Food Chemistry 122 (2010) 161–166

Contents lists available at ScienceDirect

Food Chemistry
journal homepage: www.elsevier.com/locate/foodchem

Development and evaluation of a novel biodegradable film made from chitosan

and cinnamon essential oil with low affinity toward water
Seyed Mahdi Ojagh a, Masoud Rezaei a,*, Seyed Hadi Razavi b, Seyed Mohamad Hashem Hosseini b
a
Dept. of Fisheries, Faculty of Marine Sciences, Tarbiat Modares University, Noor, Iran
b
Dept. of Food Science and Engineering, Faculty of Engineering and Agricultural Technology, University of Tehran, Iran

a r t i c l e i n f o a b s t r a c t

Article history: Combining antimicrobial agents such as plant essential oils directly into a food packaging is a form of
Received 9 August 2009 active packaging. In this work chitosan-based films containing cinnamon essential oil (CEO) at level of
Received in revised form 18 January 2010 0.4%, .0.8%, and 1.5% and 2% (v/v) were prepared to examine their antibacterial, physical and mechanical
Accepted 11 February 2010
properties. Scanning electron microscopy was carried out to explain structure–property relationships.
Incorporating CEO into chitosan-based films increased antimicrobial activity. CEO decreased moisture
content, solubility in water, water vapour permeability and elongation at break of chitosan films. It is pos-
Keywords:
tulated that the unique properties of the CEO added films could suggest the cross-linking effect of CEO
Chitosan film
Cinnamon
components within the chitosan matrix. Electron microscopy images confirmed the results obtained in
Antimicrobial this study.
Physical Ó 2010 Elsevier Ltd. All rights reserved.
Mechanical
Scanning electron microscopy

1. Introduction the antimicrobial efficacy of chitosan is its positively charged ami-

Microbial growth on food surfaces is a major cause of food
spoilage. There have been remarkable developments in recent
years in the polymeric packaging films incorporated with antimi-
crobial agents for improving the preservation of packaged foods.
These films possess the potential for improving microbial stability
of foods by acting on the food surface, upon contact (Cha, Choi,
Chinnan, & Park, 2002).
Chitosan is the deacetylated (to varying degrees) form of chitin
which is the second most abundant natural biopolymer after cellu-
lose, largely widespread in living organisms such as crustacean,
insects, and fungi. It is a linear binary heteropolysaccharide com-
posed of b-1,4-linked glucosamine and N-acetylglucosamine, and
is determined as a non-toxic, a biodegradable and a biocompatible
polymer (Beverlya, Janes, Prinyawiwatkula, & No, 2008; Shahidi,
Arachchi, & Jeon, 1999). Interestingly some antibacterial and anti-
fungal activities have been described for chitosan and modified
chitosan such as more soluble and biodegradable polymers with
the knowledge that this aminopolysaccharide was able to reduce
microbial growth (Tsai, Wu, & Su, 2000). One of the reasons for
* Corresponding author. Address: Dept. of Fisheries, Faculty of Marine Sciences,
Tarbiat Modares University, P.O. Box 46414-356, Noor, Iran. Tel.: + 98 122 6253101/
3; fax: +98 122 6253499.
E-mail address: rezai_ma@modares.ac.ir (M. Rezaei).
no group which interacts with negatively charged microbial cell
membranes, leading to the leakage of proteinaceous and other
intracellular constituents of the microorganisms (Jeon, Kamil, &
Shahidi, 2002; Shahidi et al., 1999). Chitosan-based films are excel-
lent oxygen barriers; however, due to their hydrophilic nature,
they have poor moisture barrier properties (Caner, Vergano, &
Wiles, 1998).
Many spices and herbs and their extracts possess antimicrobial
activity, which minimise questions regarding their safe use in food
products. Essential oils and their constituents have wide spectra of
antimicrobial action. The composition, structure as well as func-
tional groups of the oils play an important role in determining their
antimicrobial activity (Holley & Patel, 2005). Usually compounds
with phenolic groups are most effective (Dorman & Deans, 2000).
Among these, the oils of clove, thyme, cinnamon, rosemary, sage
and vanillin have been found to be most consistently effective
against microorganisms. Because of the effect of direct addition
of essential oils to food on sensory characteristics of added food,
incorporation of essential oils to films may have supplementary
applications in food packaging (Ojagh, Rezaei, Razavi, & Hosseini
2010; Seydim & Sarikus, 2006).
The overall objective of the current study was to improve anti-
microbial efficacy of chitosan-based films by incorporating cinna-
mon essential oils (CEO). Mechanical and physical properties
were characterised, and antimicrobial efficacy was assessed
against five pathogenic and spoilage bacteria. The effects of CEO

0308-8146/$ - see front matter Ó 2010 Elsevier Ltd. All rights reserved.
doi:10.1016/j.foodchem.2010.02.033
162 S.M. Ojagh et al. / Food Chemistry 122 (2010) 161–166
on the antimicrobial, physical and mechanical properties of films
were evaluated and explained in terms of their microstructures.
2. Materials and methods
2.1. Bacterial strains and maintenance
The bacterial strains used in this study were Listeria monocytog-
enes PTCC1163, Escherichia coli PTCC1399, Lactobacillus plantarum
PTCC1058, Lactobacillus sakei PTCC1272 and Pseudomonas fluores-
cens ATCC 17482. These bacteria were obtained from Persian Type
Culture Collection (Tehran, Iran) on nutrient agar slants and kept at
4 °C. Subculturing was carried out each 14 days to maintain bacte-
rial viability. Overnight cultures of L. plantarum and L. sakei were
grown in MRS (Man-Rogosa-Sharpe) broth at 37 and 30 °C at CO2
chamber (New Brunswick Scientific Co, Edison, NJ), respectively,
and L. monocytogenes, E. coli, and Ps. fluorescens were grown and
agitated at 140–150 rpm in an incubator shaker for 24 h in BHI
(brain heart infusion) broth at 37 °C. All the media were purchased
from Merck Co (Darmstadt, Germany). The bacterial population in
all the inoculated media was more than 1  109 CFU/ml after 24 h
incubation.
2.2. GC–MS analysis of cinnamon essential oil
Cinnamon (Cinnamomum zeylanicum) essential oil was obtained
from Zardband Co. (Tehran, Iran). GC/MS analysis was carried out
on a Hewlett Packard 6890 (II) coupled to an HP 5973 mass spec-
trometer detector with electron impact ionisation (70 eV) (Agilent,
Palo Alto, CA, USA). The analysis was carried out using DB-5 fused
silica capillary column (60 m, 0.25 mm I.D.; 0.25 lm film thickness,
J&W Scientific Inc., Rancho Cordova, CA, USA). The temperature
program was 10 min at 60 °C, then to 250 °C at 5 °C/min, held for
10 min. Other operating conditions were as follows: carrier gas, he-
lium (99.999%), with a flow rate of 1.1 ml/min; injection volume of
1 ll and split ratio, 1:50. Essential oil components were identified
by comparison of their retention indices (RI) relative to (C6–C24)
n-alkanes with those of authentic compounds under the same con-
ditions (Davies, 1990). Further identification was made by match-
ing their mass spectra fragmentation patterns with those stored in
the Wiley/NBS mass spectral library and other published mass
spectra (Adams, 2001). Moreover, relative percentage of the com-
ponent cinnamon oil was calculated by normalisation of the base
GC–mass peak.
2.3. Determination of MIC and MBC of CEO against bacterial strains
Minimum inhibitory concentration (MIC) and minimum bacte-
ricidal concentration (MBC) of CEO were determined according to
the method of Kim, Marshall, and Wei (1995).
2.4. Preparation of antimicrobial films
Chitosan-based film was prepared by dissolving crab shell
chitosan (medium molecular weight (190–310 kDa), 75–85%
deacetylated, Sigma Chemical Co., St. Louis, MO., USA) in an aque-
ous solution (1% v/v) of glacial acetic acid to a concentration of
2% (w/v) while stirring on a magnetic stirrer/hot plate. The solution
was stirred with low heat (at 40 °C) which typically required 6 h
stirring. The resultant chitosan solution was filtered through a
Whatman No. 3 filter paper to remove any undissolved particles.
After filtration the solution was returned to the magnetic stirrer/
hot plate and glycerol (Sigma Chemical Co., St. Louis, MO, USA)
was added to a level of 0.75 ml/g chitosan as a plasticiser. The plas-
ticiser was mixed into the solution for 30 min. Then Tween 80 at
level of 0.2% (v/v) of essential oil was added as an emulsifier to as-
sist essential oil dissolution in film forming solutions. After 1 h of
stirring, CEO was added to chitosan solution to reach a final con-
centration of 0.4%, 0.8%, 1.5% and 2% (v/v) as essential oil concen-
tration per film in emulsifying equipment (IKA T25-Digital Ultra
Turrax, Staufen, Germany) at 7000 rpm for 2 min. After cooling to
room temperature, the film forming solutions were degassed under
vacuum for 5 min. The film forming solutions (160 ml) were casted
on the centre of 27  27 cm2 glass plates, and then dried for 30 h at
ambient conditions (25 °C). Dried films were peeled and stored in a
desiccator at 25 °C and 51% relative humidity until evaluation. Sat-
urated magnesium nitrate solution was used to meet required rel-
ative humidity.
2.5. Determination of antimicrobial effects of films
The agar diffusion method was used for determining the anti-
bacterial effects of films on bacterial strains. The films were cut
into 15 mm diameter discs with a circular knife. Film cuts were
placed on BHI agar for L. monocytogenes, E. coli and Ps. fluorescens
and on MRS agar for L. plantarum and L. sakei. Agar plates had pre-
viously been seeded with 0.1 ml of an overnight broth culture of
indicator strains. The plates were incubated at appropriate temper-
ature for 24 h in the appropriate incubation chamber. The diameter
of the zone of inhibition was measured with a caliper to the nearest
0.02 mm. The whole zone area was calculated then subtracted
from the film disc area and this difference in area was reported
as the ‘‘zone of inhibition” (Seydim & Sarikus, 2006).
2.6. Determination of physical properties of films
2.6.1. Thickness
Thickness of the films was determined using a digital coating
thickness gauge (Elcometer A300 FNP 23, Elcometer Instrument
Ltd., Manchester, England) to the nearest 0.001 mm. Informed val-
ues are an average of at least ten random locations of the film
sheets. The means were calculated and used in the determination
of mechanical and physical properties.
2.6.2. Moisture content
Moisture content of films was determined measuring weight
loss of films, upon drying in an oven at 110 °C until a constant
weight was reached (dry sample weight).
2.6.3. Water vapour permeability coefficient (WVPC)
Standard method E 96 was used to determine WVPC with a 75%
relative humidity (RH) gradient at 25 °C (ASTM, 1995). Diffusion
cells containing anhydrous calcium chloride desiccant (0% RH, as-
say cup) or nothing (control cup) were sealed by the test film
(0.00287 m2 film area). To maintain a 75% RH gradient across the
film, a sodium chloride-saturated solution (75% RH) was used in
the desiccator. The RH inside the cell was always lower than out-
side. This difference in RH corresponds to a driving force of
1753.55 Pa, expressed as water vapour partial pressure. Water va-
pour transport was determined from the weight gain of the diffu-
sion cell at a steady state of transfer. Changes in the weight of the
cell were recorded to the nearest 0.0001 g and plotted as a function
of time. The slope of the weight vs time plot was divided by the
effective film area to obtain the water vapour transmission rate
(WVTR). This was multiplied by the thickness of the film and di-
vided by the pressure difference to obtain the water vapour perme-
ability coefficient (WVPC). All WVPC values were corrected for air
gap distance between calcium chloride and film surface by correct-
ing the values of WVTR, according to the equations of Gennadios,
Weller, and Gooding (1994).
 S.M. Ojagh et al. / Food Chemistry 122 (2010) 161–166 163

2.6.4. Surface properties measurements 2.9. Statistical analysis

Contact angle measurements were performed with water using
a goniometer (Kruss G23, Germany). A small drop of distilled water
was deposited on the film surface. The contact angle is defined as
the angle between the film surface and the tangent line at the point
of contact of the water droplet with the surface. For each film
type, at least five measurements were made and the average was
taken.
The triplicate data were performed to an analysis of variance for
the significance of added essential oils using MSTATC programs
(version 2.10, East Lansing, MI, USA). Duncan’s multiple range tests
was used to compare the difference among means at the level of
0.05.

3. Results and discussion

2.6.5. Film solubility in water
Pieces of film of 1  3 cm2 were cut from each film and weighed 3.1. Identification of volatile components from CEO
to the nearest 0.0001 g. The solubility in water of the different
chitosan films was measured from immersion assays under con- GC–MS analytical data of major compounds in CEO are shown
stant agitation in 50 ml of distilled water for 6 h at 25 °C. The in Table 1. E-Cinnamaldehyde was a predominant component
remaining pieces of film after immersion were dried at 110 °C to and accounted for 60.41% of the total peak area. The other compo-
constant weight (Final dry weight). The initial dry weight was nents were linalool (6.46%), ortho-methoxycinnamaldehyde
determined by thermal processing at 110 °C to constant weight. (3.63%), b-caryophyllene (3.5%), 1,8-cineole (3.32%), and eugenol
Solubility in water (%) was calculated by using the following equa- (3.19%).
tion (Eq. (1)):
3.2. MIC and MBC of CEO
Solubility in water ð%Þ
 
Initial dry weight  Final dry weight MIC and MBC values (antibacterial properties) of CEO are pre-
¼  100 ð1Þ
Initial dry weight sented in Table 2. The lower concentration of the essential oil could
fully inhibit the growth or could almost kill L. sakei (MIC 250 lg/
mL and MBC 1000 lg/mL) and Ps. fluorescens (MIC 500 lg/mL
2.6.6. Surface colour measurements and MBC 1000 lg/mL). However, MIC and MBC of the essential
Film colour was determined by a colourimeter (Minolta CR 300 oil for the other three bacteria were 500 and more than 1500 lg/
Series, Minolta Camera Co., Ltd., Osaka, Japan). The CIELab scale mL, respectively.
was used, lightness (L) and chromaticity parameters a (red–green) This is not supported by many other reports on the greater sus-
and b (yellow–blue) were measured. Measurements were per- ceptibility of gram-positive bacteria to inhibitory effect of essential
formed by placing the film sample over the standard white plate oils and their components (Oussalah, Caillet, Saucier, & Lacroix,
(L* = 93.49, a* = 0.25 and b* = 0.09). Colour differences (DE) 2007; Shan, Cai, Brooks, & Corke, 2007). Similar results have been
were calculated by the following equation (Eq. (2)):
qffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi

DE ¼ ðL  LÞ2 ða  aÞ2 ðb  bÞ2 ð2Þ Table 1
Essential oil composition of Cinnamomum zeylanicum.
where L*, a* and b* are the colour parameter values of the standard No. Compound RI % of the total peak area
and L, a and b are the colour parameter values of the sample.
1 a-Pinene 936 2.18
2 Camphene 950 0.14
3 Myrcene 982 1.03
2.7. Determination of mechanical properties of films
4 a-Phellandrene 1001 0.54
5 d-3-Carene 1010 0.94
The mechanical properties of films including tensile strength 6 P-cymene 1016 2.85
(MPa) and elongation at break (%) were performed at 25 °C and 7 1,8-Cineole 1026 3.32
51% RH. Tests were performed using a Testometric Machine 8 Linalool 1086 6.46
9 Z-Cinnamaldehyde 1188 0.29
M350-10CT (Testometric Co. Ltd., Rochdale, Lancs., England) 10 E-Cinnamaldehyde 1247 60.41
according to the ASTM method D882-91 (ASTM, 1996). In prepar- 11 Hexagerman 1312 0.48
ing samples, films were cut into 1  10 cm2 strips. The films were 12 Eugenol 1334 3.19
held parallel with an initial grip separation of 5 cm and then pulled 13 a-Copaene 1386 0.34
14 Coumarine 1398 0.43
apart at a head speed of 25 mm/min. Elongation at break (deforma-
15 E-Cynnamyl acetate 1413 2.01
tion divided by initial grip separation and multiplying by 100) and 16 b-Caryophyllene 1431 3.50
maximum force were obtained from force versus deformation 17 a-Humulene 1463 0.27
curves. Tensile strength was calculated by dividing maximum force 18 O-methoxycinnamaldehyde 1492 3.63
by film cross section (thickness  width). 19 Caryophyllene oxide 1585 0.34
20 Benzyl benzoate 1736 0.65
21 Equilin 2049 0.91
2.8. Scanning electron microscopy

The chitosan-based films were mounted on the specimen

Table 2
holder with aluminium tape and then sputtered with gold in
MIC and MBC values of CEO against five tested bacterial strains.
BAL-TEC SCD 005 sputter coater (BAL-TEC AG, Balzers, Liechten-
stein). All the specimens were examined with a Philips XL 30 scan- Bacteria L. E. coli Ps. L. L.
monocytogenes fluorescens plantarum sakei
ning electron microscope (Philips, Eindhoven, Netherlands) under
high vacuum condition and at an accelerating voltage of 20.0 kV. MIC (ppm) 500 500 500 500 250
Samples were photographed at tilt angles of 60–90° to the electron MBC >1500 >1500 1000 >1500 1000
(ppm)
beam for the views in the cross section.
164 S.M. Ojagh et al. / Food Chemistry 122 (2010) 161–166

reported by Kim et al. (1995) who found that L. monocytogenes was migration of active agents. Chitosan does not diffuse through the
more resistant to the inhibitory effects of eleven essential oil con- adjacent agar media in agar diffusion test method; so that only
stitutes than the gram-negative bacteria tested under the same organisms in direct contact with the active sites of chitosan are
condition, including E. coli, E. coli O157:H7, Salmonella typhimurium inhibited (Coma et al., 2002).
and Vibrio vulnificus. It seems that the variability of the inhibitory
effect of essential oil may be due to differences between strains 3.4. Physical properties of films
of the same bacterial species. This hypothesis was confirmed by
Sivropoulou et al. (1996) with two strains of Staphylococcus aureus The effects of incorporating CEO on the physical properties of
in the presence of carvacrol and thymol. chitosan films are shown in Table 4. Thin films were easily re-
moved from the cast plate. Thickness of films varied between
0.095 and 0.107 mm, as shown in Table 4.
3.3. Antimicrobial properties of films
In general, the moisture content value decreased as CEO was
incorporated into chitosan-based film, which is attributed to com-
Effects of CEO addition on antimicrobial properties of chitosan-
pactness of film network. CEO caused to formation covalent bonds
based films are shown in Table 3. When antimicrobial agents are
between the functional groups of chitosan chains, leading to a de-
incorporated into films, these materials diffuse through agar gel
crease in the availability of hydroxyl and amino groups and limit-
and result in clear zone around the film cuts.
ing polysaccharide–water interactions by hydrogen bonding and
Incorporation of CEO into chitosan-based films at higher than
resulting in a decrease of moisture content value of edible films
0.4% (v/v) exhibited a clear inhibitory zone by the absence of bac-
(Park & Zhao, 2004). As CEO concentration increased, the moisture
terial growth around the film cuts. At CEO concentration of 0.4% (v/
content of films decreased significantly (p < 0.05).
v) the clear zone of inhibition was not observed with L. plantarum.
Chitosan control films were compact, and the film surface had a
As the concentration increased, the zone of inhibition also in-
smooth surface without pores or cracks (Fig. 1A and B). Control
creased significantly (p < 0.05). However, increasing level of CEO
film had water vapour permeability coefficient (WVPC) value
at its high concentration did not reveal significant an increased
2.250  1010 g s1 m1 Pa s1. Same results were obtained by
inhibitory. It was generally caused by the compactness of chitosan
Wong, Gastineau, Gregorski, Tillin, and Pavlath (1992), Caner
film network containing CEO because of the occurrence of func-
et al. (1998) and Seydim and Sarikus (2006). Incorporation of
tional groups interaction phenomenon. The active component of
CEO into chitosan-based films decreased WVPC of films. Beside
CEO is cinnamaldehyde (Matan et al., 2006). As shown in Table 1,
the hydrophobic nature of CEO which could affect the hydro-
cinnamaldehyde constitutes more than 60% of CEO. Sublethal con-
philic/hydrophobic property of the films, the physical factors had
centrations of cinnamaldehyde have been found to inhibit produc-
an influence on the WVPC of the films enriched with CEO. When
tion of amylase and proteases by Bacillus cereus (Thoroski, Blank, &
CEO was incorporated into the chitosan film formulation, the sur-
Biliaderis, 1989).
Chitosan control films did not show inhibitory zone in bacterial
strains tested. Despite antimicrobial activity of chitosan because of Table 4
its innate characteristic, this effect of chitosan occurred without Physical properties of chitosan films incorporated with CEO.

Physical properties* CEO conc. (v/v) in

film solution (%)
Table 3
Antibacterial activity (inhibitory zone) of chitosan films incorporated with CEO Thickness (mm) Control 0.095 ± 0.0025b
against gram-positive and gram-negative bacteria. 0.4 0.098 ± 0.0024b
0.8 0.104 ± 0.0029a
Bacteria CEO conc. (v/v) in film Inhibitory zone* 1.5 0.105 ± 0.0032a
solution (%) (mm2) 2 0.107 ± 0.0039a

L. monocytogenes (Gram +) Control 0c Moisture content (%) Control 20.82 ± 1.82a

0.4 38.17 ± 1.00b 0.4 18.72 ± 2.11a
0.8 39.80 ± 0.94b 0.8 14.04 ± 0.93b
1.5 53.05 ± 1.84a 1.5 10.82 ± 1.47c
2 52.31 ± 2.65a 2 8.47 ± 1.53c

L. plantarum (Gram +) Control 0c Water vapor permeability coefficient Control 2.250 ± 0.074a
0.4 0c (g s1 m1 Pa s1  1010) 0.4 1.352 ± 0.152b
0.8 43.6 ± 0.94b 0.8 1.234 ± 0.040b
1.5 49.80 ± 2.77a 1.5 1.014 ± 0.040c
2 53.64 ± 3.07a 2 1.003 ± 0.067c

L. sakei (Gram +) Control 0c Water contact angle (°) Control 37.3 ± 2.25d
0.4 39.60 ± 2.36b 0.4 39.6 ± 1.89d
0.8 41.08 ± 1.44b 0.8 45.3 ± 1.38c
1.5 56.69 ± 2.82a 1.5 59.2 ± 2.77b
2 57.23 ± 1.50a 2 70.3 ± 1.42a

Ps. fluorescens (Gram ) Control 0c Solubility in water (%) Control 23.2 ± 1.09a
0.4 34.68 ± 1.71b 0.4 21.6 ± 0.65a
0.8 36.91 ± 1.32b 0.8 16.8 ± 0.85b
1.5 41.60 ± 2.09a 1.5 13.6 ± 1.55c
2 43.60 ± 1.21a 2 10.4 ± 0.94d

E. coli (Gram ) Control 0d Total colour difference (DE) Control 17.35 ± 0.98d
0.4 36.31 ± 0.77c 0.4 17.83 ± 0.47d
0.8 38.90 ± 0.94b 0.8 21.29 ± 0.55c
1.5 51.72 ± 1.15a 1.5 23.35 ± 0.87b
2 51.04 ± 2.12a 2 25.62 ± 0.58a
* *
Means in each column with different superscript letters are significantly different Means in each column with different superscript letters are significantly different
(p < 0.05). Control is a film disc containing no essential oil. (p < 0.05).
 S.M. Ojagh et al. / Food Chemistry 122 (2010) 161–166 165

Fig. 1. Scanning electronic microscopic images of chitosan control film (A) and (B), and film containing CEO at level of 1.5% (C) and (D) surfaces and cross sections,
respectively.

face view suggested a sheet like and dense structure, while the results agreed with visual observations. Nevertheless, the sensory
cross section revealed the sheets stacked in compact layers studies should be conducted for evaluating consumer acceptability
(Fig. 1C and D), which showed CEO incorporated uniformly in the of this colour change.
matrix.
The value of the contact angle with water indicates how hydro-
phobic the surface is. It is well-known that the water contact angle 3.5. Mechanical properties of films
will increase with increasing surface hydrophobicity. The higher
hydrophilicity of the control film is attributed to the water binding The effect of incorporating CEO on mechanical properties of
capacity of plasticiser (glycerol) and functional groups of chitosan. chitosan films is presented in Table 5. Chitosan control film had
According to Table 4, addition of CEO increased water contact angle tensile strength value 10.97 MPa. Incorporation of CEO into chito-
of films and resulted in decreasing hydrophilicity of the chitosan san films increased tensile strength values significantly (p < 0.05).
films which might be due to the loss of free functional groups (ami- As previously mentioned, a strong interaction between the poly-
no and hydroxyl groups). mer and the CEO produced a cross-linker effect, which decreases
Chitosan control film had low solubility in water below 24% at the free volume and the molecular mobility of the polymer. This
25 °C after 6 h dipping. Same result was reported by García, Pinotti, phenomenon led to a sheet like structure (Fig. 1C). Arrangement
Martino, and Zaritzky (2004). Control film in general was very of stacking layers of CEO added chitosan sheets (Fig. 1D) means
hydrophilic thus absorbed water quickly and resulted in swelling. that in these films a compact structure was present thus increased
Incorporation of CEO into the chitosan film formulation at level continuities within the polysaccharide network leading to decrease
of 1.5% and 2% (v/v) led to 41% and 55% reduction in solubility in in elongation at break. There are also possibilities such as decrease
water respectively. This phenomenon is due to the cross-linking ef- in moisture content of films incorporated with CEO during the film
fects of CEO components leading to esters and/or amide groups. production, thus leading to the decrease in strain and the increase
Cross-linking in the chitosan film matrix resulted in a decrease in
solubility in water and produced films with low affinity toward
Table 5
water which is beneficial when product integrity and water resis- Mechanical properties of chitosan films incorporated with CEO.
tance are intended. Cross-linking of chitosan-hydroxy propyl
Mechanical properties* CEO conc. (v/v) in film solution (%)
methyl cellulose based films using citric acid (polycarboxylic acid)
as the cross-linking agent, was reported by Möller, Grelier, Pardon, Tensile strength (MPa) Control 10.97 ± 0.54e
and Coma (2004). 0.4 13.35 ± 1.23d
0.8 17.43 ± 1.08c
The colour of the films may influence the consumer acceptabil- 1.5 24.10 ± 1.47b
ity of a product (Sivarooban, Hettiarachchy, & Johnson, 2008). Total 2 29.23 ± 2.25a
colour differences (DE) of chitosan films are shown in Table 4. Elongation at break (%) Control 24.73 ± 1.86a
Visually, chitosan films had a slightly yellow appearance. Control 0.4 16.57 ± 0.77b
film had DE value 17.35. Its transparency was reduced as the 0.8 11.26 ± 1.39c
CEO was incorporated. Incorporating CEO revealed DE values, 1.5 6.42 ± 0.63d
2 3.58 ± 0.35e
which were significantly higher than that of the control. The values
of chromaticity parameter b of chitosan films incorporated with *
Means in each column with different superscript letters are significantly different
CEO were higher than that of control film (data not shown). These (p < 0.05).
166 S.M. Ojagh et al. / Food Chemistry 122 (2010) 161–166
in tensile strength. The results of moisture content measurements
are shown in the Table 4. According to Gontard, Guilbert, and Cuq
(1993) and Pouplin, Redl, and Gontard (1999) water is the most
ubiquitous and uncontrollable plasticiser for most hydrocolloid-
based films because of its ability to modify the structure of natural
polymers. Thus, the plasticisation effect of water could not be neg-
ligible for these films, and their plasticisation efficiency does not
only come from intrinsic plasticiser action of glycerol. Differences
between data of chitosan films obtained in the current study and
those of reported in literature (Caner et al., 1998; García et al.,
2004) may be attributed to chitosan composition and suppliers,
plasticiser presence and film preparation.
4. Conclusions
Chitosan is a promising biopolymer for active food packaging.
Its sensitivity to moisture can be offset by blending it with CEO.
The results of this study showed that a unique compatibility can
be achieved between chitosan and CEO. The incorporation of CEO
improved the antibacterial properties of chitosan. Films containing
CEO are useful for coating of highly perishable foods such as fish
and poultry. The other edible films may be ruptured upon contact
with wet surfaces. Functional groups interaction phenomenon in
edible films has critical effect on their antibacterial, physical and
mechanical properties which are important in food packaging
applications. Moreover, the antimicrobial effect of CEO enriched
films should be determined on an entire model food.
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