LWT - Food Science and Technology 61 (2015) 484e491

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LWT - Food Science and Technology

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

Physicochemical and antimicrobial properties of nanoencapsulated

Eucalyptus staigeriana essential oil
^nia A.T. de Figueiredo b,
Emanuele D. Herculano b, Haroldo C.B. de Paula a, *, Eva
Flayanna G.B. Dias b, Vanessa de A. Pereira a
a
b

, Fortaleza, 60455-760, Ceara

, Brazil
Department of Analytical and Physical Chemistry, Building 940, Federal University of Ceara
, Fortaleza, 60356-000, Ceara
, Brazil
Department of Food Science and Technology-DETAL, Building 858, Federal University of Ceara

a r t i c l e i n f o

a b s t r a c t

Article history:
Received 25 June 2014
Received in revised form
7 October 2014
Accepted 1 December 2014
Available online 9 December 2014

In this work Eucalyptus staigeriana essential oil (ESO) was nanoencapsulated using cashew gum (CG) as
wall material. The nanoparticles had their antimicrobial activity against Listeria monocytogenes (Grampositive) and Salmonella Enteritidis (Gram-negative) evaluated by determining their Minimum Bactericidal Concentration, in addition to being characterized by infrared spectroscopy, thermal analysis, particle size distribution and zeta potential. Data from MBC showed greater activity against Gram-positive
bacteria, due to a likely synergistic effect between the CG and ESO. The nanoparticles had sizes ranging
from 27.70 nm to 432.67 nm and negatively charged surfaces. The ESO content varied between 4.76% and
7.12% and the encapsulation efciencies from 24.89% to 26.80%. The aforementioned data suggest that
ESO nanoparticles have potential for use as a natural food preservative.
2014 Elsevier Ltd. All rights reserved.

Keywords:
Nanoencapsulation
Cashew gum
Eucalyptus staigeriana
Salmonella Enteritidis
Listeria monocytogenes

1. Introduction
The nanoencapsulation of labile bioactive compounds represents an efcient alternative to increase their physical stabilities,
protecting them from deleterious interactions with food components and the environment, besides increasing their bioactivity
mainly owning to their nanometer size range (Dons, Annunziata,
Sessa, & Ferrari, 2011). Upon encapsulation, sample can now be
easily handled because the active principle is protected against
oxidation and other environmental harms. Moreover, its volatile
ingredients are now retained in a particular matrix, exhibiting
prolonged sensory perception, bioavailability and improved efcacy (Neethirajan & Jayas, 2011).
The extracts obtained from leaves of Eucalyptus have been
approved as food additives, being also used in cosmetic formulations, although most studies have been focused on their functional
properties (Gilles, Zhao, An, & Agboola, 2010).
Several properties have been assigned to the genus Eucalyptus,
such as antimicrobial (Tyagi & Malik, 2011), antihyperglycemic

* Corresponding author. Tel.: 55 85 3366 9961.

E-mail address: hpaula@ufc.br (H.C.B. de Paula).
http://dx.doi.org/10.1016/j.lwt.2014.12.001
0023-6438/ 2014 Elsevier Ltd. All rights reserved.

(Gray & Flatt, 1998), antioxidant (Santos et al., 2012) and anthelmintic (Macedo et al. 2010). A number of researches have been
done to demonstrate the antimicrobial properties of the Eucalyptus
essential oils. However, these studies are concentrated in a few
species, especially Eucalyptus citriodora that has a broad spectrum
of antifungal activity. Only a few studies have been conducted to
assess the activity against pathogenic and food spoilage bacteria, as
well as yeast (Gilles et al., 2010).
Cashew gum is a heteropolysaccharide extracted from the
exudate of Anacardium occidentale, an abundant tree in the Brazilian Northeast region, whose structure resembles gum Arabic.
Cashew gum is able to interact with water and thus act as stabilizer,
emulsier and adhesive, and may be a suitable substitute for gum
 & Rao, 2000).
Arabic, which is more expensive (Mothe
Taken into consideration the need for more research on the
formulation and application of nanoparticles with natural antimicrobials in food, this work aimed at the preparation of nanoparticles of cashew gum (CG) loaded with Eucalyptus staigeriana
essential oil (ESO), aiming to preserve the active ingredient and
prolong its release to the medium. Moreover, Minimum Bactericidal Concentration of ESO, CG and ESO loaded nanoparticles on
Listeria monocytogenes and on Salmonella Enteritidis bacteria
were determined, in order to evaluate their antimicrobial actions.

E.D. Herculano et al. / LWT - Food Science and Technology 61 (2015) 484e491

Furthermore, the nanoparticles were characterized regarding

their physicochemical properties and the effect of the wall material and surfactant ratio on zeta potential, particle size, loading
and encapsulation efciency.
2. Materials and methods
2.1. Materials
E. staigeriana essential oil (FERQUIMA, Brazil), ethanol 99% and
Tween 80 surfactant (VETEC, Brazil) were used as received.
Cashew gum (1.1  105 g/mol) was extracted from native trees of
Cear
a, and puried as described by De Paula, Heatley, and Budd
(1998).
2.2. Nanoparticle preparation
An organic phase was obtained by mixing Tween 80 (T80) and
E. staigeriana essential oil in a vortex mixer (PHOENIX, Model AP 56,
Brazil) under continuous stirring. The above mixture was dripped
into an aqueous solution of cashew gum (CG) 2% (w/v) under
constant stirring (18,000 rpm) in an ultrasonic homogenizer Turratec (TECNAL, model TE-101, Brazil) for 2 min. Three formulations
were established: F.1, with ratios (m/m) for CG:ESO 2:1 and
ESO:T80 2:1; F.2, with CG:ESO 4:1 and ESO:T80 2:1 and F.3,
with CG:ESO 2:1 and ESO:T80 1:1.
The emulsion obtained was fed into a Mini Spray Dryer B-290
(BCHI, Switzerland), at an inlet temperature of 160 10  C and an
outlet temperature of 70 10  C, pump feed ow of 5 mL/min, air
volume ow of 35 m3/h, and aspirator ow 84 L/h (Paula, Sombra,
Abreu, & De Paula, 2010). The nanoencapsulated sample was stored
in amber glass at room temperature (25  C).
2.3. Minimum Bactericidal Concentration
The Minimum Bactericidal Concentration (MBC) of the nanoparticles was determined in order to evaluate its antimicrobial
action. The nanoparticle concentrations tested ranged from 0.5 g/L
to 7 g/L of the active ingredient (D-limonene). It was also determined the MBC of bare essential oil, and of cashew gum solutions.
As controls were employed culture media and culture
media bacterial inoculum. The experiments were performed in
duplicate on three different days (n 6) according to Sahm, &
Washington (1991). Briey, selected strains of Gram-positive
pathogenic bacteria, L. monocytogenes (ATCC 19115- Microbiologics) and Gram-negative, Salmonella Enteritidis (IAL 1132- Adolfo
Lutz Institute, Brazil) were used. The lyophilized strains were
activated as recommended by the manufacturer.
In test tubes containing 3.95 mL of Tryptone Soy Broth (TSB/
Difco) sterile for L. monocytogenes and Infusion Brain Heart Broth
(BHI/Difco) for S. Enteritidis was added 50 mL of bacterial suspension in concentration of 105 UFC/ml. For each culture, were added
separately the E. staigeriana essential oil, the nanoparticle solution
and cashew gum solution. The tubes were incubated at 35  C for
24 h under stirring at 200 rpm.
After incubation, 100 mL of culture were inoculated on plates
containing the Listeria Oxford Medium (OXA/Oxoid) for
L. monocytogenes and Hecktoen Enteric Agar (HE/Difco) for S.
Enteritidis, and spread with handle Dringalsky. The plates were
incubated at 35  C for 24 h and observed for the presence of bacteria growth. The lowest concentration at which no growth
occurred was taken as the MBC. Determination of Minimum
Inhibitory Concentration was not possible due to the occurrence of
turbidity of the medium when the samples were added.

485

2.4. Nanoparticles characterization

2.4.1. GCeMS analysis
The composition of ESO was determined by gas
chromatography-mass spectrometry (GCeMS) using an SHIMADZU
equipment model QP2010 SE, (Japan), equipped with a capillary
column Rtx-5MS, Restek, Germany (30 m, ID 0.25 mm, lm thickness 0.25 mm) and auto injector AOC-20i. Helium was used as carrier gas (1 mL/min). The initial column temperature was kept at
60  C for 3 min and then was increased to 280  C at a rate of 3  C/
min. The mass selective detector (SHIMADZU) was used in the
electron ionization mode (70 eV), the mass range between 35 and
500. Ion source (260  C) and transfer line (300  C) temperatures
were employed throughout analyses.
2.4.2. Essential oil loading and encapsulation efciency
The encapsulated oil content was determined by absorption
spectroscopy in the UVeVis region (Micronal, model B582, Brazil),
at a wavelength of 210 nm, according to Paula et al. (2010). Oil
concentration was determined by crushing a 10 mg sample in
ethanol and calculating its concentration using a calibration curve
(Eq. (1)). All analysis were performed in triplicate and conventional
statistical methods (ANOVA, followed by comparison with the
Tukey test) were used to calculate average and standard deviations.
Differences between means were considered to be signicant when
p < 0.05.

abs 0:02602conc 0:0807

R2 0:991

(1)

Where abs signies absorbance and conc represents the oil concentration in ppm. Oil loading is given by the ratio of corrected ESO
mass obtained from the calibration curve of Eq. (1) and the sample
mass.
The encapsulation efciency (E.E) was calculated according to
Eq. (2):

E:E% M=M0  100

(2)

Where M is the amount of ESO in the loaded sample (mg), as

determined from Eq. (1) and M0 is the initial added ESO amount
(mg).
2.4.3. FT-IR spectroscopy
The particles were characterized by Fourier transform infrared
spectroscopy (FT-IR) using KBr pellets on a Perkin Elmer device,
using wavenumbers in the range 400e4000 cm1, with a resolution
of 4 cm1.
2.4.4. Thermal analysis
The thermal stabilities of the nanoparticles was determined by
thermogravimetric analysis (TGA) in a Shimadzu-TGA-50 (Japan)
under a nitrogen atmosphere, applying a heating rate of
10  C min1, from 25 to 900  C, and by differential scanning
calorimetry (DSC) in a Shimadzu-DSC-50 (Japan), with a heating
rate of 10  C min1, from 25 to 400  C according to Paula et al.
(2010).
2.4.5. Particle size distribution and zeta potential
Particle size distribution and zeta potential of the sample
solution were determined by a Zetasizer Nano (Malvern, model
3600) using a red light beam with a wavelength of 633 nm and
measuring an angle of 175 . Data were expressed as the averages
of 3 readings (Paula, Sombra, Cavalcante, Abreu, & De Paula,
2011).

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E.D. Herculano et al. / LWT - Food Science and Technology 61 (2015) 484e491

2.5. In vitro release kinetics

Release kinetics was carried out by dissolving 100 mg of
sample in 10 mL distilled water and transferring to a dialysis
membrane with pore size of 14 kDa (Sigma), which was then
immersed in 200 mL distilled water. Aliquots were taken at certain
time intervals and analyzed by spectrophotometry in the UVeVis
region. Oil concentration was determined using a calibration curve
(Eq. (3)):

Abs 0:02354conc 0:0091

R2 0:998

(3)

Kinetic parameters were calculated in order to characterize the

various types of release. The models used were the zero, rst and
second order, Higuchi, and Korsmeyer-Peppas (Dash, Murthy, Nath,
& Chowdhury, 2010; Paula et al., 2010). Ritger and Peppas (1987)
used this value to characterize different release mechanisms. According to the model kinetic theory for spherical geometry, n, a
diffusion coefcient, can take different values.
2.6. Nanoparticle storage stability
ESO contents in nanoparticles was monitored as a function of
time (365 days), in order to determine the ability of the samples to
maintain the oil within the polymer matrix, at storage. The powder
samples were stored in amber glass containers with a capacity of
30 mL, headspace of about 0.5 cm and kept protected from exposure to light on racks at 25 2  C. At certain time intervals, dry
samples were smashed in ethanol, aliquots were taken and
analyzed by spectrophotometry in the UVeVis region, as described
in Section 2.4.2.
3. Results and discussion
3.1. GCeMS analysis
Table 1 shows the results for GCeMS. The main constituent was
found to be limonene (23.20%), followed by geranial (7.98%), methyl
geranate (6.29%) and 1,8-Cineole (6.03%). Gilles et al. (2010) reported as main constituents 1,8-Cineole (34.8%), neral (10.8%),
geranial (10.8%) and a-phellandrene (8.8%). A higher limonene
concentration (72.9%) was described by Ribeiro et al., 2013. The
abovementioned data is in agreement with the fact that essential

Table 1
Composition of the Eucalyptus staigeriana essential oil (ESO).
Retention time (min)

Component

(%)

5.636
6.834
8.362
8.565
8.635
11.099
12.471
14.348
14.721
14.944
16.576
17.190
17.809
18.548
19.828
20.836
22.037
22.568
23.448

a e Pinene
b e Pinene

2.62
1.54
4.95
23.20
6.03
2.59
0.37
1.29
2.55
1.22
1.70
5.20
5.16
7.98
1.19
6.29
0.43
2.49
5.90
17.28

o e Cymene
Limonene
1,8-Cineole
Linalool
cis-Limonene oxide
Terpinen-4-ol
p e Cymen-8-ol
a e Terpineol
Nerol
Neral
Geraniol
Geranial
Geranyl formate
Methyl geranate
Citronellyl acetate
Neryl acetate
Geranyl acetate
Other constituents

oil composition depends on several factors such as climate and soil,

therefore it is expected that oils from different sources present
discrepancies in their component contents.
3.2. Minimum Bactericidal Concentration
Table 2 shows the results for MBC. For nanoparticles, the MBC
for S. Enteritidis (Gram-negative) was 4 g/L while for
L. monocytogenes (Gram-positive) was 3 g/L, pointing out to
bactericidal action being more effective to Gram-positive than to
Gram-negative.
As regard to the free oil, the bactericidal effect was shown to be
more competent for the Gram-positive bacteria, where a lower oil
concentration (2 g/L) is required to the L. monocytogenes compared
to the Gram-negative (5 g/L). These data are in agreement with
Tyagi and Malik (2011) and Jouki, Yazdi, Mortazavi, and Koocheki
(2014). MBC data also show that the nanoencapsulated oil exhibited enhanced activity against S. Enteritidis, being more efcient
than pure oil. However, for L. monocytogenes the opposite effect was
observed.
Divalent cations act by reducing the repulsion between molecules of lipopolysaccharide (LPS), which are highly anionic, present
in the outer membrane of the cell wall of Gram-negative bacteria.
Mg2 has the important role to form complexes between the proteins of the outer membrane and LPS. Treatment with chelating
agents such as ethylenediamine tetraacetic acid (EDTA) affects the
properties of the outer membrane of bacteria (Coughlin, Tonsager,
& Groarty Mc, 1983; G
anzle, Weber, & Hammes, 1999).
As cashew gum contains carboxylate (-COO) groups and hence
a negative charged surface, it may acts as a coupling agent for
magnesium ions of the lipopolysaccharide present in the cell wall
of Gram-negative bacteria such as S. Enteritidis, causing it to
become sensitive. Moreover, the nanoencapsulated essential oil in
cashew gum seem to exhibit a synergistic effect against those
bacteria, while the free oil had a lesser ability to penetrate the
bacteria outer membrane. As L. monocytogenes does not have an
outer membrane, the free oil was able to easily penetrate the
bacteria cell wall and sensitize the phospholipid bilayer of the cell
membrane, with consequent increased permeability and leakage of
vital intracellular compounds.
Dons et al. (2011) obtained for D-limonene MBC values greater
than 25 g/L for bacteria tested, whereas for the same compound
encapsulated in nanoemulsions, data were always equal to or lower
than that gure. For CG solutions used in this work, all plates
showed bacterial growth, revealing no action on bacteria. Campos
et al., (2012) determined the MBC for puried and raw CG,
reporting that it was not possible to determine the MBC for puried
cashew gum, for all tested bacteria. However, for the raw gum, it
was reported an MBC of 5 g/L. The authors attributed this result to
the gum purication process.
3.3. Nanoparticles characterization
3.3.1. Essential oil loading and encapsulation efciency
Table 3 shows data for encapsulated oil content and E.E. for the
different nanoparticle formulations. It can observed that a
Table 2
Minimum Bactericidal Concentration (MBC) of CG-ESO nanoparticles and ESO
against S. Enteritidis and L. monocytogenes.
MBC (g/L)

CG-ESO nanoparticles
ESO

S. Enteritidis

L. monocytogenes

4
5

3
2

E.D. Herculano et al. / LWT - Food Science and Technology 61 (2015) 484e491
Table 3
Oil loading and encapsulation efciency (E.E) for different CG-ESO nanoparticle
samples.
Sample

CG:ESO (m/m)

ESO:T80 (m/m)

E.Ea (%)

Oil loadingb (%)

F.1
F.2
F.3

2:1
4:1
2:1

2:1
2:1
1:1

24.89
26.16
26.80

7.12
4.76
6.70

a
b

Error: 0.56e1.90 %.
Error: 0.42e1.94 %; T80 Tween 80.

reduction of ESO and the consequent increase in the ratio of surfactant led to an increase in encapsulation efciency from 24.89% to
26.80% (samples F.1 and F.3), although there was a slight loading
reduction from 7.12% to 6.70%.
Comparing F.1 and F.2 samples it can be seen an increase in E.E
to 26.16% and a reduction in the loading to 4.76%, the latter being
explained by the fact that for F.2 sample, loading is related to a
higher CG content. Thus, it can be inferred that augmenting the
proportion of the matrix (CG) there was an increase in E.E, although
with lower loading. Similar conclusions were obtained by Paula
et al. (2010). Chitosan nanoparticles loaded with oregano essential oil present E.E between 5.45% and 24.72% and loading from
1.32% to 2.12% (Hosseini, Zandi, Rezaei, & Farahmandghavi, 2013).
3.3.2. FT-IR spectroscopy
Structural characterization by FT-IR spectroscopy is shown in
Fig. 1. Spectra reveal that CG showed a broad band at 3379 cm1 due
to the stretching vibration of OeH, a band at 2933 cm1 assigned to
the CeH stretching vibration and absorption at 1649 cm1, due to
vibrations OeH from the water molecules. Strong bands at 1150,
1080 and 1030 cm1 are due to stretching vibrations of CeOeC of
glucosidic bonds and OeH bending of alcohols (Da Silva, Feitosa,
Paula, & De Paula, 2009). The ESO spectrum showed a broad
band at 3438 cm1 corresponding to OeH stretching vibration from
alcohols; a band at 2969 cm1, due to stretching vibration of the
methyl groups (eCH3); at 2929 cm1 owing to methylene groups
(eCH2e); at 1721 cm1 related to the carbonyl C]O stretching; at
1672 cm1 from to C]C stretching of aromatic groups and at
1445 cm1 due to the CeH deformation (Esteves, Marques,
Domingos, & Pereira, 2013; Sheet, Saleh, & Hamed, 2007). The
presence of the ESO in the nanoparticles can be mainly observed in
spectra through appearance of new band (at 1735 cm1) and
changes (broadening) in bands of CG spectrum (in the region

Fig. 1. FT-IR spectra of cashew gum (CG), Eucalyptus staigeriana essential oil (ESO) and
CG/ESO nanoparticles.

487

2852e2904 cm1, CeH stretching; at 1649 cm1 due to CG COO

and water). It is also worth notice a relative increase in the intensity
at 2933 cm1 and a decrease at 1080 cm1. Changes were more
noticeable for sample F.3.
3.3.3. Thermal analysis
3.3.3.1. Thermogravimetric analysis. The thermogravimetric curves
are presented in Fig. 2. The data of mass losses and peak temperatures for the thermal events are shown in Table 4. The gum
decomposition occurs in three stages, where the rst is related to
moisture loss and the others are associated with the CG degradation through depolymerization and subsequent formation of water,
CO and CH4 (Da Silva et al., 2009). Degradation occurred at 310  C
and 497  C. Lower values were reported for other polysaccharide
such as dextran, guar gum and chitosan (Silva, Feitosa, Maciel,
Paula, & De Paula, 2006). At temperatures up to 200  C the CGESO nanoparticles were found to exhibit greater stability than CG,
(Fig. 2a) although being less stable at temperatures above 400  C.
F1 sample was found to be the least heat labile, up to 400  C. CGESO nanoparticles exhibit a rst decomposition event in the
range 313e321  C, the second in the range of 400e510  C. Ash
contents at 600  C follow the order CG > F1 > F3 > F2, where the
low ash content for F.2 is likely to be due to the fact that a greater
CG:ESO ratio yield a gum excess which seem to form more heat
labile complexes with ions such as sodium and magnesium which
would decompose at temperatures below 600  C.
3.3.3.2. Differential scanning calorimetry. Fig. 3 and Table 5 show
the data obtained by DSC for CG and nanoparticles. CG showed an

Fig. 2. (a) Thermogravimetric analysis (TGA) and (b) dTG of CG and CG/ESO
nanoparticles.

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E.D. Herculano et al. / LWT - Food Science and Technology 61 (2015) 484e491

Table 4
Thermogravimetric analysis (TGA) for CG and different CG-ESO nanoparticles.
Sample

Events

Maximum temperature ( C)

Weight loss (%)

CG

I
II

310
497
Residual
319
400
Residual
313
510
Residual
321
392
Residual

46.50
34.80

F.1

I
II

F.2

I
II

F.3

I
II

mass at 600  C 10.56%

Table 5
Differential Scanning Calorimetry (DSC) for CG and different CG-ESO nanoparticle
samples.
Sample

51.44
29.36

CG
F.1
F.2

56.21
33.02

F.3

mass at 600  C 8.56%

mass at 600  C 3.52%

56.22
33.24


mass at 600 C 8.00%

endothermic peak at 152.6  C, corresponding to its melting temperature and a single exothermic peak at 307.3  C due to the
decomposition process. These temperatures were higher than
those values obtained by Okoye, Onyekweli, and Kunle (2012). The
events of nanoparticle endothermic transitions ranged from
137.9  C to 167  C whereas F.1 shows the highest melting temperature compared to F.2 and F.3, a clear indication of a stronger solventesolute interaction.
Regarding to exothermic transitions, nanoparticles showed a
slight reduction (in comparison to pure CG) at degradation temperature for the rst event, indicating that the presence of oil led to
a decrease of thermal stability at temperatures of approximately
300  C. Moreover, the second decomposition temperature was less
substantial for F.2 and F.3 and could not be detected for F.1.
3.3.4. Particle size distribution and zeta potential
Table 6 and Fig. 4 shows the particle size distribution by volume
and zeta potential. It can be seen that the samples F.1 and F.2
showed unimodal distribution, whereas F.3 had a bimodal one.
Regarding to particle size, it is observed that for samples with
unimodal distribution, F.1 has higher size (153.80 nm) than F.2
(27.70 nm). A similar result was obtained by Paula et al. (2012)
where it is reported that an increase in the proportion of cashew
gum in nanoparticle formulation leaded to a decrease in particle
size. Chitosan nanoparticles as a glazing material for cryogenically
frozen shrimp showed particle sizes ranging from 179.7 nm to
234.2 nm (Solval, Rodezno, Moncada, Bankston, & Sathivel, 2014).
Ilyasoglu and El (2014) reported particle size of 232.3 nm for
eicosapentaenoic acid and docosahexaenoic acid nanoparticles
with sodium caseinate-gum Arabic complex.

Exothermic transition

Endothermic transition

Temperature ( C)

DH (J/g)

Temperature ( C)

DH (J/g)

307.3
300.1
303.2
415.7
305.1
408.8

141.8
101.9
125.0
38.5
111.0
47.4

152.6
167.0
137.9

356.9
164.1
104.1

147.6

101.1

The zeta potential is an important parameter that characterizes

the net electric charge on a particle surface. Table 6 shows the
values of the zeta potential for the nanoparticles. It is observed that
all samples present negative zeta potential values. According to Da
Silva et al. (2009) negative zeta potentials are due to CG carboxylic
acid groups in the carboxylated form (eCOO) which make it a
slightly acidic polysaccharide. The zeta potential is indicative of the
stability of a nanoparticle and is considered good when it exceeds
the value of 30 mV (Silva et al., 2011). Therefore, F.1 is likely to the
be the most stable, while both F.2 and F.3 present reduced negative
surface charges and decreased stabilities in a water solution.
Jasmine essential oil loaded gelatin/gum Arabic nanocapsules
presented values ranging from 9.45 mV to 16.2 mV, at pH 7 (Lv,
Yang, Li, Zhang, & Abbas, 2014).
3.3.5. In vitro release kinetics
CG-ESO nanoparticles release kinetics are presented in Fig. 5.
F.3 sample shows a release prole similar to F.1, being faster in
the early hours, and gradually becoming distinct, after 20 h. F.1
presented a more prolonged release prole with only 67% of oil
released after 144 h, while F.3 released about 93% in 87 h. The increase in the proportion of CG (F.2 sample) favored a rapid oil
release, likely due to the increased nanoparticle hydrophilicity,
whereby within only 7 h of experiment around 90% of the oil was
released. Keawchaoon and Yoksan (2011) investigated the in vitro
release of carvacrol doped in chitosan nanoparticles, reporting a
22.5% release within 60 days, at pH 7. Data obtained from release
prole of F.1, F.2 and F.3 samples were applied to kinetic models and
the correlation coefcients are shown in Table 7. It can be observed
that the best correlation coefcients were obtained for KorsmeyerPeppas and Higuchi models, for all matrices, being followed by the
order zero model. A more detailed study on the Korsmeyer-Peppas
model was performed and its calculated kinetic parameters are
shown in Table 7. In accordance with the kinetic theory for spherical geometry model (Ritger & Peppas, 1987), when the coefcient n
is equal to 0.43, release is ruled by molecular diffusion due to a
chemical potential gradient (Diffusion Fickian or Case I). When it is
between 0.43 and 0.85, release has a non-Fickian behavior
(Anomalous transport), occurring through joint mechanisms,
diffusion and relaxation (Case II transport). When it is 0.85, equation corresponds to the release kinetics of zero order (Case II
transport), that is, the phenomenon of polymer relaxation. When n
is greater than 0.85 the diffusion is controlled by polymer swelling
and relaxation mechanisms (Super Case II transport) (Prajapati,
Richhaiya, Singh, Kumar, & Chaudhary, 2012).
Table 6
Particle size and zeta potential for different CG-ESO nanoparticles samples.

Fig. 3. Differential Scanning Calorimetry (DSC) of cashew gum (CG) and CG/ESO
nanoparticles.

Sample

Particle size (nm)

Volume (%)

Zeta potential (mV)

F.1
F.2
F.3

153.80 8.20
27.70 3.42
432.67 41.47
29.66 23.36

100.00
100.00
85.50
14.50

24.50 0.45
14.47 1.42
10.45 0.21

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489

Fig. 4. Particle size distribution by volume for the three nanoparticle formulations doped with E. staigeriana essential oil.

Fig. 5. Release kinetics of CG/ESO nanoparticles (a) F.1 and F.3; (b) F.2.

Data obtained for F.1 and F.3 showed release coefcients outside
the boundaries of Korsmeyer-Peppas model, whereas F.2 showed n
greater than 0.85, indicating a release mechanism by means of super
Case II transport. According to Shoaib, Tazeen, Merchant, and Yousuf
(2006) this law gives only a limited view about the exact mechanism
of release of the active principle. However, Gierszewska-Druzynska
and Ostrowska-Czubenko (2012) argue that this case is called the
Less Fickian behavior and can also be classied as Fickian diffusion.

to higher CG proportion in matrix, which probably provided the

formation of a more stable complex for oil stabilization. On the
other hand, F.1 and F.3 showed greater oil reduction amounting to
26.12% and 14.20%, respectively, indicating that these samples are
less capable of maintaining the oil trapped in their polymeric shell.
Gupta and Ghosh (2012) obtained reduction of 8.8% and 7.4% to bcarotene/ester and a-lipoic acid/ester nanocapsules, respectively,
for a period of 90 days.

3.3.6. Nanoparticle storage stability

Fig. 6 shows nanoparticle storage stability over 365 days. F.2 is
the best formulation for preserving the essential oil within matrix.
The initial oil content decreased circa 8%, which is likely to be due

4. Conclusions
Nanoparticle bactericidal action was shown to be more effective
to Gram-positive than to Gram-negative bacteria, where the

490

E.D. Herculano et al. / LWT - Food Science and Technology 61 (2015) 484e491

Table 7
Kinetic parameters for oil release from CG-ESO nanoparticles.
Samples

F.1
F.2
F.3

Correlation coefcient (R2)

Zero
order

First
order

Second
order

Higuchi

Korsmeyer e
Peppas

0.627
1.000
0.670

0.507
0.860
0.429

0.363
0.530
0.217

0.835
0.968
0.837

0.933
1.000
0.874

0.27
0.14
0.26

0.31
1.00
0.35

Fig. 6. Storage stability of CG/ESO nanoparticles.

nanoencapsulated oil exhibited enhanced activity against S.

Enteritidis. Data from different nanoparticle formulations revealed
that augmenting the proportion of the matrix (CG) there was an
increase in encapsulation efciency and a decrease in particle size.
The presence of the ESO in the nanoparticles was observed in FTIR
spectra through appearance of new bands characteristic of encapsulated oil and relative increase in band intensities. The nanoparticle stability upon storage was found to be dependent on the
CG:ESO ratio, preserving the essential oil properties within one
year. Data obtained suggests that CG-ESO nanoparticles are a
promising alternative to the use of chemical preservatives, being a
good candidate for natural food preservation.

Acknowledgments
 Foundation for
The authors wish to thank the FUNCAP (Ceara
Scientic and Technological Development) for the nancial
support.

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