refined kappa-carrageenan (SRC).

 The influence of sorbitol and glycerol as plasticizers was investigated.

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 Plasticized SRC films exhibited improved mechanical and oil barrier properties.

 Films with sorbitol were more effective oxygen barriers.

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 SRC has a high potential to produce edible packaging films.

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1 Characterization of edible packaging films based on semi-refined kappa-

2 carrageenan plasticized with glycerol and sorbitol

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4

5 Abdulaal Farhan a, b,* and Norziah Mohd Hani a

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6

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7 a Food Technology Department, School of Industrial Technology, Universiti Sains Malaysia,

8 11800 Penang, Malaysia. (Norziah Mohd Hani, E-mail: norziah@gmail.com)

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9 b College of Agriculture, University of Wasit, Main Campus, Rabee District, University City, Kut,

10 Wasit, Iraq.

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12

13 * Corresponding author: E-mail: abdulaalfarhan@yahoo.com

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14 Tel: +964 7714714176

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15 ABSTRACT

16 Semi-refined kappa-carrageenan (SRC) based edible films plasticized with glycerol or sorbitol at

17 20, 25 and 30% (w/w) were prepared and characterized. SRC films containing plasticizers had

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18 higher tensile strength and elongation at break than the control (unplasticized) SRC film.

19 Increased moisture content, water solubility and water vapor permeability were found in films

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20 plasticized with 25‒30% of glycerol and 30% of sorbitol as compared to the control SRC film.

21 Water sorption kinetics indicated that all plasticized films reached the equilibrium rate. Films

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22 were more transparent and their seal strength increased significantly, whilst, their oil

23 permeability was reduced when the plasticizers concentration increased from 20‒30%. Films

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24 plasticized with sorbitol were more effective oxygen barriers than films plasticized with glycerol.

25 Scanning electron microscopy and atomic force microscopy showed that SRC films plasticized
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26 with sorbitol were more homogeneous and smoother than the control SRC film. This study

27 showed that the inexpensive SRC is a promising packaging material and has good potential to be

28 used as an alternative polysaccharide material for the production of edible biodegradable

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29 packaging films with improved mechanical and barrier properties.

30
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31 Keywords:
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32 Semi-refined kappa-carrageenan; biodegradable films; glycerol; sorbitol; mechanical properties;

33 barrier properties
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34

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35 1. Introduction

36 Petroleum-based synthetic polymers (plastics) which are non-biodegradable polymers have

37 been used widely in food packaging due to their low cost, durability and water resistance

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38 properties. However, concerns over the disposal of these non-biodegradable plastic-based

39 packaging materials, growing environmental problems and safety issues have led to an increase

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40 interest in the development of biodegradable and eco-friendly materials for use in food

41 packaging (Callegarin, Gallo, Debeaufort, & Voilley, 1997; Dick et al., 2015; Maniglia, de

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42 Paula, Domingos, & Tapia-Blácido, 2015; Spotti, Cecchini, Spotti, & Carrara, 2016; Sukhija,

43 Singh, & Riar, 2016). In the development of edible and biodegradable materials, carrageenans

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44 are among the most important biopolymers due to their great processability and abundance

45 (McHugh, 2003; Imeson, 2000). Carrageenans are natural and water soluble sulfated
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46 polysaccharides extracted from seaweeds of the Rhodophyceae family. They are composed of

47 alternating 3-linked β-ᴅ-galactopyranose (G-units) and 4-linked α-ᴅ-galactopyranose (D-units) or

48 4-linked 3,6-anhydro-α-ᴅ-galactopyranose (DA-units), forming the disaccharide repeating unit of

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49 carrageenans (Campo, Kawano, Silva Jr, & Carvalho, 2009). These sulfated polysaccharides are

50 grouped according to the presence or absence of the 3,6-anhydrobridge on the 4-linked-galactose

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51 residue and to the position and number of sulfate groups. On the basis of this classification, there
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52 are three main types of these sulfated polysaccharides exploited on a commercial scale including:

53 kappa (mono-sulfate), iota (di-sulfate) and lambda (tri-sulfate) carrageenans (Raveendran,

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54 Yoshida, Maekawa, & Kumar, 2013). Also, the 3,6-anhydrobridges are present in kappa (κ) and

55 iota (ι) carrageenans, whereas lambda (λ) carrageenan does not have the 3,6-anhydrobridges.

56 These differences in the chemical structure of carrageenans are essential for their

57 physicochemical properties and the helical structure formation which resulted in varied

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58 applications (De Ruiter & Rudolph, 1997). Among these carrageenans, κ- and ι- carrageenans

59 have strong potential to form a gel. However, the resulting gels are varied in their properties.

60 Typically, κ-carrageenan forms hard, strong and brittle gels, while ι-carrageenan forms elastic,

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61 soft and weak gels. Thus, the gelling power of κ-carrageenan enhances its good film forming

62 ability. Depending on the industrial production process, carrageenan is extracted from raw

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63 seaweed species of Eucheuma cottonii (Kappaphycus alvarezii) or Eucheuma spinosum

64 (Eucheuma denticulatum) and is manufactured by two distinct processes: The refined process

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65 and semi refined process. In the refined process, the carrageenan is dissolved completely out of

66 the cellulose matrix of the seaweed leaving a very clean and clear product. The semi refined

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67 process leaves the cellulose intact, thus enhancing the gel strength of the carrageenan within the

68 seaweed itself. The final semi refined product has excellent gelling and binding properties at a
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69 lower cost, but forms a gel that is less clear than the refined.

70 The only difference between refined κ-carrageenan (RC) and semi-refined κ-carrageenan

71 (SRC) is in the refining process in which extra steps are used in the processing and production
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72 steps (including an alkali treatment at high temperature, several precipitation, filtration and

73 concentration steps) to remove most of the cellulose to obtain RC that results in its high
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74 production cost. However, in producing SRC, milder processing steps are used and thus resulting
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75 in remaining residual cellulose presents in SRC obtained and this product is called either

76 carrageenan by FDA, Philippines natural grade (PNG-carrageenan) or Philippines processed

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77 grade regulated by EU as carrageenan E407a. As a result, SRC product produces a slight cloudy

78 solution whilst RC gives a clearer solution when dissolve in water (Ghosh, Siddhanta, Prasad,

79 Meena, & Bhattacharya, 2006; Prajapati, Maheriya, Jani, & Solanki, 2014). SRC may serve as an

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80 alternative and cheaper raw material for use as a film forming material in the development of

81 edible films.

82 In order to produce efficient packaging films, the properties of films must be optimized for

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83 commercial applications. A good approach to optimize such properties is the plasticization

84 process. Plasticizers are generally small molecules such as polyols like sorbitol and glycerol that

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85 intersperse and intercalate among and between polymer chains, disrupting hydrogen bonding and

86 spreading the chains apart, which not only increases flexibility, but also water vapor and gas

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87 permeabilities (McHugh & Krochta, 1994). However, the type of plasticizer and its

88 concentration are critical factors affecting the properties of films (Jost, Kobsik, Schmid, &

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89 Noller, 2014; Vieira, da Silva, dos Santos, & Beppu, 2011). Thus, in order to improve film

90 properties i.e. adequate mechanical strength and good barrier properties, the suitable
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91 concentration and type of plasticizer must be added to overcome brittleness caused by the high

92 intermolecular forces. Therefore, the objective of this study was to produce SRC based edible

93 films and to investigate the effect of plasticizers (glycerol and sorbitol) incorporated at different
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94 concentrations on the films’ physico-mechanical and barrier properties.

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96 2. Materials and methods

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97 2.1. Materials

98 Semi-refined κ-carrageenan (SRC), extracted from Kappaphycus alvarezii seaweed was

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99 kindly donated by TACARA company (Sabah, Malaysia). Food grade glycerol was purchased

100 from SIM company (Penang, Malaysia). Sorbitol was supplied by Rhodia Malaysia company

101 (Selangor, Malaysia). Osmium tetroxide (OsO4) and sodium cacodylate were procured from

102 Sigma Chemical Co. (St Louis, MO, USA). All chemicals used were of analytical grade.

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103 2.2. Preparation of SRC films

104 Prior to film preparation, semi-refined κ-carrageenan powder (SRC) was dissolved in distilled

105 water (2%, w/w) and heated at 90 °C for 30 min using hot plate magnetic stirring (Model: 11-

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106 102-50SH, Fisher Scientific, Iowa, USA). After dissolution, plasticizers, glycerol or sorbitol

107 were added into carrageenan solution at different concentrations (20, 25 and 30%, w/w based on

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108 dry weight of carrageenan). The total weight of film forming solution (FFS) was adjusted to 100

109 g with hot distilled water followed by filtration through cheesecloth to remove undissolved

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110 matter (the weight of undissolved matter was obtained by drying to be 0.52% from initial weight

111 of SRC). The solution was cooled down to 50 °C before film casting by pouring 80 g FFS onto a

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112 polyacrylic casting plate (16 cm × 16 cm × 0.3 cm) and dried for 36 h at room temperature (25 ±

113 2 °C). Films were then manually peeled off from casting plates. The control SRC films were
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114 prepared in a similar manner but without any addition of plasticizer. Finally, films were

115 conditioned at 25 °C and 56.8% relative humidity (RH) (using a saturated NaBr solution) for 48

116 h in an environmental chamber for further analysis.

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117

118 2.3. Characterization of SRC films

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119 2.3.1. Thickness measurement

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120 A hand-held digital micrometer (Mitutoyo Co., Tokyo, Japan) was used to measure the

121 thickness of film samples to the nearest ± 0.001 mm at ten random positions for each film
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122 sample. The mean values of thickness were used in the calculations for film opacity, mechanical

123 properties, water vapor permeability, oil permeability and oxygen permeability.

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125 2.3.2. Moisture content

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126 Test specimens (2 cm × 2 cm) at three random positions of each film type were uniformly cut

127 and placed on glass petri dishes. The moisture content of the films was determined by drying test

128 specimens in an oven at 103 °C until a constant dry weight was obtained.

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129

130 2.3.3. Film solubility in water

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131 Water solubility of the film samples was determined according to the method described by

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132 Liu, Tellez-Garay, and Castell-Perez (2004) with slight modifications. Film samples were

133 uniformly cut (2 cm × 2 cm), dried at 103 °C for 24 h in a laboratory oven (Binder, FD 115,

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134 Tuttlingen, Germany) and weighed to the nearest 0.0001 g to determine their initial dry weight.

135 Film samples were then placed in 50 ml screw-capped centrifuge tubes containing 30 ml of

136 distilled water. The tubes were placed in a water bath (Memmert, D-91126, Schwabach,
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137 Germany) under constant shaking at 25 °C for 24 h. After that, undissolved films were filtered

138 through Whatman No. 1 filter paper (Whatman International Ltd., Maidstone, England) and dried
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139 at 103 °C for 24 h to determine their final dry weight. The water solubility (WS, %), expressed as

140 a percentage was calculated using the following equation:

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141 WS (%)  [(W0  Wf ) W0 ]  100 (1)

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142 where W0 is the initial dry weight of the film and Wf is the final weight of the dried undissolved

143 film.
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144

145 2.3.4. Determination of mechanical properties

146 Tensile strength (TS) and elongation at break (EAB) of the film samples were measured

147 according to ASTM D882-00 method (ASTM, 2000) using a Texture analyzer (Stable Micro

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148 Systems TA-XT plus, Surrey, UK). Six film samples were uniformly cut into strips (14 cm × 2

149 cm) and clamped between tensile grips with an initial grip separation and crosshead speed set at

150 90 mm and 60 mm/min, respectively using a load cell of 30 kg. TS was calculated using the

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151 following equation:

152 TS (MPa)  Fmax  (2)

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153 where Fmax is the maximum load and Ф is the cross-sectional area. EAB was calculated using the

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154 following equation:

155 EAB (%)  ( l l 0 )  100 (3)

156
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where ∆l is the film extension and l0 is the initial length of the film sample.
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157

158 2.3.5. Determination of water vapor permeability

159 Water vapor permeability (WVP) of the film samples was measured following the method
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160 described by Sobral, Menegalli, Hubinger, and Roques (2001). Film samples were firmly sealed

161 on top of glass permeation cups containing silica gel (0% RH). The cups were then placed in a
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162 desiccator containing distilled water (100% RH) and kept at 22 °C. The cups were weighed at 24

163 h intervals for 7 days. The weight gain was recorded to the nearest 0.0001 g. Linear regression
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164 was used to fit the data of weight vs time plot to calculate the slope of each line. WVP (g mm

165 cm−2 h−1 Pa−1) of film sample was calculated using the following equation:
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166 WVP  ( w t A)  ( X P) (4)

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167 where X is the film thickness (mm), A is the exposed area of the film (cm2), ∆P is the partial

168 water vapor pressure difference across the film (Pa), and w ∕ t was calculated from the slope of

169 weight gain vs time plot.

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170

171 2.3.6. Water sorption kinetics

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172 The water sorption kinetics of the film samples were evaluated according to the method

173 described by Lavorgna, Piscitelli, Mangiacapra, and Buonocore (2010). Dried film samples (2

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174 cm × 2 cm) were weighed and placed in beakers containing 30 ml of distilled water. Beakers

175 were covered with aluminium foil and incubated in a water bath (Memmert, D-91126,

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176 Schwabach, Germany) at 25 °C. The swelling kinetics of the film samples were periodically

177 evaluated by measuring the water gain until equilibrium was reached. The film wet surface was
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178 lightly blotted with a filter paper before weighing with a balance accurate to 0.0001 g. The water

179 gain (WG) was calculated using the following equation:

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180 WG (%)  [ (Ws  Wd ) Wd ]  100 (5)

181 where Ws is the weight of swollen film and Wd is the weight of dry film.
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182

183 2.3.7. Color measurement

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184 Color of the film samples was determined by measuring the lightness/brightness (L*),

185 redness/greenness (a*) and yellowness/blueness (b*) using a colorimeter (Minolta

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186 Spectrophotometer, CM-3500D, Osaka, Japan). Prior to the measurement, the instrument was

187 warmed up for 10 min and then calibrated with standard black and white tiles.

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189 2.3.8. Opacity measurement

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190 The opacity of the film samples was determined according to the method described by

191 Shojaee-Aliabadi et al. (2014) (with a slight modification) using a UV-Visible spectrophotometer

192 (UV-160A, Shimadzu Co., Japan). Film samples were cut into a rectangle and placed on the

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193 internal side of a spectrophotometer test cell. An empty test cell was used as the reference. The

194 opacity was calculated using the following equation:

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195 Opacity  Abs550 X (6)

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196 where Abs550 is the absorbance value at 550 nm and X is the film thickness (mm). The higher

197 opacity values indicate lower sample transparency.

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198

199 2.3.9. Seal strength measurement

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200 Film samples were uniformly cut into strips (7.62 cm × 2.54 cm). Two film strips were placed

201 on top of one another, and an area of 2.54 cm × 0.2 cm of the film strips was heat-sealed at 100

202 °C for a few seconds using an Impulse Auto Sealer (Mercier Corporation, ME-455A1, Taipei,
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203 Taiwan). Prior to seal strength determination, sealed film samples were conditioned at 25 °C and

204 56.8% RH for 48 h. The seal strength of the heat-sealed films was determined according to
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205 ASTM F-88 method (ASTM, 2005) using a TA-XT plus Texture Analyzer (Stable Micro

206 System). The sealed film strips were clamped at non-sealed parts and the tensile force needed to
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207 separate the seal vertically was measured using a load cell of 30 kg, test speed and an initial

208 distance between the grips set at 90 mm/min and 2.5 cm, respectively. The seal strength (N/m)
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209 was calculated using the following equation:

210 Seal strength  peak force film width (7)

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212 2.3.10. Oil permeability measurement

213 Oil permeability of the film samples was measured according to the method described by

214 Yan, Hou, Guo, and Dong (2012). Film samples were firmly sealed on the open end of a glass

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215 bottle containing 5 ml of soybean oil. The bottle was turned upside down, put on filter paper and

216 kept in a desiccator for two days at 25 ± 1 °C. Oil permeability (Po) was calculated using the

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217 following equation:

218 Po  (W  X ) ( A  t ) (8)

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219 where, ∆W is the weight variation of filter paper (g), X is the film thickness (mm), A is the

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220 permeation area (m2) and t is the storage period (day).

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222 2.3.11. Oxygen permeability measurement

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223 Oxygen gas transmission rate of the film samples was measured using an oxygen permeability

224 analyzer (OX-TRAN, model 2/21, MOCON, USA) based on the ASTM D3985-81 method
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225 (ASTM, 1993). The film samples were uniformly cut into specific dimensions to fit the sample

226 chamber. The sealing rim of the chamber was greased, and then each film sample was placed on
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227 the greased sample chamber with an exposed testing area of 50 cm2. Subsequently, film samples

228 were exposed to 100% O2 on one side and to a mixture of 98% N2 and 2% H2 on the other side.
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229 The analysis was performed through a computerized system using MOCON permeability system

230 software and the oxygen permeability (O2P) was calculated using the following equation:
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231 O 2 P  (OTR  X ) P (9)

232 where OTR is oxygen gas transmission rate, X is the film thickness and ∆P is the partial pressure

233 difference of oxygen across the film surface.

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234 2.3.12. Fourier transform infrared (FTIR-ATR) spectroscopy

235 The FTIR spectra of glycerol, sorbitol and selected film samples were recorded using an

236 infrared spectrometer (Thermo Scientific Nicolet iS10 FTIR Spectrometer, Massachusetts, USA)

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237 equipped with a horizontal attenuated total reflectance (ATR) trough plate crystal cell (45 ZnSe;

238 80 mm long, 10 mm wide and 4 mm thick) (PIKE Technology Inc., Madison, WI, USA). The

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239 FTIR spectra were obtained in ATR mode between 600 and 4000 cm−1 with automatic signal

240 gain collected in 64 scans at a resolution of 4 cm−1. Data analysis of each film was performed

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241 with a computerized system using OMNIC 8.3 software.

242

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243 2.3.13. Scanning electron microscopy (SEM)

244 Microstructural analysis of the surface of selected film samples was carried out using a
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245 scanning electron microscope (EVo MA10, ZEISS, Germany) following the method described

246 by Denavi et al. (2009). Our preliminary experiments showed that the film fixation using the

247 following treatments was led to obtain clearer images of the films surface as compared to the
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248 untreated film samples. Prior to SEM analysis, film samples were cut and treated with 2% (v/v)

249 glutaraldehyde in 0.1 M sodium cacodylate buffer (pH 7.2) for 2 h, followed by washing with 0.1
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250 M sodium cacodylate buffer for 10 min (repeated 3x). Then films were post fixed with 0.4%
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251 (w/v) OsO4 in 0.2 M sodium cacodylate buffer (pH 7.2) for another 2 h, followed by washing

252 with 0.1 M sodium cacodylate buffer for 10 min (repeated 3x). Then, the films were dehydrated
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253 for 15 min through a graded ethanol series: 30, 50, 70 and 90% (v/v), and lastly in 99.5% (v/v)

254 ethanol for 15 min (repeated 3x). Samples were then dried at 25 ± 2 °C for 24 h. Film samples

255 were coated with gold/palladium and examined under SEM at 500 X magnification and

256 accelerating voltage of 15 kV through a computerized system using Smart SEM software.

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257 2.3.14. Atomic force microscopy (AFM)

258 The surface morphology of selected film samples was also observed using an atomic force

259 microscope (Dimension EDGE, BRUKER, Germany). Film samples were uniformly cut and

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260 placed in the sample chamber. Measurements (50 × 50 µm2 scan size) were taken from several

261 zones of the film surface to capture images. Images obtained were analyzed using NanoScope

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262 analysis software version 1.40 to transform into three-dimensional images. The roughness values

263 of the film surface can be evaluated from two statistical parameters: Rq (root-mean square

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264 average of height deviations taken from the mean data plane) and Ra (average of the absolute

265 value of the height deviations from a mean surface) (Shojaee-Aliabadi et al., 2014).

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266

267 2.4. Statistical Analysis

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268 Data obtained were analyzed using SPSS software (SPSS for Windows, SPSS Inc., Chicago,

269 IL, USA). Analysis of variance (ANOVA) was performed and significant differences between

270 mean values were determined by Duncan’s test. The results obtained (samples run in triplicates
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271 or more) were expressed as means ± standard deviation. Differences were considered significant

272 at p < 0.05.

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273
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274 3. Results and discussion

275 3.1. Film appearance, thickness and moisture content

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276 All SRC films plasticized with different concentrations of glycerol (SRC-20G, SRC-25G,

277 SRC-30G) and sorbitol (SRC-20S, SRC-25S, SRC-30S) had smooth surfaces with good

278 appearance, were flexible and transparent. All SRC films were easily peeled off from casting

279 plates. However the control SRC film was slightly opaque and less flexible than the glycerol and

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280 sorbitol plasticized films. Thickness and moisture content of SRC films plasticized with glycerol

281 (G) and sorbitol (S) are shown in Table 1. The results showed that the film thickness increased

282 significantly (p < 0.05) with an increase in plasticizer concentration. The control SRC film had

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283 significantly lowest thickness; whilst the SRC-30G film had significantly (p < 0.05) highest

284 thickness compared to the other films produced. The interstitial spacing between polymer chains

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285 in the film matrix could be increased due to the dispersed plasticizer molecules in the film matrix

286 (Jongjareonrak, Benjakul, Visessanguan, & Tanaka, 2006; Tong, Xiao, & Lim, 2013), which

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287 could contribute to the increased thickness of resulting films. The results also showed that a

288 significant increase in moisture content was found in SRC-25G (10.2%), SRC-30G (14.5%) and

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289 SRC-30S (11.0%) films compared to the control SRC film (8.8%). The increase in moisture

290 content of these films could be attributed to the increase in their hydrophilic characteristics
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291 compared to the control SRC film. At the same plasticizer concentration, SRC films plasticized

292 with glycerol contained higher moisture content than SRC films plasticized with sorbitol. This

293 result could be attributed to the smaller molecular weight and higher hydrophilicity of glycerol
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294 compared with sorbitol (Mali, Sakanaka, Yamashita, & Grossmann, 2005; McHugh & Krochta,

295 1994; Tong et al., 2013; Vieira et al., 2011). In general, the increase in moisture content of
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296 plasticized films could be attributed to the hydrophilic nature of glycerol and sorbitol. However,
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297 the moisture content of SRC films plasticized with glycerol and sorbitol (at any plasticizer

298 concentration) produced in this study was lower than those of RC films reported by other
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299 authors. Martins et al. (2012) had reported a higher moisture content of 19.28% in 30% glycerol

300 plasticized κ-carrageenan film. Shojaee-Aliabadi et al. (2014) had also reported a higher

301 moisture content of 20.15% in 50% glycerol plasticized κ-carrageenan film.

302

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303 3.2. Film solubility

304 A significant increase in water solubility was found in SRC-25G (45.9%), SRC-30G (49.5%)

305 and SRC-30S (46.3%) films compared to the control SRC film (42.8%) (Table 1). An increase in

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306 film solubility with increasing plasticizer concentration could be briefly explained by hydrophilic

307 plasticizers enhancing film solubility in water. Highest solubility (49.5%) was found in the SRC-

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308 30G film. This might be due to the smaller molecular weight of glycerol (compared to sorbitol)

309 which may possibly interact easily with SRC polymer chains causing increased affinity towards

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310 water (Mali et al., 2005; Tong et al., 2013). In general, SRC films were able to maintain more

311 than 50% (w/w) of their integrity in water at 25 °C over 24 h, suggesting that films were partially

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312 soluble in water irrespective of plasticizer type. The water solubility of SRC films produced in

313 the present study is comparable to the other edible films produced by other authors such as
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314 protein films (49.99%) (Liu et al., 2004) and chitosan films (42.05%) (Bourbon et al., 2011).

315

316 3.3. Mechanical properties

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317 Mechanical properties (TS and EAB) of SRC films plasticized with glycerol (G) and sorbitol

318 (S) are shown in Table 2. The control SRC film showed the lowest TS (40.3 MPa) and EAB
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319 (1.77%) values compared to glycerol and sorbitol plasticized films. These results indicated that
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320 the addition of both plasticizers significantly affected the mechanical properties of SRC films

321 produced. SRC films plasticized with glycerol and sorbitol (at any plasticizer concentration) had
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322 higher TS (p < 0.05) than the control SRC film. The increased TS of SRC films plasticized with

323 glycerol or sorbitol compared to the control SRC film could be attributed to the strong polymer-

324 plasticizer interaction formed via hydrogen bonding between carrageenan matrix chains and

325 plasticizer (glycerol or sorbitol) molecules. An increase in TS of edible films as a result of the

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326 increase in the plasticizers concentration has also been reported in cassava flour films (Suppakul,

327 Chalernsook, Ratisuthawat, Prapasitthi, & Munchukangwan, 2006) and in curdlan film (Prodpran

328 & Benjakul, 2014). However, increasing glycerol and sorbitol concentration did not significantly

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329 affect TS of SRC films produced. This result could be attributed to the narrow range of

330 concentrations used (20‒30%) of both plasticizers. A similar result was reported in edible films

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331 made from kefiran when the plasticizers (glycerol and sorbitol) concentration increased from 15

332 to 35% (Ghasemlou, Khodaiyan, & Oromiehie, 2011). Also, SRC films plasticized with glycerol

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333 and sorbitol (at any plasticizer concentration) exhibited good flexibility and stretchability,

334 presenting therefore higher EAB values (p < 0.05) than the control SRC film with a drastic

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335 increase in EAB when 30% of glycerol or sorbitol was added. In addition, the highest value of

336 EAB (10.01%) was obtained for the SRC-30S film compared to all films produced. The
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337 differences in composition, size, structure and shape of different plasticizers (glycerol and

338 sorbitol) influence their ability to function in the film network (Jongjareonrak et al., 2006) and

339 thus might determine their distribution and interaction with the SRC network in the films
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340 produced. Plasticizers are added to polymers to overcome brittleness, impart flexibility and

341 improve toughness (Mekonnen, Mussone, Khalil, & Bressler, 2013). Therefore, the addition of
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342 plasticizers to improve the mechanical properties of edible films is highly required (Dick et al.,
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343 2015). In food packaging, packaging film materials that can exhibit good mechanical properties

344 are extensively desirable in order to maintain and to protect the packaged food integrity from
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345 deterioration during storing and transporting conditions (Hu, Chen, & Gao, 2009). In the present

346 study, TS and EAB were measured to explore the effect of plasticizers (glycerol or sorbitol) on

347 the mechanical properties of SRC films in order to develop customized packaging materials.

348 From the results obtained in the present study, the SRC based films exhibited higher TS values

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349 than those reported for RC based films produced by several other authors. Savadekar, Karande,

350 Vigneshwaran, Bharimalla, and Mhaske (2012) found that the TS and EAB of κ-carrageenan

351 film plasticized with 10% glycerol were about 25 MPa and 13%, respectively. Rhim (2012)

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352 reported that TS and EAB of κ-carrageenan film plasticized with about 50% glycerol were 39.34

353 MPa and 19.5%, respectively. While, TS and EAB of κ-carrageenan film plasticized with 30%

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354 glycerol were 19.95 MPa and 16.18%, respectively (Martins et al., 2012). These differences in

355 mechanical properties between RC films produced by other authors and SRC films produced in

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356 the present study could be attributed to the difference in the type and concentration of plasticizer.

357 Also, it could be attributed to the difference in the composition between SRC and RC (Chan,

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358 Mirhosseini, Taip, Ling, & Tan, 2013). In addition, TS values of SRC based films produced in

359 the present study are comparable to those of commonly used plastic films such as ethylene vinyl
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360 alcohol (39.2‒68.7 MPa) (Chick & Ustunol, 1998), low-density polyethylene (8.2‒31.4 MPa),

361 polyvinylidene chloride (19.3‒34.5 MPa), polyvinyl alcohol (44‒64 MPa) (Kumar, Sandeep,

362 Alavi, Truong, & Gorga, 2010), high-density polyethylene (22‒31 MPa), polypropylene (31‒38
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363 MPa) and polystyrene (45‒83 MPa) (Rhim, 2012). Overall, SRC films formulated with different

364 concentrations of glycerol or sorbitol had good tensile strength, indicating potential applications
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365 as edible packaging materials for a variety of food products.

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366

367 3.4. Water vapor permeability

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368 Water vapor permeability (WVP) of SRC films plasticized with glycerol (G) and sorbitol (S)

369 is shown in Table 2. A significant increase in WVP was found in SRC-25G (3.7 × 10−8 g mm

370 cm−2 h−1 Pa−1), SRC-30G (4.32 × 10−8 g mm cm−2 h−1 Pa−1) and SRC-30S (4.25 × 10−8 g mm cm−2

371 h−1 Pa−1) films compared to the control SRC film (3.26 × 10−8 g mm cm−2 h−1 Pa−1). This increase

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372 in WVP could be attributed to the hydrophilic nature of glycerol or sorbitol. This tendency could

373 be explained by structural modifications of the polymer network in which the network becomes

374 less dense. It must be noted that an increase in the concentration of these hydrophilic plasticizers

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375 effected reorganization of the polysaccharide network and increase in the free volume and

376 segmental motions, allowing water molecules to diffuse more easily and giving a higher WVP

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377 (Khazaei, Esmaiili, Djomeh, Ghasemlou, & Jouki, 2014). The increase in WVP with increasing

378 plasticizer concentration with hydrophilic properties is common in edible films such as those

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379 formed from gelatin (Sobral et al., 2001) or starch (Yan et al., 2012).

380

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381 3.5. Water sorption kinetics

382 Water sorption kinetics of SRC films plasticized with glycerol (G) and sorbitol (S) were
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383 evaluated by measuring the water uptake vs. time at 25 °C as shown in Fig. 1 A and B. It was

384 observed that the behavior of the control SRC film was totally different from glycerol and

385 sorbitol plasticized films. The water uptake kinetic of the control SRC film was divided into
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386 three zones; whilst, only two zones were observed in all the plasticized films i.e. water

387 absorption zone followed by equilibrium zone. Fig. 1A shows that, among the glycerol-
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388 plasticized films, SRC-20G and SRC-25G films displayed a similar behavior and reached the
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389 equilibrium rate after 40 min of immersion in water. Beyond that, no additional water uptake was

390 observed. However, in SRC-30G film; water absorption was highest and continued for about 80
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391 min, beyond which no further uptake of water was noted. Among the sorbitol-plasticized films,

392 all films rapidly absorbed water in the first 20 min, and reached equilibrium at about 40 min

393 (Fig. 1B). The results show that SRC-30G film absorbed highest water content which could be

394 attributed to the more hydrophilic characteristic of glycerol than that of sorbitol. Nevertheless,

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395 SRC films plasticized with different concentrations of glycerol or sorbitol absorbed large

396 amounts of water and were swollen to a large extent. However, all the plasticized films remained

397 intact and stable even after water uptake for the entire period of time (120 min), indicating a

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398 stable film network induced by good interaction (hydrogen bonds) between SRC matrix and both

399 plasticizers, as indicated from the improved mechanical properties of plasticized films (Table 2).

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400 Lavorgna et al. (2010) have found that the glycerol-plasticized chitosan films were more stable

401 than unplasticized films when those films were in direct contact with water, suggesting that the

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402 presence of glycerol confers a higher dimensional stability to the films. Thus, the presence of

403 glycerol or sorbitol in SRC films has led to form more stable film network. This property might

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404 enhance the applications of SRC films plasticized with glycerol and sorbitol as water absorbent

405 pads used in trays for packaged retail meats, poultry, and fish to soak up their exudates, and
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406 could also be used for the packaging of fresh-cut fruits and vegetables for the same purpose

407 (Rhim, 2012).

408
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409 3.6. Color and opacity

410 Color and opacity values of SRC films plasticized with glycerol (G) and sorbitol (S) are
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411 shown in Table 3. All the SRC films produced exhibited good color properties as indicated by
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412 high L* values (93.47‒93.97) with a slight decrease in L* values at higher plasticizer

413 concentration. Both types of plasticized films exhibited lower opacity (higher degree of the
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414 transparency) compared to the control SRC film. This property could be attributed to increased

415 polymer chain mobility and intermolecular spacing in the carrageenan matrix with added

416 glycerol or sorbitol (Jongjareonrak et al., 2006; Tong et al., 2013; Yan et al., 2012), which could

417 facilitate the permeability of the light through SRC films, resulting in low values of the opacity.

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418 However, the sorbitol-plasticized films showed markedly lower opacity compared to the

419 glycerol-plasticized films.

420

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421 3.7. Heat seal strength

422 Fig. 2 shows the seal strength of SRC films plasticized with glycerol (G) and sorbitol (S). All

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423 films produced in this study were heat-sealable. The seal strength of the glycerol-plasticized

424 films ranged from 109 to 181 N/m, and that of sorbitol-plasticized films from 94 to 174 N/m;

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425 while the seal strength of the control SRC film (76 N/m) was significantly lower than those of

426 plasticized films. In both types of plasticized films, the seal strength significantly increased with

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427 increasing plasticizer concentration, with both SRC-30G and SRC-30S films recording the

428 highest seal strength. Heat sealing of a polymer is a combination of mass and heat transfer
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429 process. Glycerol plasticized films (SRC-20G and SRC-25G) showed significantly higher seal

430 strength compared to sorbitol plasticized films with the same plasticizer concentration. This

431 could possibly be due to the higher heat transfer mediated through the contact surfaces of the
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432 glycerol-plasticized films, resulting in better melting properties (Abdorreza, Cheng, & Karim,

433 2011), which in turn led to enhance the seal strength. In general, the high value of seal strength is
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434 desirable during packaging process of food products due to the high capacity to resist separation
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435 or seal tearing, while a lower value of seal strength might be desirable in other applications

436 where easy peel opening of the package seal is required (Voon, Bhat, Easa, Liong, & Karim,
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437 2012). The highest seal strength values obtained in the present study were 181 N/m and 174 N/m

438 for the SRC-30G and SRC-30S films, respectively. These values are lower than that for heat-

439 sealed synthetic polymers (≥730 N/m) (Abdorreza et al., 2011; Kim & Ustunol, 2001). However,

440 SRC-30G and SRC-30S films are comparable in seal strength to lactic-acid-casein-based films

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441 (153‒247 N/m), whey protein isolate/lipid emulsion films (105‒323 N/m) (Kim & Ustunol,

442 2001) and sago starch films (about 20‒375 N/m) (Abdorreza et al., 2011).

443

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444 3.8. Oil permeability

445 Oil permeability (Po) of SRC films plasticized with glycerol (G) and sorbitol (S) is shown in

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446 Table 4. Oil permeability values of glycerol and sorbitol plasticized SRC films were significantly

447 lower than the control SRC film. Among the glycerol-plasticized films, the oil permeability

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448 significantly reduced (p < 0.05) with increasing glycerol concentration. A similar trend was

449 found among the sorbitol-plasticized films. Among all the plasticized films, SRC-30G and SRC-

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450 30S films exhibited good anti-permeate property for oil with minimum oil permeability (0.37 g

451 mm m−2 d−1) observed in the SRC-30S film. The presence of higher amount of plasticizers
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452 (glycerol or sorbitol) in the film increased the total number of hydroxyl groups (oleophobic) in

453 the film network, which might prevent the passage of oil molecules through the film, thus

454 resulting in reduced film oil permeability. Hu et al. (2009) reported that oxidized potato starch
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455 films have many oleophobic hydroxyl and carboxyl groups, which could prevent the adsorption

456 of oil molecules on the film surface, resulting in good anti-permeate properties for oil.
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457
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458 3.9. Oxygen permeability

459 Oxygen permeability (O2P) of SRC films plasticized with glycerol (G) and sorbitol (S) is
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460 shown in Table 4. No significant difference (p > 0.05) in O2P values between the glycerol-

461 plasticized films and the control SRC film was observed. However, O2P values for the sorbitol-

462 plasticized films were significantly lower than the control and glycerol-plasticized SRC films.

463 These results showed that sorbitol as a plasticizer had a distinct influence on O2P of films

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464 compared to the glycerol-plasticized films and this may be attributed to the high compatibility

465 between the SRC network and sorbitol which led to the formation of a continuous and compact

466 network, resulting in lower O2P values. Other authors had also reported similar results in casein

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467 based edible films (Chick & Ustunol, 1998) and alginate based films (Jost et al., 2014)

468 containing sorbitol as a plasticizer that produced films with a better oxygen barrier than films

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469 containing glycerol.

470

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471 3.10. Fourier transform infrared (FTIR) spectroscopy

472 Since the greater plasticizing effect could clearly be observed in films plasticized at higher

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473 concentrations (30%), SRC-30G and SRC-30S films were selected for FTIR analysis and

474 compared with the control SRC film to highlight the chemical interactions between the SRC
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475 matrix and plasticizers in the films as shown in Fig. 3 (C and D). In FTIR analysis, the specific

476 chemical groups present in the plasticizers (glycerol and sorbitol) were also characterized (Fig. 3,

477 A and B). The main functional groups of glycerol (Fig. 3A) were O-H stretching at 3350 cm−1 to
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478 3310, C-H stretching at 2930 cm−1 and 2880 cm−1, H2O bending at 1650 cm−1, C-O-H bending

479 at1400 to 1460 cm−1, C-O stretching from 1450 cm−1 (primary alcohol) to 1100 cm−1 (secondary
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480 alcohol) and O-H bending 920 cm−1 (Kongjao, Damronglerd, & Hunsom, 2010). The main
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481 functional groups of sorbitol (Fig. 3B) were O-H stretching at 3335 cm−1 (this peak was broader

482 due to the large number of hydroxyl groups present compared to glycerol, which has less number
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483 of hydroxyl groups), C-H stretching at 2938 cm−1 and C=O stretching at 1645 cm−1. The FTIR

484 spectra of the control SRC, SRC-30G and SRC-30S films showed similar patterns in the

485 positions of characteristic bands indicating that SRC had a good miscibility with both

486 plasticizers. Fig. 3C shows the broad band between 3200 and 3400 cm−1 attributed to O–H

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487 stretching vibration formed by the hydroxyl groups of carrageenan and water; and higher peak

488 intensities were observed in this region for films plasticized with glycerol or sorbitol (compared

489 to the control SRC film). A broad band around 2800‒3000 cm-1 was attributed to C–H stretching

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490 (Pereira, Amado, Critchley, van de Velde, & Ribeiro-Claro, 2009). Fig. 3D shows the bands

491 corresponding to characterised groups of κ-carrageenan at 1220 cm−1 (ester sulfate groups); a

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492 sharp peak at 1034 cm−1 (glycosidic linkages); 920 cm−1 (3,6-anhydrogalactose ring) and 843

493 cm−1 (galactose-4-sulfate) (Pereira et al., 2009).

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494

495 3.11. Morphology of films

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496 Since the SRC-30S film had a better mechanical property (highest EAB) and showed good

497 oxygen and oil barrier properties which could also be linked to the surface properties, it was
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498 selected for SEM and AFM studies. Scanning electron micrographs of the control SRC film (A)

499 and the SRC-30S film (B) are shown in Fig. 4. SEM analysis clearly showed that the addition of

500 sorbitol in the SRC based film induced a significant change in the structural arrangement of the
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501 film surface compared to the control film surface. The control SRC film showed an irregular and

502 rough surface with tangled, branched and aggregated lines (Fig. 4A), which may possibly be due
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503 to the high degree of intermolecular forces (naturally occurring) between SRC matrix chains
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504 which led to the formation of complex entanglements. Overall, SEM micrographs of SRC-30S

505 film revealed a relatively compact and uniform surface compared to the control SRC film. This
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506 result suggested that an enhanced film network via the plasticization process, in turn led to

507 increased film flexibility with improved mechanical properties (Table 2) and reduced opacity

508 (Table 3).

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509 Atomic force micrographs showing 3D images of the control SRC film (a) and the SRC-30S

510 film (b) are shown in Fig. 4. The surface features of the SRC film were influenced by the

511 addition of sorbitol. The control SRC film displayed a rough surface with Rq and Ra values of

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512 0.396 µm and 0.313 µm, respectively, while, the SRC-30S film possessed a smoother surface, as

513 indicated by lower values of Rq (0.145 µm) and Ra (0.111 µm). The plasticizing effect (by

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514 sorbitol) reduced the intermolecular forces between polymer chains which could possibly cause

515 much difference in the roughness parameters between the SRC-30S film and SRC control film.

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516 These results correlated with visual examinations (flexibility and smoothness). AFM results

517 confirmed that the plasticized film showed a homogeneous and continuous network, indicating

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518 ordered network as was also observed in SEM micrographs.

519
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520 4. Conclusions

521 The results obtained suggest that the semi-refined carrageenan; an inexpensive seaweed

522 polysaccharide can be used as an alternative packaging material to the expensive refined
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523 carrageenan for the production of edible films with improved mechanical and barrier properties

524 that can be applied in food packaging systems. The tensile strength and elongation at break of
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525 films were significantly improved by the addition of glycerol and sorbitol with a drastic increase
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526 in the elongation at break was found in the film plasticized with 30% sorbitol. The heat seal

527 strength of films significantly increased with increasing plasticizers concentration. Furthermore,
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528 the oil permeability of films was considerably reduced with increasing glycerol and sorbitol

529 concentrations, and the minimum oil permeability was observed in the film plasticized with 30%

530 sorbitol. The sorbitol-plasticized films showed excellent barrier properties to oxygen as

531 compared to the control and glycerol-plasticized films, and therefore could be excellent

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532 packaging films for food products sensitive to oxygen. Further studies will be focused on the

533 incorporation of active compounds from a natural source into the SRC film plasticized with

534 sorbitol to produce edible active films to be exploited in active food packaging systems.

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535

536 Acknowledgements

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537 The authors would like to acknowledge Universiti Sains Malaysia for the facilities given for

538 this research work and funding given by the Ministry of Agriculture and Agro-based Industry,

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539 Malaysia (grant no: 304-PTEKIND-650539).

540

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541 References

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657 48, 870‒878.

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658 Vieira, M. G. A., da Silva, M. A., dos Santos, L. O., & Beppu, M. M. (2011). Natural-based
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659 plasticizers and biopolymer films: A review. European Polymer Journal, 47, 254‒263.

660 Voon, H. C., Bhat, R., Easa, A. M., Liong, M. T., & Karim, A. A. (2012). Effect of addition of
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661 halloysite nanoclay and SiO2 nanoparticles on barrier and mechanical properties of bovine

662 gelatin films. Food and Bioprocess Technology, 5, 1766‒1774.

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663 Yan, Q., Hou, H., Guo, P., & Dong, H. (2012). Effects of extrusion and glycerol content on

664 properties of oxidized and acetylated corn starch-based films. Carbohydrate Polymers, 87,

665 707‒712.

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Table 1
Thickness, moisture content and water solubility of semi-refined κ-carrageenan (SRC) based
films plasticized with glycerol (G) or sorbitol (S) at different concentrations (20, 25 and 30%).
Film samples Thickness (mm) Moisture content (%) Water solubility (%)
SRC-Control 0.057 ± 0.001a 8.8 ± 0.6ab 42.8 ± 1.6a
SRC-20G 0.066 ± 0.001bc 9.8 ± 0.3bc 44.4 ± 1.3ab

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SRC-25G 0.073 ± 0.002d 10.2 ± 1.4cd 45.9 ± 1.7b
SRC-30G 0.079 ± 0.001e 14.5 ± 1.0e 49.5 ± 1.9c
SRC-20S 0.063 ± 0.002b 7.7 ± 0.2a 42.6 ± 1.4a

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SRC-25S 0.066 ± 0.001bc 9.6 ± 0.6bc 44.9 ± 1.2ab
SRC-30S 0.073 ± 0.002d 11.0 ± 0.5d 46.3 ± 0.6b

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a‒eValues with different letters within the same column are significantly different (p < 0.05). Data
reported are means ± standard deviation.

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Table 2
Tensile strength (TS), elongation at break (EAB) and water vapor permeability (WVP) of semi-
refined κ-carrageenan (SRC) based films plasticized with glycerol (G) or sorbitol (S) at different
concentrations (20, 25 and 30%).
Film samples TS ( MPa) EAB (%) WVP × 10−8 (g mm cm−2 h−1 Pa−1)
SRC-Control 40.30 ± 3.95a 1.77 ± 0.33a 3.26 ± 0.06a

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SRC-20G 44.63 ± 2.49b 3.80 ± 0.52b 3.30 ± 0.07a
SRC-25G 47.07 ± 2.95b 4.68 ± 0.55b 3.70 ± 0.16b
SRC-30G 47.42 ± 2.72b 8.09 ± 1.34c 4.32 ± 0.41c

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SRC-20S 45.54 ± 2.75b 4.02 ± 1.03b 3.26 ± 0.13a
SRC-25S 48.00 ± 3.93b 4.68 ± 0.24b 3.66 ± 0.12ab
SRC-30S 48.74 ± 2.84b 10.01 ± 1.44d 4.25 ± 0.28c

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a‒dValues with different letters within the same column are significantly different (p < 0.05). Data
reported are means ± standard deviation.

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Table 3
Color parameters (L*, a*, b*) and opacity values of semi-refined κ-carrageenan (SRC) based
films plasticized with glycerol (G) or sorbitol (S) at different concentrations (20, 25 and 30%).
Film samples L* a* b* Opacity
SRC-Control 93.88 ± 0.13bc −0.23 ± 0.01d 4.00 ± 0.18a 4.82 ± 0.14f
SRC-20G 93.97 ± 0.21c −0.34 ± 0.03c 4.76 ± 0.30b 4.47 ± 0.26e

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SRC-25G 93.90 ± 0.22bc −0.36 ± 0.01bc 5.12 ± 0.26bcd 4.28 ± 0.05d
SRC-30G 93.62 ± 0.59ab −0.39 ± 0.02ab 5.51 ± 0.19d 4.02 ± 0.08c
SRC-20S 93.89 ± 0.11bc −0.36 ± 0.05bc 4.67 ± 0.36b 3.83 ± 0.15b

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SRC-25S 93.72 ± 0.53ab −0.40 ± 0.01ab 4.94 ± 0.11bc 3.76 ± 0.06ab
SRC-30S 93.47 ± 0.24a −0.41 ± 0.02a 5.31 ± 0.39cd 3.63 ± 0.12a

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a‒fValues with different letters within the same column are significantly different (p < 0.05). Data
reported are means ± standard deviation.

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Table 4
Oil permeability (Po) and oxygen permeability (O2P) of semi-refined κ-carrageenan (SRC) based
films plasticized with glycerol (G) or sorbitol (S) at different concentrations (20, 25 and 30%).
Film samples Po (g mm m−2 d−1) O2P × 10−4 (cm3 mm m−2 d−1 kPa−1)
SRC-Control 1.12 ± 0.05e 1.82 ± 0.10b
SRC-20G 0.97 ± 0.05d 1.84 ± 0.10b

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SRC-25G 0.81 ± 0.04c 1.85 ± 0.10b
SRC-30G 0.49 ± 0.08b 1.67 ± 0.03b
SRC-20S 0.90 ± 0.02d 0.89 ± 0.20a

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SRC-25S 0.77 ± 0.04c 0.88 ± 0.20a
SRC-30S 0.37 ± 0.03a 0.76 ± 0.09a

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reported are means ± standard deviation.

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A 2500 Control SRC-20G SRC-25G SRC-30G

Water gain (%)

2000

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1500

1000

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500

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0
0 20 40 60 80 100 120
Time (min)

B 2500 Control
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SRC-20S SRC-25S SRC-30S
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Water gain (%)

2000

1500
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1000
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500
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0
0 20 40 60 80 100 120
Time (min)
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Fig. 1.

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200 f
f
180
Film seal strength (N/m)

160 e

140 d

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120 c
b
100

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a
80

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60

40

20

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Control SRC-20G SRC-25G SRC-30G SRC-20S SRC-25S SRC-30S
Film type
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Fig. 2.
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T (%)
A

Glycerol

4000 3500 3000 2500 2000 1500 1000 500

Wavenumber (cm)

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T (%)

B
Sorbitol

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4000 3500 3000 Wavenumber
2500 2000 (cm)
1500 1000 500

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T (%)

SRC-
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SRC-30G

SRC-30S
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4000 3500 3000 2500 2000 1500 1000 500

Wavenumber (cm)
1034

920

843

D
1
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T (%)

SRC-
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SRC-30G

SRC-30S
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1500 1400 1300 1200 1100 1000 900 800 700 600 500
Wavenumber (cm)

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A a

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Rq = 0.396 µm
Ra = 0.313 µm

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B b

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Rq = 0.145 µm
Ra = 0.111 µm
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Fig. 4.
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List of Figure captions

Fig. 1. Water sorption profiles of semi-refined κ-carrageenan (SRC) based films plasticized with
different concentrations (20, 25 and 30%) of glycerol (A) and sorbitol (B). Error bar
shows standard deviation.

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Fig. 2. Seal strength of semi-refined κ-carrageenan (SRC) based films plasticized with different
concentrations (20, 25 and 30%) of glycerol (G) and sorbitol (S). Different letters
represent significant differences (p < 0.05). Error bar shows standard deviation.

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Fig. 3. Fourier transform infrared (FTIR) spectra of glycerol (A), sorbitol (B), semi-refined κ-
carrageenan (SRC) control film, SRC films plasticized with 30% glycerol (SRC-30G)
and 30% sorbitol (SRC-30S) in the wave numbers range between 4000 and 600 cm−1 (C)

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and between 1500 and 600 cm−1 (D).

Fig. 4. Scanning electron micrographs (A, B) and atomic force microscopy images (a, b) of
semi-refined κ-carrageenan (SRC) based films: (A, a) SRC-control film, (B, b) SRC film

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plasticized with 30% sorbitol. Rq: root-mean square average of height deviations taken
from the mean data plane, Ra: average of the absolute value of the height deviations
from a mean surface.
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