International Journal of Biological Macromolecules 106 (2018) 834–839

Contents lists available at ScienceDirect

International Journal of Biological Macromolecules

journal homepage: www.elsevier.com/locate/ijbiomac

Development and characterization of cassava starch films

incorporated with blueberry pomace
Cláudia Leites Luchese a , Tania Garrido b , Jordana Corralo Spada a , Isabel Cristina Tessaro a ,
Koro de la Caba b,∗
a
Department of Chemical Engineering, Federal University of Rio Grande do Sul (UFRGS), Rua Ramiro Barcelos, 2777, Anexo I Campus Saúde, ZC: 90035-007,
Porto Alegre, RS, Brazil
b
BIOMAT Research Group, University of the Basque Country (UPV/EHU), Escuela de Ingeniería de Gipuzkoa, Plaza de Europa 1, 20018 Donostia-San
Sebastián, Spain

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

Article history: This work is focused on the development of renewable and biodegradable films by the valorisation of
Received 27 May 2017 wastes from food processing industries, with the aim of contributing to the development of more sustain-
Received in revised form 11 July 2017 able films. In this context, different contents of blueberry pomace (BP) were incorporated into cassava
Accepted 13 August 2017
starch (CS) film forming solutions and the functional properties of the films prepared by solution casting
Available online 18 August 2017
were investigated, specifically, thermal, optical and physicochemical properties. BP-incorporated films
showed good barrier properties against light, indicating their beneficial effect to prevent food deterio-
Keywords:
ration caused by UV radiation when these films are used for food packaging applications. These results
Cassava starch
Blueberry pomace
were related to the presence of aromatic compounds in BP, which can absorb light at wavelengths below
Film 300 nm. Furthermore, all films maintained their structural integrity after immersion in water (24 h) and
UV barrier the maximum swelling displayed was lower than 300%. Additionally, the release of active compounds
Swelling from BP into food simulants (after 10 days) showed higher migration into the acetic acid medium in
Migration comparison with the ethanol medium. Therefore, the incorporation of BP into CS film forming solution
resulted in the improvement of film performance, suggesting the potential application of these films as
active packaging.
© 2017 Elsevier B.V. All rights reserved.

1. Introduction
Currently, approximately 40% of food packaging is manufac-
tured by using plastic materials [1]. In this regard, concerns related
to improper disposal, non-biodegradability and recycling difficulty
of these materials have attracted increasing attention in order to
avoid serious environmental problems, particularly because they
are used as one-time use packaging materials [2–4]. Besides, there
has been an increasing consumers’ awareness concerning their
health, food nutritional value and food safety [5,6]. To overcome
these issues, biopolymers from renewable sources are considered
as an attractive alternative since they are abundant, inexpensive,
environmentally friendly and biodegradable [7–10].
Among them, starch is considered as one of the most promising
materials because of its low cost, abundance, variety of botanical
sources, good film-forming properties [11–13]. In particular, cas-
∗ Corresponding author.
E-mail address: koro.delacaba@ehu.eus (K. de la Caba).
sava starch (CS) can be considered of great potential due to its
odourless, tasteless, colourless, non-toxic and biodegradable char-
acteristics [14,15]. Furthermore, starch can be used as carrier of
antioxidants and antimicrobials to develop active packaging [16].
This active compounds could be released into food or the surround-
ing environment in the package (e.g. head space) so as to extend
food shelf life and, thus, improve food quality and safety [17].
In this context, Brazil is the world’s third largest fruit producer
with an annual productivity of over 43 Mt, of which about 47%
were consumed fresh and 53% were processed [18], increasing sig-
nificantly the generation of agro-industrial waste. In particular,
blueberry juice processing wastes, including skins and seeds, are
usually discarded, but they are rich in vitamins, minerals, fibres, and
phenolic compounds, such as anthocyanins, antioxidant substances
that can help preventing degenerative diseases [19,20]. Moreover,
anthocyanins are compounds with the ability to change colour
when subjected to different pH values and this change could be cor-
related with the pH changes in some food products [21,22]. Hence,
the incorporation of blueberry pomace (BP) into filmogenic formu-
lations can be an interesting alternative to produce biodegradable,

http://dx.doi.org/10.1016/j.ijbiomac.2017.08.083
0141-8130/© 2017 Elsevier B.V. All rights reserved.
 C.L. Luchese et al. / International Journal of Biological Macromolecules 106 (2018) 834–839
active and intelligent food packaging, adding value to this waste
through the development of environmentally friendly products as
well as contributing to minimize waste generation.
The objective of this work was to develop and characterize
cassava starch films with blueberry pomace for their use as food
packaging. For this, different contents of blueberry pomace were
incorporated into the starch film formulations in order to evaluate
the effect of BP addition in the film performance; in particular, ther-
mal and optical properties, as well as physicochemical behaviour
concerning swelling and release of phenolic compounds.
2. Materials and methods
2.1. Materials
Native cassava starch (moisture content of 14.8 ± 0.05%)
acquired in Natal Public Market (Natal, Brazil) was used to prepare
film forming solutions. Sorbitol obtained from Panreac Química
SA (Barcelona, Spain) was used as plasticizer. All chemicals were
used as received without further purification. Blueberries (Vac-
cinium corymbosum L.), provided by Italbraz Ltda (Vacaria, Brazil),
were bleached for 3 min with water vapour in an autoclave
(Phoenix Luferco) and afterward, cooled with water according to
the methodology of Rossi et al. [23]. After producing juice, the
pomace was separated for filtration, freeze-dried, milled and sieved
(100 mesh) to obtain the BP in powder form.
2.2. Film preparation
Film forming solutions were prepared by mixing 4 g of cas-
sava starch (CS), sorbitol (30 wt% based on CS) and blueberry
pomace (4, 8 and 12 wt% based on CS) in 125 mL of distilled water.
First, solutions were hydrated for 5 min under continuous stir-
ring and subsequently, they were heated in an oil bath at 90 ◦ C
for 40 min under stirring. Then, solutions were casted into Petri
dishes (0.2 g cm−2 ) and left for drying at ambient conditions. The
obtained films were easily removed from Petri dishes and condi-
tioned at 25 ◦ C and 55% relative humidity for 48 h before testing.
Films were designated as CS0BP, CS4BP, CS8BP, and CS12BP as a
function of BP content. Film thickness was measured to the near-
est 0.001 mm with a QuantuMike Mitutoyo hand-held digimatic
micrometer (Neurtek, Spain). The values obtained for each sample
at five different locations were averaged.
2.3. Differential scanning calorimetry (DSC)
DSC measurements were performed using a Mettler Toledo
DSC822 (Mettler Toledo S.A.E., Barcelona, Spain). The samples
(approximately 3.5 mg) were hermetically encapsulated in alu-
minium capsules to prevent mass loss during heating from −50 to
250 ◦ C at a rate of 10 ◦ C/min under nitrogen conditions (10 mL/min).
2.4. Thermo-gravimetric analysis (TGA)
TGA was performed in a Mettler Toledo TGA SDTA 851 (Mettler
Toledo S.A.E., Barcelona, Spain). The assays were running from 30 ◦ C
up to 800 ◦ C at a heating rate of 10 ◦ C/min under nitrogen atmo-
sphere (10 mL/min) in order to avoid thermo-oxidative reactions.
2.5. Ultraviolet-visible (UV-vis) spectroscopy
The light-barrier properties of the films were determined by
using an UV–vis spectrophotometer Jasco (Model V-630, Tokyo,
835
Japan) in the range of 200–800 nm. The samples transparency was
calculated as Eq. (1):
A600
Transparency =
x
where A600 is the absorption at 600 nm and x is the film thickness.
Higher values of transparency indicate lower transparency of films
[24].
2.6. Colour measurements
Colour parameters (L*, a*, b*) were determined using a CR-400
Konica Minolta Chroma-Meter (Multi-Gloss 268 Plus). Films were
placed on the surface of a white standard plate (calibration plate
values: L* = 97.39, a* = 0.03, and b* = 1.77). Colour parameters were
measured using the CIELAB colour scale: L* = 0 (black) to L* = 100
(white), − a* (greenness) to + a* (redness), and − b* (blueness) to
+b* (yellowness) and colour difference (AE*) was calculated as Eq.
(2):
 2  2  2
E ∗ = ∗
Lcontrol ∗
− Lsample + a∗control − a∗sample + b∗control − b∗sample (2)
where CS0BP sample was considered as control. Ten measurements
were taken for each sample.
2.7. Gloss measurements
The gloss was measured at 60◦ incidence angle according to
the ASTM D523-14 using a CR-400 Konica Minolta Chroma-Meter
(Multi Gloss 268 Plus). Measurements were taken ten times for each
composition.
2.8. Swelling measurements
Swelling was gravimetrically measured according to ASTM
D570-98 using three specimens of each film formulation. Films
were weighed (Wo ) and immersed in distilled water. Samples were
removed at fixed times (each 1 h in the first 10 h of analysis and
after 24 h), wiped with paper and reweighed (Wt ). The percentage
of swelling was calculated as Eq. (3):
(Wt − Wo )
Swelling (%) = x100 (3)
Wo
The experimental points were fitted using an exponential regres-
sion.
2.9. Migration assays
Migration assays were also performed according to EU
2016/1416. Ethanol (96 %) and 3 % acetic acid (v/v) were selected
as food simulant solutions for fatty and aqueous foods, respectively
[25]. Total immersion migration tests were performed as follows: a
6 cm2 piece of each sample and 10 mL of the simulant were placed
in a glass vial. Then, samples (film and simulant) were introduced in
a thermostatic oven, set at 20 ◦ C, for 10 days. Immediately after the
exposure of films to simulants, samples were removed and UV–vis
measurements were performed for the simulant solutions. Samples
in triplicate were investigated for each film formulation and each
simulant.
2.10. Statistical analysis
Analysis of variance (ANOVA) was used to determine the sig-
nificance of differences among the samples. The analysis was
performed with a Statistic computer program (STATISTIC 8.0, Stat-
soft Inc., Tulsa, USA) and Tukey’s multiple range test was used for
836 C.L. Luchese et al. / International Journal of Biological Macromolecules 106 (2018) 834–839

Fig. 1. DSC thermograms of cassava starch (CS) films with different contents of Fig. 2. TGA results as derivative curves for cassava starch (CS) films with different
blueberry pomace (BP). contents of blueberry pomace (BP).

multiple comparisons. Differences were statistically significant at

the p < 0.05 level.

3. Results and discussion

At macroscopic scale, all films were homogenous, transparent,

odourless and easy to handle. BP addition significantly affected
(p < 0.05) film thickness, which ranged from 91.3 to 124.3 ␮m mean
values, as shown in Table 1.

3.1. Thermal properties

DSC measurements were carried out and thermograms are dis-

played in Fig. 1. As stated by Alvani et al. [26], enthalpy values (H)
can be regarded as an indicator of the loss molecular order due
to the disruption of hydrogen bonds. This was confirmed by the
H values determined for cassava starch films: 35.3, 30.5, 29.1,
and 25.3 J/g for CS0BP, CS4BP, CS8BP, and CS12BP, respectively. In
addition to enthalpy values, the maximum temperature of the DSC
peaks was also analyzed. For cassava starch film, the peak presented
the maximum temperature at 80 ◦ C, while this peak shifted to
higher temperatures, up to 96 ◦ C, for the films prepared with blue-
berry pomace. These results indicated that an increased amount of
thermal energy is needed to dissociate interactions among compo-
nents in composite films compared to cassava starch films without
blueberry pomace. Similar trend was observed for other biopoly-
mers, such as gelatin, when compounds rich in polyphenols, such
as seaweed extracts, were incorporated into film forming solu-
tions [27,28]. This behaviour is related to the changes promoted
in inter- and intra-molecular interactions by the incorporation of
compounds rich in polyphenols into biopolymeric matrices.
Concerning thermal degradation of the films, DTGA curves are
shown in Fig. 2, where two weight loss stages are displayed. As can
be observed, there was a slight weight loss close to 100 ◦ C, which
can be attributed to the moisture content of the samples. This first
stage is associated with the loss of free and bound water in the film.
After that, the samples exhibited a high thermal stability, maintain-
ing their mass almost unaltered up to 150 ◦ C (weight retention > 85
%). The weight loss due to the thermal decomposition of cassava
starch at 307 ◦ C was the main phase [29,30], and remained con-
stant for all the formulations under study. Similar results have been
found in recent research works on cassava starch films [31–33].
Fig. 3. UV–vis absorbance curves of the cassava starch (CS) films with different
contents of blueberry pomace (BP).
3.2. Optical properties
UV-vis absorption was measured at the wavelength of
200–800 nm and the results are displayed in Fig. 3. It can be
observed that there was a considerable difference between cassava
starch films with and without blueberry pomace. However, there
was no significant variation in the spectra of cassava starch films
regardless of blueberry pomace content. The addition of blueberry
pomace caused high absorption below 300 nm, indicating that films
with blueberry pomace were able to protect against UV light, pre-
venting the deterioration of food due to UV radiation. Moreover, the
addition of blueberry pomace promoted the formation of a shoul-
der at 280 nm due to the presence of aromatic compounds in the
pomace [6,34]. It is important to highlight that the UV light bar-
rier values presented in this work for cassava starch films prepared
with blueberry pomace were higher than those found for synthetic
polymers, such as oriented polypropylene (OPP) and low density
polyethylene (LDPE) [3].
However, the addition of blueberry pomace reduced the film
transparency, as also shown by other authors when adding blue-
berry pomace in biopolymeric film forming solutions [35,36], due
 C.L. Luchese et al. / International Journal of Biological Macromolecules 106 (2018) 834–839 837

Table 1
Optical properties of cassava starch (CS) films with different contents of blueberry waste (BW).

Film Thickness (␮m) Transparency (A600 /mm) L* a* b* E* Gloss60◦ (◦ )

CS0BP 91.3 ± 10.C

9.2 96.7 ± 0.1A
0.05 ± 0.03 D
2.3 ± 0.C
– 18.6 ± 0.7 A
CS4BP 115.0 ± 4.8 B 3.5 85.8 ± 0.5 B 6.6 ± 0. C 2.5 ± 0.1 B 12.6 ±0.6 C 15.5 ± 0.8 B
CS8BP 122.6 ± 10.9 AB 6.0 71.6 ± 0. C 18.1 ± 0.6 B 2.6 ± 0.1 B 30.9 ± 0.7 B 9.6 ± 0. C
CS12BP 124.3 ± 8.6 A 5.8 61.1 ± 0.5 D 26.7 ±0.4 A 5.4 ± 0.1 A 44.6 ± 0.6 A 6.9 ± 0.3 D
A–D
Mean ± standard deviation followed by the same letter in the same column are not significantly (p > 0.05) different through the Tukey’s multiple range test.

Fig. 4. (a) Visual aspect of cassava starch (CS) films with different contents of blueberry pomace (BP) after 24 h of immersion in water and (b) swelling behaviour of cassava
starch (CS) films with different contents of blueberry pomace (BP). Symbols represent experimental data, while lines correspond to the exponential fit.

Fig. 5. Visual aspect of the cassava starch (CS) films with blueberry pomace (BP) as a function of BP after 10 days of immersion in simulants: (a) acetic acid (3%) and (b)
ethanol (96%) and UV absorbance curves of the simulant solutions after extracting cassava starch (CS) films with different contents of blueberry pomace (BP) in (c) acetic
acid and (d) ethanol.
838 C.L. Luchese et al. / International Journal of Biological Macromolecules 106 (2018) 834–839
to the fact that anthocyanin-derived pigments present in blueberry
pomace are involved in the absorption at this wavelength [37].
Transparency values are shown in Table 1, where colour parameters
have been also displayed. It is known that blueberry pomace can
provide a characteristic reddish colour [37]. This fact was confirmed
by the values of the a* parameter displayed in Table 1.
According to the results presented in Table 1, the addition of
blueberry pomace film forming formulations promoted a decrease
in L* parameter and an increase in a*, b*, and E* values. More-
over, the greater the amount of blueberry pomace added, the
greater its influence on the colour. As expected, control sample
(CS0BP) presented coloration closer to yellow, whereas the films
with blueberry pomace showed reddish colour tonality due to
those anthocyanin-derived pigments present in blueberry, which
also caused the decrease in transparency, as previously shown by
UV–vis spectroscopy.
The addition of blueberry pomace promoted a decrease in gloss
values. It is known that the lower the gloss value, the higher the
surface roughness [16]. Therefore, the incorporation of blueberry
pomace powder in film formulations promoted an increase in the
surface roughness. Additionally, it is important to remark that
roughness is required when intending to print or heat-seal the film
in order to improve the adhesion of inks onto the film surface or
the adhesion between surfaces, respectively.
3.3. Physicochemical properties
Swelling tests were carried out and it is worth noting that all
samples maintained their structural integrity after 1440 min (24 h)
of immersion in water, as can be seen in Fig. 4a. In addition, swelling
kinetics was determined, as shown in Fig. 4b. In the first hour, the
swelling range for all films varied between 157 and 177%. As can be
seen in Fig. 4b, swelling values increased up to 300 min (5 h); after
that time, swelling values remained constant, being lower than 300
% in all cases. Similar values have been reported for starch films
in the literature [38]. After curve fitting, the calculated regression
values were higher than 0.9 (R2 > 0.9). The values for the films with
blueberry pomace were slightly higher than those for the film with-
out blueberry, indicating that blueberry pomace affected the water
uptake capacity of the films, probably due to the fibre present in
blueberry pomace.
This behaviour can be considered as an advantage for the release
of active compounds. In order to analyse in vitro release rate of
the phenolic compounds present in blueberry pomace, migration
assays were performed. This analysis was carried out by the total
immersion of the film in food simulating solvents, liquids that can
simulate the behaviour of real food, such as aqueous simulants
(acetic acid 3%) or fatty simulants (ethanol 96%). As can be seen
in Fig. 5a and b, at the end of the migration assay (10 days), films
did not show any deterioration and all samples remained intact.
As can be observed by the change in the colour of the solutions in
Fig. 5a, the release of phenolic compounds from blueberry pomace
was higher when films were immersed into acetic acid (Fig. 5c).
In contrast, lower amount of phenolic compounds was detected
when ethanol was used as fatty food simulant (Fig. 5d). According
to Buonocore et al. [5], some factors can influence the migration
behaviour of active compounds from biopolymer films, includ-
ing their chemical composition, matrix structure, and surrounding
media, among others.
The films submerged in ethanol or acetic acid presented differ-
ent coloration due to the fact that the blueberry pomace is rich in
anthocyanin, an active compound capable of changing colour when
subjected to different pH values. In acidic conditions (pH 1.0–3.0),
the predominant form is the flavylium cation and colour ranges
from orange to purple; at pH ∼4.5, colourless structures (chalcone)
are formed; in basic conditions (pH > 7), the formation of quinoidal
base causes blue coloration [39]. UV absorbance curves of the sim-
ulant solutions after immersion of all the films in acetic acid and
ethanol are displayed in Fig. 5c and 5d, respectively. As can be seen
in Fig. 5, there was an increase of the release of aromatic compounds
(␭ = 280 nm) into both food simulants when the content of blue-
berry pomace increased; however, the release of blueberry pomace
was greater into acetic acid. Furthermore, when acetic acid was
used as simulant, there was also an increase in the absorbance at
525 nm due to the presence of anthocyanins in blueberry pomace.
4. Conclusions
The thermal, optical and physicochemical properties of cassava
starch films incorporated with blueberry pomace from juice pro-
cessing were investigated. The results highlight the potential of
cassava starch and blueberry pomace to prepare renewable and
biodegradable products to promote a sustainable production of
food packaging. The films with blueberry pomace were found to
be UV resistant due to the presence of aromatic compounds in
the blueberry pomace and, thus, able to prevent food deteriora-
tion caused by UV radiation, extending food shelf-life. Furthermore,
migration assays showed the release of active compounds into the
acetic acid medium, indicating the feasibility of these films as food
packaging for aqueous foods.
Acknowledgements
The authors acknowledge the financial support received from
CAPES (Coordenadoria de Aperfeiçoamento de Pessoal para o
Ensino Superior), CNPq (Conselho Nacional de Desenvolvimento
Científico e Tecnológico) and FAPERGS (Fundação de Amparo
à Pesquisa do Estado do Rio Grande do Sul, Brazil). In partic-
ular thanks to the CAPES CSF-PVE’s Project, process number:
88881.068177/2014-01. BIOMAT thanks the University of the
Basque Country UPV/EHU (PPG17/018). Tania Garrido thanks
Euskara Errektoreordetza (UPV/EHU) for her fellowship. Thanks
also Advanced Research Facilities (SGIker) from the UPV/EHU.
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