Food Biophysics

https://doi.org/10.1007/s11483-018-9522-y

ORIGINAL ARTICLE

Fabrication and Characterization of Native and Oxidized Potato Starch

Biodegradable Films
Laura Martins Fonseca 1,2 & Angélica Karine Henkes 1 & Graziella Pinheiro Bruni 2 & Lorena Aparecida Nunes Viana 1 &
Catarina Motta de Moura 1 & Wladimir Hernandez Flores 1 & Alexandre Ferreira Galio 1

Received: 12 September 2017 / Accepted: 27 February 2018

# Springer Science+Business Media, LLC, part of Springer Nature 2018

Abstract
The need to replace conventional polymers due to environmental pollution caused by them has led to increased production of
biodegradable polymers such as starch. Thus, the application possibilities of starch have increased. In this study, we produced and
characterized biodegradable films derived from native and oxidized potato starch. The film-forming solution was prepared with
different concentrations of extracted starch (native or oxidized) and a plasticizer (glycerol or sorbitol). Then, the mechanical,
barrier, morphological, and structural properties of the films were characterized. The moisture content of the films varied from
15.35 ± 1.31 to 21.78 ± 0.49%. The elastic modulus of the films ranged from 219 ± 14.97 to 2299 ± 62.91 MPa. The film of
oxidized starch plasticized with sorbitol in the lowest content was the most resistant and flexible; moreover, this film also
presented lower water vapor permeability and low solubility in water. Fourier-transform infrared spectroscopic analysis of the
biodegradable films indicated the presence of same functional groups as those of starch with bands in the same regions. The film
thickness was lower for the films plasticized with glycerol whereas the color variation (ΔΈ) was lower for the ones plasticized
with sorbitol. In case of both plasticizers, the increase in their content decreased the ΔΈ value. All the biodegradable films
presented stability against water absorption owing to their low solubility in water. Morphological evaluation revealed the
presence of partially gelatinized starch granules in the films. The roughness parameter (Rq) of the films varied from 3.39 to
10.9 nm, indicating that their surfaces are smooth. X-ray diffraction studies showed a B-type pattern for the starches, which is
representative of tubers. Further, the films present higher relative crystallinity (RC) compared to the starches. The biodegradable
starch films are uniform, transparent and with low solubility in water. The oxidation of starch and use of sorbitol as a plasticizer
resulted in improved properties of the starch films, which is suitable for application.

Keywords Potato starch . Oxidized starch . Biodegradable films . Mechanical properties . Barrier properties

Introduction
The environmental damage caused by waste from convention-
al polymers used in food packaging continues to grow.
Because of this issue, natural polymeric films are being widely
investigated as potential polymer substitutes for food packag-
ing. Starch-based films show great potential as substitutes
because starch is a readily available, versatile, and low cost
* Laura Martins Fonseca
laura_mfonseca@hotmail.com
1
Federal University of Pampa, Bagé/RS, Brazil
2 rite, under a controlled pH and temperature. Oxidation reac-
Department of Agroindustrial Science and Technology, Federal
University of Pelotas, Pelotas/RS, Brazil
material, and the resulting films are colorless, flexible, and
non-toxic [1, 2].
Wide-ranging research is conducted using accessible and
easily extractible sources such as potato starch, to produce
biodegradable films. However, native starch films are weaker
and less resistant than conventional plastics, rendering it nec-
essary to chemically modify starch to improve the mechanical
and barrier properties of their films. The modification of starch
diversifies its applications, rendering its introduction in the
production chain possible, with the challenge of developing
economic packages that are efficient in food protection as well
as environmentally friendly [3–5].
Chemical modification of starch is accomplished mainly by
its reaction with an oxidizing agent such as sodium hypochlo-
tions with sodium hypochlorite include the polymer chain

cleavage and the oxidation of hydroxyl groups to carbonyl
and carboxyl groups, altering the molecular structure of starch
[6]. During this process, partial depolymerization of starch
occurs, thereby decreasing its molecular mass and paste vis-
cosity, resulting in improved stability and clarity of the starch
solution as well as its film-forming ability [7, 8]. Berski et al.
[8] evaluated the pasting and rheological properties of oxi-
dized oat starch and reported that the modification of starch
changed its composition (mostly, the amylose and lipid con-
tents decrease) and lowered its apparent viscosity and retro-
gradation. Compared with the native form, the mechanical
properties are also improved due to starch oxidation, preserv-
ing the packaged food safety and integrity. Zamudio-Flores
et al. [7] studied films from oxidized banana starch and eval-
uated their mechanical properties. The authors obtained the
lowest tensile strength for the native starch film, and reported
that the oxidized starch improves the resistance of the film.
The tensile strength was improved as the degree of modifica-
tion increased.
Another issue that should be addressed is that, in the ab-
sence of plasticizers the films are brittle which is undesirable
for film production. Adding plasticizers to the film matrix is
an established method to improve the properties of the film,
such as their mechanical and barrier properties, and diversify
their applicability. The plasticizer interacts with the starch
chains and increases the molecular motility of starch. To be
compatible with the polymer, it should be polar and its effec-
tiveness is directly linked to the molecular size and depends
on the concentration at which it is used relative to the biopoly-
mer content.
In this context, the use of oxidized starch combined with an
appropriate concentration of the plasticizer can generate bio-
degradable films with superior quality and better mechanical
and barrier properties to be applied in food packaging. In this
study, we produced biodegradable films, from native potato
starch and oxidized potato starch (oxidized using different
active chlorine concentrations) with the addition of two types
of plasticizers, sorbitol or glycerol, at different concentrations.
Material and Methods
Potatoes from the asterix (Solanum tuberosum L.) cultivar
were acquired from the local market in Bagé city, Rio
Grande do Sul, Brazil. All the chemicals and reagents used
were of analytical grade.
Isolation of Starch and its Oxidation
The starch was extracted according to a method reported by
Liu et al. [9], with some modification (drying conditions).
Potato samples were peeled and soaked in 0.1% sodium bi-
sulfite (NaHSO3) for 2 h at pH of 6.0. The dispersion was then
Food Biophysics
triturated, sieved, and decanted. The obtained starch was
washed at least thrice with distilled water and dried in an oven
with air circulation at 40 °C until the moisture content was
between 7 and 10%. The extraction efficiency was 7.5%.
Oxidation was carried out according to the method reported
by Dias et al. [10]. The starch was treated in a reactor made
from a glass beaker with sodium hypochlorite (NaClO) at
different active chlorine concentrations (0.5% and 1.5%) un-
der controlled stirring and pH.
Preparation of Biodegradable Starch Films
Biodegradable starch films were prepared by a casting tech-
nique, in which the suspension was prepared with 2.0% starch
using distilled water and glycerol or sorbitol as plasticizer at
the gplasticizer/gstarch ratio of 0.20 and 0.30. The suspension was
heated under stirring at 90 °C for 15 min and then 25 g of it
was poured into acrylic plates (8.5 cm diameter) and dried in
an oven with air circulation at 30 °C for 16 h. The film was
dried in an oven for 16 h at 40 °C [11].
The optimum plasticizer concentration required for forming
films was verified in preliminary tests and an appropriate con-
centration was chosen according to the film forming ability. At
lower concentrations, (gplasticizer/gstarch < 0.20) the films were
brittle whereas they were to elastic at higher concentrations
(gplasticizer/gstarch = 0.40) and present pores. The nomenclature
of the biodegradable starch films is provided in Table 1 as well
as the films moisture content. The analyses were performed on
the bright side of the biodegradable films.
Characterization of Biodegradable Starch Films
Dynamic Mechanical Analysis (DMA)
The mechanical properties were analyzed using a Dynamic
Mechanical Analyzer Q800™ (TA Instruments, USA) at
25 °C with a ramp force initiated at 1 N/min and reaching a
maximum force of 18 N. Using the same test parameters, the
synthetic materials were evaluated and the results were com-
pared with those obtained for the biodegradable films pro-
duced in this study.
Water Vapor Permeability (WVP)
WVP test was conducted as described by Fonseca et al. [3],
using the standard method, E96–95 (ASTM, 1995). The sam-
ples were placed in permeation cells containing anhydrous cal-
cium chloride (CaCl2) (0% relative humidity (RH)) and condi-
tioned in a desiccator containing 30% (w/w) sulfuric acid solu-
tion (H2SO4) (75% RH). The WVP was calculated by Eq. 1.

m e 
WVP ¼
ab
ð1Þ
t A∙ΔP
Food Biophysics

Table 1 Biodegradable starch

films nomenclature and moisture Treatment Nomenclature Active chlorine (%) Plasticizer (gplasticizer/ gstarch) Moisture content (%)
content
1 N G0.20 Native Glycerol 0.20 17.23 ± 2.37 bcd
2 N G0.30 Native Glycerol 0.30 15.35 ± 1.31 d
3 0.5% G0.20 0.5 Glycerol 0.20 15.82 ± 0.74 cd
4 0.5% G0.30 0.5 Glycerol 0.30 18.79 ± 1.75 abc
5 1.5% G020 1.5 Glycerol 0.20 18.84 ± 0.79 abc
6 1.5% G0.30 1.5 Glycerol 0.30 15.83 ± 0.76 cd
7 N S0.20 Native Sorbitol 0.20 20.00 ± 0.92 ab
8 N S0.30 Native Sorbitol 0.30 19.94 ± 0.89 ab
9 0.5% S0.20 0.5 Sorbitol 0.20 21.78 ± 0.49 a
10 0.5% S0.30 0.5 Sorbitol 0.30 20.03 ± 0.21 ab
11 1.5% S0.20 1.5 Sorbitol 0.20 20.87 ± 0.52 a
12 1.5% S0.30 1.5 Sorbitol 0.30 20.03 ± 0.52 ab
a,b,c,d
Values with different letters in the same column are significantly different (p < 0.05)

Were WVP is the water vapor permeability (g/m·s·Pa), mab  

the absorbed moisture mass (g), t the incubation time period
(s), e the film thickness (m), A the exposed film surface area
(m2) and ΔP the difference of partial pressure trough the film
(Pa).
Fourier-Transform Infrared (FT-IR) Spectroscopy
The FT-IR spectroscopy was used to identify the functional
groups of starch and biodegradable starch films. The spectra
were obtained using a spectrometer (PerkinElmer, Spectrum
Two, USA) equipped with an attenuated total reflection (ATR)
accessory. The samples were placed in the ATR crystal at room
temperature. An average of 30 scans was made for each sample.
Thickness, Color Variation (ΔΈ), and Water Solubility
The film thickness was measured using a digital micrometer
(Inside, China) at five different points on the sample. The ΔΈ
was determined with a Konica Minolta Colorimeter (Japan)
by the CIEL *a *b system; the measurement was made at five
different points on the sample and the value calculated accord-
ing to Eq. 2.
qffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
 2  2
Δ0 E ¼ ðΔa* Þ2 þ Δb* þ ΔL*
Were Δa∗ = (a∗ − a0∗), Δb∗ = (b∗ − b0∗)eΔL∗ = (L∗ − L0∗);
being L*, a* and b* the color values of the films and; L0*, a0*
and b0* the color values of the control.
The solubility in water was calculated as the percentage of
dry matter of the solubilized film after immersion for 24 h in
water at 25 °C, using Eq. 3, according to the method described
by Araujo-Farro et al. [12].
M i −M f
SOL ¼  100 ð3Þ
Mi
Were SOL is the percentage of dry matter of the solubilized
film after immersion (%), Mi the initial dry matter (g) and Mf
the final dry matter (g).
The moisture content of the films was expressed as a per-
centage and measured within the solubility in water analysis
by the weight loss of the film, after drying until the weight was
constant.
Scanning Electron Microscopy (SEM)
Surface micrographs were obtained using a Scanning Electron
Microscope (JEOL, JSM 6010LV, Japan). The samples were
coated with gold in a vacuum chamber (Denton Vacuum in
Sputtering DESK V, USA) and examined at 10 kV accelera-
tion voltage and magnifications of 100 and 500 x.
Atomic Force Microscopy (AFM)
The surface topography of the films was evaluated using an
Atomic Force Microscope (Agilent 5500, USA). The roughness
(Rq, root mean square) parameter was calculated with the data of
the topographic micrographs, using the equipment software.
ð2Þ
X-Ray Diffraction (XRD)
The relative crystallinity (RC) of the starch samples and
starch films were investigated by X-ray diffraction. The
XRD patterns were recorded using an X-ray
Diffractometer (Rigaku ULTIMA IV, Japan), in the
Bragg-Brentano geometry. The RC was calculated using
Eq. 4, where Ac is the area of the crystalline signals and
 Food Biophysics

Aa is the area of the amorphous signal in the X-ray values was made by Tukey’s test to a significance level of 5%
diffractogram. (p ≤ 0.05) using the analysis of variance (ANOVA).
AC
RC ð%Þ ¼  100 ð4Þ
AC þ Aa
Results and Discussion

Dynamical Mechanical Analysis (DMA)

Statistical Analysis
Typical stress-strain curves of biodegradable starch films are
The analyses were made in triplicate and standard deviations shown in Fig. 1. The starch films plasticized with glycerol
are reported, except for Rq and RC. A comparison of the mean showed different behaviors from sample to sample; curves

Fig. 1 Stress-strain typical curves

for potato starch films plasticized
with (a) glycerol and (b) sorbitol
Food Biophysics

with less deformation represent a rigid material and those with
large deformation represent a ductile material. In contrast,
films plasticized with sorbitol presented similar behaviors
for replicate samples with the same plasticizer content, show-
ing less deformation for the films with less content of sorbitol
and more deformation for the ones with a higher content.
The elastic modulus (E) and tension at break (Tb) data show
significant variation between different samples (Table 2). For
both E and Tb, starch films plasticized with higher content of
glycerol presented the highest values for native starch films (N G
0.30), followed by those of starch oxidized with 0.5% and 1.5%
of active chlorine (0.5% G 0.30 and 1.5% G 0.30). These me-
chanical properties are influenced by the films moisture con-
tent, that was reported in Table 1 and varied from 15.35 ±
1.31 to 21.78 ± 0.49%. Halal et al. [6] also reported the
moisture content of barley starch (Hordeum sativum)
films with values ranging from 16.6 ± 0.5 to 24.3 ± 0.3%.
The desired properties of starch films are determined by its
application, For example, in food packaging, a greater flexibil-
ity or rigidity of the film is desired. Thus, for this application
intermediate E values might be the best, as the films with this
characteristic are resistant as well as flexible (Table 2). The
elastic modulus E is a property that renders the material stiff,
and therefore, the higher E is the more rigid the film is. Halal
et al. [6] evaluated the E value of films of barley starch plasti-
cized with glycerol (gglycerol/gstarch = 0.30) with 3% of native
starch and oxidized starch with 1.0, 1.5, and 2.0% of active
chlorine and reported E = 25, 70, 288, and 45 MPa, respective-
ly; these values are considerably lower than those of our films,
suggesting that the films produced in this study are more rigid.
The plasticizer type influenced the Tb values. The most
resistant films were the native starch plasticized with
glycerol and the oxidized starch films (obtained using
both concentrations of active chlorine) plasticized with
sorbitol. The difference in Tb results can be explained
by the molecular structure differences of the plasticizers.
In addition, glycerol has a greater capacity to penetrate
the starch polymeric matrix because of its small chain
length [13].
The films with lower plasticizer contents presented the
highest values of E and Tb, for all samples. This might be
because the use of plasticizers reduces the intermolecular
forces and increases the polymeric chain mobility by increas-
ing the free space in the chains. When the film is subjected to
mechanical stress, the movement of the polymer chains is
facilitated by the addition of plasticizer. This is because the
plasticizer modifies the starch network, generating a film with
a less dense matrix. The plasticizer incorporation increases the

Table 2 Elastic modulus (E), tension at break (Tb), maximum tension for synthetic polymers (σ18N), water vapor permeability (WVP), thickness, color
variation (ΔΈ) e water solubility from the biodegradable potato starch films

Samples* E (MPa) Tb (MPa) e σ18N** WVP (g/m·s·Pa) Tickness (mm) ΔΈ (%) Water solubility (%)

N G0.20 2299 ± 62.91 a 47.35 ± 4.04 a 1.40.10−11 ± 9.49.10–13 bc 0.054 ± 0.004 b 14.98 ± 0.94 a 17.07 ± 0.75 a
N G0.30 1005 ± 50.47 d 23.69 ± 2.27 cd 1.78.10−11 ± 5.67.10–13 a 0.056 ± 0.004 b 9.26 ± 0.30 c 13.10 ± 1.56 c
0.5% G0.20 1651 ± 74.57 b 29.89 ± 0.57 bc 1.39.10−11 ± 9.36.10–13 bc 0.052 ± 0.004 b 6.48 ± 0.41 e 7.44 ± 0.89 f
0.5% G0.30 646 ± 86.03 ef 15.88 ± 1.60 e 8.85.10−12 ± 7.58.10–13 e 0.058 ± 0.004 b 5.54 ± 0.08 f 14.75 ± 0.55 b
1.5% G020 443 ± 28.02 f 5.61 ± 0.40 f 8.41.10−12 ± 1.72.10–12 e 0.054 ± 0.004 b 7.39 ± 0.36 e 13.01 ± 0.27 c
1.5% G0.30 219 ± 14.97 g 5.44 ± 0.42 f 1.47.10−11 ± 3.00.10–12 b 0.058 ± 0.004 b 6.16 ± 0.56 ef 14.97 ± 1.12 b
N S0.20 1306 ± 51.58 c 25.98 ± 1.49 c 1.11.10−11 ± 2.15.10–12 cd 0.072 ± 0.004 a 11.07 ± 0.56 b 8.64 ± 0.12 e
N S0.30 635 ± 25.57 ef 17.17 ± 1.51 de 1.26.10−11 ± 7.56.10–13 c 0.077 ± 0.004 a 8.37 ± 0.38 d 8.83 ± 0.42 e
0.5% S0.20 1389 ± 65.28 c 31.92 ± 1.94 b 1.02.10−11 ± 1.58.10–12 d 0.072 ± 0,004 a 5.70 ± 0.32 f 3.05 ± 0.36 g
0.5% S0.30 764 ± 42.48 e 19.89 ± 1.40 d 5.04.10−12 ± 1.83.10–12 h 0.076 ± 0.005 a 6.20 ± 0.30 ef 9.18 ± 2.13 e
1,5% S0.20 1210 ± 57.56 c 19.70 ± 1.41 d 7.52.10−12 ± 7.73.10–13 g 0.074 ± 0.005 a 6.34 ± 0.23 e 4.55 ± 0.75 fg
1.5% S0.30 762 ± 47.20 e 17.33 ± 2.12 de 8.20.10−12 ± 8.04.10–13 f 0.076 ± 0.005 a 5.33 ± 0.37 f 9.89 ± 0.15 d
PET 2513 ± 14.00 σ18N = 26.27 ± 0.04 – – – –
HDPE 610 ± 42.57 σ18N = 21.77 ± 1.53 – – – –
LDPE 171 ± 3.26 σ18N = 20.82 ± 0.76 – – – –
*
N G0.20: native starch film with 0.20 gglycerol/ gstarch. N G0.30: native starch film with 0.30 gglycerol/ gstarch. 0.5% G0.20: oxidized with 0.5% of
active chlorine starch film with 0.20 gglycerol/ gstarch. 0.5% G0.30: oxidized with 0.5% of active chlorine starch film with 0.30 gglycerol/ gstarch. 1.5%
G0.20: oxidized with 1.5% of active chlorine starch film with 0.20 gglycerol/ gstarch. 1.5% G0.30: oxidized with 1.5% of active chlorine starch film with
0.30 gglycerol/ gstarch. N S0.20: native starch film with 0.20 gsorbitol/ gstarch. N S0.30: native starch film with 0.30 gsorbitol/ gstarch. 0.5% S0.20:
oxidized with 0.5% of active chlorine starch film with 0.20 gsorbitol/ gstarch. 0.5% S0.30: oxidized with 0.5% of active chlorine starch film with 0.30
gsorbitol/ gstarch. 1.5% S0.20: oxidized with 1.5% of active chlorine starch film with 0.20 gsorbitol/ gstarch. 1.5% S0.30: oxidized with 1.5% of active
chlorine starch film with 0.30 gsorbitol/ gstarch. All the results correspond to the films moisture content shown in Table 1
**
σ18N: Maximum tension for synthetic polymers
a,b,c,d,e,f,g
Values with different letters in the same column are significantly different (p < 0.05)

film elongation by increasing the molecular space in the ma-
trix and reducing the hydrogen bonds. As E represents the
material stiffness, films with a high plasticizer content are
more flexible and thus present lower E values [14]. Similar
reduction in the E value with an increase in the plasticizer
content was observed for films of rice starch [13], quinoa
starch (C. quinoa, Willdenow) [12], and cassava starch
(Manihote sculenta) [14].
Synthetic polymers used in the food packaging industry
(polyethylene terephthalate – PET, high-density polyethylene
– HDPE, and low-density polyethylene – LDPE) were ana-
lyzed by DMA technique to compare their properties with
those of the biodegradable starch polymers produced in this
study. Owing to the equipment constraints, the samples did
not reach Tb, and therefore, the test was finalized after the
application of a maximum load force of 18 N (σ18N) on the
materials (stress-strain curves not shown) (Table 2).
Through the evaluation of the mechanical properties of
synthetic polymers (Table 2), it is, by analogy, better to iden-
tify an application for the biodegradable starch films which
show intermediate elastic modulus compared to PET (more
rigid) and LDPE (more flexible).
Water Vapor Permeability (WVP)
The WVP values if the films are presented in Table 2. The
starch films plasticized with glycerol showed a higher mass
gain (data not shown) and consequently greater WVP than
those plasticized with sorbitol. Sorbitol has more hydroxyls
to interact with water; however, glycerol has a higher chemical
affinity to it, which increases its plasticizing effect. The struc-
ture of the films plasticized with sorbitol is more compact and
less capable of interacting with water because of its cyclic
conformation [15].
In comparison with the native starch films, the starch films
derived from starch oxidized with 1.5% of active chlorine
presented lower WVP when plasticized with a lower content
of glycerol and sorbitol. Meanwhile, the films oxidized with
0.5% of active chlorine showed lower WVP for both plasti-
cizers at higher concentrations. Thus, the oxidized starch films
led to lower WVP regardless of the type and concentration of
the plasticizer as well as the active chlorine concentration.
These results demonstrate that the oxidized biodegradable
starch films can potentially be applied in food packaging, once
less permeability and less migration of water through the film
is achieved, which is essential for food packaging [16].
In this study, the oxidation of starch was effective in reduc-
ing the WVP. The same was reported previously [17] for bio-
degradable films derived from sorghum starch (Sorghum
bicolor) chemically modified by oxidation with 1.5% of active
chlorine. Except for the oxidized starch films with 0.5% of
active chlorine, the increase in the plasticizer content contrib-
uted to the increase in WVP. Still, these films showed lower
Food Biophysics
WVP than those of native starch for the same plasticizer con-
centration. Higher plasticizer concentrations lead to higher
interaction with water, rendering the films more hydrophilic,
and thus increasing their water absorption, and consequently,
their WVP [15]. In another study on potato starch films plas-
ticized with sorbitol, glycerol, and xylitol [18] the same char-
acteristic was observed.
All samples plasticized with sorbitol showed lower WVP
than those plasticized with glycerol. This is because, plasti-
cizers are hydrophilic additives; however, glycerol has a
higher capacity to interact with water and therefore, the
WVP of films plasticized with it is increased. The highest
moisture content found was for the films plasticized with sor-
bitol which can also influenced in the WVP, since the water is
also know as a film plasticizer.
It has been reported that between rice starch films plasti-
cized with sorbitol and glycerol (gplasticizer/gstarch = 0.20 and
0.30), the films produced with glycerol show higher WVP.
According to the authors, the increase in the plasticizer con-
centration increases the water migration [13]. The same was
observed by other authors who evaluated rice starch [19] and
cassava films [20].
FT-IR Analysis
The spectra shown in (Fig. 2a) illustrate similar functional
groups in the three different starch samples, with bands in
the same region, differing only in their intensity. The
broad band at 3276 cm−1 is attributed to the stretching
of the hydroxyl (-OH) groups [21]. In this region, the
band intensity decreases according to the treatment of
the starch (native > 0.5% > 1.5%). This might be because,
during the oxidation, the hydroxyl groups are replaced by
carbonyl and carboxyl groups.
The vibrations of the -CH groups appear in the 2900 to
3000 cm−1 region; therefore, the band at 2927 cm−1 indicates
the presence of -CH group [21]. The band located at
1748 cm−1 corresponds to carbonyl groups (C=O) [22] and
the one located at 1646 cm−1 is related to absorbed water
(H2O) in the granules of the amorphous region. The spectral
region of 1550–1750 cm−1 corresponds to the amorphous
H2O. This H2O band is directly related to the crystallinity
and its position can change according to the polysaccharide
crystallinity pattern. The bands at 1570 cm−1 and 1457 cm−1
might correspond to the carboxyl groups (-COO). The band at
1335 cm−1 represents the -CH2 groups which are located be-
tween 1200 and 1350 cm−1 or the O-H groups of H2O [21, 23].
Bands at 1163, 1150, 1124, and 1103 cm−1 indicate the C-
O and C-C stretch with the contribution of C-OH stretch. The
band at 995 cm−1 is also described as the amorphous part of
the structure, due to a water that interacts with the intermolec-
ular hydrogen bonds [23]. As this interaction occurs different-
ly between each type of starch, the band intensity changes, and
Food Biophysics

Fig. 2 1 FT-IR spectra of (a) 995

native and oxidized potato (A)
starches and (b) biodegradable
potato starch films

1150

1335

Absorbance
3276 2927 1457
1646
1748
Native starch

Oxidized starch 0.5%

Oxidized starch 1.5%

4000 3000 2000 1000

-1
Wavenumbers (cm )

(B)
997
1150

3294 1337
2929 1645 1457
N G0.20
N G0.30
0.5% G0.20
0.5% G0.30

1.5% G0.20
Absorbance

1.5% G0.30

N S0.20
N S0.30
0.5% S0.20
0.5% S0.30
1.5% S0.20
1.5% S0.30

4000 3500 3000 2500 2000 1500 1000 500

-1
Wavenumber (cm )

may vary by the amylose and amylopectin content in each
type of starch.
Kizil et al. [24], who analyzed the FT-IR spectra of potato
starch and also reported bands at 1642 cm−1, attributed it to
water absorption, relating to starch crystallinity. The same
authors found similar bands to those in this study in the re-
gions of 3740, 2933, 1344 and 1163 cm−1. In a study that also
evaluated oxidized potato starch, similar bands to those of the
present study were observed [25].
The infrared spectra of native and oxidized starch did not
show large differences. This could be because of the formation
of discrete functional groups during the oxidation. In the spec-
tra of biodegradable starch films (Fig. 2b), the disappearance
of some bands as well as small displacements in bands are
obvious, compared to those of the raw material.
Thickness, Color Variation (ΔΈ), and Water Solubility
The thickness, ΔΈ and water solubility are presented in
Table 2. The biodegradable starch films are uniform, transpar-
ent, and have low solubility in water. The films plasticized
with glycerol showed lower thickness than those plasticized
with sorbitol. Kefiran films were evaluated by Ghasemlou
et al. [15] and the films plasticized with glycerol had a lower
thickness than those plasticized with sorbitol, with the thick-
ness ranging from 0.064 ± 0.0098 mm (kefiran+ glycerol) to
0.074 ± 0.0045 mm (kefiran+ sorbitol). According to the au-
thors, sorbitol penetrates the polymer matrix more easily and
quickly, forming stronger and thicker films.
The films were found to be transparent and translucent with
low ΔΈ values (Table 2). For both types of films, i.e., those

plasticized with glycerol and sorbitol, the ΔΈ was lower at
higher concentration of the plasticizer. Between the films with
different plasticizers, the ones with sorbitol presented lower
ΔΈ. Tajik et al. [1] evaluated the color variation of soluble soy
protein films and found that the addition of glycerol to the
matrix lightened the films The use of plasticizers in a
filmogenic matrix results in superior optical properties.
The oxidized starch films present lower ΔΈ relative to
native starch films. Thus, it can be stated that the use of sor-
bitol as a plasticizer and chemical modification by oxidative
treatment reduced the transparency, since greater the ΔΈ, the
less transparent the biodegradable film. A study that evaluated
two different sweet potato starch films and bean starch films,
all plasticized with glycerol (gglycerol/gstarch = 0.25) reported
that bean starch films followed by sweet potatoes starch films
showed lower ΔΈ [26].
The films were found to be transparent and translucent with
low ΔΈ values (Table 2). For both types of films, i.e., those
plasticized with glycerol and sorbitol, the ΔΈ was lower at
higher concentration of the plasticizer. Between the films with
different plasticizers, the ones with sorbitol presented lower
ΔΈ. The use of plasticizers in a filmogenic matrix resulted in
superior optical properties [1]. The authors evaluated the color
variation of soluble soy protein films and found that the addi-
tion of glycerol to the matrix lightened the films.
The oxidized starch films present lower ΔΈ relative to
native starch films. Thus, it can be stated that the use of sor-
bitol as a plasticizer and chemical modification by oxidative
treatment reduced the transparency, since greater the ΔΈ, the
less transparent the biodegradable film. A study that evaluated
two different sweet potato starch films and bean starch films,
all plasticized with glycerol (gglycerol/gstarch = 0.25) reported
that bean starch films showed lower ΔΈ followed by sweet
potatoes starch films [26].
The films remained intact after 24 h of immersion in water
under agitation and showed low water solubility and stability
against water absorption. The solubility in water results cor-
respond to the films with moisture content showed in Table 1.
As shown in Table 2, upon comparing the type of plasticizer at
the same content, the films plasticized with glycerol showed
greater solubility in water than those plasticized with sorbitol.
In case of rice starch films plasticized with different amounts
of glycerol and sorbitol [19], greater solubility was observed
for the films plasticized with higher glycerol concentration.
The authors explain that this occurs because glycerol has a
higher hydrophilic character than sorbitol. Therefore, the in-
crease in the plasticizer concentration leads to an increase in
the solubility of the film, causing reduction in the interaction
between the biopolymer molecules and their interaction with
water is increased because of the hydrophilicity of the
plasticizer [15].
The water solubility of potato starch films was analyzed
and a value of approximately 35% was reported [27]. The
Food Biophysics
films produced in the present study presented lower solubility
compared to the cited study. Other authors [4] evaluated po-
tato starch films oxidized with sodium hypochlorite contain-
ing 0.5% of active chlorine and obtained reduced solubility for
the oxidized starch films compared to the native films. The
same authors reported that oxidation decreased the solubility,
because of the increase in the interaction between amylose and
amylopectin, and the stretching of intramolecular bonds. In
the present study, lower solubility is observed for oxidized
starch films when compared to the native ones at a low plas-
ticizer concentration (gplasticizer/gstarch = 0.20); however, the
water solubility is higher for the films plasticized at the higher
plasticizer concentration. The water solubility and thickness
influenced the properties of the films, as observed by DMA
and WVP evaluation.
Scanning Electron Microscopy (SEM)
The biodegradable films with the lowest plasticizer concentra-
tion were analyzed by SEM as well as AFM and XRD, owing
to their higher mechanical resistance and lower WVP. In the
micrographs (Fig. 3), partially gelatinized starch granules are
visible. It is possible to observe that the modification affects
the morphology of the film. As the extent of oxidation is
increased (from native to 1.5% of active chlorine), the shape
of the granule changes and a more homogeneous morphology
is observed. This is because of the depolymerization of starch
molecules due to the oxidation process, which also influences
the interaction between the starch and plasticizer [6].
The partially gelatinized granules can be attributed to the
incomplete rupture of hydrogen bonds between amylose and
amylopectin, which results in partial gelatinization of the
granules. During the heating of the film-forming solution,
the amylose molecules leach into the intergranular space.
The granules begin to break due to the rupture of the hydrogen
bonds and solubilization of the amylose and amylopectin mol-
ecules. Upon decreasing the temperature of the solution, ret-
rogradation occurs and the amylose and amylopectin chains
tend to re-approximate [28].
In general, the films with plasticizers in their composition
present a compact structure and smooth surfaces with no pores
[29]. Despite the partially gelatinized granules, a filmogenic
solution is formed, generating resistant, flexible films with
good barrier properties.
Atomic Force Microscopy (AFM)
The surface topographies of biodegradable starch films, with
5.0 × 5.0 μm2 area are shown in Fig. 4. The roughness param-
eter, Rq, of the biodegradable films showed variation, ranging
from 3.39 nm to 10.9 nm. All the analyzed films presented low
roughness, although the films with the lowest Rq are consid-
ered extremely smooth. The images indicate an irregular
Food Biophysics

Fig. 3 SEM micrographs from (A) (B)

potato starch films (a) N G0.20
(b) N S0.20 (c) 0.5% G0.20 (d)
0.5% S0.20 (e) 1.5% G0.20 (f)
50 µm
1.5% S0.20 50 µm

100 µm 100 µm

(C) (D)

50 µm 50 µm

100 µm 100 µm

(E) (F)

50 µm 50 µm

100 µm 100 µm

structure with darker and lighter regions in the z height scale.
However, owing to the low roughness, the biodegradable films
can be considered smooth and homogeneous, because the
roughness is directly related to the irregularity of the film sur-
face [30]. In general, the surfaces of materials present irregu-
larities, even the smooth ones [31]. The films showed no pores,
although the 0.5% S0.20 (Fig. 4d) and 1.5% S0.20 (Fig. 4f)
showed grooves which can be related to the mold used to
fabricate the films.
The surface topography of tapioca starch films was evaluat-
ed by Resa et al. [32] and they reported low roughness values
(10.00 ± 1.02 nm), similar to those observed in this study.
Another study also revealed low roughness (11.26 ± 2.56 nm)
with a smooth and homogenous surface topography, although
for sodium alginate and sodium alginate mixed with chitosan,
the roughness was significantly higher (Rq = 68.37 ± 16.17 and
69.59 ± 27.21 nm, respectively), indicating the formation of
heterogeneous films [33].
X-Ray Diffraction (XRD)
X-ray diffractograms of the starch samples and biodegradable
starch films are presented in Fig. 5. The diffractograms of
native and oxidized starch samples with 0.5 and 1.5% of ac-
tive chlorine (Fig. 5a) present a B-type crystallinity pattern,
which is typical of tubers. The X-ray diffractograms show
differences between the starch types that are determined by
the peaks. In case of B-type pattern, main peaks appear at 2θ
 Food Biophysics

(A) Rq= 3.39 nm (B) Rq= 4.93 nm

(C) Rq= 7.02 nm

(D) Rq= 10.9 nm

(E) Rq= 3.82 nm (F) Rq= 4.47 nm

Fig. 4 Topographic images of AFM from potato starch films (a) N G0.20 (b) N S0.20 (c) 0.5% G0.20 (d) 0.5% S0.20 (e) 1.5% G0.20 (f) 1.5% S0.20

diffraction angles of 5.6°; 14.4°; 17.2°; 22.2° and 24° [34].
The three types of starch presented similar patterns, indicating
a semi-crystalline structure with different peak intensities and
low relative crystallinity (RC).
A difference in the crystallinity pattern was observed
when the effect of the acid modification and oxidation
in sorghum starch was analyzed [17]. As in the present
study, other authors have reported a B-type diffraction
pattern for potato starch derived from different cultivars
[9, 35].
The RC of starch can show large variation according to the
raw material, extraction method, and humidity. For potato
starch obtained from different cultivars, an RC of approxi-
mately 30% has been reported, which is approximately 10%
higher than that found in the present study for native starch
(Fig. 5a) [35].
Food Biophysics

Fig. 5 XRD diffractograms from 19.74

(a) native and oxidized potato 17.04 (A)
starches and (b) biodegradable 22.22
potato starch films plasticized 15.04
with glycerol and sorbitol in the 24.14
less concentration

5.62

Intensity (cps)
Native RC= 18.64%

Oxidized 0.5% RC= 12.69%

Oxidized 1.5% RC= 17.53%

0 10 20 30 40 50
Diffraction angle

16.96

15.0 19.22
(B)
22.36

5.24

N G0.20 RC= 28.80%

Intensity (cps)

0.5 % G0.20 RC= 25.61%

1.5% G0.20

N S0.20 RC= 31.59%

0.5% S0.20 RC= 32.42%

1.5% S0.20 RC= 35.66%

0 10 20 30 40 50
Diffraction angle

Biodegradable films (Fig. 5b) show similar peaks with Conclusions

lower intensities than those of the raw material (Fig. 5a).
The diffractograms reveal that the films are crystalline.
This indicates that after the gelatinization and retrograda-
tion, the molecules could reassociate and present crystal-
linity. The RC of biodegradable starch films is higher than
that of the raw material. The films plasticized with sorbitol
present higher RC than those plasticized with glycerol. The
addition of a plasticizer to the polymer matrix helps in
maintaining the crystalline arrangement of starch with the
formation of hydrogen bonds between the polymer chains
and the plasticizer molecules [7].
Changes in the granule structure, such as their crystallinity,
influence the properties such as WVP, since the crystalline
region is generally impermeable owing to the denser and more
compact structure [36]. The films plasticized with sorbitol
(gsorbitol/gstarch = 0.20) present higher RC, and only the ones
produced with oxidized starch with lower concentration of the
plasticizer present lower WVP (0.5% S0.20 shows lower
WVP and higher RC than 0.5% G0.20).
In general, biodegradable starch films produced in different
compositions were found to be flexible, resistant, and transpar-
ent, with low solubility in water. The films produced with native
starch at lower concentrations of both plasticizers were relative-
ly more rigid and those produced with oxidized starch at higher
concentration of the plasticizer were more ductile. The oxida-
tion of starch resulted in improved barrier properties of the
films, and films with relatively lower WVP values were obtain-
ed from oxidized starch with 0.5% of active chlorine plasticized
with sorbitol. The films of oxidized starch plasticized with low-
er concentrations of sorbitol showed lower water solubility.
The films of oxidized starch plasticized with sorbitol
showed relatively better results with respect to the mechanical
properties, WVP, and water solubility, and are the best choice
to be applied in food packaging. Therefore, potato starch is a
good raw material for producing biodegradable films, because
the properties of their films such as WVP and mechanical
properties are satisfactory.
 Food Biophysics

Acknowledgements We would like to thank CAPES (process N° 17. B. Biduski, F.T. Da Silva, W.M. Silva, S.L.D.M. Halal, V.Z. Pinto,
1381538), UNIPAMPA, CEME SUL – FURG and UNIPAMPA A.R.G. Dias, E.D.R. Zavareze, Food Chem. 214, 53–60 (2017)
Professor’s André Gündel and Flávio André Pavan for the AFM and 18. R.A. Talja, H. Helén, Y. Roos, K. Jouppila, Carbohydr. Polym. 71,
FT-IR, respectively. 269–276 (2008)
19. N. Laohakunjit, A. Noomhorm, Starch-Starke 56, 348–356 (2004)
20. C.M.O. Müller, F. Yamashita, J.B. Laurindo, Carbohydr. Polym.
72, 82–87 (2008)
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