International Journal of Biological Macromolecules 141 (2019) 713–720

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International Journal of Biological Macromolecules

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

Ozonation of cassava starch to produce biodegradable films☆

Carla I.A. La Fuente a,⁎, Andressa Tamyris de Souza b, Carmen C. Tadini b,c,d, Pedro Esteves Duarte Augusto a,d
a
Department of Agri-food Industry, Food and Nutrition (LAN), Luiz de Queiroz College of Agriculture (ESALQ), University of São Paulo (USP), Piracicaba, SP, Brazil
b
University of São Paulo, Escola Politécnica, Department of Chemical Engineering, Main Campus, 05508-010, SP, Brazil
c
University of São Paulo, Food Research Center (FoRC/NAPAN), SP, Brazil
d
Food and Nutrition Research Center (NAPAN), University of São Paulo (USP), São Paulo, SP, Brazil

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

Article history: In this study, biodegradable films were produced from cassava starch modified by ozone at different levels. The
Received 15 August 2019 films were produced by casting technique using native and ozonated cassava starch, glycerol as the plasticizer,
Received in revised form 29 August 2019 and water as the solvent. Films were characterized in term of their mechanical, barrier and functional properties,
Accepted 4 September 2019
morphology, crystallinity, colour, and opacity. The morphology of the ozonated films was more homogeneous in
Available online 05 September 2019
comparison to the films produced with the non-modified starch and enhanced properties were achieved. Films
Keywords:
produced with ozonated cassava starch presented higher tensile strength, Young's modulus and lower elonga-
Biodegradable films tion. The water vapour permeation and the oxygen permeation were increased by increasing the ozonation
Cassava time. Moreover, ozone processing resulted in films with a more hydrophilic surface and lower solubility after
Starch 24 h. Possible explanations and applications were discussed. In conclusion, the ozone processing showed to be
Ozonation a good alternative for starch based packaging production.
© 2019 Elsevier B.V. All rights reserved.

1. Introduction achieved due to complex relation among molecular modifications on

While hundreds of millions of tons of petroleum-based plastics are
produced annually all over the world, the concern about their environ-
mental impact is increasing each day. Consequently, different renew-
able and biodegradable source are being studied to produce
biodegradable plastics. Starch is one of the promising biopolymeric ma-
trix for the development of biodegradable packaging [1].
Starch is one of the most important and abundant polysaccharides in
nature [2,3]. They are obtained from renewable sources, presenting rel-
atively low cost and a wide possibility of application. Consequently,
starch is a very versatile material, being used in several industries
such as the food, paper, textile, chemical and pharmaceutical [2].
Being the native sources limited by nature, different starch modification
techniques are available in order to improve the processability, mechan-
ical and/or barrier properties of biodegradable films.
Among different possibilities of starch modification, ozone process-
ing has gaining attention due to the demand for methods that meet
safety and health standards for both consumers and the environment
[4].
Ozone is a powerful oxidizing agent, which reacts with starch, in-
creasing its carboxyl and carbonyl contents, and also reducing its molec-
ular size and distribution [5]. Consequently, different properties are
size, charge and chemical affinity. In fact, different starch sources were
already modified by ozonation, such as corn, sago, wheat, potato and
tapioca/cassava [5–10].
However, different starch sources have different molecular and
granule structure, being differently affected by ozonation. For instance,
cassava starch had both polygonal and spherical shaped granules with
size b50 μm, while potato starch contained both large and small gran-
ules, which had spherical to oval shapes [11].
Therefore, different ozonated starches present different molecular
sizes and electrical charges, which affect their behaviour for re-
association. These molecular changes are competitive in relation to
film production and properties. Consequently, the properties of films
produced with ozone modified starches are unpredictable and, depen-
dent on the relations among the source, reactor and processing
conditions.
Hence, the aim of this study was to produce films from ozone-
modified cassava starch, considering different process condition and
evaluating the most important mechanical, barrier and functional prop-
erties, as well as their morphology, crystallinity, colour, and opacity.
2. Material and methods

☆ A patent related with this work was filed (BR1020190112166).

This work was developed in three parts, as summarized on Fig. 1.
⁎ Corresponding author at: Avenida Pádua Dias, 11, Piracicaba, SP 13418-900, Brazil. Firstly, cassava starch was ozonated, in two conditions. Then, films
E-mail address: carlalfa@usp.br (C.I.A. La Fuente). based on the starches (native and the two ozonated conditions) were

https://doi.org/10.1016/j.ijbiomac.2019.09.028
0141-8130/© 2019 Elsevier B.V. All rights reserved.
714 C.I.A. La Fuente et al. / International Journal of Biological Macromolecules 141 (2019) 713–720

Nomenclature dry basis; 1500 mL) and then ozonated for different periods under con-
stant stirring in the same system reported by Castanha [5]. The ozone
a⁎ redness/greenness CIELab parameter (−) rich stream was produced from industrial oxygen by the coronal dis-
A area (m2) charge method using an ozone generator unit (Ozone & Life, O&G
b⁎ yellowness/blueness CIELab parameter (−) Model RM 3.0, São Jose dos Campos, Brazil). This stream was dispersed
C chroma (−) in the sample, which was placed in a glass reactor, as small bubbles
E Young's Modulus (MPa) through a gas disperser. A gas flow of 1 L∙min−1 was used, with an
E* CIELab total color change (−) ozone concentration in the gas current of 41 mg O3∙L−1. The amount
L⁎ brightness/darkness CIELab parameter (−) of ozone was measured using an ozone monitor (2B Technologies
m mass (g) Model 106-H, USA). After passing through the samples, the gas stream
OTR oxygen transmission rate (cm3·m−2·d−1) was led out of the reactor and converted to oxygen in a thermal ozone
p pressure (kPa) destroyer (Ozone & Life, São José dos Campos, Brazil). The procedures
P’O2 oxygen permeability (cm3/m·day·Pa) were performed at 25 ± 1 °C. Fig. 1 presents a schematic representation
RC relative crystallinity of the ozone processing system. Three samples were evaluated: the
S solubility (%) non-modified starch and those ozonated for 15 and 30 min.
t time (h) After processing, the excess of water was removed through decanta-
TS Tensile Strength (MPa) tion and centrifugation, and the starch samples were dried in an air cir-
WVP Water Vapour Permeability (g·mm/m2·day·kPa) culation oven for ~ 12 h at 35 °C until moisture content of ~12 g/100 g w.
x thickness (mm) b.
Y opacity (%) Glycerol P.A. grade (Sigma-Aldrich, Brazil) and distilled water were
also used for film production.
Subscripts
0 initial 2.2. Film preparation
b black standard
f final The films were produced by casting technique, as shown in Fig. 1. A
w white standard solution containing 5 g/100 g of cassava starch was homogenized in dis-
tilled water with a magnetic stirrer (GEHAKA, HU-30/2, Brazil) for
Greek 10 min. Then, the solution was heated for 45 min in a reactor with exter-
Δ delta nal recirculation of water at 75 °C. Glycerol was added as the plasticizer
ε elongation at break (%) (25 g/100 g of starch) according to Souza [12] and, the suspension was
heated for more 15 min under agitation. To eliminate the bubbles, the
filmogenic solution was placed in an ultrasonic bath for 30 min at 70
formed. Finally, the obtained films were evaluated through their struc-
°C (154 W, 25 kHz; UNIQUE, model USC-1850, Brazil). The solution
ture and the most important properties.
was then poured into acrylic Petri dishes (0.15 g/cm2) for ~12 h to
rest, and then dried at 35 °C and 45% HR for ~10 h in a climatic chamber
2.1. Materials (TIRACLIMA, model TCC 7034, Germany).
The films were conditioned for at least 48 h in desiccators containing
The native cassava starch (Amilogill 1500) was donated by Cargill a saturated solution of NaCl (75% RH) before characterization at room
Company. This starch was suspended in distilled water (10 g/100 g in temperature (~25 °C).

Fig. 1. Schematic representation of the ozone processing system and the film elaboration.
 C.I.A. La Fuente et al. / International Journal of Biological Macromolecules 141 (2019) 713–720
2.3. Films characterization
2.3.1. Mechanical properties
The film thickness was determined using a digital micrometer
with a precision of 0.001 mm (MITUTOYO, Japan) at five random
positions.
The mechanical properties were evaluated through a uniaxial tensile
assay, according to ASTM D882-09 [13]. Tensile strength (TS) [MPa] and
percent elongation at break (ε) [%] were evaluated on a texture analyzer
(TAXT2i e Stable Micro Systems, UK) with a load cell of 30 kgf (294 N),
using the A/TGT self-tightening roller grips fixture. The initial grip sep-
aration and the crosshead speed were set at 50 mm and 1 mm·s−1, re-
spectively. Young's modulus (E) was calculated as the slope of the initial
linear portion of the stress versus strain curve. At least ten strips
(100 mm × 25 mm) were evaluated for the non-modified and modified
films.
Furthermore, the force and the deformation at the breaking point of
the film was determined in a punctuation test [14], on a texture ana-
lyzer (TAXT2i e Stable Micro Systems, UK) with a load cell of 30 kgf
(294 N). The films were fixed in a circular permeation cells (d =
0.05 m) and perforated by a 3 mm diameter probe. The puncture force
(F) and the displacement of the probe (D) were obtained from force-
deformation curves, according to Fig. 2.
2.3.2. Barrier properties
Water vapour permeability (WVP) of films was measured gravi-
metrically according to the ASTM E96-80 [15]. The films were placed
in circular permeation cells containing silica gel, and then placed in a
climatic chamber (TIRACLIMA, model TCC 7034, Germany) main-
tained at 75% RH and temperature of 25 °C. The cell weight was re-
corded hourly for 8 h. Water vapour permeability was calculated
according to Eq. (1).
Δm  x  24
WVP ¼
Δt  A  Δp
wherein: Δm/Δt is the moisture gain per unit of time (g·h−1); x is the
film thickness (mm); A is the film area exposed to permeation
(1.96·10−3 m2); Δp is the vapour pressure difference (kPa) of the at-
mosphere over silica gel and pure water (3.168 kPa at 25 °C).
Fig. 2. Diagram of the system used to determine the punctuation deformation. Adapted from Sobral [14].
715
The oxygen permeability coefficient (P’O2) of the ozonated films was
measured at 23 °C and 75% RH on a 50 cm2 circular films using an oxy-
gen permeation system (OXTRAN 2/21, Mocon, USA), according with
ASTM F1927-14 [16]. The oxygen permeability reported is the mean of
four films. The oxygen permeability coefficient was calculated according
to Eq. (2).
OTR
P’O2 ¼ X ð2Þ
p
wherein: P’O2 is the oxygen permeability coefficient (cm3·m−1·d−1·-
Pa−1); OTR is the oxygen transmission rate (cm3·m−2·d−1); p is the
partial pressure of oxygen, and X is the average thickness of the speci-
men (m).
2.3.3. Functional properties
The film wettability was determined by the measurement of the
contact angle with water, according to ASTM D7334 − 08 [17] using
an OCA15- Dataphysiscs (OCA 15, Dataphysiscs, Germany). Two
angle measurements were carried out (one on each drop edge)
quickly to avoid changes due to water vaporization on each drop.
The contact angle was defined as the average of six angles measures.
The film solubility in water was evaluated with discs (diameter of
20 mm) immersed in 50 mL of water at 25 °C for 24 h, under mechanical
stirring at 120 RPM, according to Gontard [18] The solubility in water
was calculated as the percentage of dry matter of the solubilized film,
as described in Eq. (3).
m f −mo
S¼  100 ð3Þ
mo
wherein: mo and mf are the initial and the final discs' mass (g),
respectively.
The films moisture was determined through the gravimetric method
ð1Þ in an oven, without recirculation, at 105 °C until a constant mass
(~24 h), according to Gontard [18].
2.3.4. Colour and opacity
The film colour was determined using a colorimeter (HUNTERLAB,
model ColorQuest XE, USA) with the CIELab scale (L⁎, a⁎, b⁎). The colour
measurements were expressed in terms of L* (L* = 100 brightness and
716 C.I.A. La Fuente et al. / International Journal of Biological Macromolecules 141 (2019) 713–720

Fig. 3. Scanning electron micrographs of the surfaces and cross-sections of the films prepared with non-modified and ozonated cassava starch. Surface images at and, the cross-sections
images at and, respectively. The surface and the cross-section images were obtained at 1000× and 3100×, respectively.

L* = 0 darkness); a* (+a* = redness and −a* = greenness) and b* The scanning rate was 0.02°/s with 2ϴ angles varying between 2° to
(+b* = yellowness and −b* = blueness). The change of each of the pa- 40°. The software Origin 2018b was employed for calculation (OriginLab
rameters was calculated according to Eqs. (4), (5) and (6). Corporation, Massachusetts, USA).

ΔL ¼ L −L0 ð4Þ

2.3.6. Morphology

Δa ¼ a 
−a0 ð5Þ The surface and cross-section
morphology of the films were visualized by a scanning electron mi-

Δb ¼ b −b0
 
ð6Þ croscope (SEM). The films were dried in silica gel for 48 h to ensure an
accurate and finer cut. They were then horizontally and vertically placed
wherein: L0* = 94.04; a0* = −0.86 and b0* = 1.53 determined from a on an adhesive tape placed over circular stubs. The films were then
white standard pattern. The total colour change (ΔE⁎), the opacity (Y) coated with a 10-nm platinum in a high-vacuum coating system (BAL-
and the Chroma (C) [19,20] were calculated using Eqs. (7), (8) and (9) TEC, model MED020, Switzerland) and the side which was not in con-
respectively. tact with the Petri dish was observed using the microscope (FEI,
model Quanta 600FEG, Netherlands) with an acceleration voltage of

h i1 15 kV and magnification ranged from 500× to 3100×.
 2
ΔE ¼ ðΔL Þ þ ðΔa Þ2 þ ðΔb Þ
2 2
ð7Þ

2.4. Experimental design and statistical analyses

Yb
Y¼
Yw
 obtained data was evaluated through analysis of variance (ANOVA), ap-
 1
2 2
C ¼ a2 þ b
wherein: Yb is the opacity on the black standard and, Yw is the opacity on
the white standard. Three measurements were performed per film for-
mulation to determine all the colour parameters.
2.3.5. Crystallinity
The relative crystallinity of the films was calculated as the ratio be-
tween the crystalline area and the total area of the X-ray diffractogram
(plotted from 2° to 40°), as described by Nara & Komiya [21]. An X-ray
diffractometer (PANalytical, model X'Pert PRO, Netherlands), with de-
tector X'Celerator operating at 45 mA, and voltage of 40 kV was used.
ð8Þ
A completely randomized design was applied in three replicates. The
plying Tukey's test as a post-hoc test through Statgraphics Centurion XV
ð9Þ
software (StatPoint, Inc., USA).
3. Results and discussion
Fig. 3 presents the morphology of the films. In general, the ozonated
films present a compact structure and smooth surfaces, in comparison
to the non-modified starch films. Heterogeneity and roughness were al-
ready observed on the surface of non-modified cassava films [22]. For
the ozonated films, smoother surface is clearly evident in the surface
of the films, while, in the cross-section, some cracks are visible, probably
appeared after the film was broken. In the study performed by Biduski
 C.I.A. La Fuente et al. / International Journal of Biological Macromolecules 141 (2019) 713–720
[23] although oxidized sorghum films were more homogeneous, similar
fissures were also observed.
According to the literature, films produced with oxidized starches
allow a higher penetration of the plasticizer molecules in the starch
chains, and improve the interaction between them [23–25]. Moreover,
low retrogradation is expected in oxidized starches [24], promoted by
the repulsion between molecules, increasing the free space between
the molecules, and also increasing the homogeneity of the film.
However, ozonation is more than only an oxidation process and this
process also changes the molecular charges and chemical affinity, which
affect the interaction among the molecules and altering the morphology
of the films.
Fig. 4 shows the X-ray diffractograms for the non-modified and
ozonated films. Semi-crystalline characteristic (presence of amorphous
and crystalline zones) was observed, with peaks located at 5.6°, 16.8°,
19.2°, 21.8°, and 25.6° (2θ). The film produced with the non-modified
cassava starch showed peaks at 16.8° and 19.2°, in the same regions
than those observed in the study of Fama [26] and, Jaramillo [27].
Ozonated films resulted with a B-type crystalline structure [28]. The
introduction of complexing agents, like glycerol, into starch prepara-
tions disrupts double helix conformations by forming stable single
chain V-conformation helices [26].
During film formation, both amylose and amylopectin macromole-
cules are rearranged and their linear fractions form re-associations by
hydrogen bonds. Therefore, the crystallinity of starch-based films de-
pends on several factors, such as drying and storage conditions: temper-
ature and relative humidity and also the content of plasticizer [29]. It is
important to point out that the drying and storage conditions and glyc-
erol content were similar for all the treatments in this study.
On the other hand, according to Rindlav-Westling [30] crystallinity is
also dependent on the ability of the chains to form crystals as well as the
mobility of the chains during the crystallization process. In fact, an in-
crease in the Relative Crystallinity (RC) was also observed due to the
ozonation process. The ozonation results in the disruption of the starch
polysaccharide chains resulting in a greater number of molecules with
decreased sizes and weight and, thereby, is possible to result in a facility
to form crystals, thus, increasing the RC.
Fig. 4. X-ray diffraction patterns of the films prepared with non-modified and ozonated cassava starch (15 and 30 min) and their respectively Relative Crystallinity (RC).
717
As no statistical difference was observed for the film thickness,
their production procedure was considered adequate and their me-
chanical properties could be evaluated being presented in Table 1.
The results obtained for the non-modified cassava film were close to
those reported in the literature for native cassava starch [12,31–33]
while the films obtained from ozonated starch were stronger.
The tensile strength (TS) increased according to the ozone process-
ing time, showing that starch ozonation made stronger films. An in-
crease of ~44% in the TS was observed between the non-modified and
the 30 min ozonated films, whose Young's modulus (E) was also in-
creased. Potato starch oxidized with sodium hypochlorite [34] and ba-
nana starch oxidized with active chlorine [35] also resulted in films
with increased TS and E.
According to the literature, films more rigid resulted with less elon-
gation at break (ε) [34]. However, no statistical difference between the
non-modified film and the 30 min ozonated film was observed, and
only the 15 min ozonated film resulted with less elongation at break,
in comparison to the non-modified film – which can reinforce the
unique characteristics of ozone processing.
Regarding the punctuation assay, the maximum displacement of
the probe (D) (Fig. 2) was observed for the non-modified film.
Moreover, an increase in the punctuation force (F) was observed
due to the ozonation time, confirming that native starch films
were weaker than those produced with ozonated starches. The
punctuation test is not as common as the uniaxial tensile assay to
determine the mechanical properties of films. As observed in this
study, both analyses are related and complement each other. There-
fore, this assay can be used as an alternative to evaluate the me-
chanical properties of this type of materials.
As detailed by Maniglia [36], the ozonation results in the cleavage of
the glycosidic bonds of both amylose and amylopectin molecules [5,37]
and the replacement of the hydroxyl groups by carbonyl and carboxyl
groups [5,24,35,38]. The cleavage of the glycosidic bonds results in a
greater number of molecules with decreased sizes, which may facilitate
the tendency of their re-association [37]. Therefore, a new polymeric
matrix with potentially more interaction between molecules is formed,
which partly explain the observed behaviour.
718 C.I.A. La Fuente et al. / International Journal of Biological Macromolecules 141 (2019) 713–720

Table 1 Herein, ozone processing improved the strength of cassava starch

Mechanical properties: Tensile strength (TS), Elongation at break (ε), Young's modulus films, resulting in films with higher TS, E, F and, minimum D (Fig. 2)
(E), Distance (D) and, Force (F). Funtional Properties: Moisture Content (MC), WVP, and
Contact angle and, parameters of colour (L*, a*, and b*), colour difference (ΔE*), chroma
and lower ε which was evident by both assays (the tensile and punctu-
(C) and opacity (Y) of ozone –modified cassava starch films. ation). Moreover, according to Rindlav-Westling [30], films with in-
creased TS and E and decreased ε properties also presents increased
Property Non modified 15 min 30 min
RC – thus confirming the results obtained herein. It is worth mention
starch ozonated ozonated
this result is very interesting in different applications of packages, as
Thickness μm 0.08 ± 0.01a 0.08 ± 0.00a 0.07 ± 0.00a
well as for the confection of plastic bags.
TS MPa 2.42 ± 0.75c 3.75 ± 1.10b 5.49 ± 1.27a
ε % 43.83 ± 6.95a 37.39 ± 6.05b 39.45 ± Table 1 shows the results for Water Vapour Permeation (WVP)
6.95ab and Oxygen Permeation (P’O2). Regarding the films made with na-
E MPa 51.75 ± 71.18 ± 82.04 ± tive cassava starch, the values obtained for P’O2 were similar to the
25.76b 30.59ab 26.52a reported by Llanos [32], while the WVP was in the same order of
D mm 12.36 ± 2.48a 7.07 ± 0.53b 7.55 ± 1.45b
F* N 1.72 ± 0.09a 3.55 ± 0.95b 3.68 ± 1.04b
magnitude of those reported by Souza [12]. As shown in Table 1, an
WVP g·mm/day·m2·kPa 23.77 ± 2.94b 26.84 ± 4.56ab 29.95 ± 1.72a increase in the WVP and the P’O2 due to the ozonation time was ob-
P’O2 × cm3/m·day·Pa 1.27 ± 0.45b 2.03 ± 0.52ab 2.22 ± 0.14a served. An increasing in both WVP (26%) and P’O 2 (75%) was ob-
10−7 served for the 30 min ozonated film in comparison to the non-
MC g H2O/g w.b. 0.16 ± 0.016a 0.192 ± 0.025a 0.182 ±
modified cassava film.
0.021a
L* – 96.85 ± 0.14b 97.21 ± 0.12a 96.85 ± 0.05b The functional properties of ozonated cassava films, expressed in
a* – 0.06 ± 0.04ab 0.10 ± 0.04b 0.03 ± 0.02a terms of contact angle, solubility and moisture content, are presented
b* – 0.16 ± 0.08b −0.12 ± 0.08a 0.14 ± 0.04b in Fig. 5 and Table 1. Similar values are reported in the literature for
ΔE* – 3.15 ± 0.14b 2.80 ± 0.12a 3.16 ± 0.05b non-modified cassava film, regarding the contact angle [22,31,39], the
C – 0.19 ± 0.06a 0.16 ± 0.08a 0.15 ± 0.04b
solubility [39] and the moisture content [33,39].
Y – 9.32 ± 3.99b 16.12 ± 3.97a 3.69 ± 1.20c
As shown in Fig. 5, ozonation time decreased the solubility and the
Average ± standard deviation. Same letters in the same column are not significantly dif-
water contact angle. Therefore, films less soluble, but with a more hy-
ferent (p N 0.05). *Statistical analyses performed at p N 0.1.
drophilic surface, were achieved. No statistical difference was observed
for the moisture content (Table 1).
Moreover, according to Zhang [38] the carbonyl groups are available The permeability and water absorption of edible films depends on
to form strong hydrogen bonds with the hydroxyl groups on starch, several factors, such as their thickness and the glycerol content [40], be-
resulting in films more rigid with reduced elongation. sides of the starch molecules. As both thickness and the glycerol content
On the other hand, the formed carboxylic groups are known to be were similar for all the treatments, the changes on barrier and func-
electronegatively charged and, thus, they cause an electrostatic repul- tional properties can be related with the starch molecular changes due
sion between the molecules, making difficult their association [37]. to ozonation process – and the consequent new polymeric matrix.
Even so, the resulted network seems to be stronger, which means that The complex behaviour of ozonation here observed is highlighted
the ozonation is a complex process and, the results obtained are the bal- again. The molecular charge induced by the presence of carboxyl groups
ance between both molecular size and charge, with different tendency results in distance among molecules, increasing the network permeabil-
of the molecules to re-association. ity by water vapour or oxygen [23,24]. On the other hand, the smaller

Fig. 5. Functional Properties: Solubility (S) and Contact angle of ozone –modified cassava starch films. Average ± standard deviation. Same letters in the same column are not significantly
different (p N 0.05).
 C.I.A. La Fuente et al. / International Journal of Biological Macromolecules 141 (2019) 713–720
molecules (also with a different distribution) rearrangement makes a
different network, whose connections are stronger (maybe due to the
interaction between carbonyl and hydroxyl groups), as revealed in the
mechanical properties.
The electrostatic repulsion in general increases the solubility of
starch [5,37] which is contrary to the results observed in this study for
the films. A possible explanation could be that, once the molecules re-
associate between them, it might reduce the ability of these molecules
to linkage with water molecules of the surrounding, thus, reducing the
solubility. However, the location of polar groups in the film surface
can favour bonds with the first drop of water deposited onto the film, in-
creasing its hydrophilicity. Both results are not contradictory, since this
last essay (solubility) is performed for 24 h with the whole film im-
mersed into water and not only the surface.
Hence, the ozonation technology resulted in cassava films with in-
creased strengthens, more permeable to vapour water and oxygen,
less soluble, which could be an interesting or limiting result, depending
on the films practical application. A possible application can be the coat-
ing of surfaces/products, in which, degradation reaction due to the
water or oxygen diffusion are not important.
The film colour parameters (L*, a*, b* and ΔE*), Chroma (C) and
Opacity (Y) are presented in Table 1. In general, the films presented
good transparency. For native cassava films, similar values of L*, a*, b*
and opacity were already reported [39,41,42].
The ozonated starch films presented higher opacity (15 min of pro-
cessing) or lower (30 min of processing) than the native cassava film.
Once again, the different charge and size distribution of starch mole-
cules must be highlighted, which result in the observed complex
behaviour.
The film opacity is an important property for packaging and coating
applications. Through the results obtained herein, starch ozonation can
produce films with both higher or smaller opacity. Ozonation of cassava
starch for 30 min can be an interesting alternative to enhance its film
transparency, which is, in general, the most desired result.
4. Final remarks
The ozonation process results in the replacement of the hydroxyl
groups by carbonyl and carboxyl groups, as well as the cleavage of the
glycosidic bonds, which results in the depolymerisation of parts of the
amylose and amylopectin molecules. Consequently, a polymeric matrix
with different charge distribution was achieved.
The cleavage of the glycosidic bonds results in a greater number of
molecules with decreased sizes, which may facilitate the tendency of
their re-association. Moreover, the carbonyl groups formed strong hy-
drogen bonds with the hydroxyl groups on starch. Thus, resulting in
the increase of the tensile strength.
On the other hand, the formed electronegative carboxyl groups pro-
mote repulsive forces among the polymer chains, resulting in an in-
creased inter chain spacing. Thus, the permeability was increased.
Therefore, a more open, but stronger polymeric matrix, was formed.
The new charge distribution, might favour somehow that some polar
groups migrate to the surface of the film favouring the linkage with
the first drop of water, thus increasing film hydrophilicity.
The results obtained in this work expand the applications of starch to
produce bioplastics.
5. Conclusions
This work produced and evaluated films based on ozonated cassava
starch. Ozonation resulted in films enhanced mechanical properties,
with increased tensile strength and Young's modulus, although de-
creased elongation and deformation during punctuation. This processes
also increased the water vapour permeation and the oxygen perme-
ation, also resulting in films with a more hydrophilic surface and less
soluble. In general, the films presented good transparency. The
719
ozonated films presented higher (15 min of processing) or lower
(30 min of processing) opacity than the native cassava film, highlighting
ozonation as a versatile alternative for this purpose. Finally, the mor-
phology of the ozonated films was more homogeneous, with increased
relative crystallinity.
In conclusion, the ozone processing showed to be a great alternative
for packaging production.
Acknowledgements
The authors are grateful to the São Paulo Research Foundation
(FAPESP, Brazil), for funding the project n° 2016/18052-5, the post-
doctoral fellowship of CIA La Fuente (2017/05307-8) and the B.Sc.
scholarship of Andressa Tamyris de Souza (2018/24291-8); and the Na-
tional Council for Scientific and Technological Development (CNPq,
Brazil) for funding the project of CIA La Fuente (429043/2018-0) and
the productivity grants of PED Augusto (306557/2017-7) and CC Tadini
(306414/2017-1).
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