LWT - Food Science and Technology 54 (2013) 346e352

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LWT - Food Science and Technology

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Cassava starch composite films incorporated with cinnamon essential

oil: Antimicrobial activity, microstructure, mechanical and barrier
properties
A.C. Souza a, G.E.O. Goto a, J.A. Mainardi a, A.C.V. Coelho b, C.C. Tadini a, c, *
a
Department of Chemical Engineering, Escola Politécnica, University of São Paulo, SP, Brazil
b
Department of Metallurgical and Materials Engineering, Escola Politécnica, University of São Paulo, SP, Brazil
c
NAPAN e Food and Nutrition Research Center, University of São Paulo, SP, Brazil

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

Article history: Antimicrobial activity of cinnamon and clove essential oils is widely known; however their application to
Received 3 March 2013 polymeric materials is already limited. Based on results of their minimum inhibitory concentration
Received in revised form against Penicillium commune and Eurotium amstelodami, cinnamon was chosen to be incorporated into
20 May 2013
cassava starch films in order to develop an active packaging. The effect of cinnamon essential oil were
Accepted 20 June 2013
evaluated on antimicrobial activity, mechanical and barrier properties of films and the results were
compared with those of control films (without antimicrobial agent). ANOVA (P < 0.05) showed that the
Keywords:
essential oil content influenced significantly the properties of the films. The release of antimicrobial
Cassava starch
Antimicrobial activity
agent and the microstructure of cassava films incorporated with cinnamon essential oil were also
Microstructure studied. Furthermore, all films, containing different amounts of essential oil, showed effective antimi-
Mechanical properties crobial activity against P. commune and E. amstelodami, fungi commonly found in bread products.
Barrier properties Ó 2013 Elsevier Ltd. All rights reserved.

1. Introduction this regard (Bertuzzi, Castro Vidaurre, Armanda, & Gottifredi, 2007;
Chen & Lai, 2008; Chillo et al., 2008; Famá, Goyanes, &
In order to keep food quality and freshness, it is necessary to Gerschenson, 2007; Famá, Flores, Gerschenson, & Goyanes, 2006;
select correct materials and packaging technologies. In this way, Kaisangsri, Kerdchoechuen, & Laohakunjit, 2012; Kechichian et al.,
current tendencies include the development of packaging materials 2010; Mali, Grossmann, García, Martino, & Zaritsky, 2006; Müller,
that interact with the product. One of the several possibilities, Yamashita, & Laurindo, 2008; Pelissari et al., 2012; Souza et al.,
which are being extensively studied, is the incorporation of active 2012; Veiga-Santos, Ditchifield, & Tadini, 2011; Vercelheze et al.,
substances within the package material, as films based on cassava 2012; Veiga-Santos, Suzuki, Nery, Cereda, & Scamparini, 2008). Films
starch (Kechichian, Ditchifield, Veiga-Santos, & Tadini, 2010). developed from starch are described as isotropic, odorless, tasteless,
Although many types of new polymers are being industrially colorless, non-toxic and biodegradable (Souza et al., 2010).
produced (PLA, PHA, PCL, PEA and others), polymers from agri- In a previous study (Souza et al., 2012), it was described the
cultural sources are the most studied by researchers, especially development of cassava starch films plasticized with sugars and
polysaccharides. Among the films made from polysaccharides, glycerol and reinforced with clay nanoparticles. Results reported
those obtained from starch are the most important because it is one were important for the continuity of the research because they gave
of the most commonly used agricultural raw materials, since it is a information about optimal formulation to produce composites
renewable source, inexpensive and widely available (Souza, films with better mechanical and barrier properties. Now, authors
Ditchifield, & Tadini, 2010). Beyond this, it has good film-forming are trying to incorporate antimicrobial agents in the formulation of
properties. cassava starch films since carrying natural additives could be
Cassava starch has been extensively used to produce films and the considered as a new tendency of functional food packaging in the
results indicated that these carbohydrates are promising materials in near future. Active packaging provides microbial safety for con-
sumers, reducing, inhibiting or retarding the growth of microor-
* Corresponding author. Department of Chemical Engineering, Escola Politécnica,
ganisms, and then, could extend the shelf life of the packaged food.
University of São Paulo, SP, Brazil. Tel.: þ55 11 30912258; fax: þ55 11 30912255. Based on results presented by Kechichian et al. (2010), cinna-
E-mail address: catadini@usp.br (C.C. Tadini). mon essential oil and clove essential oil were chosen to continue

0023-6438/$ e see front matter Ó 2013 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.lwt.2013.06.017
 A.C. Souza et al. / LWT - Food Science and Technology 54 (2013) 346e352 347

their research, which was developed by the same research group of solution was prepared according to the following procedure: firstly,
the present work. Other authors also demonstrated the antimi- 0.1 g of clay nanoparticles were suspended in 25 g of distilled water
crobial efficacy of these agents in literature (Goñi et al., 2009; Kim, for 1 h, under stirring (500 rpm), and, after rest for 24 h, they were
Park, & Park, 2004; Nielsen & Rios 2000; Oussalah, Caillet, Saucier, blended with a suspension of 5.0 g of starch and 70 g of distilled
& Lacroix, 2006; Oussalah, Caillet, Saucier, & Lacroix, 2007). Cin- water. After that, cinnamon essential oil (0.40 g, 0.60 g or 0.80 g)
namon and clove has been used as spices for thousands of years. was mixed with emulsifier (0.010 g, 0.015 g or 0.020 g), corre-
The main constituents of their oils are cinnamaldehyde and spondent to 0.025 for emulsifier content/essential oil content
eugenol, respectively, two well known agents due to their antimi- proportion; and glycerol (0.75 g, 1.13 g or 1.50 g) at (38  2)  C,
crobial activities. correspondent to 1.88 for glycerol content/essential oil content
Oussalah et al. (2006) reported that cinnamon essential oil proportion, using a magnetic stirrer (200 rpm). Both mixtures
showed a strong antimicrobial activity against Pseudomonas putida prepared were then homogenized and heated in a domestic mi-
strain isolated from meat. Kim et al. (2004) suggested that the crowave oven (Panasonic, model Family Plus, Brazil) until starch
antimicrobial activity of cinnamaldehyde is bactericidal against gelatinization, which occurs at (69  2)  C. After cooling, the fil-
Escherichia coli O157:H7. Scanning electron microscopic observa- mogenic solution was diluted with 14.25 g of ethanol, and, for each
tions revealed that the bacterial cells treated with cinnamaldehyde formulation, a specific content of filmogenic solution was poured
suffered severe damages in their surface structure. Nielsen & Rios onto rectangular plates (97.5 cm2 of area) of polytetrafluoro-
(2000) tested the effect of essential oils against the most impor- ethylene (TeflonÒ) to obtain a constant thickness of (100  10) mm,
tant spoilage fungi of bread and demonstrated that cinnamon followed by drying at (35  2)  C for approximately (18e24) h, in a
essential oil had high activity. Results obtained by Oussalah et al. conventional chamber dryer with forced air circulation (Nova Ética,
(2007) showed that one of the most active essential oil against series N480, Brazil).
four pathogenic bacteria was the cinnamon. Moreover, Goñi et al. The quantities of glycerol, emulsifier and cinnamon essential oil
(2009) tested a combination of cinnamon and clove essential oils were defined according preliminary tests and based on previous
against a wide range of bacteria in the vapor phase as a preservative work (Souza et al., 2012), taking into account the maximum levels
method to prevent microorganism proliferation. of cinnamon essential oil which could be incorporated into the
In the present work, the minimum inhibitory concentration matrix without oil phase separation during film drying. Composite
(MIC) of two essential oils, cinnamon (Cinnamomum cassia) and film without cinnamon essential oil and emulsifier was also pro-
clove (Eugenia caryophyllata), were established. In a second step, duced and considered as control.
cinnamon essential oil was incorporated into cassava starch films After drying, all films were placed in a controlled relative hu-
elaborated by casting. The main goal was to develop active com- midity of 75% and at ambient temperature of (23  2)  C and stored
posite films, and to verify the influence of cinnamon essential oil prior to testing.
addition on microstructure, mechanical (tensile strength and
percent elongation at break) and barrier (water vapor permeability 2.3. Antimicrobial activity e disk diffusion method
and oxygen permeability coefficient) properties of produced films.
Also, the antimicrobial activity against fungi commonly found in Using the pour plate method, inoculums (1 mL) of P. commune
bread was tested by two different techniques: disk diffusion and E. amstelodami were spread on the surface of Petri dishes
method and release mass experiments by UVevis spectroscopy. prepared with the selected medium for fungi. The inoculums were
adjusted to each microorganism to yield a cell concentration of
2. Materials and methods 108 CFU/mL.
The minimum inhibitory concentration (MIC) was defined as the
2.1. Materials lowest essential oil concentration resulting in the lack of visible
microorganism growth using the disk diffusion method.
Native cassava starch, kindly supplied by Cargill Agrícola, Brazil Circular disk samples of filter paper (25 mm of diameter) were
(amylose: 19.7 g/100 g; moisture: 12.5 g/100 g) was used as the absorbed with alcoholic solutions of each essential oil at different
film-forming component to provide a continuous matrix of films. concentrations: (0.0, 0.5, 1.0, 2.0, 4.0, 8.0, 16.0 and 32.0) g/100 g, and
Glycerol (Synth, Brazil) and natural -Na montmorillonite clay placed on the solidified medium surface. Petri dishes were then
(commercial product Argel T, used as received, without purifica- incubated at (25  2)  C for 5 days and the inhibitory zone was
tion, Bentonit União, Brazil) were used as plasticizer and rein- determined measuring its diameter.
forcement filler, respectively. Cinnamon essential oil (Ferquima, In order to evaluate the efficiency of each cinnamon essential
Brazil) with 82.5 g/100 g of cinnamaldehyde and clove essential oil oil contents incorporated into composite films, circular disk sam-
(Ferquima, Brazil) with 75.0 g/100 g of eugenol were used as ples of these films were placed, instead of filter paper, on the so-
antimicrobial agents. Sucrose ester of fatty acids was used as lidified medium surface. Petri dishes were then incubated at
emulsifier, specific for oil/water emulsion, in order to incorporate (25  2)  C for 5 days. Antimicrobial agent efficiency was evaluated
the cinnamon essential oil into the films (commercial name: SP70, by the formation of an inhibition zone around the disk samples,
Sisterna, Brazil). Distilled water and ethanol (Synth, Brazil) were which was characterized by surrounding clear areas. To a better
used as solvents of the filmogenic solutions. Penicillium commune presentation of the results, the diameter of inhibition zone of each
and Eurotium amstelodami were obtained in a lyophilized form Petri dish was measured and, knowing the total area of the Petri
(André Tosello Foundation, Brazil). All growth experiments were dish, inhibition results were converted to percentage of inhibition
carried out on a medium for fungi prepared with Czapek Dox (Difco, areas. All tests were performed in triplicate, seven days after the
USA) and agar (Synth, Brazil). film elaboration.

2.2. Film preparation 2.4. Antimicrobial agent release

The films were produced by casting technique, using the The amount of antimicrobial agent incorporated in cassava
methodology and optimum contents of cassava starch, glycerol and starch films was quantified by UVevis spectroscopy (Spectropho-
clay nanoparticles proposed by Souza et al. (2012). The filmogenic tometer JASCO, model 550, Japan) measuring at 289 nm,
348 A.C. Souza et al. / LWT - Food Science and Technology 54 (2013) 346e352

corresponding to the maximum absorption wavelength of cinna-

mon essential oil, and using a pre-determined calibration curve.
The release experiments were carried out at room temperature
with the films immersed in distilled water (150 mL) for 2 h. After
the first 2 h, tested samples were once more immersed in distilled
water and a new spectroscopy quantification was done at 289 nm in
order to ensure that this time was enough to guarantee full release
of antimicrobial agent from the films (in this case, the content
quantified should be zero). Assays were performed in duplicate.

2.5. Film thickness

The thickness (t) [mm] was measured using a flat parallel sur-
face micrometer (MITUTOYO Sul Americana Ltda., model 103-137,
Brazil, precision 0.002 mm), at five random positions of the films.

2.6. Scanning Electron Microscopy (SEM)

Fig. 1. Typical curve of weight gain (w) of the samples as a function of time (t) in
Small strips (5 mm  5 mm) of cassava starch films were comparison with weight gain of the samples in the cell without silica gel.
mounted on aluminum stubs, coated with a thin layer of gold and
observed on a Scanning Electron Microscope (Philips, model XL-30
w  
FEG), at an accelerate voltage of 5 kV. 24  t
WVP ¼  (1)
q A  Dp
2.7. Mechanical properties
wherein:
Tensile strength (TS) [MPa] and percent elongation at break (E) WVP is the water vapor permeability [g mm m2 d1 kPa1]; w is
[%] were evaluated by a tensile test performed on a texture the weight gain (from the straight line) [g]; q is the time during
analyzer (TA.XT2i e Stable Micro Systems, UK) with a load cell of which w occurred [h]; t is the average film thickness [mm]; A is the
5 kg, using the A/TGT self-tightening roller grips fixture, according test area (cell top area) [m2] and Dp is the vapor pressure difference
to ASTM D882-09 (2009). Twenty strips (130 mm  25 mm) were [kPa]. All formulations were evaluated in triplicate.
cut from each formulation of preconditioned films and each one
was mounted between the grips of the equipment for testing. 2.9. Oxygen permeability coefficient
Initial grip separation and test speed were set to 50 mm and
0.8 mm s1, respectively. Tensile strength (nominal) was calculated Oxygen transmission rate (OTR) of the films was measured at
dividing the maximum load by the original minimum cross- 23  C and 75% RH on a 50 cm2 circular films using an oxygen
sectional area of the specimen (related to minimum thickness). permeation system (OXTRAN 2/21, MOCON, USA), in accordance
Percent elongation at break (nominal) was calculated by dividing with ASTM F1927-07 (2007). A starch based film was sealed be-
the extension at the moment of rupture of the specimen by its tween two chambers (each one with two channels), the lower one
initial gage length and multiplying by 100. All formulations were supplied with O2 at a controlled flow rate (20 mL min1) to keep the
evaluated in triplicate. pressure constant in that compartment, and the other one was
purged by a stream of nitrogen carrier gas (98% of nitrogen and 2%
of hydrogen), at controlled flow rate (10 mL min1). A coulometric
2.8. Water vapor permeability
sensor determined the amount of oxygen transmitted through the
film into the carrier gas. The oxygen transmission rate was deter-
Water vapor transmission (WVT) was determined by a gravi-
mined for all formulations in duplicate. The permeance (PO2) of the
metric method based on ASTM E96/E96M-05 (2005), using the
films was calculated according to Equation (2):
Desiccant Method. This property was reported as water vapor
permeability (WVP), which is the rate of water vapor transmission OTR
(WVT) through a unit area of flat material of unit thickness induced PO2 ¼ (2)
p
by unit vapor pressure difference between two surfaces, under
specified humidity condition of 75%. Each film sample was sealed wherein:
with paraffin over a circular opening of 44 cm2 at the permeation PO2 is the permeance of the films [cm3 m2 d1 Pa1]; OTR is the
cell (PVA/4, REGMED, Brazil) that was stored, at ambient temper- oxygen transmission rate [cm3 m2 d1]; and p is the partial
ature, in a desiccator. To maintain 75% of relative humidity (RH) pressure of oxygen, which is the mol fraction of oxygen multiplied
gradient across the film, a constant mass of silica gel was placed by the total pressure (nominally, 1 atm) in the test gas side of the
inside the cell and a sodium chloride saturated solution (75% RH) diffusion cell. The partial pressure of O2 on the carrier gas side is
was used in the desiccator. Two cells without silica gel were pre- considered to be zero.
pared and submitted to the same conditions to account for weight The oxygen permeability coefficient (P0 O2) was calculated as
changes occurring in the film, since it is a hydrophilic material. The follows:
RH inside the cell was always lower than the outside, and water
vapor transport was determined from the weight gain of the P ’ O2 ¼ PO2  t (3)
permeation cell. After steady state conditions were reached (about
2 h), ten weight measurements were made over 48 h Fig. 1 shows a wherein:
typical curve indicating that the weight gain from the straight line P0 O2 is the oxygen permeability coefficient [cm3 m1 d1 Pa1];
was 3.15  102 g h1. WVP was calculated according Equation (1): and t is the average thickness of the specimen [mm].
 A.C. Souza et al. / LWT - Food Science and Technology 54 (2013) 346e352 349

2.10. Statistical analysis inhibition began with 0.5 g/100 g of cinnamon essential oil solu-
tion (diameter: 4 mm) and with 4.0 g/100 g of clove essential oil
Analysis of variance (ANOVA) was applied on the results using solution (diameter: 6 mm) and was completed (100% of inhibition)
the statistical program Statgraphics Centurion program v.15.2.06 with 2.0 g/100 g and 16 g/100 g, respectively. For E. amstelodami,
(StatPointÒ, Inc., Warrenton, USA) and the Tukey test was used to inhibition was completed with only 0.5 g/100 g of cinnamon
evaluate average differences (at a 95% of confidence interval). essential oil solution and began with 4.0 g/100 g of clove essential
oil solution (diameter: 14 mm) and was completed with 16 g/
3. Results and discussion 100 g.
With these results, it can be concluded that cinnamon essential
The study was conducted in two steps: firstly, antimicrobial oil was more effective against the fungi selected for this work, since
activities of cinnamon and clove essential oils were evaluated, us- it presented a better inhibition with lower concentration. In this
ing the disk diffusion method, against P. commune and E. amstelo- way, cinnamon essential oil was chosen to be incorporated in
dami, fungi commonly found in bread products (Saranraj & Geetha, composite films based on cassava starch.
2012). It was possible to quantify the minimum amount of each
essential oil necessary to be incorporate in cassava starch films in 3.2. Antimicrobial activity of active cassava starch films
order to develop films with antimicrobial properties. incorporated with cinnamon essential oil
In the second step, cinnamon and clove essential oils were
incorporated in cassava starch films. In preliminary assays, it was Inhibition areas yielded by cassava starch film disks with
noted that the amount of clove essential oil necessary to provide different contents of cinnamon essential oil against each studied
films with effective antimicrobial activity against fungi tested was microorganism are shown in Table 2. ANOVA indicated that there
too high and, therefore, it became infeasible to obtain films with were significant differences among antimicrobial activity of films
suitable visual and handling properties. Thus, it was decided to with different cinnamon essential oil contents (P < 0.05). As pre-
produce the active films with only cinnamon essential oil, since this dictable, no inhibition zone against the microorganisms was
agent presented more promising results in the first step. observed for film disks without incorporation of essential oil
Despite initial results of microbiological inhibition were quite (control films). Comparing the microorganisms, it can be concluded
satisfactory, indicating an almost complete inhibition of fungi, that E. amstelodami is more sensitive for cinnamon essential oil
materials produced showed a compromised surface because films because its inhibition was greater, reaching approximately 91% of
became more and more brittle with the increase of essential oil inhibition with the highest concentration used. Fig. 2 shows the
content in the formulation. To overcome this hurdle, it was inhibition of P. commune caused by active films produced with
necessary to vary the plasticizer content in accordance with the three different contents of cinnamon essential oil. As expected, a
increase of essential oil content in the formulation. better inhibition was observed with higher content of cinnamon
Since it is known that it is impossible to make homogeneous essential oil (Fig. 3).
suspensions of oil in water (that was used as the solvent of the Even at minimum concentration applied into the film formula-
filmogenic solution), an emulsifier in the formulation of cassava tion, cinnamon essential oil showed inhibition against both mi-
starch films was added in order to avoid a phase separation. croorganisms, which was considered an important result since that
Therefore, active cassava starch films with emulsifier and plasti- higher concentrations could imply a sensorial impact, altering the
cizer in contents varying according to the amount of cinnamon natural taste of the food packaged by exceeding the acceptable
essential oil were developed and characterized. flavor thresholds.
After preliminary results and based on previous work (Souza A great number of studies on the antimicrobial characteristics of
et al., 2012), proportions of 1.88 for glycerol content/essential oil films made from starch have been carried out earlier. Nevertheless,
content; and 0.025 for emulsifier content/essential oil content, were no information has been presented about the effect of cinnamon
chosen to provide films with good visual and tactile characteristics. essential oil on P. commune and E. amstelodami, which plays an
important role in the spoilage of bread products.
3.1. Determination of minimum inhibitory concentration (MIC) of
cinnamon and clove essential oils
3.3. Release of cinnamon essential oil
Different results of inhibition were obtained for each essential
Cinnamon essential oil (CEO) release profiles from cassava
oil and for each microorganism studied (Table 1). For P. commune,
starch films, for a monitoring period of 2 h, are shown in Fig. 2.
Table 1
Inhibition areas [%] for each essential oil evaluated against P. commune and
E. amstelodami. Table 2
Inhibition areas [%] against P. commune and E. amstelodami of active cassava starch
Essential oil Inhibition area [%]
films produced with different contents of glycerol, emulsifier and cinnamon
solution [g/100 g]
P. Commune E. amstelodami essential oil.

Cinnamon Clove Cinnamon Clove Formulation Glycerol Emulsifier Cinnamon Inhibition area [%]
essential oil essential oil essential oil essential oil [g/100 g] [g/100 g] essential oil
P. commune E. amstelodami
[g/100 g]
0.0 0.0 0.0 0.0 0.0
0.5 10.4 0.0 100.0a 0.0 Control 0.75 0 0 0.00  0.00a 0.00  0.00a
1.0 32.1 0.0 100.0 0.0 A 0.75 0.010 0.40 1.93  0.74b 14.83  3.79b
2.0 100.0a 0.0 100.0 0.0 B 1.13 0.015 0.60 10.77  5.31c 47.02  4.69c
4.0 100.0 11.9 100.0 18.7 C 1.50 0.020 0.80 25.94  5.72d 91.06  15.48d
8.0 100.0 40.1 100.0 62.3 Tukey 10.25 21.50
16.0 100.0 100.0a 100.0 100.0a HSD 5%
32.0 100.0 100.0 100.0 100.0
*Means in the same column with the same letter are not significantly different
a
MIC: minimum inhibitory concentration. (P > 0.05).
350 A.C. Souza et al. / LWT - Food Science and Technology 54 (2013) 346e352

Fig. 4. Cassava starch film incorporated with 0.4 g of cinnamon essential oil/100 g of
filmogenic solution (from Eduardo de Oliveira, with permission).

3.4. Appearance and microstructure of cassava starch films

Fig. 2. Petri dishes with circular disks of films incorporated with three different
contents of cinnamon essential oil (CEO), showing the inhibitory zone against Peni-
cillium commune in comparison with Petri dish without the active films. Homogeneous, thin and flexible cassava starch films were ob-
tained. They could be easily removed from the TeflonÒ plates after
drying. Visually, all films were colorless and slightly opaque (Fig. 4).
Released amounts of CEO varied from (0.88  0.10) mg CEO/g film Fig. 5 shows SEM micrographs of the surface of active cassava
to (1.19  0.02) mg CEO/g film for films incorporated with different starch films with remarkable differences. A continuous matrix was
contents of antimicrobial agent. It can also be observed that in 2 h observed for active films elaborated with emulsifier (Fig. 5a).
all added CEO was released, since the results for the new quanti- Smooth, uniform and regular surface was observed in all samples.
fication done after 2 h were zero. Moreover, it should be pointed On the other hand, the absence of the emulsifier caused a discon-
out that cassava starch film samples did not dissolve in the water tinuous structure, with lipid droplets embedded in the polymer
after 2 h, but their volume were increased, demonstrating that films network (Fig. 5b).
were susceptible to water uptake.
The pronounced initial increase of mass released content sug- 3.5. Mechanical and barrier properties of cassava starch films
gests that it is necessary to incorporate the antimicrobial agent into
matrix by another technique, like as supercritical solvent impreg- Data of tensile strength, elongation at break, water vapor
nation, if a slower release is desired. permeability and oxygen permeability coefficient obtained from

Fig. 3. Mass released of cinnamon essential oil from cassava starch films.
 A.C. Souza et al. / LWT - Food Science and Technology 54 (2013) 346e352 351

Compared to most commonly used synthetic polymers, TS and E

were rather low, but sufficient for use in many food applications. In
previous work, Souza et al. (2012) tested films based on cassava
starch reinforced with 1.0 g/100 g of clay, at the same conditions of
this work, and found that the increase of glycerol content from
(0.75e1.25) g/100 g, decreased the TS from (3.96  0.60 to
2.07  0.33) MPa and increased E from (123.61  19.57 to
200.24  33.50) %. Considering these previous results, the increase
of the glycerol content in cassava starch films elaborated in this
present work can also contributed with the decrease of TS.
When comparing films prepared according formulation A with
the control ones, it can be observed that the presence of emulsifier
plus cinnamon essential oil also decreased significantly the TS from
(3.75  0.70 to 2.32  0.40) MPa and increased the E from
(128.81  18.67 to 256.13  48.57) %. Analyzing these values, we can
conclude that the introduction of essential oil reduced the inter-
molecular interaction between polymeric chains, resulting in ma-
terials with lower tensile strength.
Concerning barrier properties, a rise of contents of glycerol,
emulsifier and cinnamon essential oil caused an increase in both
permeabilities. Water vapor permeability (WVP) and oxygen perme-
ability coefficient (P0 O2) of films with cinnamon essential oil incor-
porated varied from (9.78  1.40 to 14.79  2.76) g mm m2 d1 kPa1
and from (27.50  0.60 to 143.47  8.30)  109 cm3 m1 d1 Pa1 ,
respectively.
In previous work, Souza et al. (2012) tested films based on
cassava starch reinforced with 1.0 g/100 g of clay, at the same
conditions of this work, and found that the increase of glycerol
content from (0.75e1.25) g/100 g also decreased barrier properties:
WVP from (3.81  0.58 to 5.38  0.80) g mm m2 d1 kPa1 and
P0 O2 from (22.51  0.79 to 94.00  3.90)  109 cm3 m1 d1 Pa1).
Considering these results, the increase of the glycerol content in
cassava starch films elaborated in this work can also contributed
with the decrease of the studied barrier properties.
Glycerol is a relatively small hydrophilic molecule, which can be
entrapped between adjacent polymeric chains, decreasing inter-
Fig. 5. Micrographs of surface of cassava starch films: active film elaborated (a) with molecular attractions and increasing molecular mobility, facili-
emulsifier and (b) without emulsifier. tating migration of water vapor molecules (Rodríguez, Osés, Ziani,
& Maté, 2006). However, in this work, when comparing films
elaborated according formulation A with the control ones, both
cassava starch films produced with cinnamon essential oil as with the same glycerol content, it can be verified that the WVP
antimicrobial agent are shown in Table 3. All data were analyzed by increased significantly from (3.81  0.58 to 9.78  1.40)
ANOVA and the results indicated there were significant differences g mm m2 d1 kPa1.
among films properties with different cinnamon essential oil con- It is known that the addition of a lipidic component in the
tents (P < 0.05). formulation could act as a barrier in the films. Therefore, it is ex-
Tensile strength (TS) and elongation at break (E) of films with pected that cinnamon essential oil was not the responsible agent
cinnamon essential oil incorporated varied from (2.32  0.40 to for the elevation of permeabilities values.
1.05  0.16) MPa and from (264.03  35.06 to 191.27  22.62) %, Based on previous observations, the emulsifier, as a hydrophilic
respectively, therefore an increase of cinnamon essential oil, glyc- agent, probably it is the most component responsible for the WVP
erol and emulsifier contents lowered the TS and the E of the films, increasing. However, it should be pointed that its presence in the
indicating a loss of macromolecular mobility. From presented data, film elaboration process is necessary, because it promotes the
it was realized that control films (without essential oil) presented incorporation of the antimicrobial agent into the aqueous solution,
higher TS (3.96  0.60) MPa and lower E (123.61  19.57) % resulting in a homogeneous polymer matrix.

Table 3
Tensile strength (TS), elongation at break (E), water vapor permeability (WVP) and oxygen permeability coefficient (P0 O2) of active cassava starch films produced with different
contents of glycerol, emulsifier and cinnamon essential oil.

Formulation TS [MPa] E [%] WVP P0 O2  109 [cm3 m1 d1 Pa1]

[g mm m2 d1 kPa1]

Control 3.75  0.70d 128.81  18.67a 3.61  0.69a 21.50  0.89a

A 2.32  0.40c 256.13  48.57c 9.78  1.40b 27.50  0.60a
B 1.36  0.18b 264.03  35.06c 10.11  1.63b 67.44  0.81b
C 1.05  0.16a 191.27  22.62b 14.79  2.76c 143.47  8.30c
Tukey HSD 5% 0.12 14.02 3.52 19.95

*Means in the same column with the same letter are not significantly different (P > 0.05).
352 A.C. Souza et al. / LWT - Food Science and Technology 54 (2013) 346e352

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This research was supported by FAPESP (The State of São Paulo Saranraj, P., & Geetha, M. (2012). Microbial spoilage of bakery products and its
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(2012). Cassava starch biodegradable films: influence of glycerol and clay
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