Journal of Food Engineering 111 (2012) 343–350

Contents lists available at SciVerse ScienceDirect

Journal of Food Engineering
journal homepage: www.elsevier.com/locate/jfoodeng

A novel active bionanocomposite film incorporating rosemary essential oil and nanoclay into chitosan
Mehdi Abdollahi a, Masoud Rezaei a,⇑, Gholamali Farzi b
a b

Department of Fisheries, Faculty of Marine Sciences, Tarbiat Modares University, P.O. Box 46414-356, Noor, Iran Department of Material and Polymer Engineering, Faculty of Engineering, Sabzevar Tarbiat Moallem University, Sabzevar, Iran

a r t i c l e

i n f o

a b s t r a c t
Montmorillonite (MMT) nanoclay and rosemary essential oil (REO) were incorporated into chitosan film to improve its physical and mechanical properties as well as antimicrobial and antioxidant behavior. The MMT weight percent relative to chitosan was varied from 1 to 5 and was activated by three REO levels (0.5%, 1%, and 1.5% v/v), and their impact on physical, mechanical, and barrier properties of the chitosan films was investigated. Total phenolic and antimicrobial activity were also evaluated. Microstructure of chitosan/MMT–REO nanocomposites was characterized through X-ray diffraction (XRD) and Fourier transform infrared spectroscopy (FTIR). The results showed that incorporating MMT and REO into chitosan improves water gain, water vapor permeability, and solubility of the chitosan film by more than 50%. It was also shown that the combined effect of clay and REO improves significantly the tensile strength and elongation of chitosan (p < 0.05). The XRD and FTIR results confirmed that the improvements are related to the MMT exfoliation and good interaction between chitosan and MMT in the presence of REO. Antimicrobial properties of the films also improved by REO incorporation in 1.5% v/v. Ó 2012 Elsevier Ltd. All rights reserved.

Article history: Received 9 August 2011 Received in revised form 4 February 2012 Accepted 7 February 2012 Available online 16 February 2012 Keywords: Bionanocomposite Chitosan Montmorillonite Rosemary essential oil Food packaging

1. Introduction Biopolymers have been widely investigated over the last two decades because they can be a viable solution to the waste disposal of foods’ plastic packaging materials. Furthermore, biopolymer films are excellent vehicles for incorporating a wide variety of additives, such as antioxidants, antifungal agents, antimicrobials, colours, and other nutrients (Rhim and Ng, 2007; Wong et al., 1994), and so these biodegradable materials can also improve food quality and extend shelf life by minimizing microbial growth in the product. Chitosan, a deacetylated derivative of chitin, is the second most abundant polysaccharide found in the nature after cellulose. It has been found to be non-toxic, biodegradable, biofunctional, and biocompatible (Rinaudo, 2006; Shahidi et al., 1999; Srinivasa and Tharanathan, 2007). Chitosan brings some advantages over other biomolecule-based active polymers used as packaging materials because of its antibacterial behavior and bivalent minerals chelating ability (Aider, 2010). However, poor mechanical and gas barrier properties, and weak water resistance limit its application particularly in the presence of water and humidity (Wang et al., 2005; Xu et al., 2006). As for other biopolymers, many strategies have been explored to improve the barrier and mechanical properties of chitosan⇑ Corresponding author. Tel.: +98 122 6254986; fax: +98 122 6253499.
E-mail address: rezai_ma@modares.ac.ir (M. Rezaei). 0260-8774/$ - see front matter Ó 2012 Elsevier Ltd. All rights reserved. doi:10.1016/j.jfoodeng.2012.02.012

based biodegradable films. These include the addition of plasticizers and salt, chemical modification of hydroxyl groups, cross-linking of polysaccharides, the use of suitable solvent, the change of pH, the addition of different polysaccharides, and blending with other polymers (Bourtoom and Chinnan, 2008; Ghosh et al., 2010; Hoagland and Parris, 1996; Park and Zhao, 2004; Srinivasa et al., 2007; Li et al., 2010; Gómez-Estaca et al., 2011). Recently, polymer–clay nanocomposites have received significant attention as an alternative to conventional filled polymers, because of their ability for nano-scale dispersion, which brings significant improvement in mechanical and physical properties compared to micro-scale polymer composites. Several studies have reported amelioration of mechanical properties (Lavorgna et al., 2010; Xu et al., 2006), thermal stability (Darder et al., 2003; Wang et al., 2005), functional properties (Rhim et al., 2006), barrier properties (Casariego et al., 2009; Rhim et al., 2006), and water solubility (Casariego et al., 2009) of chitosan films via incorporation of nanoclay into chitosan in the range of 1–5 wt.%. Montmorillonite is the most widely studied type of clay which is a hydrated alumina-silicate layered clay consisting of an edge-shared octahedral sheet of aluminum hydroxide between two silica tetrahedral layers. The imbalance of the surface negative charges is compensated by exchangeable cations (typically Na+ and Ca2+) (Azeredo, 2009). On the other hand, antibacterial and antioxidant activities of essential oils (EOs) have long been acknowledged, but the food industry has recently been paying more attention to their

Mizonokuchi.5%. .. using a magnetic stirring plate at 90 °C and 1250 rpm for 20 min and then cooled to room temperature. XRD patterns were taken with a Philips X’Pert MPD Diffractometer (Eindhoven. and DP is the partial water vapor pressure difference (Pa) between the two sides of the film. 2. was added to the mixture and stirred at 40 °C for 30 min.2.1. 1. as an emulsifier. the combined effects of montmorillonite nanoclay and rosemary essential oil on the physico-chemical and antimicrobial properties of chitosan composite films have been investigated. 2. and the antimicrobial/antioxidant properties contributed by the natural antimicrobial/antioxidant agents impregnated within. USA) and Na+montmorillonite (MMT) from Southern Clay Products. Sample preparation 2. Selected amounts of clay (1.89% relative humidity until evaluation. bornyl acetate (0–17%). 2004). Iran. The slope of weight loss vs. Sigma–Aldrich Chemical Co. Nanocomposite samples were prepared according to methods reported by Xu et al. 2009. Netherlands). Materials Crab shell chitosan (medium molecular weight. In the present study.3. The initial dry matter of samples (4 Â 4 cm) was determined by drying films at 105 °C for 24 h. The WVP was then calculated as follows: WVP ¼ ðWVTR Â LÞ=DP ð2Þ Where WVTR (water vapor transmission rate. 2007).001 mm.. and 1. 2. Germany) in the range of 400–4000 cmÀ1 at a resolution of 4 cmÀ1. rosmadial. 3.. Burt.. 2007). etc. Then. and stored in dark container at 4 °C until used. 2010a).5% v/v.5.3. Measurements were repeated in ten different regions of each sample. Staufen. on the basis of neat chitosan solution.544 nm. carnosoic acid. Water resistance The solubility of the film was determined as the percentage of soluble matter to initial dry matter of film sample (Tunc et al. Although several reports on chitosan/clay nanocomposites have appeared in the literature. Average values were calculated and used in water vapor permeability and tensile properties calculations. Calculation of water vapor permeability The water vapor permeability (WVP) of the films was measured gravimetrically according to the ASTM E96–92 method as described by Casariego et al. epirosmanol. 1 filter paper. rosmarinic acid.3. with Co Ka radiation at a wavelength of 1. 2. 2. The films were then immersed in 50 mL distilled water and then placed in a shaker incubator at 25 °C and stirred for 24 h at 250 rpm. then dried for 72 h at ambient conditions (T = 25 °C and RH = 55% ± 2%) to prepare nanocomposite films. rosmadiphenol. Preparation of chitosan/clay-rosemary solutions Aqueous solution of chitosan was prepared by dissolving 20 g of chitosan powder in 1000 mL of aqueous acetic acid solution (1%. v/ v) to obtain a 2% w/v chitosan solution. After cooling the resulting mixture at room temperature. 2007). Sánchez-González et al.2. Then Tween 80 at a level of 0. 75–85% deacetylated. (2006). Germany) at 7000 rpm for 2 min. The cell contained distilled water (100% RH. Materials and methods 2. The main advantage of EOs’ application is their greater activity as compared with the effects of the individual active compounds. Mitutoyo.2. kg/m2 s1) is the measured slope. / Journal of Food Engineering 111 (2012) 343–350 application as natural antimicrobials and antioxidants (Du Plooy et al. Measurement of film thickness The thickness of the samples was determined with a manual digital micrometer (0. and air was stirred in the desiccators. Among essential oils the largest number of reports on the effective antioxidant properties pertains to extracts from plants belonging to family Labiatae.1.. 1%.) (Waszkowiak.3.. Preparation of nanocomposite films The chitosan/clay-rosemary solutions (160 mL) were cast in simple cubic molds made from Teflon-coated steel with dimension of 25. Dried films were then peeled and stored in a desiccator containing saturated magnesium nitrate solution at 25 °C and 52. and 5 w% on solid chitosan) were dispersed in 100 mL of 1% (v/v) aqueous acetic acid solution and vigorously stirred for 24 h. 2004). rosmanol. to reach a final concentration of 0.5 cm. Glacial acetic acid and Tween 80 were purchased from Merck. the samples were filtered through Whatman No. 2008). Rosemary essential oil was obtained from Barij Essence Pharmaceutical Co. 2. with a scan speed of 1°/min at room temperature. camphor (2– 14%). are considered to have antioxidant ability (Bozin et al. Germany. In addition.5 Â 28..5% RH (28.3. 2. Chitosan/clay-rosemary films were scanned over the range of diffraction angle 2h = 1–12°. 2010b. USA were used as received. The film solubility (%) was calculated using the following equation: Solubility in waterð%Þ ¼ ðInitial dry weight À Final dry weightÞ Â 100=Initial dry weight ð1Þ 2.8-cineole (3–89%) (Burt. Afterward 150 mL of chitosan solution was added to the clay solutions and the mixture was stirred for 4 h. Major components of REO which have showed antimicrobial properties are a-pinene (2–25% of composition). Japan). The weight loss of the cell was considered equal with the transferred water through the film and adsorbed by the desiccant.1. Kashan. L is the mean film thickness (m).044 Pa water vapor pressure) with silica gel. to the best of our knowledge there are no reported data on the combined effect of nanoclay and antioxidant/antimicrobial compounds like EOs. Sample characterization 2. Abdollahi et al. it was degassed under vacuum for 5 min in order to remove all bubbles.3. 2008. The tested film was sealed on the top of a glass permeation cell (internal diameter = 3 cm). This test was repeated three times for each specimen to confirm its repeatability.2.4. probably due to the synergistic effects (Bakkali et al. (2009).2. and homogenized with Ultra-Turrax (IKA T25-Digital Ultra-Turrax. X-ray diffraction (XRD) characterization The structure of nanoparticle and its nanocomposites was evaluated by XRD measurement. Phenolic compounds such as carnosol.3.344 M..2% w/v of essential oil. Weight loss of the permeation cell was determined at intervals of 2 h for 10 h. The papers containing any insolubilized film were dried at 105 °C for 24 h. Sánchez-González et al.337 Â 103 Pa vapor pressure at 20 °C) placed in a desiccator. 2. 190–310 kDa. time was obtained by linear regression. Finally appropriate amounts of rosemary essential oil (REO) were added to the solution. It seems that the combination of nanoclay and an antimicrobial/antioxidant compound in chitosan films can be suitable due to its acceptable structural integrity and barrier properties imparted by the nanocomposite matrix. Fourier transform infrared (FTIR) spectra FTIR spectra were collected in transmission mode by using a Bruker Equinox 55 FTIR spectrometer (Ettlingen. particularly rosemary (Rosmarinus officinalis L. It was maintained at 20 °C and 1. such nanocomposite films prepared based upon natural biopolymers are environmentally friendly with all the benefits one may expect from biopolymer and nanocomposite packaging materials (Rhim and Ng. at 40 kV and 30 mA.

CA. The weighed samples were placed in closed beakers containing 30 mL of water (pH = 7) and stored at 25 °C. 2. Water Gainð%Þ ¼ ðWeight of wet film À Weight of dry filmÞ Â 100=Weight of dry film ð 5Þ 2. Escherichia coli (PTCC 1533). Streptococcus agalactiae (PTCC 1768).0001 g.% gave a broad peak at 2h = 5. 2010).82° (d0 0 1 = 17.59 Å). These results are in good agreement with those obtained by other investigators (Lavorgna et al. (2010). the data reported in Fig. The procedure was repeated three times for each sample to confirm its repeatability. desiccated overnight (containing silica gel). 2. Surface colour measurement Colour properties of films were measured using a colour meter from BYK Gardner.3. 30 lL of REO. +++: totally inhibited. Spain) at 37 °C. The color development is due to the transfer of electrons at basic pH to reduce the phosphomolybdic/phosphotungstic acid complexes (Curcio et al.1 ml of inoculums by swab containing approximately 106–107 CFU/ml of the indicated bacteria. such as its degree of deacetylation and molecular weight (Rhim et al.88. Hiwa Engineering Co. MMT solution. Phenolic compounds undergo a complex redox reaction with phosphotungstic and phosphomolybdic acids present in the Folin–Ciocalteu reaction. Germany). 1 shows that the structure of nanocomposites is not affected by the presence of the REO. and film disks Antibacterial properties of REO.3.9 ml of distilled water were added to it. The film samples were cut in rectangular specimens (2. after their plates had been seeded with 0. With the addition of 1 and 3 wt. in the text. This agrees with previously reported results of Lavorgna et al. The crystalline structure of chitosan is strongly dependent on its processing treatment. (3) and (4)..63. and different film-forming solutions were poured into Mueller Hinton (Scharlua.5 ml of Folin–Ciocalteu reagent (Merck Company. À: no inhibition as explained by Gómez-Estaca et al. Duncan’s multiple range tests were used to compare the means to identify which groups were significantly different from other groups (p < 0.8. Initial grip separation was set at 50 mm. the insertion of polymer into the organoclay galleries forces the platelets apart and increases the d-spacing. 3. .10. A calibration curve was drawn using gallic acid in specific concentrations and the total phenolic content of the films was expressed as Gallic acid equivalents. Fig. and b⁄ (yellow/blue).. However.3. Total phenolic assay Total phenolic content of the films was studied using the Folin– Ciocalteu method as described by Siripatrawan and Harte (2010) with some modification. the plates were incubated in chamber at 37 °C for 24 h. 50 mg of each film sample were dissolved in 3 ml of methanol. After incubation the microbial growth was observed and the degree of inhibition was expressed as follows.9. Bacterial strains were cultured overnight in Brain Heart Infusion Broth (Scharlua. 2006).56° (d0 0 1 = 11. ++: partially inhibited.3. The films’ wet surface was gently blotted with a tissue before weighing with a balance accurate to 0. Strong polar interactions. During intercalation. It was shown that inorganic and hydrophilic MMT in low concentration (1–5%) could easily be dispersed in the chitosan solution to form a favorable interaction with hydrophilic chitosan. Fremont. In the same way. film solutions.4. 2006). Darmstadt. Results of XRD characterization of chitosan and chitosan/MMT nanocomposite are shown in Fig. The absorbance values were then measured at 765 nm using a spectrophotometer (Lambda 25. Further increasing the amount of MMT to 5 wt.6.1 ml of film extract solution was mixed with 7 ml distilled water and 0. USA). 1. the reflection peak disappeared. The mixture was stored in darkness and at room temperature for 2 h... and cross-head speed was set at 50 mm/min.5 ml of saturated sodium carbonate solution and 0. Five different pathogenic and spoilage bacteria including Listeria monocytogenes (Persian Type Culture Collection (PTCC) 1163). The water gain of each sample was calculated as follows: The difference between factors and levels was evaluated by the analysis of variance (ANOVA). Perkin–Elmer. Spain) agar wells (5 mm diameter). and Lactococcus lactis (PTCC 1336) were used for testing. 1. The parameters were determined by placing film samples on a standard plate (L⁄ = 94. Next. which means the crystallinity of chitosan is slightly reduced by the addition of the MMT. 1 show lower intensity for the characteristic crystallinity peak of chitosan for composite films at 2h = 10. indicating formation of an exfoliated structure.% MMT to chitosan. After preserving the mixture for 8 min at room temperature. The weighing was continued until equilibrium state. suggesting that intercalation occurs together with some exfoliation (Xu et al. a⁄ (red/green). +: slightly inhibited.05).18°. Colour difference (DE) and whiteness index (WI) were calculated with respect to standard plate parameters by using following Eqs.54  10 cm). Pseudomonas putida (PTTC 1694). This test was repeated five times for each specimen and the Gallic acid equivalent value was reported as mean ± SD of triplicate. Results and discussion 3. 2006). 2010. Rhim et al. as well as its origin and molecular constitution. (2010).. This was much lower than that of pristine MMT. MMT had a characteristic peak at 2h = 8. Mechanical properties Tensile strength (TS) and elongation at break (E%) of the film samples were determined according to ASTM standard method D 882–02 with an Instron Universal Testing Machine (Model 200. resulting in a shift of the diffraction peak to lower angles (Xu et al. a⁄ = À0. MMT. / Journal of Food Engineering 111 (2012) 343–350 345 2. The sample was cut into small pieces (2  2 cm).7.M.. Swelling Swelling of the clay and REO containing chitosan films was evaluated by determining their water sorption according to the method explained by Lavorgna et al. films were punched into discs of 6 mm diameter. XRD analysis of chitosan nanocomposites Wide-angle X-ray diffraction is a classical method for determining the gallery height (d-spacing distance) in clay particles.. Folin–Ciocalteu phenol reagent was used to obtain a crude estimate of the amount of phenolic groups present in the films.. respectively.93 Å). Xu et al. 2006. Statistical Analysis 2.3. Antibacterial activity of the essential oil. USA. which was disordered and not detectable by XRD. 2009). and 0..65). and then placed on the plates. The kinetic of swelling was evaluated by periodically measuring the weight increment of the samples. film-forming solution and disks were studied using the agar diffusion method (Gómez-Estaca et al. Abdollahi et al. and b⁄ = 0. qffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi à DE ðDaà Þ2 þ ðDb Þ2 þ ðDLà Þ2 qffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi Ã2 ð100 À Là Þ2 þ aà 2 þ b ð 3Þ WI ¼ 100 À ð 4Þ 2.1. Measurements are expressed as L⁄ (lightness). This test was repeated five times for each specimen to confirm its repeatability. All data are presented as mean ± SD. 2006). and weighed to determine their dry mass. Iran). (2010).

2b) compared to those incorporated with clay (Fig. Film appearance characterization The colour of the packaging is an important factor in terms of general appearance and consumer acceptance (Bourtoom and Chinnan.346 M. with a slight yellowish tint according to the amounts observed for b⁄ of the films. two strong bands at 1541 and 1403 cmÀ1. 1997). These peaks flattened more with incorporating clay and REO. The results were in good agreement with the results of Rhim et al. 2c) and REO (Fig. (2006). and 1403 cmÀ1. Abdollahi et al. 2008) which could act as hydrogen-bonding sites for functional groups of chitosan. 2008). 2010).. yellow/blue (b⁄) coordinates. 2008). chitosan-MMT3% (c).. chitosan (b).. Fig. associated with OH in-plane bending. especially hydrogen bonding. These characteristic peaks have shifted to lower frequencies in nanocomposites which recommend some interaction between functional groups of chitosan and MMT (Katti et al. 2d). 3. chitosan.3. The bands appearing between 2750 and 3000 in the spectrum of chitosan film are due to stretching vibrations of C–H bond in –CH2 (m = 2930 cmÀ1) and –CH3 (m = 2870 cmÀ1) groups (Paluszkiewicz et al. / Journal of Food Engineering 111 (2012) 343–350 Fig. These are stronger in neat chitosan film (Fig. FTIR spectra of MMT (a). In addition. 2010).. Casariego et al. The peaks at 3426 and 1639 cmÀ1 were the H–O–H stretching and bending vibrations of the adsorbed water. Red/green (a⁄). A shift towards lower frequency in the absorption bands at 3209 in the FTIR spectra was coincidental with the decrease in peaks at 3500–3000. The result of colour measurements for chitosan films are shown in Table 1. 1541. FTIR analysis of chitosan/clay nanocomposites Fig. It is further shifted to lower frequency (m = 3130 cmÀ1) for chitosan/clay 1% nanocomposite containing 1% REO. 2009). respectively. and total colour differences (DE) significantly increased by incorporating REO and increased as REO increased. 2010) respectively. and shifted towards lower frequency (m = 3202 cmÀ1) in nanocomposite with 3% MMT. The peak of Al–Al–OH stretching vibration at 3626 cmÀ1 found in the spectrum of MMT is typical for smectites with high amount of Al in the octahedral layer. and chitosan/ MMT nanocomposite films. 2008). and chitosanMMT3%–REO1% (d) films.2. In the spectra of the nanocomposites. . which shows that hydrogen-bonding interaction between chitosan and clay is enhanced in the presence of REO in accordance with other results (Tang et al. (2009) reported a similar result about incorporating MMT in chitosan film. the N–H bonded to the O– H vibration band at 3209 cmÀ1 in chitosan. The results are in agreement with the results of Ojagh et al.. Chitosan film lightness increased significantly by incorporating MMT but REO did not affect the nanocomposites’ lightness. All of the films were transparent. (2010) and Siripatrawan and Harte (2010). This observation presents an assumption that there could be a particular arrangement in the films due to the interactions of MMT and REO ingredients’ functional groups with hydroxyl and amino groups in chitosan matrix (Siripatrawan and Harte. It is also important to note that the peaks between 3500 and 3000 cmÀ1 correspond to the stretching vibration of free hydroxyl and to asymmetric and symmetric stretching of the N–H bonds in the amino group (Siripatrawan and Harte. critically affect the formation of intercalated and exfoliated hybrids (Vaia and Giannelis. This may be due to hydrogen bonding between the –OH group in MMT and functional groups in REO ingredients and the –NH and –OH groups in chitosan (Wang et al. 3. This fact indicates that ÀNH2 and ÀOH groups of chitosan formed hydrogen bonds with the ÀOH group of MMT (Wang et al. 1. XRD patterns of MMT powder and chitosan-based nanocomposite films. respectively. are less discernible in the films incorporated with clay and REO. 2006). Whiteness (WI) increased in the presence of MMT and decreased when REO was incorporated. 2 shows the FTIR spectra of MMT. The sharp peak at 1034 cmÀ1 was attributed to Si–O stretching frequency (Tunç and Duman. 2.

71bcd ± 7.004 0.47 12...051 ± 0. Fig.99 83.24 86.33bcd ± 2.31 16.40fg ± 1.43 81.77 10.37 11.02 86.61 87.65 12.00 0.4 80. As the results presented in Table 2. 07 À1.21b ± 1.12abce ± 1. water could not break hydrogen bonds between MMT layers and chitosan sufficiently which caused the lower solubility in the nanocomposites (Tunç and Duman.005 0.96 80. Swelling Knowledge of the swelling is also important for predicting stability and quality changes during packaging and storage of food product (Srinivasa et al. Swelling of chitosan and its nanocomposites are displayed in Fig.52 80.005 0.42de ± 1.004 0. The result was in contrast with result of Lavorgna et al.78). Montmorillonte (MMT).28bc ± 0.10a ± 0.17bce ± 1. as confirmed by FTIR analysis.60 11.94abcd ± 0.43c ± 0.09b ± 0.86def ± 0.90bcd ± 0.004 0.006 0.65 L⁄ a⁄ b⁄ 347 DE WI 80.45 13.85 80.10abcd ± 2.61abcd ± 0.4. In nanocomposites containing 1 and 5% MMT.31bc ± 0.54 9. (2010) that showed chitosan/clay nanocomposite had more water sorption compared to neat chitosan film.94a ± 1.003 0.Tensile Strength (TS).44bcd ± 0.6) similar to that reported by Ojagh et al.01 60.68 85.83cde ± 0.24a ± 0.50b ± 1.20 À1.44f ± 0.052 ± 0.52 4.74 82.99 11.98 74.50a ± 1.005 17.47 Thickness (mm) Solubility (%) WVP (g/msPa) 10À10 TS (MPa) E (%) a-g Different superscripts within the same column indicate significant differences among formulations (p < 0.38 9.39de ± 0.18 8.15 À1.11 4.5 0. 3 A).05). to avoid exudation of fresh or frozen products (Bourtoom and Chinnan.51ab ± 0.36 À0. 3 B.10 a-e Different superscripts within the same column indicate significant differences among formulations (p < 0.81abc ± 0.35b ± 0.5 1 1.01 0.96ab ± 0. 3 B).17de ± 1.052 ± 0.46 73. Water resistance Water resistance is an important property of biodegradable or edible films for applications in food protection where water activity is high.28a ± 1.21 À1. This fact is highly attributable to cross-linking effects of REO components to esters and/or amide groups (Ojagh et al.37def ± 1.96 85.39 71.76cd ± 0.09 4. Table 2 Summary of water vapor permeability.02bc ± 6.78 12.30g ± 0.36f ± 0. Colour difference (DE) and Whiteness index (WI).19 15. 2010).49 À1..31bc ± 0.36ef ± 0.97 ± 0.54 3. 3B and D) but in films with 3% MMT it was detected in 1% REO (Fig. Higher cross-linking density in chitosan leads to a matrix with lower water solubility results.20 12. Rosemary Essential oil (REO).54f ± 0. Moreover.60abc ± 2.5 0 0. Abdollahi et al.47b ± 0.049 ± 0.57 79. indicates.91bc ± 0. C. the lowest amount of water sorption was observed in 1.48abc ± 4.67a ± 1. As can be seen in these figures. (2009).60 11.5 0 0.42c ± 0.5 0 0.93ab ± 0.23 10.11 À1. and mechanical and physical properties for chitosan-based nanocomposite films. / Journal of Food Engineering 111 (2012) 343–350 Table 1 Surface colour parameters of chitosan-based nanocomposite films. 3.04 À0.02 85.74cd ± 3.75efg ± 1.M. and D show swelling of REO-incorporated chitosan nanocomposites.04 À0.96 12.87bc ± 0.051 ± 0.24 À0.68 16.5 0 0.01 0.03 13.10bc ± 0.79 cd ± 0.71ce ± 2.54 13. When MMT was added to the film.22 83.00 10.01 0.57ab ± 0.55g ± 0.37a ± 0.5 82.050 ± 0.42a ± 1.41cd ± 0.66ab ± 2.70c ± 0.01 0.83ab ± 0.5 1 1.70 9.01cd ± 1.27 4.51a ± 0.5 1 1.22bce ± 0. This can be justified by the fact that MMT nanoclay brings significant barrier properties due to its disk-like morphology and there are of course some specific interactions between chitosan and MMT that stabilize the film structure. a significant (p < 0.86bcd ± 1.03bc ± 0.45bcd ± 3.33bcd ± 5. It was supported by FTIR spectra that showed the interaction of chitosan-functional groups with MMT and further interactions in the presence of REO.62a ± 1. Rosemary Essential oil (REO).006 0.90abcd ± 1.75 16.01 13.31g ± 0.66 10.01 0..05). chitosan film absorbed large amounts of water (about 1400%) in the first few minutes.19a ± 2.55 ± 0. 2007b).20 16.56cd ± 0.56 cd ± 1. As can be seen from this figure.051 ± 0.28 82.56a ± 0.42cd ± 0.31def ± 0.78 71.70b ± 0.050 ± 0.51b ±0.33bcd ± 5.01bcd ± 0.18 15.51 7.005 0. Similar results were reported by Casariego et al. the hydrophobic nature of REO may increase film hydrophobicity.95 81.54 4. In the other word.74 5. chitosan film showed a low solubility value of (17.11efg ± 1. which is beneficial when product integrity and water resistance are intended (Hosseini et al. e. and swelled and crumpled before reaching equilibrium (Fig. As explained previously. (2010)..15a ± 1.53 3.50a ± 1.052 ± 0. or when the film must be in contact with water permanently as long as it acts as food protective film.16cd ± 1.00a ± 1. The lowest water solubility was observed for nanocomposite films containing 3% clay and 1% REO (9.20ab ± 1.04 84.75 15.16 79.54abc ± 0.01d ± 0. 2010).11b ± 0.12b ± 1.14 À2. When REO was added to the chitosan clay nanocomposite matrix the solubility of the film further decreased.66 12. this low water sorption is also most likely to be due both to the formation of the cross-link network induced by the hydrogen bonds between the chitosan and REO and good intercalation of chitosan molecules into the silicate galleries (Lavorgna et al.80d ± 8.95b ± 1.22c ± 1.43a ± 1.33 15.59 84.54bc ± 1. 3.003 0.5% REO (Fig.63b ± 1.49 3. Elongation €.67c ± 0.05) decrease in water solubility was observed (Table 2).60 63.47 65.38 72.26a ± 2.25 82.050 ± 0.29 0.5 1 1.77 15.71a ± 0. 2010).70defg ± 1.71 3.53de ± 1.049 ± 0.051 ± 0. Water Vapor Permeability (WVP).60abc ± 2.5 1 1.22 4.051 ± 0.02 0.64 82.39d ± 0.36 À1.22b ± 0.02 11.35f ± 0.38d ± 1.14 85.43 7.57 82.5 1 1.48 74.84 3.02 0.47a ± 0.00a ± 3.34 69. .5.23 14.30 13.01 0.77bc ± 2.00 0.97a ± 0.005 0. The lowest amount of water sorption relates to chitosan nanocomposite containing 3% MMT.g.004 0.11bc ± 0. 3.90 11. water sorption of REO containing nanocomposites is 85% less than films not containing REO. 2009).01 0.02 0.15 À2. Concentration MMT (%w/w chitosan) Control 1 1 1 1 3 3 3 3 5 5 5 5 REO (%v/v) 0 0 0.47 72.90 6. 2008).45b ± 0.17d ± 0.21bcd ± 1.71ab ± 4.90 64.15 82. But all nanocomposites reached equilibrium and the amount of sorbed water was less than chitosan film.09a ± 2.Montmorillonte (MMT).61 3.96abc ± 0. Concentration MMT (%w/w chitosan) Control 1 1 1 1 3 3 3 3 5 5 5 5 REO (%v/v) 0 0.

.5% REO. (N) CS/MMT 3%. 2006). when the amount of MMT increased to 5 w%. and (d) CS/MMT 5%. This hypothesis was supported by FTIR spectra in which the interaction of chitosan functional groups with MMT in the presence of REO is shown. Abdollahi et al. exfoliation of chitosan MMT occurs together with some intercalation (Fig. (Ç) CS/MMT 3/REO 0. As can be seen from Table 2. This can be seen from the XRD pattern of the chitosan/MMT. for similar samples Elongation (E%) decreased slightly. even at a low amount of REO. 2009).7.. The decrease in TS at high concentration of MMT may be caused by some aggregation of MMT particles with high surface energy (Xu et al. 3.5%. (N) CS/MMT 5/REO 1%. Further decreasing WVP in the presence of REO relates to the hydrophobic nature of REO. subsequently leading to a decrease in the affinity of chitosan film to water and decreasing WVP (Siripatrawan and Harte. 3. TS value for . As mentioned previously. and (d) CS/MMT3/REO 1. (Ç) CS/MMT 1%. This parameter was studied to evaluate the combined effect of clay and REO on the barrier properties of the chitosan film. 2010). As can be seen from this table. The WVP of chitosan-based films is shown in Table 2. 1997). The highest TS was observed for nanocomposite with 5% of MMT and 0.5%. Mechanical properties Mechanical properties of chitosan-based films are presented in Table 2.1988. (Ç) CS/MMT 1/REO 0. This can be attributed to the quality of clay dispersion in chitosan. TS increased about 20% and 15% when the concentration of MMT was 3 and 5 w%. (Ç) CS/MMT5/REO 0. As the XRD pattern showed under this condition. This is the case for decreasing the WVP of chitosan/clay nanocomposites compared to pure chitosan.348 M. (B) film containing 1% MMT: (j) CS/MMT 1%. It seems that orientation of nanoclay changes once its concentration reaches a certain level and/or some aggregation of nanoclay probably occurs (Casariego et al. A significant decrease of WVP relates to the presence of ordered dispersed nanoparticle layers with relatively large aspect ratios in chitosan (Cussler et al..5% of REO. Water sorption profiles of (A) chitosan film without REO: (j) CS. 2006). The presence of REO resulted in further decreasing of WVP. and 1% or 1. It was demonstrated that for chitosan nanocomposite containing 3 w% of clay.. WVP decreased about 40 or 45%.6. However. it is believed that clay improves the barrier properties of polymers due to its disk-like morphology and its creation of a tortuous path for the molecule diffusion (Rhim et al. / Journal of Food Engineering 111 (2012) 343–350 Fig..5%. This improvement in the TS of nanocomposite in a low amount of MMT may be attributed to the uniform dispersion of MMT in chitosan matrix and a strong interaction between chitosan and MMT. (2010) also found that the TS of the chitosan film containing various concentrations of MMT was significantly higher than that of chitosan. It may be that the hydrogen and covalent interactions between the chitosan network and REO ingredients limit the availability of hydrogen groups to form hydrophilic bonding with water.5%. The result showed that the tensile strength (TS) of chitosan film increased significantly by incorporating MMT into chitosan up to 3 w% but adding more MMT could not improve it. WVP decreases by 58%. Despite the low percentages (1 and 3 w%) of MMT could decrease WVP of chitosan/MMT nanocomposite. and (d) CS/MMT 1/REO 1. (N) CS/MMT3/ REO 1%. when 1 or 3 w% MMT was incorporated into chitosan. nanocomposite did not improve. respectively. Yano et al.5%. 1). (D) film containing 5% MMT: (j) CS/MMT 1%. 3. and (d) CS/MMT 5/REO 1. WVP of chitosan/MMT. Xu et al.5%. The TS of nanocomposite improved significantly by incorporating REO. respectively. which could affect the hydrophilic/ hydrophobic property of the films. (2006) and Lavorgna et al. (N) CS/MMT 1/REO 1%. (C) film containing 3% MMT: (j) CS/MMT 3%. Water vapor permeability WVP is one of the most important parameters for biodegradable films.

F. agalactiae ++ À À À À + + ++ À À + + E. 2009. this nanocomposite is about 36.. Abdollahi et al. 2010.M.. Conclusions It was shown that incorporation of very low amounts of nanoclay into chitosan significantly improves its physical and mechanical properties. L. and film discs. Total phenolic content of chitosan-based films was shown in Fig. 2009.. which make them very interesting for food preservation. the high antimicrobial properties of REO are mainly related to phenol diterpenes.. MMT. (%) 0 0 0 Control 0. Biological effects of essential oils .. our group will report the results of the antimicrobial and antioxidant aspects of chitosan nanocomposites. Azeredo. total phenol content of chitosan film increased significantly by incorporating REO and the highest value (2. References Aider.Food Science and Technology 43 (6). Chitosan/clay nanocomposite without REO did not show antibacterial properties. 4. H. LWT .. 2007. (2005) about chitosan film. Antibacterial activity and total phenolic content Antibacterial activity of REO. The E% increased significantly by incorporating REO up to 44%.. In this regard. coli) was also notable and interesting. (2010) and Zivanovic et al.23 mg galic acid/g sample) was for the film formulated with clay 3% and REO1. which was in agreement with the results of Rhim and Ng (2007). coli strain. S. Good interaction between chitosan and MMT could improve WVP. (2007) also reported good inhibitory activity for REO against E. D..5% REO could show antibacterial activity in comparison to neat chitosan. Clay con.5% higher than pure chitosan film. The results are in agreement with the results of other published data (Bozin et al. (%) Pure REO MMT MMT MMT FFS well 1 3 5 3 3 3 Film disk 3 3 3 REO con. water sensitiveness. Films solution and discs containing 1% and 1. As described for swelling. and mechanical properties of the chitosan films. These results coincide with the results of Ojagh et al. Idaomar. S. 2008). 2010).. Averbeck.A review. Food and Chemical Toxicology 46 (2). Norajit et al.. 3. Zarei for graciously providing some raw materials. +: slightly inhibited. MMT. film-forming solutions (FFS). the essential oil showed its best antibacterial activity in the disc diffusion test on gram-positive bacteria (i. such as carnosic acid.5 1 1.5% which was in agreement with other reported results (Gómez-Estaca et al. We also thank to Dr. M. Gómez-Estaca et al. film forming solutions... Averbeck.. coli ++ À À À À À + + À À À + 349 L. 837–842. 4. chitosan nanocomposites containing REO exhibit antimicrobial properties and more phenol content. like the number of phenolic hydroxyl or methoxyl groups. monocytogenes ++ À À À À À + ++ À À + + À: No inhibited. and film discs is shown in Table 3. Regarding their extra protective outer membrane. Chitosan application for active bio-based films production and potential in the food industry: Review. Badiei for preparing some experimental facilities for this research. ++: partially inhibited. Türe et al. isorosmanol. Bakkali. keto groups. its inhibitory activity on gram-negative bacteria (especially on E. In the near future.5 L. 2002). Fig. gram-negative bacteria are usually considerably more resistant to antibacterial agents than their gram-positive counterparts. MMT did not show antibacterial activity in the well test but we could not find reported results about this test for comparison. Siripatrawan and Harte.5 1 1. monocytogenes. M. Bozin et al. so that only organisms in direct contact with the active sites of chitosan are inhibited (Coma et al. 2010. agalactiae). However. and other structural features (Jayabalan et al. Food Research International 42 (9). carnosol. The effect of phenolic compounds on lipid molecules may depend on structural factors. As expected. In general. flavone hydroxyl. XRD pattern showed MMT exfoliation and FTIR spectra demonstrated good interaction between chitosan and MMT. Acknowledgments We are grateful to Dr.8. . rosmanol..e. 446–475. In addition.5 Control 0. free carboxylic groups. 1240–1253. Total phenol content of chitosan-based films. / Journal of Food Engineering 111 (2012) 343–350 Table 3 Antibacterial activity of REO. 2008). This effect of chitosan may be related to the fact that chitosan does not diffuse through the adjacent agar media in the agar diffusion test method. 2007. Nanocomposites for food packaging applications. perhaps the presence of REO in low amounts changes the hydrogen-bonding network within the material and allows better interaction between nanofiller and matrix.. As explained before. 2008. The compatibility of REO with chitosan/MMT nanocomposite was confirmed in order to produce an active bionanocomposite for food packaging. 4. lactis + À À À À À À + À À À À P. Neat chitosan did not show antimicrobial properties in neither solution form nor in film form. and rosmarinic acid (Bozin et al. putida + À À À À À + + À À À À S. 2010). and it seems that REO has shown a plasticizing ability. it has been showed in many recent studies that the antioxidant activity of plants is caused mainly by phenolic compounds.

Usuki. 065707. 2010. J. C... J. I. Draughon. (1994). 93–99. Park.. Polymer Degradation and Stability 94 (1). Mimica-Dukic. 2005. López de Lacey. Chemistry of Materials 15 (20).. International Journal of Food Microbiology 94 (3). Chitosan/clay films’ properties as affected by biopolymer and clay micro/nanoparticles’ concentrations. Ghosh. Colilla.... González-Martínez. Sánchez-González... 1437–1442. Physical properties of edible chitosan films containing bergamot essential oil and their inhibitory action on Penicillium italicum. 1996. H. P. P. Incorporation of a high concentration of mineral or vitamin into chitosan-based films. K. 443–452. 727–743.. N. P. Y. 2007... food quality.. Edible coatings and films to improve. O. Picci. R.... Eroglu. 2008. Parris. D. C... The effect of different molecular weight of poly (ethylene glycol) on the electrokinetic and rheological properties of Na-bentonite suspensions..E. S. Carbohydrate Polymers 82 (2). Chi.H. X..M. Tharanathan.. A. M. Regnier. Abdollahi et al.. Walls. Burt. Changes in free-radical scavenging ability of kombucha tea during fermentation. Siripatrawan.. Food Microbiology 27 (7). P. 2009. S. Kim. Polymer Degradation and Stability 90 (1).. ecofriendly packaging materials with multiple potential uses. 1915–1919. Lattice model of polymer melt intercalation in organically-modified layered silicates. S. Journal of Agricultural and Food Chemistry 55 (19).. M... Wang. Lavorgna. 2009. Shahidi. Food applications of chitin and chitosans. 2006. W.. bovine-hide and tuna-skin. Critical Reviews in Food Science and Nutrition 47 (4). Sánchez-González. Ward III. 1633–1641.. Effect of plasticizers and fatty acids on mechanical and permeability characteristics of chitosan films. R. Carbohydrate Polymers 82 (2). Parisi. 3774–3780. Stodolak. Zhang. Ng.. Journal of Food Engineering 98 (3). 484–490. 161–166. Food Chemistry 122 (1).. K. O. Yano. Okada. Biopolymer.. Chiralt. Y. Gómez-Guillén. Chitosan/pectin laminated films.. Katti. Preparation and characterization of chitosan-based nanocomposite films with antimicrobial activity. 2005.A. Antioxidant properties of tuna-skin and bovine-hide gelatin films induced by the addition of oregano and rosemary extracts... Shen. Biopolymer chitosan/montmorillonite nanocomposites: preparation and characterization.. Deschamps. Journal of Polymer Science Part A: Polymer Chemistry 35 (11). A.. / Journal of Food Engineering 111 (2012) 343–350 Park. Chinnan. M. B. X. Buonocore. D. T. M. 2010.. P.C. 1684–1691. Wang.. Journal of Food Science 70 (1). 45–51. A. Jovin.. Rhim... S. Chalier. 2008. Hong.. Razavi.. Ojagh. 603–632. E.. J. 2009. C.G. Y. Carbohydrate Polymers 81 (2). Dash.. Journal of Membrane Science 38 (2). Essential oils: their antibacterial properties and potential applications in foods-a review.. E. Li. U. S. 2004.... F. Özen. 2010. (2002). Curcio. A.. Hosseini. Lim... Physical properties and antioxidant activity of an active film from chitosan incorporated with green tea extract.. J. Fu. Covalent insertion of antioxidant molecules on chitosan by a free radical grafting procedure. E. Food Hydrocolloids 21 (7). Development and evaluation of a novel biodegradable film made from chitosan and cinnamon essential oil with low affinity toward water. characterization and antimicrobial activity of chitosan/layered silicate nanocomposites. Modification of microstructural morphology and physical performance of chitosan films.. 159–168. LWT ... López-Caballero. Antifungal activity of biopolymers containing natamycin and rosemary extract against Aspergillus niger and Penicillium roquefortii. Copinet.. Journal of Agricultural and Food Chemistry 57 (13).. International Journal of Biological Macromolecules 46 (2).. 223–253. F. W. H. A.. J. 2010. Aris.. A. Gómez-Estaca. Azam Ali. Fernández-Martín. Preparation and properties of chitosan nanocomposites with nanofillers of different dimensions. Cirillo.. Food Reviews International 23 (1). Food Hydrocolloids 24 (8). S. Cháfer.H.. G.. T. S.S. Food Hydrocolloids 23 (7).Food Science and Technology 41 (9). Díaz. Hosseini. Preparation and properties of rice starchchitosan blend biodegradable film. Spizzirri.. 1162-1169. Hu. L.. Mangiacapra. Food Chemistry 109 (1).. 7990–7999.. K. Q.. X.. M. M... Souza. Journal of Agricultural and Food Chemistry 54 (16). Synthesis and properties of polyimide-clay hybrid films... 117–122.. Pavlath.G.. P.. Harte. 1999. 414–424.... L. Gómez-Guillén. Samojlik. N.R. Tharanathan. E. 2010.Clay Nanocomposites Based on Chitosan Intercalated in Montmorillonite.C. 161–174.. Zhang.. Gómez-Estaca.. Rinaudo. P. Rhim. Montero. Tong. 37–51.C. Montero.. Coma. Barrier membranes. on the properties of compound gelatin–chitosan films. M. 2008. Preparation. Giannelis... Razavi. Liu. Srinivasa. Combrinck.. Du Plooy. F. Gastaldi. Hasik... Synthesis and characterization of a novel chitosan/montmorillonite/hydroxyapatite nanocomposite for bone tissue engineering. 277–283. Tunç. M. M. A. W. Yang. Bravo.. Camirand... Journal of Applied Polymer Science 99 (4). 1997... E. Piscitelli. J. and Salvia officinalis L. Study of the combined effect of both clay and glycerol plasticizer on the properties of chitosan films. R.. Journal of Membrane Science 289 (1–2). G.C. S. 123–131.. Mousavi. P. Antimicrobial. Wang. Ren.. Progress in Polymer Science 31 (7). Subathradevi. Xu. Cháfer. R. F. S. Colloids and Surfaces A: Physicochemical and Engineering Aspects 317 (1–3). Darder. Vaia. 5814–5822.. 2006.. E. R. Jeon. K... Y. 1461–1469. S. K. Y.S. 5933– 5938. González-Martínez. Casariego. Soyer. 2009. Journal of Agricultural and Food Chemistry 52 (7). Ng.. Angellier.. Blazewicz. 2008. Journal of food science 67(3).. Trends in Food Science & Technology 10 (2). Nanotechnology 19 (6). M. 2010... Cussler.. Song. I. K. physical and mechanical properties of chitosan-based films incorporated with thyme. Salin. 1997. 124–131. 2004. P. Lamiaceae) essential oils. Chitin/chitosan—Safe. Cahyana. Sathishkumar. Preparation and functional properties of blend films of gliadins and chitosan. Ryu.. Arachchi.. J.. S. M. Y. M. N.. Food Chemistry 112 (1)... H. Edible antimicrobial films based on chitosan matrix. 2009. Tunç.. 179–186. L. R. Postharvest Biology and Technology 53 (3). 2003. 1988. 2010.M. Chitin and chitosan: properties and applications. 53–72. Biodegradable gelatin-chitosan films incorporated with essential oils as antimicrobial agents for fish preservation.. E. Chen. Journal of Food Engineering 98 (4). Antimicrobial activity of chitosan films enriched with essential oils. B. M. Shi. Bozin.. 2026–2032.A. 2008. 770–775. Duman.. 2007.. 2011. Puoci. X. G. Phang. L.. S.. Hoagland. 18–25. F.W. Bourtoom. J. Macromolecules 30 (25).I. M. P. Lin. Paluszkiewicz. Polymer 47 (19). 2010. Vicente. 2010a.. Effects of gelatin origin... X. Journal of Food Processing and Preservation 33 (6).S. 2289–2294. 1–12. FTIR study of montmorillonite-chitosan nanocomposite materials... 2008.. M. S... Swaminathan. Wang. Chitosan/clay nanocomposite film preparation and characterization. International Journal of Food Science & Technology 43 (8). M. Y. Zhao. M. 2010b. Du.. Marimuthu.. Alemán. X.. Y. 889–896. 2007. Q. Ruiz-Hitzky. 291–298. P. Guo.350 M.. 1933–1939.. L. Y.. Ramesh. Waszkowiak. Tang. M. Iemma. Teixeira. International Journal of Food Science & Technology 43 (11). Comparative studies on the characterization and antioxidant properties of biodegradable alginate films containing ginseng extract. Luo. Functional properties of wheat gluten/montmorillonite nanocomposite films processed by casting. M. J.. A.H.N. Hughes. B. B. Gómez-Guillén. P. 2006. Zivanovic. Applied Clay Science 48 (3). drug-controlled release property and cytotoxicity.. 2007. Duman. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 79 (15). Chiralt.. Development of edible coatings for minimally processed fruits and vegetables. S. 1895–1902. Biopolymer/montmorillonite nanocomposite: preparation. 2008... V. 65–88. Türe. 411–433. Natural biopolymer-based nanocomposite films for packaging applications. A. 784–788. Antimicrobial and antioxidant properties of rosemary and sage (Rosmarinus officinalis L. A.. Cruz.. Food Hydrocolloids 25 (6).. K. 377–384.R.. 227–234. Chen. C. N.. F. A. Martial Gros.. Garreau.. P. F. M. L.. 2010. R. Fan. 6738–6744. Biomedical Materials 3. .F. A. O. S.N. 2009... Gómez-Estaca.. N. Hanna. M. Jayabalan... A. M. Rezaei.. Wang. Preparation and characterization of biodegradable methyl cellulose/montmorillonite nanocomposite films. Wong.. 1113–1122.. R. U.M. Physical and antimicrobial properties of chitosan-tea tree essential oil composite films. X.. Norajit. Srinivasa. Cerqueira.. A.. Essential oil amended coatings as alternatives to synthetic fungicides in citrus postharvest management. Montero.. T. S. clove and cinnamon essential oils. Y... Journal of Agriculture and Food Chemistry 44 (7). S. Katti. 2006. Antioxidative activity of rosemary extract using connective tissue proteins as carriers..A. C. 7879–7885.... Gontard. Tunc. 2007. Du.

Sign up to vote on this title
UsefulNot useful