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Food Hydrocolloids 38 (2014) 20e27

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Food Hydrocolloids
journal homepage: www.elsevier.com/locate/foodhyd

Evaluation of edible lms and coatings formulated with cassava


starch, glycerol, carnauba wax and stearic acid
Marcela Chiumarelli*, Miriam D. Hubinger
Dept. of Food Engineering, School of Food Engineering, University of Campinas (UNICAMP), P.O. Box 6121, Campinas, SP 13083-862, Brazil

a r t i c l e i n f o

a b s t r a c t

Article history:
Received 16 March 2013
Accepted 19 November 2013

This study aimed at characterizing four formulations of cassava starch, glycerol, carnauba wax and
stearic acid-based edible coatings/lms, evaluating their water vapor resistance, the respiration rate of
coated apple slices, surface solid density, solubility, mechanical properties, thermal properties and
microstructure. Among the formulations evaluated, only the formulation with higher wax content did
not promote an effective barrier to oxygen and water vapor and resulted in more rigid lms. Formulations with higher glycerol content promoted lower respiration rate of apple slices, good water vapor
resistance and exible lms, but the microstructure analysis showed a non-homogenous surface of the
lms with higher wax content. The glycerol added in formulations increased the solubility and reduced
the melting temperature of the lms. Formulation with 0.2% carnauba wax showed intermediate values
of analyzed properties and promoted lms with a more regular surface. Formulation containing 3% (w/
w) cassava starch, 1.5% (w/w) glycerol, 0.2% (w/w) carnauba wax and 0.8% (w/w) stearic acid, presented
lms with a cohesive matrix, resulting in better mechanical properties, barrier to moisture and gas
exchange.
2013 Elsevier Ltd. All rights reserved.

Keywords:
Cassava starch
Respiration rate
Water vapor resistance
Microstructure
Fresh-cut apple

1. Introduction
The market for minimally processed fruits and vegetables has
increased in recent years. These products are ready to eat, convenient and maintain the freshness and nutritional quality of the
whole product. On the other hand, the peeling and cutting operations can accelerate the metabolic activities of plant tissue, making
the minimally processed product more perishable than intact fruits
and vegetables (Olivas & Barbosa-Cnovas, 2005). So, new strategies and technologies to maintain quality and extend shelf life are
required.
The use of edible coatings is a promising technology to preserve the quality of whole and minimally processed fruits (Han &
Gennadios, 2005; Talens & Krochta, 2005; Vargas, Pastor, Chiralt,
McClements, & Gonzlez-Martnez, 2008; Zahedi, Ghanbarzadeh,
& Sedaghat, 2010). The coatings act as barriers to water loss and
gas exchange, controlling the transfer of moisture, oxygen, carbon dioxide, lipids and avor components, with similar effect to
that promoted by storage under controlled or modied atmosphere (Vargas et al., 2008). A range of polymers can be used in
edible coatings formulation. However, their mechanical and

* Corresponding author. Tel.: 55 19 3521 4036; fax: 55 19 3521 4027.


E-mail address: chiumarelli@gmail.com (M. Chiumarelli).
0268-005X/$ e see front matter 2013 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.foodhyd.2013.11.013

barrier properties are intrinsically linked to physical and chemical characteristics of their constituents. The polymers most
commonly used in the preparation of edible lms and coatings
are proteins (gelatin, casein, wheat gluten and zein), polysaccharides (starch, pectin, cellulose, alginate and carrageenan)
and lipids (stearic acid, waxes and fatty acid esters), which can
be used alone or in combinations (Sothornvit & Krochta, 2001;
Vargas et al., 2008).
Polysaccharides-based coatings have low oxygen permeability,
which can reduce the respiration rate of minimally processed
products. Starch is the most important polysaccharide used in the
formulation of biodegradable lms and edible coatings. Cassava
starch-based coatings are tasteless, odorless and transparent, not
changing the taste, aroma and appearance of the product
(Chiumarelli, Pereira, Ferrari, Sarantpoulos, & Hubinger, 2010;
Garcia, Pereira, Sarantpoulos, & Hubinger, 2010; Pareta &
Edirisinghe, 2006). Although the starch is a cheap and abundant
material, able to form a continuous polymer matrix, it exhibits a
strong hydrophilic character, constituting poor barriers to water
vapor (Averous & Boquillon, 2004; Ghanbarzadeh, Almasi, &
Entezami, 2011; Karbowiak, Debeaufort, & Voilley, 2007; Liu,
2005; Vargas et al., 2008). The addition of lipids can reduce water
vapor permeability, but it can affect coatings transparency and
mechanical properties, despite the lipid offers after taste, which
may impair the sensory characteristic of food (Bourlieu, Guillard,

M. Chiumarelli, M.D. Hubinger / Food Hydrocolloids 38 (2014) 20e27

Valls-Pamis, Guilbert, & Gontard, 2009; Fakhouri et al., 2007;


Rhim & Shellhammer, 2005).
The most used lipids in coatings for minimally processed
products are stearic acid, palmitic acid, and some vegetable oils
such as soybean and sunower ones (Bourlieu et al., 2009; Colla,
Sobral, & Menegalli, 2006; Garcia, Martino, & Zaritzky, 2000;
Martn-Belloso, Soliva-Fortuny, & Baldwin, 2005). Despite the
good results obtained in some formulations, these materials have
lower water vapor permeability than that of natural waxes, such as
carnauba wax (Rhim & Shellhammer 2005; Rojas-Argudo, del Rio, &
Prez-Gago, 2009; Talens & Krochta, 2005).
Glycerol is a plasticizer used to enhance the lms/coatings
exibility, but can affect the water vapor and gases permeability,
since it is very hydrophilic and hygroscopic (Sothornvit & Krochta,
2001). Glycerol has low molecular weight, modifying the interactions between the macromolecules, which increase mobility of
the polymer chains and reduce the glass transition temperature of
the system (Krochta, 2002).
In this context, the objective of this work was to characterize
four formulations containing cassava starch, glycerol, carnauba wax
and stearic acid-based edible coatings/lms, through the evaluation of their water vapor resistance, respiration rate of coated apple
slices, solubility, mechanical properties (tensile strength, elongation at break, elastic modulus and coating strength), thermal
properties
(transition
temperature
and
enthalpy)
and
microstructure.
2. Material and methods
2.1. Material
Cassava starch (17% amylose content, Pilo Amidos Ltda., Guara,
Brazil), carnauba wax type 1 (Cresal Comrcio e Representaes
Ltda., So Paulo, Brazil), stearic acid (Synth, Diadema, Brazil) and
glycerol as plasticizer agent (ECIBRA, So Paulo, Brazil) were used
for coating/lm formulations.
Apples (Malus domestica Borkh cv Gala) were obtained from
the local market (Ceasa, Campinas, Brazil). A lot of 150 apples was
bought and the fruits of uniform weight (120.59  5.24 g), maturity
stage (based on skin color and rmness) with no physical damage
were selected to the experiments.
2.2. Methods
2.2.1. Coating/lm preparation
Cassava starch water suspensions were prepared at 75  C with
constant stirring. Glycerol was added after starch gelatinization.
Carnauba wax and stearic acid were melted at 85  C and homogenized into the cassava starch suspensions with a rotorestator homogenizer (model MA 102, Marconi Ltda., Piracicaba, Brazil) for
3 min at 12,000 rpm. The temperature of homogenization was
85  C and emulsions were then cooled until room temperature. The
coatings/lms formulations are presented in Table 1. The cassava

Table 1
Cassava starch (CS), glycerol (GLY), carnauba wax (CW) and stearic acid (SA) concentrations on coatings/lms formulations.
Variables

CS (% w/w)
GLY (% w/w)
CW (% w/w)
SA (% w/w)

starch, glycerol, carnauba wax and stearic acid concentrations were


chosen through an experimental design, and these formulations
were obtained in optimized area, as described in a previous work
(Chiumarelli & Hubinger, 2012). The stearic acid was used to reduce
the carnauba wax melting point.
Apple slices and cylinders were dipped into cassava starchecar
nauba waxestearic acid emulsions for 2 min and used in respiration
rate and water vapor resistance analyses. For the other analyses, lms
were prepared by weighing an emulsion amount that provided
130  2 mm of lm thickness on a Teon casting plate resting on a
leveled surface. Films were dried until constant moisture content at
40  C and 60% RH to avoid cracking, peeled off the casting surface and
conditioned at 25  C and 58% RH in desiccators with a saturated NaBr
solution for 72 h prior to analysis.
2.2.2. Respiration rate
The respiration rate of coated apple slices was measured by
static method (Bierhals, Chiumarelli, & Hubinger, 2011; Garcia et al.,
2010). Apples were peeled and cut into 8 slices, which were
immersed in coating formulations for 2 min, placed in a sieve and
left at room temperature (15  C and 80% RH) for 1 h to dry the
coating material. The samples (around 50 g) were placed in sealed
180 mL glass jars with silicon septum. The jars were closed and kept
at 5  C during 1 h. After this time, gas sampling was carried out
using an O2/CO2 Dual Head Space Analyzer (Model PAC CHECK 325,
Mocon, Minneapolis, USA), measuring the CO2 production. Uncoated apple slices were used as control in order to calculate the
coatings efciency. The respiration rate was determined in triplicate and expressed in mL CO2 kg1 h1.
2.2.3. Water vapor resistance
The determination of coatings water vapor resistance (WVR)
was carried out according to the method described by AvenaBustillos, Krochta, Saltveit, Rojas-Villegas, and Sauceda-Prez
(1994), using a modied Ficks equation proposed by BenYehoshua, Burg, and Young (1985). This method was also used by
Garcia, Martino, and Zaritzky (1998), Rojas-Gra, Tapia, Rodrguez,
Carmona, and Martn-Belloso (2007), Tapia et al. (2008) and Garcia
et al. (2010) to calculate the WVR of different edible coatings.
Apple cylinders with 25 mm of diameter and 10 mm of thickness were used for coatings application. The exposed area, taken as
the upper surface and the lateral area of the samples, was
12.76 cm2. Coated and uncoated apple cylinders were equilibrated
for 24 h at 20  1  C in desiccators maintained at 98.9% RH with a
0.6 M NaCl solution. Then, samples were placed in small test cups,
weighed in an analytical balance, and placed in desiccators equilibrated at 33.3% RH with saturated MgCl2$6H2O at 20  C. Weight
was taken periodically during a 24 h period and the water vapor
ux was calculated according to Eq. (1):


WF

  
dP
1
$
dt
A

"
2

2.50
2.82
0.10
0.90

2.50
2.61
0.38
0.62

3.50
1.00
0.40
0.60

3.00
1.50
0.20
0.80

(1)

where: WF is the water vapor transferred per unit of area


(g s1 cm2), dP/dt is the water vapor transferred (g s1) and A is the
exposed area (cm2).
WVR was calculated using Eq. (2), proposed by Ben-Yehoshua
et al. (1985):

Formulation
1

21

WVR

!
# 

RH
aw  100
1
$Pwv $
WF
R$T

(2)

where: WVR is the resistance of the coating to water diffusion


(s cm1); aw water activity of the sample; RH 33.3% relative

22

M. Chiumarelli, M.D. Hubinger / Food Hydrocolloids 38 (2014) 20e27

humidity of environment; Pwv saturated water vapor pressure at


20  C (17.4 mmHg); R is the gas constant (3464.629 mmHg cm3
K1 g1) and T is the temperature (293.15 K).
The resistance factor of the uncoated sample to water vapor was
evaluated and the coatings efciency was calculated. Water activity
of samples was measured with an Aqualab Series 3TE (Decagon
Devices Inc, Pullman, USA), at 20  C.
2.2.4. Surface solid density
An estimation of coating thickness was calculated by means of
the quantication of the surface solid density, SSD, according to the
methodology described by Villalobos-Carvajal, Hernndez-Munz,
Albors, and Chiralt (2009), Vargas, Albors, Chiralt, and GonzlezMartnez (2006) and Vargas et al. (2008). Apple cylinders with
approximately 2.0 cm of diameter and 0.5 cm of thickness, with a
total surface area of 9.42 cm2 were used. Apple cylinders were
weighed, immersed in different emulsions for 2 min and placed in a
sieve to drain the coating excess. Then, samples were weighed and
the coating surface solid density (SSD) was calculated using Eq. (3).

SSD

Xs $Mcoating
As

(3)

where: SSD is the surface solid density (g cm2), Xs is the mass


fraction of solid of each lmogenic solution (%), Mcoating is the mass
of the coating solution adhered to the fruit surface (g) and As is the
sample area (cm2).

2.2.8. Thermal properties


Thermal properties of lms were determined by using differential scanning calorimetry DSC 2920 (TA Instruments, New Castle,
USA). Approximately 5 mg of lms were stored in a NaBr solution
(58% RH) for three weeks and, after this period, samples were
placed in hermetically sealed aluminum pans and heated into the
DSC from 0 to 130  C at 10  C/min and subsequent cooling from 130
to 30  C. The reference was an empty pan and the equipment was
calibrated with indium (Tm 156.60  C and DH 28.71 J/g). The
melting temperature (Tm) was calculated as the temperature where
the endothermic peak occurs, and the transition enthalpy (DH) was
calculated as the area over the endothermic peak. Analyses were
performed in triplicate and two scans were performed on each
sample to eliminate the relaxation enthalpy.
2.2.9. Scanning electron microscopy (SEM)
The scanning electron microscopy was performed using methodology described by Andrade-Mahecha (2009). Films were frozen
in liquid nitrogen and fractured. Samples (3  3 mm) were xed on
aluminum supports with conductive copper tape and were maintained on a desiccators with silica gel (0% RH) for 7 days. Then,
samples were coated with a gold layer of 92 
A, using a sputter
coater (Balzers SCD050 Sputter Coater, Bal-Tec, Wetzlar, Germany).
The surface and cross section of the lms were observed in a
scanning electron microscope JEOL JSM-5800 LV (JEOL, Peabody,
MA, USA), at an accelerating voltage of 10 kV.
2.3. Statistical analysis

2.2.5. Water solubility of lms


Film solubility was determined according to the methodology
described by Colla et al. (2006). Three discs with 2 cm diameter of
each lm were cut, weighed, immersed in 50 mL of distilled water.
Then, they were slowly and periodically agitated during 24 h at
25  C. The dry mass content of initial and nal samples was
determined by drying the samples at 105  C for 24 h. The measurements were made in triplicate.
2.2.6. Film thickness measurements
Film thickness was measured using a caliper micrometer (Tesa
Technology, Renens, Switzerland), taking measurements at 15
random positions on the lm. The average of these values was used
to determine the mechanical properties of each lm.
2.2.7. Mechanical properties
Mechanical properties were determined by tensile tests (tensile
strength, elongation at break and elastic modulus) using a Universal Testing Machine (model TA-TX plus, Stable Micro Systems,
Surrey, England). Measurements were made following the American Society for Testing and Materials standard method D882-02
(ASTM, 2005). Sample lms were cut into 25 mm wide strips
each one at least 115 mm in length, using a scalpel. The initial grip
separation was set at 80 mm and the crosshead speed at 1.0 mm/s.
The tensile strength (MPa) and elongation at break (%) were
calculated using the software Texture Expert (Stable Micro Systems,
Surrey, England). The elastic modulus was calculated as the slope of
the initial linear portion of this curve. At least, ve samples from
each lm were evaluated.
The coating strength was also estimated. According to Alcantara,
Rumsey, and Krochta (1998), an ideal coating should be quite
strong, but also provide good exibility when subjected to tension.
Thus, Joshi, Kral, and Topp (1989) proposed a coating strength indicator by the ratio of tensile strength and elastic modulus of the
lms. For the indicator calculation, tensile strength and elastic
modulus values obtained in the tensile test were used.

The results were statistically evaluated by analysis of variance


(ANOVA) and Tukeys test at 5% signicance level, using the software Statistica 8 (StatSoft Inc., Tulsa, USA).
3. Results and discussion
3.1. Respiration rate of coated apple slices, water vapor resistance
and surface solid density of coatings
The results obtained for barrier properties and coatings surface
solid density are presented in Table 2. According to Vargas et al.
(2008), the barrier properties of edible coatings can be evaluated
by the water vapor resistance and by respiration rate of coated
product. The interaction between the product surface and the
coating can modify the barrier properties of the lm. Therefore, it is
important to measure these properties of coatings applied to the
plant tissue (Morillon, Debeaufort, Blond, Capelle, & Voilley, 2002;
Vargas et al., 2008).
The average respiration rate and water vapor resistance of apple
slices without coatings were 8.92  0.34 ml CO2 kg1 h1 and
22.92  2.34 s cm1, respectively. Coatings promoted lower
Table 2
Respiration rate (RR) of coated apples, water vapor resistance (WVR) and surface
solid density (SSD) of coatings applied on apple cylinders and lm solubility (sol).
Formulationa

RR (mL CO2
kg1 h1)

WVR (s cm1)

SSD (103
g cm2)

Sol (%)

1
2
3
4

5.75  0.22B
4.99  0.27C
9.13  0.66A
6.13  0.12B

36.91  2.26B
50.75  5.29A
25.95  2.04C
44.51  2.15A

2.25  0.16A
2.39  0.33A
3.42  0.21B
3.45  0.48B

43.14  0.70A
42.96  2.07A
27.50  2.67C
37.87  1.74B

Values with the same letter within the column are not signicantly different
p  0.05.
a
Formulation 1 2.50% CS; 2.82% GLY; 0.10% CW; 0.90% SA; formulation
2 2.50% CS; 2.61% GLY; 0.38% CW; 0.62% SA; formulation 3 3.50% CS; 1.00% GLY;
0.40% CW; 0.60% SA; formulation 4 3.00% CS; 1.50% GLY; 0.20% CW; 0.80% SA.

M. Chiumarelli, M.D. Hubinger / Food Hydrocolloids 38 (2014) 20e27

respiration rates than the uncoated sample, except for formulation


3 that promoted the highest respiration rate (RR) of apple slices
among treatments (Table 2). Han, Seo, Park, Kim, and Lee (2006)
reported that oxygen molecules can penetrate through wax/
starch interfaces that are connected between wax particles,
providing oxygen penetration channels when the lm contains
high wax content, which probably may have occurred in formulation 3.
Formulation 2 resulted in the lowest apple slices RR. Glycerol
lls the empty spaces of the matrix and reduces pores and cracks in
the surface of lms, promoting a more effective barrier to gas exchange (Garcia et al., 2000; Krochta, 2002; Ribeiro, Vicente,
Teixeira, & Miranda, 2007). Respiration rate of apple slices coated
with formulations 1 and 4 did not differ statistically and showed
low respiration rate values, promoting a good gas barrier (Table 2).
Glycerol and lipids concentrations strongly inuenced on water
vapor resistance (WVR). Glycerol percentage above 1.50% and
carnauba wax concentrations below 0.38% resulted in better WVR.
Formulation 2 presented the highest WVR, and did not differ statistically from formulation 4, showing an excellent moisture barrier
(Table 2). Formulation 3 showed the lowest WVR due to the polymer matrix discontinuity, resulting from the large amount of
carnauba wax used in this coating. According to Farris, Introzzi, and
Piergiovanni (2009) and Anker, Berntsen, Hermansson, and Stading
(2002), the water vapor barrier of emulsied lms is impaired by
hydrophilic phase (glycerol, protein or polysaccharide), that allows
the permeation of water molecules. However, the authors reported
that increase on lipid content did not improve WVR, which can be
observed in formulation 3.
Vargas, Albors, Chiralt, and Gonzlez-Martnez (2009) reported
that oleic acid addition in chitosan-based coatings reduced the
WVR, behavior also observed in this study. However, some authors
reported different results, which showed that the interactions between the ingredients used in the coatings formulations led to
different barrier properties. Navarro-Tarazaga, Massa, and PrezGago (2011) observed that the water vapor permeability of carboxymethyl cellulose and oleic acid-based lms has signicantly
increased by glycerol addition. Garcia et al. (2010) concluded that
higher cassava starch concentrations lead to higher water vapor
resistance of edible coatings applied in strawberries. However, the
authors observed that polysaccharides-based coatings without
lipids did not promote a large increase in water vapor resistance,
due to its hydrophilic nature. Auras, Arroyo, and Selke (2009) also
observed that water vapor permeability of cassava starch lms have
increased as a function of glycerol and beeswax content.
Although the glycerol is a hydrophilic component, it affects the
glass transition temperature and crystallinity of the lm. According
to Auras, Harte, and Selke (2004) and Cava, Gimnez, Gavara, and
Lagaronauras (2006), the increase of crystallinity promotes the
permeability reduction of water and oxygen molecules. Due to this
mechanism, the formulations with higher concentrations of glycerol showed higher WVR and lower RR values of apple slices.
However, formulation 1 showed poor water vapor barrier, probably
because the cassava starcheglycerol ratio was not adequate and,
associated with low carnauba wax percentage, did not result in a
polymeric matrix strong enough to promote higher WVR values.
Coating thickness is straightly related to barrier properties.
Precise measurement of this parameter is very complex, since
cutting or freezing the sample may change their values (Vargas
et al., 2008). Thus, the thickness can be estimated by surface solid
density (SSD). Higher SSD values indicate the coating is thicker
(Vargas et al., 2006; Villalobos-Carvajal et al., 2009). Formulations 3
and 4, with higher cassava starch concentration, gave higher SSD
values (p < 0.05). The surface solid density is related to the thickness of coatings, therefore, higher solids percentages in

23

formulations result in SSD increase and promote thicker coating,


which may have contributed to the higher resistance to water vapor
and good barrier to gases promoted by formulation 4 (Vargas et al.,
2006; Villalobos-Carvajal et al., 2009). Although formulation 3
resulted in SSD high value, the high lipid content may have created
a non-homogenous structure, harming the barrier properties of
coating.
3.2. Water solubility
Water solubility of cassava starch, glycerol, carnauba wax and
stearic acid lms ranged from 27.50 to 43.14% (Table 2), which are
good values for their application in fresh-cut fruits. Formulations 1
and 2 showed higher solubility due to their glycerol and lipids
content. Glycerol is very hygroscopic, increasing the solubility of
coating. Formulation 3, prepared with low glycerol concentration
and high carnauba wax content, showed the lowest solubility.
Formulation 4 showed intermediate solubility values.
Mller, Yamashita, and Laurindo (2008) reported that the
biodegradable lms solubility is inuenced by the plasticizer type
and concentration used in its preparation. According to Matta,
Sarmento, Sarantpoulos, and Zocchi (2011), plasticizer addition,
in particular of glycerol, has a great inuence on the solubility of
starch lms, due to its hydrophilic character. Glycerol interacts with
the lm matrix by increasing the space between the chains, facilitating the water diffusion and, consequently, increased lm
solubility.
3.3. Mechanical properties
The tensile strength, elongation at break and elastic modulus of
lms produced with cassava starch, glycerol, carnauba wax and
stearic acid are presented in Table 3. Formulations 1 and 2 (with
same cassava starch content, 2.50% w/w) did not differ statistically
regarding tensile strength, with the lowest values for this response.
Percentages of cassava starch around 2.6% combined with higher
glycerol concentrations resulted in lower tensile strength. Films
prepared with formulation 3 showed higher stress values, since this
treatment has the highest cassava starch (3.50%) and carnauba wax
(0.40%) concentrations, which have contributed to the formation of
a more rigid polymer matrix, as evidenced by the highest elastic
modulus among the formulations (Table 3). Formulation 4 showed
intermediate values, not resulting in highly exible or extremely
hard lms.
Al-Hassan and Norziah (2012) reported that proteins and
polysaccharides-based lms with glycerol addition had lower
values of tensile strength, indicating that lms were more exible
due to the plasticizing effect, which increased the mobility of

Table 3
Tensile strength (TS), elongation at break (Elo), elastic modulus (EM) and coating
strength (TS/EM) of lms with cassava starch, glycerol, carnauba wax and stearic
acid.
Formulationa TS (MPa)

1
2
3
4

0.220  0.015A
0.211  0.011A
1.067  0.099B
0.729  0.015C

Elo (%)

EM (MPa)

Coating
strength
(TS/EM)

17.673  0.342A
18.618  1.012A
24.217  1.704B
31.074  2.278C

0.197  0.019A
0.077  0.007B
0.504  0.023C
0.221  0.029A

1.182  0.058A
1.446  0.207A
2.009  0.120B
4.047  0.221C

Values with the same letter within the column are not signicantly different
p  0.05.
a
Formulation 1 2.50% CS; 2.82% GLY; 0.10% CW; 0.90% SA; formulation
2 2.50% CS; 2.61% GLY; 0.38% CW; 0.62% SA; formulation 3 3.50% CS; 1.00% GLY;
0.40% CW; 0.60% SA; formulation 4 3.00% CS; 1.50% GLY; 0.20% CW; 0.80% SA.

24

M. Chiumarelli, M.D. Hubinger / Food Hydrocolloids 38 (2014) 20e27

polymer chains. Mali, Karam, Ramos, and Grossmann (2004) and


Mali, Sakanaka, Yamashita, and Grossmann (2005) reported that
cassava starch lms with high glycerol concentration showed low
tensile strength and low elastic modulus, as it was observed in this
study (Table 3). Flexibility increase as a function of plasticizer
content may be related to structural changes in the starch network,
because the matrix becomes less dense and, under tension, the
movements of the polymer chains are facilitated.
Regarding the elongation at break, lower concentrations of
starch (2.50%) have decreased elongation values (formulations 1
and 2), while higher starch and glycerol contents furnished an
elongation increase (Table 3). Formulation 4 showed the highest
elongation at break, since it has higher glycerol content (1.50%)
than the formulation 3, although the cassava starch concentration
was lower (3.00%). The higher carnauba wax concentration (0.40%)
of formulation 3 could have contributed to decrease lms elongation when compared to formulation 4. Auras et al. (2009) observed
that the beeswax addition in cassava starch lms increased the
tensile strength and decreased the elongation, resulting in more
rigid lms. Han et al. (2006) reported that the addition of beeswax
emulsion in pea starch lms decreased tensile strength and elongation, and increased elastic modulus. In present work, wax
addiction increased tensile strength and elastic modulus. However,
the glycerol and cassava starch content were determinant on lm
rigidity: low glycerol concentration and high wax and cassava
starch content promoted rigid lms (formulation 3).
The coating strength was signicantly (p < 0.05) bigger in
formulation 4 (Table 3), indicating that this lm was more exible,
but with sufcient mechanical strength to protect the food. According to Alcantara et al. (1998), higher glycerol concentrations in
whey protein lms promoted bigger exibility, increasing the indicator value. Films with a high plasticizer concentration present
tensile strength and elastic modulus reduction. However, the
plasticizer effect is more pronounced on the elastic modulus, as
seen in this study.
3.4. Thermal properties
All formulations showed similar thermograms with two endothermic peaks, which can be associated to the use of lipid components (Fig. 1). The rst peak indicates the fatty acids melting and the

Table 4
Transition temperature (Tm) and enthalpy of transition (DH) of carnauba wax, stearic
acid and lms with cassava starch, glycerol, carnauba wax and stearic acid.
Samplea

1st Peak
Tm1 [ C]

Pure carnauba wax


Pure stearic acid
Formulation 1
Formulation 2
Formulation 3
Formulation 4

2nd Peak

DH1 [J g1]d.b.b Tm2 [ C]

58.13  0.20
0.58  0.11
60.48  0.82 134.02  10.42
A
56.18  0.39
19.83  0.92A
54.59  0.63B
10.30  0.81B
55.64  0.80AB 11.49  0.48B
56.05  0.42A 20.19  0.54A

83.38  1.10
e
71.46  0.25A
74.73  0.39B
75.80  0.50C
73.87  0.14B

DH2 [J g1]d.b.
171.83  4.74
e
0.36  0.05A
2.91  0.54B
4.29  0.20C
1.84  0.05D

Values with the same letter within the column are not signicantly different
p  0.05.
a
Formulation 1 2.50% CS; 2.82% GLY; 0.10% CW; 0.90% SA; formulation
2 2.50% CS; 2.61% GLY; 0.38% CW; 0.62% SA; formulation 3 3.50% CS; 1.00% GLY;
0.40% CW; 0.60% SA; formulation 4 3.00% CS; 1.50% GLY; 0.20% CW; 0.80% SA.
b
Dry basis.

second peak represents the melting of long carbon chain components of carnauba wax. In formulations containing higher carnauba
wax content (Formulations 2 and 3), the rst endothermic peak was
smaller and the second peak had become more prominent when
compared with other formulations.
The glass transition temperature could not be observed in these
thermograms. According to Ghanbarzadeh, Almasi, and Entezami
(2010) and Ghanbarzadeh and Almasi (2011), the glass transition
of carbohydrate lms with plasticizer is difcult to be determined
by DSC analysis, because the change of heat capacity is very low in
the glass transition.
The transition temperature (Tm) and the enthalpy of transition
(DH) of carnauba wax, stearic acid and lms samples are shown in
Table 4.
Transition temperature of the rst peak (Tm1) was lower than
Tm1 of lipid components alone, due to glycerol. Formulations 1 and
4 did not differ statistically with respect to Tm1, since they had a
high concentration of fatty acid and low carnauba wax content.
Formulation 3 also did not signicantly differ from these two formulations, although containing lower stearic acid concentration
and a higher wax percentage. But formulation 3 contained low
glycerol content (1%), which contributed to non-signicant reduction in Tm1. Al-Hassan and Norziah (2012) reported that the increase
in glycerol concentration reduced the Tm and DH of starch and sh

Fig. 1. Thermograms of the lms prepared with cassava starch, glycerol, carnauba wax and stearic acid storage for 3 weeks at 58% RH. Formulation 1 2.50% CS; 2.82% GLY; 0.10%
CW; 0.90% SA; formulation 2 2.50% CS; 2.61% GLY; 0.38% CW; 0.62% SA; formulation 3 3.50% CS; 1.00% GLY; 0.40% CW; 0.60% SA; formulation 4 3.00% CS; 1.50% GLY; 0.20% CW;
0.80% SA.

M. Chiumarelli, M.D. Hubinger / Food Hydrocolloids 38 (2014) 20e27

gelatin lm, indicating that the plasticizer has reduced the intermolecular forces and increased polymer chain mobility. Formulation 2 showed the lowest Tm1 due to both a low stearic acid
percentage (0.62%) and a high glycerol concentration (2.61%).
For the second peak transition temperature (Tm2), only formulations 2 and 4 did not differ statistically. This peak may be representing the fusion of carnauba wax and stearic acid mixture, which
could explain the highest Tm2 obtained by formulation 3, since this
treatment has a higher wax percentage (0.40% w/w) which would
increase the melting point of lipid mixture. The presence of two
endothermic peaks can be related to a rich lipid fraction, because
there is a polymorphic transformation of fatty acids, from a form
(rst endothermic peak) to b form (second endothermic peak). The
melting temperature of stearic acid a form of is 55  C and of b form,
73  C (Lutton & Fehl, 1970), very close to the Tm1 and Tm2 values
found for all formulations (Table 4).
These transition temperatures (Tm1 and Tm2) may be related to
mechanical properties of lms, because they are dependent on
glycerol and lipids concentrations. Films with higher values of Tm1
and Tm2 showed high values of tensile strength and elastic
modulus, as shown by formulation 3; those with low tensile
strength and elastic modulus, such as formulation 2, showed the
lowest transition temperatures. Formulation 4, with high Tm1 and
intermediate value of Tm2 formed lms with high values of elongation and coating strength. The enthalpy of transition (DH) was
strongly inuenced by carnauba wax concentration on lms. Formulations 2 and 3, with higher wax content showed low enthalpy
for the rst peak and higher values of DH2 compared to formulations 1 and 4.
3.5. Scanning electron microscopy (SEM)
The microstructure of the lms was analyzed using the scanning
electron microscopy technique (SEM). Figs. 2 and 3 show the surface and the cross section microscopy of lms, respectively.
Microstructure was analyzed qualitatively in order to verify the
features promoted by different concentrations of cassava starch,

25

glycerol, stearic acid and carnauba wax. According to Jimnez,


Fabra, Talens, and Chiralt (2010), solvent evaporation causes
changes in components concentrations and in the emulsion liquid
phase viscosity, leading to lipids aggregation and creaming, thus
affecting the inner structure and the lm surface and therefore,
barrier, mechanical and optical properties.
Film elaborated with formulation 1 showed a discontinuity in
the matrix, with very irregular surface, presence of pores and lipid
content agglomeration, indicated by arrows in micrographs
(Fig. 2(a)). However, the cross section of the lm showed a more
cohesive structure, with closed polymeric networks and without
pores, but with lipid clusters (Fig. 3(a)). The cross section microstructure of this formulation corroborates the results obtained for
mechanical properties and barrier lms analyses: cohesive polymer
network promoted effective gases and moisture barriers (Table 2),
but the presence of lipid clusters has reduced mechanical strength
and exibility of the lms (Table 3), due to the formation of fragile
points in polymer matrix. According to Jimnez et al. (2010), the
lipid addition promotes discontinuities in the polymer matrix of the
dried lm, impairing its mechanical properties.
Formulation 2 also produced lms with irregular surface, large
pores and lipid agglomeration (Fig. 2(b)). The lm cross section,
however, showed a dense and closed polymer network (Fig. 3(b)).
This microstructure contributed to the high water vapor resistance
and reduced respiration rate of coated apple slices (Table 2). But the
lipid agglomeration impaired mechanical properties of these lms,
which showed low values of elongation and coating strength
(Table 3).
The surface of the lms prepared with formulation 3 showed
great irregularity, pores and lipid accumulation in some areas
(Fig. 2(c)). This irregular surface associated to presence of pores
observed in the cross section of these lms (Fig. 3(c)), resulted in
the worst barrier properties values (Table 2). Polymer structure was
less cohesive than the other formulations matrix, possibly due to
low glycerol concentration and high carnauba wax percentage,
providing more rigid lms (Table 3). The wax content could be
higher than ideal in the emulsion, which could have caused the

Fig. 2. SEM micrographs of the lms surface produced from formulations 1 (a), 2 (b), 3 (c) and 4 (d). Scale bar 10 mm. Arrows indicate lipid cluster. Formulation 1 2.50% CS; 2.82%
GLY; 0.10% CW; 0.90% SA; formulation 2 2.50% CS; 2.61% GLY; 0.38% CW; 0.62% SA; formulation 3 3.50% CS; 1.00% GLY; 0.40% CW; 0.60% SA; formulation 4 3.00% CS; 1.50% GLY;
0.20% CW; 0.80% SA.

26

M. Chiumarelli, M.D. Hubinger / Food Hydrocolloids 38 (2014) 20e27

Fig. 3. SEM micrographs of the lms cross section produced from formulations 1 (a), 2 (b), 3 (c) and 4 (d). Scale bar 10 mm. Arrows indicate lipid cluster. Formulation 1 2.50% CS;
2.82% GLY; 0.10% CW; 0.90% SA; formulation 2 2.50% CS; 2.61% GLY; 0.38% CW; 0.62% SA; formulation 3 3.50% CS; 1.00% GLY; 0.40% CW; 0.60% SA; formulation 4 3.00% CS;
1.50% GLY; 0.20% CW; 0.80% SA.

lipid separation during lm drying and promoted the non-homogenous structure.


Formulation 4 promoted lms with more dense and regular
surface, with homogenous lipids distribution and without cracks
and pores (Fig. 2(d)), which contributed to the good barrier properties of this lm (Table 2). The cross section also showed a dense
and cohesive structure, with good lipids distribution (Fig. 3(d)).
According to Tapia-Blcido, Mauri, Menegalli, Sobral, and An
(2007), features as those observed in formulation 4 lms also results in lms with higher elongation, which was veried in this
study (Table 3).
Auras et al. (2009) reported that the microstructure of cassava
starch, glycerin and beeswax lms showed lipid droplets on lm
surface, resulting in non-homogenous lms, contrary to the ones
without wax addition. The authors also observed that glycerol
addition promoted a more compact polymer network without
pores or cracks. Bonilla, Vargas, Atres, and Chiralt (2011) reported
that chitosan lms with basil essential oil showed interruptions in
the polymer matrix due to lipid droplets, making the matrix less
dense. Kokoszka, Debeaufort, Hamblenton, Lenart, and Voilley
(2010) also observed that whey proteins lms with canola oil
showed structural discontinuity due to the components
immiscibility.
4. Conclusions
The results obtained in characterization analyses of coating/lm
formulations with cassava starch, glycerol, carnauba wax and
stearic acid showed that high wax content and low glycerol concentration forms poor lms, with rigid structure and poor gases and
water vapor barrier. Formulation 4, composed of 3% (w/w) of cassava starch, 1.5% (w/w) glycerol, 0.2% (w/w) of carnauba wax and
0.8% (w/w) of stearic acid presented coatings with good barrier
properties, and lms with good mechanical, thermal, physical and
structural properties, suitable for use as coatings. It can be potentially used in minimally processed fruit for product quality maintenance and it will be evaluated in a further work.

Acknowledgments
The authors are grateful to FAPESP (process number: 08/553431 and 09/51420-4) and CNPq for their nancial support.
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