82 Indones. J. Chem.

, 2018, 18 (1), 82 - 91

The Physico-Mechanical Properties and Release Kinetics of Eugenol

in Chitosan-Alginate Polyelectrolyte Complex Films as Active Food Packaging

Baiq Amelia Riyandari, Suherman, and Dwi Siswanta*

Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Gadjah Mada,
Sekip Utara, Yogyakarta 55281, Indonesia

Received July 14, 2017; Accepted September 9, 2017

ABSTRACT

A study of eugenol release and its kinetics model from chitosan-alginate polyelectrolyte complex (PEC) films
has been conducted. Some factors that affected the eugenol release were also studied, including the composition of
chitosan-alginate PEC and the concentration of eugenol. The chitosan-alginate-eugenol PEC films were synthesized
at pH ± 4.0, then the PEC films were characterized using a Fourier-transform infrared spectroscopy (FTIR)
spectrophotometer. An investigation of the films’ properties was also conducted, including morphology analysis
using a scanning electron microscope (SEM), differential thermal analysis (DTA) / thermogravimetric analysis (TGA),
mechanical strength, transparency testing, water absorption, and water vapor permeability. The release of eugenol
was investigated through in vitro assay in ethanol 96% (v/v) for four days, and the concentration of eugenol was
measured using an ultraviolet-visible (UV-Vis) spectrophotometer. The characterization of the films using FTIR
showed that the formation of PEC occurred through ionic interaction between the amine groups (–NH3+) of the
chitosan and the carboxylate groups (–COO–) of the alginate. The result showed that the composition of chitosan-
alginate PEC and the concentration of eugenol can affect the release of eugenol from PEC films. A higher
concentration of alginate and eugenol could increase the concentration of eugenol that was released from the films.
The mechanism for the release of eugenol from chitosan-alginate PEC films followed the Korsmeyer-Peppas model
with an n value of < 0.5, which means the release mechanism for eugenol was controlled by a Fickian diffusion
process. The antioxidant activity assay of the films using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) method resulted
in a high radical scavenging activity (RSA) value of 55.99% in four days.

Keywords: PEC chitosan-alginate; eugenol; release; antioxidant; kinetics model

ABSTRAK

Studi mengenai pelepasan eugenol dan model kinetika pelepasannya pada film kompleks polielektrolit kitosan-
alginat telah dilakukan. Beberapa faktor yang memengaruhi pelepasan seperti komposisi KPE kitosan-alginat dan
konsentrasi eugenol juga telah dipelajari pada penelitian ini. Pembuatan film kompleks polielektrolit kitosan-alginat
yang terembani eugenol dilakukan pada pH ± 4,0. Film KPE kitosan-alginat-eugenol selanjutnya dikarakterisasi
menggunakan spektrofotometer FTIR. Selain itu, dilakukan pula analisis morfologi menggunakan SEM, analisis
termal DTA/TGA, uji mekanik film, uji transparansi, uji penyerapan air, dan permeabilitas uap air. Studi pelepasan
eugenol dilakukan secara in vitro dalam etanol 96% (v/v) selama 4 hari dan konsentrasi eugenol yang terlepas
diukur menggunakan spektrofotometer UV-Vis.Hasil karakterisasi dengan FTIR menunjukkan bahwa KPE kitosan-
alginat terbentuk melalui interaksi ionik antara gugus amina (–NH3+) dari kitosan dan gugus karboksilat (–COO–) dari
alginat. Berdasarkan studi yang dilakukan, komposisi KPE kitosan-alginat dan konsentrasi eugenol akan
memengaruhi pelepasan eugenol dari film. Peningkatan kandungan alginat dan peningkatan konsentrasi eugenol
dalam film akan meningkatkan jumlah eugenol yang terlepas. Pelepasan eugenol dari film KPE kitosan-alginat
mengikuti model kinetika Korsmeyer-Peppas dengan nilai n < 0,5. Hal ini menunjukkan mekanisme pelepasannya
dikontrol oleh proses difusi Fickian. Hasil uji aktivitas antioksidan menggunakan metode DPPH menunjukkan bahwa
film memiliki aktivitas antioksidan yang baik yaitu dengan nilai RSA sebesar 55,99% dalam waktu 4 hari.

Kata Kunci: KPE kitosan-alginat; eugenol; pelepasan; antioksidan; model kinetika

INTRODUCTION The oxidative reaction is caused by free radicals that

attack the surface of the food so it can change the
Lipid oxidation is one of the factors that causes color, taste, or aroma of foodstuffs. In recent times,
degradation and reduces the quality of food products. active food packaging has been developed, which is
* Corresponding author. Tel : +62-8157951198 DOI: 10.22146/ijc.26525
Email address : dsiswanta@ugm.ac.id

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 Indones. J. Chem., 2018, 18 (1), 82 - 91 83

one of the most promising packaging systems created flexible, and have good mechanical properties.
that has been found to be very effective in maintaining Therefore, they can be used for the development of
the food safety and its shelf life [1-3]. Active packaging is food packaging.
a food packaging system that contains an antioxidant or Although some previous studies have been
an antibacterial compound so it can inhibit free-radical conducted on food packaging using chitosan, alginate,
activity, spoilage, or pathogenic microorganisms that and eugenol, no study exists regarding the
contaminate foods. The technique of adding an incorporation of eugenol into chitosan-alginate films
antioxidant into active packaging films allows a that focuses on the release mechanism for eugenol
controlled release of the antioxidant. This method is and its kinetics model. Hence, the objective of this
more efficient and effective than the addition of an research is to study the release of eugenol, including
antioxidant directly into the foods. its kinetics model, release mechanism, and some
The addition of some food additives to food factors that affect its release. Furthermore, the
packaging, such as butylated hydroxytoluene (BHT) and antioxidant activity of chitosan-alginate PEC films and
butylated hydroxylamine (BHA), has been studied. The their properties also will be studied in this research.
addition of BHT to low density polyethylene (LDPE) films
could decrease the rate of lipid oxidation and extend the EXPERIMENTAL SECTION
food shelf life [3]. However, BHT, which is a synthetic
antioxidant, can be a toxic agent and has a carcinogenic Materials
potency for the human body [4-5]. Therefore, the
development of active food packaging has been focused Chitosan (with a degree of deacetylation of 0.87)
on the use of plant extracts, such as tea extract [6-7], was purchased from CV Ocean Fresh, Bogor; alginate
grape extract [8-9], and ginger extract [8]; or active (PA) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) were
compounds, such as α-tocopherol [10], carvacrol [11- purchased from Sigma-Aldrich; eugenol was obtained
12], thymol [11], and eugenol [13]. from the Indesso factory in Bogor; and distilled water
Eugenol is a natural phenolic compound that is was also used.
extracted from clove oil. Because of its great
antibacterial, antifungal, and antioxidant activities, it has Instrumentation
been used widely in some industries including food,
pharmaceuticals, and cosmetics. A study on eugenol as This work used a pH Meter Lab 680, magnetic
an antioxidant in packaging films has been conducted by stirrer, and hot plate. The characterization of films by
Woranuch and Yoksan [13]. They investigated the effect Fourier-transform infrared spectroscopy (FTIR) was
of incorporating eugenol-loaded chitosan nanoparticles performed using a Shimadzu FTIR Prestige 21, and an
in thermoplastic flour (TPF). These films exhibited a low ultraviolet-visible (UV-vis) spectrophotometer
tensile strength (1.53 MPa) and a high oxygen- (Thermoscientific Evolution 201 UV-Vis
permeability value. However, the result shows that TPF Spectrophotometer) was used to analyze the
containing eugenol-loaded chitosan nanoparticles has concentration of the eugenol and DPPH radical
significant radical scavenging activity compared with solution. The morphology of the surface of the films
TPF without eugenol. That is why eugenol has the was observed using a scanning electron microscope
potential to be applied as an antioxidant for food (SEM) (JEOL JSM-6510LA), and thermal analysis was
packaging. performed using a Shimadzu DTG60 and a Shimadzu
Chitosan is a natural polysaccharide that is DSC-60.
obtained through the deacetylation of chitin, and is
known as a biodegradable, cheap, renewable, and non- Procedure
toxic polymer [6]. Chitosan is polycationic in an acid
medium, so it has the ability to form a polyelectrolyte Synthesis of PEC chitosan-alginate films
complex (PEC) through ionic interaction with a incorporating eugenol
polyanionic substance. The formation of a PEC film is Variation of PEC composition. Alginate solutions
one of the techniques to employed to increase the (0.125, 0.25, and 0.375%, w/v) were prepared by
mechanical properties of chitosan films [14]. Alginate is dispersing alginate powder in distilled water and stirring
also a natural polysaccharide found in brown algae, and them with a magnetic stirrer at 25 °C. Chitosan (as
it consists of ß-D-mannuronic acid (M) and α-L-guluronic shown in Table 1) was added to the alginate solution
acid (G) monomers [15]. Alginate is polyanionic so it can and it was stirred using magnetic stirrer for 20 min.
form a PEC with chitosan. Based on the previous studies After the chitosan had dissolved completely, glacial
by Yan et al. [16], and Kulig and Zimoch-Korzycka [17], acetic acid (2%, v/v) was poured into the solution.
PEC films from chitosan-alginate are transparent, Then, eugenol was added into the PEC solution to reach

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84 Indones. J. Chem., 2018, 18 (1), 82 - 91

Table 1. Variation of the PEC composition

Sample code Chitosan to alginate ratio Chitosan (% w/v) Alginate (% bwv)
A1 9:1 1.125 0.125
A2 8:2 1.000 0.250
A3 7:3 0.875 0.375

Table 2. Variation of the concentration of eugenol

Sample code Eugenol (% w/v) Chitosan (% w/v) Alginate (% w/v)
B1 0.05 1.000 0.250
B2 0.10 1.000 0.250
B3 0.20 1.000 0.250

a final concentration of 0.10% (w/v). The PEC-eugenol volume of 10 mL of distilled water was put in a Petri
solution was stirred using a magnetic stirrer for 24 h. The dish and aluminum foil was used to cover the Petri
PEC solution (10 mL) was poured onto 5.2 × 5.2 cm dish. The aluminum foil covering the Petri dish was
Petri dish and dried for 72 h at room temperature (25 perforated and had an area of 1.1 × 1.1 cm. Then, each
°C). The dried films were kept in a box containing silica. sample film (1.2 × 1.2 cm) was put in the hole and glue
Variation of eugenol concentration. Alginate solution was used to stick in each film. The WVP of each film
(0.25%, w/v) was prepared by dispersing alginate was determined three times. All Petri dishes were put
powder in distilled water and stirring it with a magnetic into the oven at a temperature of 37 ± 0.5 °C for 5 h.
stirrer at 25 °C. Chitosan (1%, w/v) was added to the Every hour, the weight of the Petri dish and its contents
alginate solution and it was stirred using a magnetic was measured. The water vapor transmission rate
stirrer for 20 min. After chitosan was dissolved (WVTR) was used to determine the WVP. The WVTR
completely, glacial acetic acid (2%, v/v) was poured into and WVP values were calculated using the following
the solution. Then, eugenol was added into the PEC equations:
solution to reach final concentrations of 0.05, 0.10, and mass water lost
WVTR  (1)
0.20% (w/v). The PEC-eugenol solution was stirred time  area
using a magnetic stirrer for 24 h. The PEC solution (10 WVTR
mL) was poured onto a 5.2 × 5.2 cm Petri dish and dried WVP  d (2)
S  R1  R 2 
for 72 h at room temperature (25 °C). The dried films
were kept in a box containing silica. where S is the saturated vapor pressure at a
FTIR analysis. The molecular interaction of the PEC temperature of 37 °C, R1 is the relative humidity in the
chitosan-alginate films incorporating eugenol was Petri dish, R2 is relative humidity at a temperature of 37
observed using FTIR. The film was dried in a desiccator °C, and d is the film thickness (in m).
containing silica for 2 weeks at room temperature
(25 °C) before analysis. The films were scanned from Water absorption
400 to 4000 cm–1. Water absorption is determined using the
Film thickness. The thickness of the films was gravimetric method. The dried film samples were kept
measured using a micrometer. Five locations on the in a desiccator for one week. The dried films were then
films were chosen randomly to determine the average immersed in 10 mL of distilled water for 1 h. Next, the
thickness of the films. films were dried, and the films were weighed before
and after immersing. The water absorption was
Mechanical properties calculated as follows:
Mo  M
The tensile strength (TS) and percentage of Water absorption  %    100% (3)
elongation at break (EAB) were measured using a M
universal testing machine. The film samples were cut where Mo is the mass of the film after immersing (in g),
into pieces measuring 100 mm × 20 mm and put into the and M is the mass of the film before immersing (in g).
machine. The test speed was set at 10 mm/min and the
measurements for the TS and EAB of each film were Film transparency value
determined with three replications. The light transmittance of the films was measured
using a UV-Vis spectrophotometer at a wavelength
Water vapor permeability range from 200 to 800 nm. The transparency value of
The water vapor permeability (WVP) was the films was calculated using the following equation:
determined using the wet cup method based on the  logT600
Tranparency value  (4)
American Society for Testing and Materials (ASTM) E96- x
95 1995 standard test method, with some modification. A

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 Indones. J. Chem., 2018, 18 (1), 82 - 91 85

where T600 is the transmittance of the films at 600 nm Morphology analysis using an SEM
and x is the film thickness (mm); a lower transparency The films’ surface morphology was examined
value indicates the greater transparency of films. using an SEM. Each film specimen was coated with
gold and observed using an accelerating voltage of 20
Study of the release of eugenol from PEC chitosan- kV.
alginate films
The profile of the release of eugenol was Thermal analysis
investigated for 4 days through an in vitro assay in The films were dried in a desiccator containing
ethanol 96% (v/v) as a fatty food simulant. The films silica for 2 weeks at room temperature (25 °C) before
were immersed in 25 mL of ethanol 96%. At specific time the analysis. The dried films were scanned using a
intervals, the concentration of eugenol was measured thermogravimetric analyzer from 30 to 300 °C at flow
using a UV-Vis spectrophotometer at a wavelength of rate of 10 °C min–1 under a nitrogen atmosphere at rate
282 nm. The concentration of eugenol that was released of 30 mL min–1.
from the films was calculated using the following
equation: RESULT AND DISCUSSION
Eugenol  released
Eugenol release  %    100% (5) Synthesis of PEC Chitosan-Alginate Films
Eugenol  total 
Incorporating Eugenol
Kinetics model for the release of eugenol The formation of the PEC films occurred through
The kinetics model for the release of eugenol was molecular interaction between polycationic groups and
determined using four models, as follows: polyanionic groups. In this research, the chitosan-
Zero-order kinetics model :  A t    A 0   kt (6) alginate PEC film was synthesized at pH ± 4.0, and it
First-order kinetics model : ln  A t   kt   A 0  (7) was formed through an ionic interaction between the
amino groups of chitosan and the carboxylate groups
Higuchi kinetics model : Q  kt 12
(8) of alginate. The pKa value of chitosan is 6.3, while the
Mt alginate pKa is 3.4–3.7 [18]. At pH ± 4.0, amino groups
Korsmeyer-Peppas kinetics model :  ktn (9)
M of chitosan and carboxylate groups of alginate are
where At is the concentration of the released active largely ionized [16]. Thus, the ionic interaction between
compound at t, A0 is the earlier concentration of the the amine groups (–NH3+) and the carboxylate groups
active compound, Q is the release percentage (%), Mt/M (–COO–) are formed strongly. When the pH of the PEC
is the cumulative fraction of the active compound at t, n solution is less than 3.4–3.7 or more than 6.3, the
is the release exponent indicating the chemical release ionization of alginate is not optimal. When the pH is
mechanism, t is the release time, and k is the rate less than 3.4–3.7, the carboxylate groups are ionized
constant of the active compound. slightly, and when the pH is more than 6.3, just a small
amount of amine groups are ionized. However, if the
Assessment of antioxidant activity pH of the PEC solution is in optimal circumstance (3.4–
The antioxidant activity of the PEC chitosan- 3.7 < pH < 6.3), both the amino groups of the chitosan
alginate films incorporating eugenol was determined and the carboxylate groups of the alginate are mostly
using the fixed reaction time method. The films ionized.
incorporating eugenol were immersed in 25 mL of
ethanol 96% (v/v). A volume of 0.5 mL of the testing FTIR Analysis
solution was added to a DPPH radical solution (75 µM,
3.5 mL) and then it was incubated in the dark at room Characterization by FTIR was used to analyze the
temperature (25 °C) for 30 min. A DPPH radical solution molecular interaction, which relates to the physical
with 0.5 mL ethanol solution (without eugenol) added to properties of the films’ matrices. The infrared spectrum
it was used as a control. At specific time intervals, the of the PEC chitosan-alginate films and PEC chitosan-
absorbance of the DPPH radical solution was measured alginate films incorporating eugenol are shown in Fig.
at 517 nm using a UV-Vis spectrophotometer. Radical 1.
scavenging activity (RSA) was defined as the decreasing The spectra of the chitosan powder (spectrum a)
absorbance of the sample compared with the DPPH shows characteristic peaks at wavenumbers of 1659
control solution. It was calculated using this equation: and 1597 cm–1. The peak at 1659 cm–1 is due to the
 Abs sample  stretching vibration of C=O groups (amide I) and the
RSA  %    1    100% (10) peak at 1597 cm–1 corresponds to the stretching
 Abs control 

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86 Indones. J. Chem., 2018, 18 (1), 82 - 91

vibration of the N−H bending from amine and amide II.

These results are similar to the observation of Peng et
al. [6]. For the alginate, there are strong peaks at 1605
cm–1, representing the stretching vibration of
antisymmetric –COO–, and at 1420 cm–1, representing
symmetric –COO–. Spectrum (c) shows that interaction
occurred between the amino groups of the chitosan
and the carboxylate groups of the alginate, so the
characteristic bands at 1597 and 1605 cm–1 shifted to
the lower wavenumber of 1566 cm–1. This shift could
be due to molecular interaction between the chitosan
and alginate, which were under the different ionic
circumstances.
When eugenol was added into the PEC chitosan-
alginate films, the interaction between the eugenol and
the PEC chitosan-alginate films is indicated by changes
in PEC films’ bands, which shifted to a lower
Fig 1. FTIR spectra for (a) A2, (b) PEC film, (c) eugenol, wavenumber (from 1566 to1558 cm–1). Other changes
(d) alginate, and (e) chitosan also occurred at a wavenumber between 1342 and
1327 cm–1. This was possibly caused by the effect of
the benzene ring in the eugenol structure. New spectra
peaks appear at wavenumbers of 1149 cm–1 (C−O) and
896 cm–1 (C-C), which indicate that a strong interaction
occurred between the PEC chitosan-alginate films and
the eugenol. Similar results are also observed by Peng
et al. [6], who report the effect of the addition of tea
extract to the chitosan composite films.

Thermal Analysis

The thermal degradation behavior of films can be

represented by a thermogravimetric analysis (TGA)
thermogram, which can be used to investigate the loss
of mass of films as a result of the decomposition
process. As illustrated in Fig. 2, a single degradation
step was observed for the A1 and B3 films. For the A1
film, a weight loss of 18.01% was obtained at a
temperature (Td) of 106.25 °C. A higher weight loss
was noted for the B3 film when a higher level
concentration of both alginate and eugenol was added
to the films. The weight loss for the B3 film was 23.61%
at a temperature (Td) of 111.35 °C. The higher weight
loss of the B3 film is possibly due to the higher content
of water because of the presence of a higher
concentration of alginate, corresponding to their
hydrophilicity. The TGA reveals that the addition of
eugenol did not significantly affect the thermal
degradation profiles of the films. This means that
eugenol could have been lost during the processing
because the observed temperature was above the
volatilization point of eugenol. A similar result was
obtained by Ramos et al. [24].
The differential thermal analysis (DTA) technique
was employed in this research; it is generally used to
Fig 2. TGA and DTA curves for films A1 (a) and B3 (b) investigate the thermal transition, glass transition (T g),

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 Indones. J. Chem., 2018, 18 (1), 82 - 91 87

Table 3. Physico-mechanical properties of PEC chitosan-alginate films

Samples Thickness Tensile Elongation at WVP Transparency
code (mm) strength (MPa) break (%) (x 10-9 g det–1 m–1 Pa–1) value
A1 0.0214 14.66 ± 0.11 4.31 ± 0.75 1.61 ± 0.02 1.86 ± 0.23
A2 0.0222 18.53 ± 1.97 3.65 ± 0.57 1.96 ± 0.01 2.57 ± 0.17
A3 0.0214 20.21 ± 7.11 3.66 ± 0.66 2.24 ± 0.03 3.58 ± 0.15
B1 0.0222 11.49 ± 1.33 3.52 ± 0.09 2.89 ± 0.03 2.01 ± 0.04
B2 0.0222 18.53 ± 1.97 3.65 ± 0.57 1.96 ± 0.01 2.57 ± 0.17
B3 0.0218 5.93 ± 0.04 4.81 ± 3.26 1.67 ± 0.01 4.08 ± 0.36

crystallization (Tc), and melting temperature of a difference was observed between the EAB values for
polymer. From the curve achieved, the A1 and B3 films A2 and A3. The effect of adding eugenol at a higher
exhibited endothermic peaks at 117.14 °C (ΔHm = concentration also increased the EAB. The same factor
801.88 J g–1) and 121.57 °C (ΔHm = 861.44 J g–1), could be used to explain this phenomenon. The effect
respectively, corresponding to their melting temperatures of a plasticizer, such as glycerol, sorbitol, or
(Tm). The higher alginate composition of the B3 film polyethylene glycol, was investigated by Bourtoom [19].
resulted in a higher melting temperature. This is due to A similar effect was found when this plasticizer was
the stronger molecular interaction that occurred between blended into rice-starch-chitosan films.
the chitosan and alginate.
Water Vapor Permeability
Thickness and Mechanical Properties
As shown in Table 3, the addition of alginate into
The data on the thickness and mechanical the composition of PEC films increases the WVP value.
properties of the PEC films incorporating eugenol are The A3 film shows the highest WVP value. A
shown in Table 3. The films incorporating eugenol show composition with a higher level of alginate increases
similar values. This means that neither the composition the free volume of chitosan, allowing for more water
of PEC nor the concentration of eugenol affected the diffusion. This could be due to the hydrophilicity of
thickness of the films. films. The addition of alginate at higher levels of
The A3 film demonstrated the highest TS and the concentration increases the amount of hydrophilic
lowest EAB. As illustrated in Table 3, the addition of the groups in the content of the film, meaning water can be
alginate to the composition of PEC film increases the TS transferred easily to the films. Similar results were
of the film (TS of A3 > A2 > A1). It implies that the higher obtained by Fakhoury et al. [20].
the amount of alginate in the composition, the stronger The WVPs of the PEC chitosan-alginate films,
the molecular interaction in the PEC chitosan-alginate, containing 0.05%, 0.10%, and 0.20% eugenol, are
which is due to the amino groups of chitosan and shown in Table 3. The WVP value of the B3 film is the
carboxylate groups of alginate being mainly ionized. The lowest compared with the others. Migration of water
incorporation of a higher amount of eugenol affected the vapor occurred on the hydrophilic site of the films.
PEC films in a different way. An increase in TS was When eugenol was added into the films at higher level
observed in the B2 film when the final concentration of of concentration, it can increase the hydrophobicity of
eugenol was 0.10%. This indicates that the ionic films, thereby decreasing the ability of water vapor to
interaction between the chitosan and alginate in the B2 migrate through the film. Rubilar et al. [12] also report
film is stronger than in the B1 film. A significant decrease that the addition of a hydrophobic agent, such as
in TS was observed in the B3 film. The addition of a carvacrol or grapeseed extract, into chitosan films
hydrophobic agent into the film causes discontinuities in reduces the WVP values of films. Similarly, the addition
the film structure and less chain mobility, so it will of a root essential oil into gelatin films also decreases
decrease flexibility and resistance to fracture. The same the WVP because of the hydrophobicity of films [21].
behavior was also found when carvacrol was mixed into
fish-skin gelatin films in a higher concentration [12]. Film Transparency Value
The highest EAB value was detected for the A1
film. When the amount of alginate in the composition One of the important factors to be considered for
increased, the EAB of films A2 and A3 decreased. This food packaging film is film transparency. A
could be due to the weaker interaction between the transparency parameter is used to explain the size of
chitosan and alginate in the A1 film. Consequently, the particles dispersed in chitosan matrix films. The
incorporation of eugenol, which can be a plasticizer, will transparency value of each film is shown in Table 3.
increase the free volume of chitosan, causing a reduced The greater the transparency value indicated, the lower
TS, but an increased EAB value. However, no significant the transparency of the film. The addition of alginate at

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88 Indones. J. Chem., 2018, 18 (1), 82 - 91

Fig 3. Profile of the release of eugenol from PEC films Fig 4. Profile of the release of eugenol from PEC films
for different PEC compositions, with films’ mass at different levels of eugenol concentration: B1
proportions of chitosan:alginate of 9:1 (A1), 8:2 (A2), and (0.05%), B2 (0.10%), and B3 (0.20%)
7:3 (A3)

a higher level of concentration into a film leads to an The release profiles for eugenol from PEC
increase in the transparency value. This implies that chitosan films are shown in Fig. 3 and Fig. 4, which
bigger particles were dispersed as a result of the higher reveal that the percentage of eugenol release was very
alginate concentration, as the light that passes through low. The A2 film had the highest percentage of eugenol
the film matrix was absorbed by the particles, causing a release, which is 1.45% in four days (Fig. 3). Fig. 4
higher transparency value. The B3 film has the highest indicates the highest percentage of eugenol release
transparency value. The results in Table 3 demonstrate occurred in the B1 film. Gargiulo et al. [10] explain that
that the incorporation of eugenol at a higher level the release of an active compound from a polymer
concentration can decrease the transparency of films. A matrix consists of two steps. First, there is diffusion of
similar result was obtained by Tongnuanchan et al. [21], the solvent from the outer solution into the film pores to
who state that the addition of essential oils from plai, dissolve the active compound. Second, there is the
ginger, and turmeric roots in higher concentrations diffusion of the dissolved active compound out of the
decreases the transparency of films. Hosseini et al. [22] film pores. Gargiulo et al. [10] report that, when the
also describe the effect of incorporating clove, thyme, hydrophobicity of the pore surface increases, it takes
and cinnamon essential oils, which also increase the more time for an ethanol solvent to diffuse from the
transparency value. outer solution to the pores [10]. Thus, the release of α-
tocopherol from a film pores depends on the
Study of the Release of Eugenol hydrophilicity of the films. The release of eugenol from
PEC chitosan-alginate films could also be related to the
Antioxidant packaging is one of the active hydrophilicity of PEC films.
packaging techniques, which allows a controlled release The hydrophilicity of the films can be reflected by
of an antioxidant from the packaging film to the surface their water absorption value. The water absorption test
area of the food. The release behavior of an antioxidant identifies that the addition of a higher level of alginate
is related to factors such as solvent, polymer solubility, increased the percentage of water absorption. The
and swelling properties. A food simulant is generally water absorption percentages for A1, A2, and A3 are
used to investigate the release behavior of an 46.25, 48.90, and 51.60%, respectively. Based on what
antioxidant in packaging films. Eugenol is a hydrophobic is shown in Fig. 3, there was a decrease in eugenol
active compound; therefore, it is suitable for high-fat release in the A3 film, although its water absorption
foods. Ethanol 96% has a similar hydrophobicity to high- value is higher than for the A2 film. This is due to the
fat foods, so it can be used as a food simulant for strong molecular interaction between the PEC
eugenol. chitosan-alginate film and the eugenol, and supported

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 Indones. J. Chem., 2018, 18 (1), 82 - 91 89

Table 4. Kinetics data from various models for PEC chitosan-alginate films
Sample Zero-order First-order Higuchi Korsmeyer-Peppas
code k r k r k r k R n
A1 0.0134 0.9272 5 × 10-5 0.9275 0.0532 0.9743 0.4307 0.9911 0.16
A2 0.0221 0.8993 8 × 10-5 0.8996 0.0895 0.9656 0.6502 0.9902 0.18
A3 0.0160 0.9611 6 × 10-5 0.9613 0.0607 0.9690 0.6070 0.9614 0.14
B1 0.0110 0.9360 8 × 10-5 0.9364 0.0868 0.9777 0.8209 0.9931 0.15
B2 0.0221 0.9005 8 × 10-5 0.9009 0.0902 0.9663 0.8209 0.9902 0.18
B3 0.0529 0.9129 9 × 10-5 0.9133 0.1050 0.9595 0.4984 0.9718 0.24

by the higher TS of A3 compared to A2 (Table 3). Thus,

it is more difficult for the eugenol in the A3 film to
dissolve in the ethanol solution, which causes a
reduction in the percentage of eugenol release. Fig. 4
reveals that the highest eugenol release occurred in the
B1 sample. This can be explained by the hydrophilicity of
each film. The water absorptions for B1, B2, and B3, are
51.48, 48.90, and 44.31%, respectively. The higher
hydrophilicity of the films also resulted in a higher
percentage of eugenol release.
The release rate of eugenol from the PEC
chitosan-alginate films is very slow. This process is
called sustained release. It indicates that the eugenol
incorporated into the PEC films has a water-soluble
character. The fast release of an active compound
usually causes toxicity because of the very high
concentration of the active compound released. For this
reason, the sustained release of an active compound
Fig 5. Antioxidant activity of PEC chitosan-alginate
from the chemical matrix is more effective than fast
films incorporating eugenol
release.
Because the n values of all films are less than 0.5, it is
Kinetics Model for Eugenol Release
revealed that the release mechanism of eugenol is
controlled by a Fickian diffusion process. This
The kinetics model for eugenol release can be
mechanism explains that the release of eugenol from
used to explain the mechanism for releasing eugenol
PEC chitosan-alginate films occurs through a diffusion
from PEC chitosan-alginate films. For this research, he
process without the disintegration of the PEC chitosan-
kinetics model for eugenol release is described using
alginate films.
four models: the zero-order kinetics model, first-order
kinetics model, Higuchi kinetics model, and Korsmeyer-
Assessment of Antioxidant Activity
Peppas kinetics model. The kinetics data for the eugenol
release in the various models are shown in Table 4, and,
The antioxidant properties of films were analyzed
based on this data, we can assume that the zero-order
and determined using a DPPH radical. This DPPH
model, first-order model, and Higuchi model do not
radical is one of the very stable nitrogen radicals, and
express the release mechanism for eugenol. Plotting the
measuring its scavenging activity is an important
regression from the zero-order model, first-order model,
method for determining the antioxidant capacity of films
and Higuchi model illustrate the correlation coefficient
[23]. Fig. 5 shows the presence of eugenol in PEC
values (r) of the films are not good. A high linearity is
chitosan-alginate films increased the activity of DPPH
expressed by a high value for the correlation coefficient
radical scavenging. The addition of eugenol at a higher
(the r value is close to 1.0).
level of concentration caused increased DPPH radical
Among all the models, the correlation coefficient
scavenging activity. The greatest DPPH radical
value for the Korsmeyer-Peppas model, for all films, is
scavenging activity of the films is about 55.99% (Fig.
close to 1.0. This indicates that the mechanism for the
5).
release of eugenol from PEC chitosan-alginate films
The incorporation of eugenol into PEC chitosan-
follows the Korsmeyer-Peppas model, and the n values
alginate films at a higher level of concentration causes
of all films are less than 0.5. The release exponent value
a higher concentration of eugenol to be released from
(n) indicates an active compound release mechanism.

Baiq Amelia Riyandari et al.

90 Indones. J. Chem., 2018, 18 (1), 82 - 91

Fig 6. SEM micrograph of films A1 (a) and B3 (b)

films. Based on the data shown in Fig. 5, the DPPH eugenol from PEC films. Higher concentrations of
radical scavenging activity values for the B1, B2, and B3 alginate and eugenol increased the concentration of
films are 35.72, 46.69, and 55.99%, respectively. The eugenol that was released from the films. The
greatest DPPH radical scavenging activity appeared in mechanism for the release of eugenol from PEC
the B3 film, which is due to the release of a higher chitosan-alginate films followed the Korsmeyer-Peppas
concentration of eugenol from the B3 film than both the model and was controlled by a Fickian diffusion
B1 and B2 films. From this release study, the process. An assessment of the antioxidant activity of
concentration of eugenol released from the B3 film is the films conducted using the DPPH method showed a
about 8.18 µg mL–1 for 96 h (four days). Meanwhile, the high RSA value of 55.99% in 96 h. Based on the
concentrations of eugenol released from the B1 and B3 mechanical properties, water permeability, and
films are 2.38 and 4.15 µg mL–1, respectively. A similar transparency value, PEC chitosan-alginate films
result was obtained by Woranuch and Yoksan [13]. The incorporating eugenol could be potentially used as
addition of eugenol-loaded chitosan nanoparticles into antioxidant packaging.
TPF resulted in a DPPH radical scavenging percentage
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