polymers

Article
Bioactive Edible Films Based on Arrowroot Starch
Incorporated with Cranberry Powder: Microstructure,
Thermal Properties, Ascorbic Acid Content and
Sensory Analysis
Farayde Matta Fakhouri 1,2, * , Gislaine Ferreira Nogueira 3 , Rafael Augustus de Oliveira 3
and José Ignacio Velasco 1
1 Centre Català del Plàstic, Dpt. of Materials Science and Metallurgy, Universitat Politècnica de Catalunya,
Carrer Colom 114, E-08222 Terrassa, Spain; jose.ignacio.velasco@upc.edu
2 Faculty of Engineering, Federal University of Grande Dourados, Dourados, MS 79804-970, Brazil
3 School of Agricultural Engineering, University of Campinas, Campinas, SP 13083-875, Brazil;
gislainefnogueira@gmail.com (G.F.N.); augustus@feagri.unicamp.br (R.A.d.O.)
* Correspondence: farayde@gmail.com




Received: 31 August 2019; Accepted: 9 October 2019; Published: 11 October 2019


Abstract: The growing global awareness about environmental preservation has stimulated the search
for alternatives to replace conventional plastics made from fossil sources. One of the advantages
is using polymers from renewable sources, such as starch and gelatin, which, in addition to being
biodegradable, may also be edible. The incorporation of cranberry into a polymeric matrix can
transfer bioactive composite films, colour and flavour to the film, which are characteristic of this
fruit, expanding its application to fruit stripes or colourful coatings for specific foods. In this context,
the aim of this work was to evaluate the influence of the incorporation of 0, 5, 15, 25, 35, 45 and 55%
(solids mass/biopolymer mass) cranberry powder on the microstructure, thermal properties, ascorbic
acid content and sensory analysis of gelatin and arrowroot starch films obtained by casting. Scanning
electron microscopy (SEM) images showed that the incorporation of cranberry made the film surface
rough and irregular. All films presented an X-ray diffraction pattern typical of a semicrystalline
material. The glass transition temperature (Tg ) decreased when increasing the concentration of
cranberry in films. All films with cranberry presented high ascorbic acid content and were well
accepted by the tasters when sensory analysis was performed.

Keywords: natural polymers; arrowroot starch; gelatin; cranberry; properties; microstructure; X-ray
diffraction; differential scanning calorimetry; sensory analysis; food packaging

1. Introduction
The use of packaging is essential, as it plays a fundamental role in controlling the interactions
between food and the environment, protecting and maintaining product quality, beyond its basic
function of containing the food [1]. However, the polymers used in this industry is made from
non-renewable synthetic materials, which, despite having excellent functional properties, are causing
serious environmental problems due to the generation of high amounts of non-degradable solid waste
in the environment [2].
One alternative to reducing the environmental impact is to use natural polymers for packaging
rather than traditional petroleum-based polymers [1–5]. Biodegradable polymers are those which,
when exposed to the bioactive environment, are degraded by the enzymatic action of living organisms

Polymers 2019, 11, 1650; doi:10.3390/polym11101650 www.mdpi.com/journal/polymers

Polymers 2019, 11, 1650 2 of 12

(such as bacteria, yeasts, fungi) and converted at the end of the process to CO2 , H2 O and biomass
under aerobic conditions and hydrocarbons, methane and biomass under anaerobic conditions [6].
In this regard, both starch and gelatin are natural polymers that have been widely used in the
preparation of polymeric matrices for applications such as edible and biodegradable films and coatings
in the food industry. In addition, these natural polymers are capable of forming odourless, tasteless,
colourless and non-toxic matrices. Starch also has the advantage of being abundant, renewable and
present in different forms depending on its origin [2,7].
Arrowroot (Maranta arundinaceae L.) starch presents good digestibility, gelling ability and high
amylose content [8–11], desirable characteristics for the formulation of films with good technological
properties [12].
Gelatin, a protein of animal origin, is obtained from collagen via acid or basic hydrolysis using a
catalyst [13]; it has the ability to form thermo-reversible gels after heating, dissolution and cooling.
The formation of gelatin gels involves ionic combinations between the amino and carboxyl groups of
its amino acids, with the support of hydrogen bonds [14,15]. Blends of starch and gelatin have been
studied for the development of edible films [15]. Composite films have the applicability of combining
the benefits of each of the biopolymers used.
Studies on the production of edible films containing fruits (liquid or powder form) have showed
that the addition of fruit into the film-forming solution results in a film with bioactive compounds,
antimicrobial and antioxidant properties, colour and flavour characteristic of the fruit, expanding its
application to fruit stripes as a source of nutritional compounds, or as colourful coatings for specific
foods such as sushi; or yet, as active food packaging or partial substitutes for non-biodegradable plastic
packaging [16–19].
Cranberry fruit (Vaccinium macrocarpon Aiton) is native to North America, and is abundant in
bioactive compounds such as flavonoids, glycosides, anthocyanins, proanthocyanidins, organic acids,
phenolic compounds [20] and ascorbic acid (high levels, that is, 200 mg/kg of berries) [21]. Anthocyanin
pigments are mainly responsible for the pink and red colour of cranberries [22] and have been studied
due to their potential to act as an antioxidant [23]. The sour taste of fresh cranberries is due to their
content of organic acids such as hippuric acid and benzoic acid [24].
The antioxidant properties of cranberry are well documented in the literature, being in the
row of common fruits, since it is the fruit with the highest antioxidant activity regarding total
oxyradical elimination capacity, followed by apple, red grape, strawberry, peach, lemon, pear, banana,
orange and pineapple [25]. Cranberries are also known for their proanthocyanidin compounds,
in particular proanthocyanidin-A, which is associated with having antibacterial properties and potent
antiadhesives [26]. The bioactive compounds of cranberry demonstrated the ability to inhibit Escherichia
coli, Helicobacter pylori, Listeria monocytogenes, Staphylococcus aureus and Salmonella typhimurium [27–29].
The incorporation of cranberry into the arrowroot starch and gelatin polymeric matrix can confer
the resulting film with modifications in the thermal, morphological, mechanical and sensorial properties,
as well as in the bioactivity, conferring bioactive compounds with antioxidant and antimicrobial activity
and expanding their application as intelligent packaging. In addition, films can function as a cranberry
encapsulant as they can form a protective barrier to their bioactive compounds and further promote
a controlled release of these compounds onto the food surface during storage, extending their shelf
life [2,3,7,18]. The aim of this study was to develop edible films based on arrowroot starch and evaluate
the influence of cranberry powder on the microstructure, thermal properties, ascorbic acid content and
sensory analysis of these films for their application in food packaging.

2. Materials and Methods

2.1. Materials
Gelatin type A (Leiner Davis Gelatin, Cotia, Brazil), cranberry (Vaccinium macrocarpon) powder
(Herbarium, Colombo, Brazil) and glycerol (Synth, Diadema, Brazil). Arrowroot starch containing
Polymers 2019, 11, 1650 3 of 12

15.24 ± 0.19% of water, 0.40 ± 0.03% of protein, 0.12 ± 0.01 % of fat, 0.33 ± 0.01% ash and 83.91 ± 0.76%
of carbohydrates [30] and amylose content of 35.20 ± 1.63% [31,32]. The arrowroot was obtained in
partnership with the Faculty of Agronomy, Federal University of Grande Dourados. All other reagents
used for the analysis were presented at analytical grade.

2.2. Methods

2.2.1. Film Preparation

Films were obtained by the casting solvent technique. Gelatin and arrowroot starch solutions were
prepared separately. For the production of the gelatin solution, 5 g of gelatin was hydrated in 100 mL
of distilled water for 1 h. After that, this solution was heated at 80 ◦ C for 10 min in a thermostatic bath
(TECNAL, Piracicaba, Brazil), without agitation, to avoid the formation of bubbles. The starch solution
was obtained by dispersing 3 g of arrowroot starch into 100 mL of distilled water and heating until
reaching 80 ◦ C in a thermostatic bath (TECNAL, Piracicaba, Brazil), with constant agitation for about
10 min. The solutions were mixed at volume ratios of 1:1 (gelatin type A solution/arrowroot starch
solution) and plasticizer (glycerol) was incorporated in a concentration of 10% (0.5 g) in relation to the
gelatin mass (w/w) and 20% (0.6 g) in relation to the starch mass (w/w) in the filmogenic solutions
(w/w). This stirring was carried out gently in order to avoid the formation of bubbles in the sample,
maintaining the natural pH of the solution [15].
The solutions were prepared by dispersing the cranberry powder into 100 mL of water in different
concentrations: 0% (0 g), 5% (0.4 g), 15% (1.2 g), 25% (2.0 g), 35% (2.8 g), 45% (3.6 g) and 55% (4.4 g)
weight of cranberry in relation to the weight of the macromolecules in the filmogenic solutions (8 g).
After, 20 mL of filmogenic solution was deposited into Plexiglas dishes (11.8 cm diameter) and
dried at 25 ◦ C for 24 h until they could be easily removed from the support. The films were conditioned
at 25 ◦ C and 52% of relative humidity for 48 h before their characterization.

2.2.2. Visual Aspect and Microstructure

A visual test was performed to select films that were flexible and presented homogeneous colour.
The morphological characteristics of the surface and cross section developed for films was observed in
a scanning electron microscope (SEM) with X-ray energy dispersive (EDS) detector bench (model of
SEM Leo 440i, model of EDS: 6070, Leo 440i-Leo Electron Microscopy/Oxford–Cambridge, England).
The film sample was placed on a double-sided carbon-adhesive tape adhered to stub, submitted to
the application of a gold layer (model K450, Sputter Coater EMITECH, Kent, UK) and observed in a
scanning electron microscope operated at 10 kV.

2.2.3. X-ray Diffractometry (XRD)

The diaphragms were obtained using an X-ray diffractometer, model X’Pert, Philips Analytical
X Ray (Almelo, Netherlands), analysis conditions: Voltage and current: 40 kV and 40 mA, Scanning
range: 2 theta from 5◦ to 40◦ , step: 0, 1◦ speed: 0.0166◦ /s.

2.2.4. Differential Scanning Calorimetry (DSC)

For the analysis of the thermal properties of the films, a differential scanning calorimeter (DSC1,
Mettler Toledo, Schwerzenbach, Switzerland) was used. 10 mg of film sample was weighed on a
microanalytical scale (MX5-Mettler Toledo, Schwerzenbach, Switzerland) using an aluminium dish
(40 µL). For reference, a standard aluminium was used. The sample was submitted to a heating
program of 25 to 160 ◦ C at the rate of 10 ◦ C/min, in an inert atmosphere (50 mL/min of N2 ). When the
temperature reached 160 ◦ C, the sample was held for 5 min at this temperature. After this first scan,
the measurement cells were cooled with liquid nitrogen to 25 ◦ C, followed by a second heating sweep
of 25 to 160 ◦ C at a rate of 10 ◦ C/min in an inert environment (50 mL/min of N2 ). The glass transition
Polymers 2019, 11, 1650 4 of 12

temperature (Tg ) was calculated as the baseline inflection point, caused by the discontinuity of specific
heat of the sample.

2.2.5. Determination of Ascorbic Acid Content

The determination of ascorbic acid content was performed by the method of Tillmans, with
titration of the sample with standardized solution of 2,6-dichlorophenolindofenol [33]. Retention of
ascorbic acid was calculated according to Equation (1):

V × F × 100
R= (1)
A
where R is the amount of ascorbic acid in the film (mg/100 g of sample), V is the volume of Tillmans
solution spent in the titration, F is the solution factor and A is the mL of the sample used. The analyses
were performed in triplicate of the duplicate, and a total of 6 values were obtained.

2.2.6. Sensory Analysis

The sensory evaluation of the edible films was performed by 56 untrained tasters. The number
of tasters and the order of presentation of samples followed the design of [34], which considers the
first-order, carry over effects. The acceptance tests were performed to assess the appearance, colour,
flavour, taste and overall acceptance attributes. The film samples were cut into square format (2 cm ×
2 cm) and served on a white plate on a white table, monadically and coded with three digits. In the
sensory evaluations, the testers evaluated how much they liked or disliked the samples, through a
hedonic scale of nine points with the corresponding extremes: “disliked extremely” (1) and “liked
extremely” (9).

2.2.7. Statistical Analysis

The results for the responses of the experimental design were evaluated using Statistica 9.0
software (StatSoft, South America). Significant differences were evaluated by analysis of variance
(ANOVA) and the Tukey test at 5% level of significance, using SAS software (SAS 9.2, Cary, NC, USA).

3. Results and Discussion

3.1. Visual Aspects and Microstructure

Figure 1 shows the photography, SEM surface and cross section of the edible films based
on arrowroot starch and gelatine incorporated with cranberry. Overall, the edible films made from
arrowroot starch and gelatine incorporated with 0, 5, 15, 25, 35, 45 and 55% of cranberry were transparent.
Arrowroot starch and gelatine-based films were colourless and odourless, with a smooth surface
and an organised polymer matrix. With the incorporation of cranberry powder, the film surface
became rough, especially in films with 35, 45 and 55% of cranberry. This is due to the protuberances
caused by suspended and agglomerated particles of cranberry powder in the polymer matrix. Similar
characteristics were observed for films with blackberry [35,36], solid lipid microparticles containing
ascorbic acid [7], lipid microparticles and starch nanoparticles [37]. The cross-section images of films
showed multilayer structure, being more evident in films with 0, 5 and 15%, as well as it was for
carrageenan-starch films with antioxidant extracts of Cuban red propolis and yerba mate prepared by
casting [3].
The addition of cranberry at a higher concentration (55%) caused changes in the matrix of arrowroot
starch and gelatine, generating cracks in the resulting film after the drying process. This confirms
what was observed in the visual aspect, where difficulty in removing the films from the backing
plates and difficulty in handling them, due to their more brittle appearance, had been reported. These
characteristics suggest that this sample is more fragile than others in this study, and that the highest
concentration of cranberry powder added to the films was 45%.
Polymers
Polymers2019,
2019,11,
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FOR PEER REVIEW 5 5ofof13
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Films with cranberry showed a slightly pinkish coloration, especially at highest concentrations,
Films with
evidencing cranberry
the possible showedofa anthocyanins.
presence slightly pinkishAnthocyanins
coloration, especially at highest
are pigments concentrations,
responsible for the
evidencing the possible presence of anthocyanins. Anthocyanins are pigments responsible
pink and red colour of cranberries [22]. Arrowroot starch films incorporated with blackberry alsofor the pink
and red colour
exhibited colourofand
cranberries [22]. Arrowroot
flavour typical starchpowder,
of blackberry films incorporated withfilms
differing from blackberry also
without exhibited
blackberry,
colour and flavour typical of blackberry powder, differing from films without blackberry, which
which were transparent and odourless [35]. This colour can be attractive for the development of food were
transparent
packages. and odourless [35]. This colour can be attractive for the development of food packages.

Figure
Figure 1.
1. Scanning
Scanningelectron
electronmicroscopy
microscopy(SEM)(SEM)images
imagesof of surface
surface (column
(column 2,2, images
images with
with 1000
1000××
magnification),cross
magnification), crosssection
section(column
(column3,3,images
imageswithwith1000 × magnification,
1000× magnification,andandcolumn
column4,4,images
imageswith
with
3000×
3000 magnification),and
× magnification), andphotographic
photographicimages
images(column
(column5)5)ofofthe
theedible
ediblefilms
filmsbased
basedon
onarrowroot
arrowroot
starchand
starch andgelatine
gelatineincorporated
incorporatedwith
with0,0,5,
5, 15,
15, 25,
25, 35,
35, 45
45 and
and 55%
55% cranberry.
cranberry.

3.2. X-ray Diffractometry (XRD)

Polymers 2019, 11, 1650 6 of 12

3.2. X-ray Diffractometry (XRD)

Polymers 2019, 11, x FOR PEER REVIEW 6 of 13
The crystallinity of the arrowroot starch and gelatine films incorporated with 0, 5, 15, 25, 35,
The crystallinity
45 and 55% of cranberryofpowder
the arrowroot starch and
was evaluated by gelatine films incorporated
X-ray diffractometry; with 0, 5, 15, is
the diffractogram 25,shown
35, 45 in
Figure 2A,B. It was possible to observe a pattern characteristic of a semi-crystalline material forinthe
and 55% of cranberry powder was evaluated by X-ray diffractometry; the diffractogram is shown
Figure 2A,B.
analysed It was
samples. Allpossible to observean
films displayed a pattern characteristic
X-ray diffraction of a semi-crystalline
pattern material
typical of a partially for the
crystalline
analysed
material samples.
with All films
two defined displayed
peaks: an X-ray
the first in thediffraction
region ofpattern
2θ = 5typical
◦ at 10 of
◦ a partially
, with crystalline
a maximum peak
2θ = 7.5 , corresponding to the crystalline triple helix structure of gelatine [38]; and the second= in
material◦ with two defined peaks: the first in the region of 2θ = 5° at 10°, with a maximum peak 2θ
the7.5°, corresponding
region of 2θ = 12◦ to the◦ , crystalline
at 30 with maximum triple helix
peak structure
2θ aroundof15 gelatine
to 20◦ , [38];
beingandthisthe second intypical
behaviour the
region of 2θ = 12° at 30°, with maximum peak 2θ around 15 to 20°, being this behaviour typical of
of semi-crystalline polymers, such as starch and gelatine (Figure 2A). Arrowroot starch is a C-type
semi-crystalline polymers, such as starch and gelatine (Figure 2A). Arrowroot starch is a C-type
crystallinity, characterized by main peaks at 2θ = 5.68, 15.42, 17.42◦ , being the most prominent the peak
crystallinity, characterized by main peaks at 2θ = 5.68, 15.42, 17.42°, being the most prominent the
at 23.14◦ [8]. A-type gelatine has a crystalline XRD peak at 2θ = 20.9◦ [39]. In the diffractograms of films
peak at 23.14° [8]. A-type gelatine has a crystalline XRD peak at 2θ = 20.9° [39]. In the diffractograms
with cranberry (Figure 2B), it was also possible to verify that the intensity of the diffraction decreased at
of films with cranberry (Figure 2B), it was also possible to verify that the intensity of the diffraction
first peak and increased at second peak by increasing the concentration of cranberry incorporated into
decreased at first peak and increased at second peak by increasing the concentration of cranberry
theincorporated
arrowroot starchinto and
the gelatine
arrowroot film. Thisand
starch behaviour
gelatineevidences
film. Thisa possible
behaviour intermolecular
evidences a interaction
possible
between the cranberry and the polymeric matrix of the film.
intermolecular interaction between the cranberry and the polymeric matrix of the film.

2000

1800

1600

0phr Cramberry
1400
5phr
15phr
Intensity (Counts/s)

1200 25phr
35phr
45phr
1000
55phr
5 por Média Móvel (0phr Cramberry)
800 5 por Média Móvel (5phr)
5 por Média Móvel (15phr)
600 5 por Média Móvel (25phr)
5 por Média Móvel (35phr)
5 por Média Móvel (45phr)
400
5 por Média Móvel (55phr)

200

0
5 10 15 20 25 30 35

Diffraction angle (2Ɵ)

(A)
1400

1200

1000
0phr Cramberry
5phr
Intensity (Counts/s)

15phr
800 25phr
35phr
45phr
5 por Média Móvel (0phr Cramberry)
600
5 por Média Móvel (5phr)
5 por Média Móvel (15phr)
5 por Média Móvel (25phr)
400 5 por Média Móvel (35phr)
5 por Média Móvel (45phr)
5 por Média Móvel (55phr)

200

0
5 6 7 8 9 10

Diffraction angle (2Ɵ)

(B)
Figure
Figure 2. 2. X-ray
X-ray diffractionofofarrowroot
diffraction arrowrootstarch
starch and gelatin
gelatin films
filmsincorporated
incorporatedwith cranberry,
with 0, 5,
cranberry, 0, 15,
5, 15,
25, 35, 45 and 55%: (A) X-axis scale from 5 to 35; (B) X-axis scale from 5
25, 35, 45 and 55%: (A) X-axis scale from 5 to 35; (B) X-axis scale from 5 to 10 .to ◦
10°.
Polymers 2019, 11, 1650 7 of 12

3.3. Differential Scanning

Polymers 2019, Calorimetry
11, x FOR PEER REVIEW (DSC) 7 of 13

DSC3.3. thermograms
Differential Scanningcan be observed(DSC)
Calorimetry in Figure 3. The thermogram of films based on arrowroot
starch Polymers 2019, 11, (0%)
and gelatin x FOR PEER REVIEWthe beginning of Tg around 121.2 ◦ C; however, the glass
indicated 7 oftransition
13
DSC thermograms can be observed in Figure 3. The thermogram of films based on arrowroot
temperature
3.3.
range decreased
Differential Scanning
when increasing
Calorimetry (DSC)
the concentration of cranberry by 5 to 55% in films,
starch and gelatin (0%) indicated the beginning of Tg around 121.2 °C; however, the glass transition
◦ C. Studies have reported that when fruit pulps are incorporated
indicating Tg around
temperature 118.8 and 101.2
DSC thermograms can be observed in Figure 3.concentration
range decreased when increasing the The thermogramof cranberry
of films by 5 toon
based 55% in films,
arrowroot
into theindicating
polymeric Tg matrix
around of118.8
the film,
and the sugars
101.2 °C. foundhave
Studies in the fruit will
reported thathave
when a possible
fruit plasticizing
pulps are
starch and gelatin (0%) indicated the beginning of Tg around 121.2 °C; however, the glass transition
effect [16,40]. In
incorporated the film,
into the the plasticizer
polymeric matrix acts
of the by
film,modifying
the sugars the
found interaction
in the fruit
temperature range decreased when increasing the concentration of cranberry by 5 to 55% in films,
between
will have athe polymers,
possible
leading plasticizing
indicating Tgeffect
to an increase [16,40].
in the
around In
118.8 the
freeandfilm,
volume theof
101.2 plasticizer
°C.the acts
system,
Studies by
haveandmodifying the interaction
consequently
reported that when to fruit between
an increase
pulps are the
in chain
motilitypolymers,
and leading
reduction to
of an
the increase
glass in the free
transition volume of
temperature the system,
(T ) ofand
the consequently
system to
[41,42].
incorporated into the polymeric matrix of the film, the sugars gfound in the fruit will have a possible an increase in
chain motility
plasticizing and[16,40].
effect reduction of the
In the glass
film, the transition
plasticizertemperature (Tg) of the
acts by modifying thesystem [41,42].
interaction between the
polymers, leading to an increase in the free volume of the system, and consequently to an increase in
chain motility and reduction of the glass transition temperature (Tg) of the system [41,42].

3. Differential
Figure Figure scanning
3. Differential calorimetry
scanning calorimetry(DSC)
(DSC) thermograms
thermograms ofof edible
edible films
films based
based on arrowroot
on arrowroot
starch
starch and and gelatin
gelatin incorporated
incorporated with
with cranberry.
cranberry.
Figure 3. Differential scanning calorimetry (DSC) thermograms of edible films based on arrowroot
3.4. Ascorbic Acid Content
3.4. Ascorbic in Films
Acid Content in Films
starch and gelatin incorporated with cranberry.
AscorbicAscorbic
acid isacid
highlyis highly sensitive
sensitive to heatto and
heatoxidation
and oxidation [43].[43].
The The incorporation
incorporation of cranberry
of cranberry powder
3.4. Ascorbic
powder into Acid Content
the film can beinaFilms
way to promote the protection of ascorbic acid. This is because the film
into the film can be a way to promote the protection of ascorbic acid. This is because the film can
can Ascorbic
incorporate acidtheisactive
highly material intotoitsheat
sensitive protective
and matrix, essentially
oxidation [43]. Theinertinert to the active
incorporation material,
of cranberry
incorporate the active material into its protective matrix, essentially to the active material, thus
thus acting as an encapsulant [44]. Consequently, the film matrix will act as a barrier for the protection
actingpowder
as
against
into the film can
an encapsulant
unfavourable
be aConsequently,
[44]. way to promote the
environmental conditions
the protection
film matrix
(light,
of ascorbic
oxygen, will actacid.
pH, etc.), as aThis is because
barrier
assisting
the protection
film
forstability
the
can incorporate the active material into its protective matrix, essentially inert to theinactive
the material, of
againstascorbic
unfavourable
acid. The environmental
values of 460 conditions
mg/100 g of (light,
ascorbic acidoxygen,
were foundpH,foretc.), assisting
cranberry powder in the
and stability
33.88 of
thus acting as an encapsulant [44]. Consequently, the film matrix will act as a barrier for the protection
ascorbic acid.
± 4.19
against The gvalues
mg/100 to 187.31
unfavourable ofmg
460± 7.91/100
mg/100
environmental ggofof
conditions ascorbic
ascorbic
(light,acid acid
werewere
oxygen, found found
for films
pH, etc.), for cranberry
containing
assisting powder
0 and
in the stability 55%,
of and
33.88 ± respectively
4.19 mg/100 (Figure
g to 4). The
187.31 results
mg ± showed
7.91/100 that
g increase
of ascorbicthe concentration
acid were
ascorbic acid. The values of 460 mg/100 g of ascorbic acid were found for cranberry powder and 33.88 of
found cranberry
for in
films the film-
containing 0
and 55%,forming
± 4.19 solution
respectively caused
(Figure
mg/100 g to 187.31 a significant
mg4). increase
The results
± 7.91/100 in
showed
g of ascorbic the ascorbic
that
acid were acid
increase content
found for thefilms embedded
concentration in
containing 0 of the gelatin
andcranberry
55%, in
and arrowroot
respectively starch
(Figure film.
The The
4).caused ascorbic
results showed acid found
that in the
increase film containing of 0%cranberry
may be derived from
the film-forming solution a significant increase inthe
theconcentration
ascorbic acid in the film-
content embedded in the
the ascorbic acid caused
contentapresent in the arrowroot starch (42.69acid
± 5.75 mg/100 g) used as inthe basis for
gelatinforming
forming
solution
and arrowroot
the films.
starch
It film.
significant
film.
is possible
The increase
ascorbic
that rhizomes,
in the
acid ascorbic
found
vitamins
in the content embedded
film containing
and containing
minerals have beenbe
0%
loaded
the
may gelatin
be derived
together
and arrowroot starch The ascorbic acid found in the film 0% may derived from
from the ascorbic
during acid content
the extraction of thepresent
starch in the arrowroot starch (42.69 ± 5.75 mg/100 g) used as the basis
the ascorbic acid content present in from arrowroot.
the arrowroot starch (42.69 ± 5.75 mg/100 g) used as the basis for
for forming the films. It is possible that rhizomes, vitamins and minerals have been loaded together
forming the films. It is possible that rhizomes, vitamins and minerals have been loaded together
duringduring
the extraction
the extractionof the starch
of the from
starch fromarrowroot.
arrowroot.

Figure 4. Ascorbic acid content of edible films from arrowroot starch and gelatin incorporated
with cranberry.
 Polymers 2019, 11, x FOR PEER REVIEW 8 of 12

3.5. Sensory Analysis

Polymers 2019, 11, 1650 8 of 12
The films were also very well accepted by the tasters when they were analysed, as shown in
Table 1. The averages for all attributes evaluated were between 6 (slightly liked) and 7 (moderately
liked)
3.5. for most
Sensory formulations of cranberry films, indicating high potential for their commercialization
Analysis
as edible packaging. For attributes such as appearance and colour, more than 77% of the testers rated
The films were also very well accepted by the tasters when they were analysed, as shown in
very positive scores for the 7 film formulations between “liked slightly” to “liked extremely” (Figure
Table 1. The averages for all attributes evaluated were between 6 (slightly liked) and 7 (moderately
5). An important quality factor for dried foods is colour, since it reflects the sensory attractiveness
liked) for most formulations of cranberry films, indicating high potential for their commercialization
[45], particularly for products like cranberry, which presents a striking reddish colour. This colour
as edible packaging. For attributes such as appearance and colour, more than 77% of the testers rated
can be attractive for the development of certain food packages. With respect to the flavour, more than
very positive scores for the 7 film formulations between “liked slightly” to “liked extremely” (Figure 5).
57 and 61% of the tasters claimed to have enjoyed “slightly” and “extremely” the films with and
An important quality factor for dried foods is colour, since it reflects the sensory attractiveness [45],
without cranberry, respectively; only 7% remarked that they had slightly disliked the flavour of the
particularly for products like cranberry, which presents a striking reddish colour. This colour can be
film with 25% (cranberry powder). From the 56 tasters, only 36% rated grades ranging from “liked
attractive for the development of certain food packages. With respect to the flavour, more than 57 and
slightly” to “liked extremely” for the taste of the film with 0%, while 80% and 82% rated these grades
61% of the tasters claimed to have enjoyed “slightly” and “extremely” the films with and without
for films with 45 and 55% cranberry, respectively. Cranberries are commonly consumed as fresh
cranberry, respectively; only 7% remarked that they had slightly disliked the flavour of the film with
fruits, dried fruits or juices [46]. Cranberry fruit itself is very acidic and has a sour taste. This is due
25% (cranberry powder). From the 56 tasters, only 36% rated grades ranging from “liked slightly” to
to their organic acids such as hippuric and benzoic acid [24]. The incorporation of cranberry into
“liked extremely” for the taste of the film with 0%, while 80% and 82% rated these grades for films
edible films can be a viable strategy for softening its sour taste and acidity and facilitate their
with 45 and 55% cranberry, respectively. Cranberries are commonly consumed as fresh fruits, dried
commercialization. The results indicated that the incorporation of cranberry into the films improved
fruits or juices [46]. Cranberry fruit itself is very acidic and has a sour taste. This is due to their organic
its taste and, consequently, pleased the consumers; the same behaviour was observed for the global
acids such as hippuric and benzoic acid [24]. The incorporation of cranberry into edible films can be a
acceptance attribute.
viable strategy for softening its sour taste and acidity and facilitate their commercialization. The results
indicated that the incorporation of cranberry into the films improved its taste and, consequently,
Table 1. Results of sensory evaluation averages.
pleased the consumers; the same behaviour was observed for the global acceptance attribute.
Sample Appearance Colour Flavour Taste Global Acceptation
0% CRY 7.48 ± 1.53
Table 1.7.32 ± 1.63 6.46 ± 1.53 5.21 ± 1.74
Results of sensory evaluation averages. 6.09 ± 1.70
5% CRY 7.29 ± 1.56 7.13 ± 1.62 6.50 ± 1.54 5.71 ± 1.72 6.25 ± 1.49
Sample Appearance Colour Flavour Taste Global Acceptation
15% CRY 7.30 ± 1.43 7.13 ± 1.36 6.58 ± 1.57 6.27 ± 1.57 6.61 ± 1.37
0% 25%
CRY CRY 7.48 ± 1.53
7.38 ± 1.41 7.20±± 1.63
7.32 1.51 6.46 6.46± ±1.61
1.53 6.665.21 ± 1.74
± 1.52 6.77 6.09 ± 1.70
± 1.57
5% CRY 7.29 ± 1.56 7.13 ± 1.62 6.50 ± 1.54 5.71 ± 1.72 6.25 ± 1.49
35% CRY 7.27 ± 1.50 7.16 ± 1.40 6.60 ± 1.40 6.70 ± 1.64 6.71 ± 1.46
15% CRY 7.30 ± 1.43 7.13 ± 1.36 6.58 ± 1.57 6.27 ± 1.57 6.61 ± 1.37
45%
25% CRY CRY 6.73 ± 1.59
7.38 ± 1.41 7.02 ± 1.41
7.20 ± 1.51 6.39 ± 1.50
6.46 ± 1.61 6.73 ± 1.51
6.66 ± 1.52 6.63 6.77
± 1.52
± 1.57
35%55%
CRYCRY 7.27 6.91 ± 1.53
± 1.50 7.07±± 1.40
7.16 1.61 6.60 6.60± ±1.48
1.40 6.826.70
± 1.57
± 1.64 6.89 6.71
± 1.45
± 1.46
45% CRY 6.73 ± 1.59 7.02 ± 1.41 6.39 ± 1.50 6.73 ± 1.51 6.63 ± 1.52
55% CRY
*Sensory 6.91
evaluation of ± 1.53
films, 7.07 ± 1.61
using a hedonic scale Mean6.60 ± 1.48
of 56 consumers6.82for ± 1.57sample. Scores
each ± 1.45
6.89 for global
Sensory evaluation of films, using a hedonic scale Mean of 56 consumers for each sample. Scores
appearance: colour, flavour, taste and overall acceptance: 1 = disliked extremely; 2 = disliked very much; 3 for global
appearance: colour, flavour, taste and overall acceptance: 1 = disliked extremely; 2 = disliked very much; 3 = disliked
= disliked moderately; 4 = disliked slightly; 5 = liked/disliked; 6 = liked slightly; 7 = liked moderately; 8 =
moderately; 4 = disliked slightly; 5 = liked/disliked; 6 = liked slightly; 7 = liked moderately; 8 = liked very much;
= liked
liked9 very much; 9 = liked extremely.
extremely.

Figure 5. Cont.
Polymers 2019, 11, 1650 9 of 12
Polymers 2019, 11, x FOR PEER REVIEW 9 of 12

Figure 5. Sensory evaluation of films, using a hedonic scale (A) colour (B) flavour, (C) taste and (D)
Figure 5. Sensory evaluation
global acceptance. Mean of 56 of films, using
consumers a hedonic
for each scale
sample. (A) for
Scores colour (B)appearance:
global flavour, (C)colour,
taste and (D)
flavour,
global acceptance.
taste and Mean of 56
overall acceptance: 1 =consumers
disliked extremely; = disliked
for each 2sample. Scores
very for 3 = appearance:
global
much; colour,
disliked moderately;
4 = disliked
flavour, tasteslightly; 5 = liked/disliked;
and overall acceptance: 16==disliked 7 = liked
extremely;
liked slightly; 2 = disliked = liked3very
very8much;
moderately; = disliked
much;
9 = liked extremely.
moderately; 4 = disliked slightly; 5 = liked/disliked; 6 = liked slightly; 7 = liked moderately; 8 = liked
very much; 9 = liked extremely.
4. Conclusions
4. Conclusions
Films produced from arrowroot starch and gelatin incorporated with cranberry showed a decrease
in TgFilms
and were flexible
produced for arrowroot
from handling; no fractures
starch or grooves
and gelatin were evident.
incorporated with The incorporation
cranberry showed of a
cranberry into the film-forming solution in concentrations of 45 and 55% made it difficult
decrease in Tg and were flexible for handling; no fractures or grooves were evident. The incorporationto detach the
films
of from the
cranberry support
into plate. Thesesolution
the film-forming high concentrations of cranberry
in concentrations may
of 45 and have
55% restricted
made the molecular
it difficult to detach
motility of the amylopectin chains, leading to an increase in T , which made the films
the films from the support plate. These high concentrations of cranberry may have restricted
g fragile and brittle
the
molecular motility of the amylopectin chains, leading to an increase in Tg, which made the films
fragile and brittle for handling. SEM images showed that the incorporation of cranberry made the
Polymers 2019, 11, 1650 10 of 12

for handling. SEM images showed that the incorporation of cranberry made the film surface rough and
irregular. The X-ray diffraction of films revealed a partially crystalline structure. The incorporation of
cranberry into the film-forming solution transferred ascorbic acid, colour and flavour to the resulting
films, which may have led to a high acceptance of the films by the tasters when the sensory analysis
was performed. The films proved to be nutritious because of the ascorbic acid content; they were well
accepted by the tasters and could be a viable alternative to traditional packaging, being applied to fruit
stripes as a source of nutritional compounds, or as colourful coatings for specific foods such as sushi;
or as active food packaging or partial substitutes for non-biodegradable plastic packaging. However,
new effects caused by this addition should be studied in other properties of the films.

Author Contributions: For research articles with several authors, a short paragraph specifying their individual
contributions must be provided. The following statements should be used “conceptualization, F.M.F.; methodology,
G.F.N. and F.M.F.; formal analysis, G.F.N. and F.M.F.; investigation, F.M.F. and G.F.N.; writing—original draft
preparation, G.F.N.; writing—review and editing, F.M.F. and J.I.V.; supervision, J.I.V. and R.A.d.O.; project
administration, J.I.V.; funding acquisition, J.I.V. and R.A.d.O.
Funding: The research leading to these results has received funding from the European Union’s Horizon
2020 research and innovation programme under the Marie Skłodowska-Curie grant agreement No 712949
(TECNIOspring PLUS) and from the Agency for Business Competitiveness of the Government of Catalonia.
Acknowledgments: The authors are grateful to the Coordination for the Improvement of Higher Education
Personnel (CAPES, Brazil-01-P-3712/2017), to the School of Agricultural Engineering–University of Campinas and
Faculty of Engineering, Federal University of Grande Dourados.
Conflicts of Interest: The authors declare no conflict of interest.

References
1. Jaramillo, C.M.; Gutiérrez, T.J.; Goyanes, S.; Bernal, C.; Famá, L. Biodegradability and plasticizing effect of
yerba mate extract on cassava starch edible films. Carbohydr. Polym. 2016, 151, 150–159. [CrossRef] [PubMed]
2. Piñeros-Hernandez, D.; Medina-Jaramillo, C.; López-Córdoba, A.; Goyanes, S. Edible cassava starch films
carrying rosemary antioxidant extracts for potential use as active food packaging. Food Hydrocoll. 2017, 63,
488–495. [CrossRef]
3. Chang-Bravo, L.; López-Córdoba, A.; Martino, M. Biopolymeric matrices made of carrageenan and corn
starch for the antioxidant extracts delivery of Cuban red propolis and yerba mate. React. Funct. Polym. 2014,
85, 11–19. [CrossRef]
4. Fakhouri, F.M.; Costa, D.; Yamashita, F.; Martelli, S.M.; Jesus, R.C.; Alganer, K.; Collares-Queiroz, F.P.;
Innocentini-Mei, L.H. Comparative study of processing methods for starch/gelatin films. Carbohydr. Polym.
2013, 95, 681–689. [CrossRef] [PubMed]
5. Bertan, L.C. Development and Charaterization of Composite Films Based on Gelatin, Fatty Acids and Breu.
Master’s Thesis, State University of Campinas, Sao Paulo, Brazil, 2008.
6. Mohee, R.; Unmar, G.D.; Mudhoo, A.; Khadoo, P. Biodegradability of biodegradable/degradable plastic
materials under aerobic and anaerobic conditions. Waste Manag. 2008, 28, 1624–1629. [CrossRef] [PubMed]
7. Sartori, T.; Menegalli, F.C. Development and characterization of unripe banana starch films incorporated
with solid lipid microparticles containing ascorbic acid. Food Hydrocoll. 2016, 55, 210–219. [CrossRef]
8. Nogueira, G.F.; Fakhouri, F.M.; de Oliveira, R.A. Extraction and characterization of arrowroot (Maranta
arundinaceae L.) starch and its application in edible films. Carbohydr. Polym. 2018, 186, 64–72. [CrossRef]
[PubMed]
9. Villas-Boas, F.; Franco, C.M.L. Effect of bacterial β-amylase and fungal α-amylase on the digestibility and
structural characteristics of potato and arrowroot starches. Food Hydrocoll. 2016, 52, 795–803. [CrossRef]
10. Moorthy, S.N. Physicochemical and functional properties of tropical tuber starches: A review. Starch 2002,
54, 559–592. [CrossRef]
11. Hoover, R. Composition, molecular structure, and physicochemical properties of tuber and root starches:
A review. Carbohydr. Polym. 2001, 45, 253–267. [CrossRef]
12. Tharanathan, R.N. Biodegradable films and composite coatings: Past, present and future. Trends Food
Sci. Technol. 2003, 14, 71–78. [CrossRef]
Polymers 2019, 11, 1650 11 of 12

13. Poppe, J. Gelatin. In Thickening and Gelling Agents for Food; Imeson, A., Ed.; Aspen Publishers: New York, NY,
USA, 1987; pp. 144–168. [CrossRef]
14. Kester, J.J.; Fennema, O.R. Edible biofilms and coatings: A review. Food Technol. 1986, 40, 47–59.
15. Fakhouri, F.M.; Martelli, S.M.; Bertan, L.C.; Yamashita, F.; Mei, I.L.H.; Queiroz, F.P.C. Edible films made
from blends of manioc starch and gelatin—Influence of different types of plasticizer and different levels of
macromolecules on their properties. LWT Food Sci. Technol. 2012, 49, 149–154. [CrossRef]
16. Azeredo, H.M.C.; Morrugares-Carmona, R.; Wellner, N.; Cross, K.; Bajka, B.; Waldron, K.W. Development of
pectin films with pomegranate juice and citric acid. Food Chem. 2016, 198, 101–106. [CrossRef] [PubMed]
17. Farias, M.G.; Fakhouri, F.M.; Carvalho, C.W.P.; Ascheri, J.L.R. Caracterização físico-química de filmes
comestíveis de amido adicionado de acerola (Malphigia emarginata D.C.). Quím Nova 2012, 35, 546–552.
[CrossRef]
18. Cerruti, P.; Santagata, G.; Gomez d’Ayala, G.; Ambrogi, V.; Carfagna, C.; Malinconico, M.; Persico, P. Effect of
a natural polyphenolic extract on the properties of a biodegradable starch-based polymer. Polym. Degrad.
Stabil. 2011, 96, 839–846. [CrossRef]
19. Wang, S.; Marcone, M.F.; Barbut, S.; Lim, L.-T. Review: Fortification of dietary biopolymers-based packaging
material with bioactive plant extracts. Food Res. Int. 2012, 49, 80–91. [CrossRef]
20. Zuo, Y.; Wang, C.; Zhan, J. Separation, characterization, and quantitation of benzoic and phenolic antioxidants
in American cranberry fruit by GC-MS. J. Agric. Food Chem. 2002, 50, 3789–3794. [CrossRef] [PubMed]
21. Hisano, M.; Bruschini, H.; Nicodemo, A.C.; Srougii, M. Review: Cranberries and lower urinary tract infection
prevention. Clinics 2012, 67, 661–667. [CrossRef]
22. Tulio, A.Z., Jr.; Jablonski, J.E.; Jackson, L.S.; Chang, C.; Edirisinghe, I.; Burton-Freeman, B. Phenolic
composition, antioxidant properties, and endothelial cell function of red and white cranberry fruits. Food
Chem. 2014, 157, 540–552. [CrossRef]
23. Seeram, N.P.; Momin, R.A.; Nair, M.G.; Bourquin, L.D. Cyclooxygenase inhibitory and antioxidant cyanidin
glycosides in cherries and berries. Phytomedicine 2001, 8, 362–369. [CrossRef] [PubMed]
24. Nowack, R.; Schmitt, W. Cranberry juice for prophylaxis of urinary tract infections—Conclusions from
clinical experience and research. Phytomedicine 2008, 15, 653–667. [CrossRef] [PubMed]
25. Sun, J.; Chu, Y.F.; Wu, X.Z.; Liu, R.H. Antioxidant and antiproliferative activities of common fruits. J. Agric.
Food Chem. 2002, 50, 7449–7454. [CrossRef] [PubMed]
26. Foo, L.Y.; Lu, Y.R.; Howell, A.B.; Vorsa, N. A-type proanthocyanidin trimers from cranberry that inhibit
adherence of uropathogenic P-fimbriated Escherichia coli. J. Nat. Prod. 2000, 63, 1225–1228. [CrossRef]
[PubMed]
27. Burger, O.; Ofek, I.; Tabak, M.; Weiss, E.I.; Sharon, N.; Neeman, I. A high molecular mass constituent
of cranberry juice inhibits Helicobacter pylori adhesion to human gastric mucus. FEMS Immunol. Med.
Microbiol. 2000, 29, 295–301. [CrossRef] [PubMed]
28. Wu, V.C.H.; Qiu, X.J.; Bushway, A.; Harper, L. Antibacterial effects of American cranberry (Vaccinium
macrocarpon) concentrate on foodborne pathogens. LWT Food Sci. Technol. 2008, 41, 1834–1841. [CrossRef]
29. Lacombe, A.; Mcgivney, C.; Tadepalli, S.; Sun, X.; Wu, V.C.H. The effect of American cranberry (Vaccinium
macrocarpon) constituents on the growth inhibition, membrane integrity, and injury of Escherichia coli O157:H7
and Listeria monocytogenes in comparison to Lactobacillus rhamnosus. Food Microbiol. 2013, 34, 352–359.
[CrossRef] [PubMed]
30. Horwitz, W.; Latimer, G.W. Official Methods of Analysis, 18th ed.; Association of Official Analytical Chemists
International: Gaithersburg, MD, USA, 2006.
31. Martinez, C.; Cuevas, F. Evaluación de la calidad culinaria y molinera del arroz. Guia de estudo; CIAT: Cali,
Colombia, 1989; p. 75.
32. Zavareze, E.R.; El Halal, S.L.M.; Pereira, J.M.; Radünz, A.L.; Elias, M.C.; Dias, A.R.G. Chemical characterization
and extraction yield of rice starch with different amylose contents. Braz. J. Food Technol. 2009, 2, 25–30.
33. Horwitz, W. Official Methods of Analysis, 3rd ed.; Association of Official Analytical Chemists: Gaithersburg,
MD, USA, 1997.
34. Macfie, H.J.; Bratchell, N.; Greenhoff, K.; Vallis, L. Designs to balance the effect of order of presentation and
first-order carry-over effects in hall tests. J. Sens. Stud. 2007, 4, 129–148. [CrossRef]
35. Nogueira, G.F.; Fakhouri, F.M.; de Oliveira, R.A. Effect of incorporation of blackberry particles on the
physicochemical properties of edible films of arrowroot starch. Dry Technol. 2018, 37, 1–10. [CrossRef]
Polymers 2019, 11, 1650 12 of 12

36. Nogueira, G.F.; Soares, C.T.; Cavasini, R.; Fakhouri, F.M.; de Oliveira, R.A. Bioactive films of arrowroot
starch and blackberry pulp: Physical, mechanical and barrier properties and stability to pH and sterilization.
Food Chem. 2019, 275, 417–425. [CrossRef]
37. Shi, D.; Nannenga, B.L.; Iadanza, M.G.; Gonen, T. Three-dimensional electron cryslallography of protein
microcrystals. Elife 2013, 2. [CrossRef] [PubMed]
38. Dammak, I.; Lourenço, R.L.; Sobral, P.J.A. Active gelatin films incorporated with Pickering emulsions
encapsulating hesperidin: Preparation and physicochemical characterization. J. Food Eng. 2019, 240, 9–20.
[CrossRef]
39. Jalaja, K.; Deboki, N.; Subhas, C.K.; Nirmala, R.J. Fabrication of cationized gelatin nanofibers by
electrospinning for tissue regeneration. RSC Adv. 2015, 5, 89521–89530. [CrossRef]
40. Otoni, C.G.; De Moura, M.R.; Aouada, F.A.; Camilloto, G.P.; Cruz, R.S.; Lorevice, M.V.; Soares, N.F.F.;
Mattoso, L.H.C. Antimicrobial and physical mechanical properties of pectin/papaya puree/cinnamaldehyde
nanoemulsion edible composite films. Food Hydrocoll. 2014, 41, 188–194. [CrossRef]
41. Ivanič, F.; Jochec-Mošková, D.; Janigová, I.; Chodák, I. Physical properties of starch plasticized by a mixture
of plasticizers. Eur. Polym. J. 2017, 93, 843–849. [CrossRef]
42. Bergo, P.V.A.; Carvalho, R.A.; Sobral, P.J.A.; Dos Santos, R.M.C.; Da Silva, F.B.R.; Prison, J.M.;
Habitante, A.M.Q.B. Physical properties of edible films based on cassava starch as affected by the plasticizer
concentration. Packag. Technol. Sci. 2008, 21, 85–89. [CrossRef]
43. Islam, M.Z.; Kitamura, Y.; Yamano, Y.; Kitamura, M. Effect of vacuum spray drying on the physicochemical
properties, water sorption and glass transition phenomenon of orange juice powder. J. Food Eng. 2016, 169,
131–140. [CrossRef]
44. Ré, M.I. Microencapsulation by spray drying. Dry Technol. 1998, 16, 1195–1236. [CrossRef]
45. Yamashita, C.; Chung, M.M.S.; dos Santos, C.; Mayer, C.R.M.; Moraes, I.C.F.; Branco, I.G. Microencapsulation
of an anthocyanin-rich blackberry (spp.) by-product extract by freeze-drying. LWT-Food Sci. Technol. 2017,
84, 256–262. [CrossRef]
46. Roopchand, D.E.; Krueger, C.G.; Moskal, K.; Fridlender, B.; Lila, M.A.; Raskin, I. Food-compatible method for
the efficient extraction and stabilization of cranberry pomace polyphenols. Food Chem. 2013, 141, 3664–3669.
[CrossRef] [PubMed]

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