Carbohydrate Polymers 157 (2017) 1874–1882

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Carbohydrate Polymers
journal homepage: www.elsevier.com/locate/carbpol

The release kinetics of ␤-carotene nanocapsules/xanthan gum coating

and quality changes in fresh-cut melon (cantaloupe)
María L. Zambrano-Zaragoza a,∗ , David Quintanar-Guerrero b , Alicia Del Real c ,
Elizabeth Piñon-Segundo d , José F. Zambrano-Zaragoza e
a
Universidad Nacional Autónoma de México, FES-Cuautitlán, Laboratorio de Procesos de Transformación y Tecnologías Emergentes en Alimentos, L-16,
Cuautitlán Izcalli, C.P. 54714, Estado de México, Mexico
b
Universidad Nacional Autónoma de México, FES-Cuautitlán, Laboratorio de Posgrado en Tecnología Farmacéutica, Av. 1o de mayo s/n, Cuautitlán Izcalli,
C.P. 54740, Estado de México, Mexico
c
Universidad Nacional Autónoma de México, Departamento de Ingeniería Molecular de Materiales, Centro de Física Aplicada y Tecnología Avanzada,
Campus Juriquilla, Querétaro, Qro., C.P. 76230, A.P. 1-1010, Mexico
d
Universidad Nacional Autónoma de México, FES-Cuautitlán, Laboratorio de Sistemas Farmacéuticos de Liberación Modificada, Cuautitlán Izcalli, Edo de
México, C.P. 54714, Mexico
e
Universidad Autónoma de Nayarit, Unidad Académica de Ciencias Químico Biológicas y Farmacéuticas, Tepic Nayarit, C.P. 63000, Mexico

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

Article history: The main aim of this work was to evaluate the effect of the ␤-carotene release rate from nanocapsules
Received 25 August 2016 incorporated into a xanthan gumcoating on the physical and physicochemical properties of fresh-cut
Received in revised form 8 November 2016 melon (var. cantaloupe). Several coatings were studied: xanthan gum alone (XG), xanthan gum combined
Accepted 26 November 2016
with nanocapsules (Ncs/XG), xanthan gum combined with nanospheres (Nsp/XG), nanocapsules (Ncs),
Available online 27 November 2016
and nanospheres (Nsp), all of which were compared to untreated fresh-cut melon in order to determine
their preservation efficiency. The ␤-carotene release profiles from the Ncs and Ncs/XG treatments corre-
Keywords:
sponded better to a Higuchi-type behavior (t1/2 ) for matrix systems (R2 > 0.95). Also observed was a good
Nanocapsules
Xanthan gum
correlation between the release of ␤-carotene by the Ncs/XG treatment and the minor changes observed
Controlled release in the whiteness index (≤10%) and firmness (≤2%). These results lead to the conclusion that incorporating
Coatings ␤-carotene nanocapsules into a polysaccharide matrix improves the properties of the coatings, thereby
Higuchi model increasing storage time to 21 days at 4 ◦ C.
© 2016 Elsevier Ltd. All rights reserved.

1. Introduction Cortez-Vega, De Souza, Prentice-Hernández, & Borges, 2013).

Edible coatings are an interesting option for the preservation
of fresh-cut fruits. A variety of materials can be used to prepare
film-forming dispersions that generate a protective barrier by pro-
ducing a modified atmosphere. The potential of polysaccharides
to serve as edible coatings has long been recognized because of
their ability to create a continuous network that is responsible
for the functional properties (Mostafavi, Kadkhodaee, Emadzadeh,
& Koocheki, 2016). Some of these materials can also be incorpo-
rated into systems for the release of active ingredients, including
antimicrobials and antioxidants, among others (Dhall, 2013; Pizato,
∗ Corresponding author.
E-mail addresses: luz.zambrano@unam.mx (M.L. Zambrano-Zaragoza),
quintana@unam.mx (D. Quintanar-Guerrero), adelreal@unam.mx (A. Del Real),
epinons@comunidad.unam.mx (E. Piñon-Segundo), jzambran@nayar.uan.mx
(J.F. Zambrano-Zaragoza).
One relatively new development in this field involves preparing
coatings that incorporate nanometric systems into a polysaccha-
ride matrix (Galindo-Pérez, Quintanar-Guerrero, Mercado-Silva,
Real-Sandoval, & Zambrano-Zaragoza, 2015; Shanmuga Priya,
Suriyaprabha, Yuvakkumar, & Rajendran, 2014; Zarei, Ramezani,
Ein-Tavasoly, & Chadorbaf, 2015) These studies have shown that
including nanoparticles contributes greatly to prolonging the shelf-
life of fresh-cut fruits and vegetables, while also helping maintain
their physicochemical and microbiological qualities. In this regard,
the elaboration of membrane systems -especially nanocapsules-
that have the capacity to control the release process represents an
innovative approach to the preservation of fresh-cut fruits (Pilon,
Spricigo, Miranda, Mattoso, & Ferrira, 2015; Zambrano-Zaragoza
et al., 2014). Given this background, the use of ␤-carotene, an
antioxidant already present in fruits like melon, is an interesting
option for maintaining product quality during refrigerated storage.
However, release systems of this kind usually require a hydrocol-
loid as a support matrix that protects and controls the release of the

http://dx.doi.org/10.1016/j.carbpol.2016.11.075
0144-8617/© 2016 Elsevier Ltd. All rights reserved.
 M.L. Zambrano-Zaragoza et al. / Carbohydrate Polymers 157 (2017) 1874–1882
active ingredient (Zandi, Mohebbi, Varidi, & Ramezanian, 2014).
Although several studies of the in vitro release of nutraceuticals
and essential oils from different platforms have been published
(Davidov-Pardo & McClements, 2014; Polowsky & Janaswamy,
2015), there is no information on this topic for coatings that come
into direct contact with foods. For these reasons, this field of inquiry
represents an important challenge for food formulators.
The term fresh-cut fruits refer to foodstuffs that have been
altered from their original state by peeling, cutting and pack-
aging. In recent years, demand for these products has increased
because of their attractive sensory characteristics. However, fresh-
cut pieces are highly-perishable and deteriorate faster than whole
fruits, principally because their inner flesh is exposed to envi-
ronmental conditions. Thus, they have a much shorter shelf-life
(Mantilla, Castell-Perez, Gomes, & Moreira, 2013).
Melon (Cucumis melo L.) is a highly-appreciated fruit because of
its attractive sensory characteristics, but fresh-cut melon pieces are
highly susceptible to softening during storage, even when chilled.
This is due to enzymatic degradation of the middle lamella of the
cell wall and the resulting loss of cell adhesion. Also, even minimal
processing causes tissue damage that increases the fruit’s respira-
tion rate (Raybaudi-Massilia, Mosqueda-Melgar, & Martín-Belloso,
2008). These changes reflect the metabolic activity that occurs in
tissues and so are useful predictors of physicochemical alterations,
such as the loss of firmness (Bett-Garber, Watson, Champagne, &
Lamikanra, 2010).
The present study evaluated the kinetic release of the antioxi-
dant ␤-carotene from nanocapsules supported in a polysaccharide
matrix and adjusted to release models, in an effort to explain phys-
ical and physicochemical changes in a model of fresh-cut fruit, in
this case melon (Cucumis melo, var. cantaloupe).
2. Materials and methods
2.1. Materials
Poly-␧-caprolactone (PCL) Mw ≈ 80,000 (␳ = 1.147 g/cm3 at
®
25 ◦ C) (Sigma Aldrich , USA), was used as the biopolymer to form
either a membrane (nanocapsules) or a matrix (nanospheres).
® ®
Pluronic , F-127 (Mw = 12,600 g/mol), (Sigma Aldrich , USA) was
utilized as the non-ionic stabilizer. ␤-carotene (99%) and sun-
®
flower oil (␳ = 0.921 g/cm3 at 25 ◦ C) (Sigma-Aldrich , USA) were
incorporated as the oily core. The partially water-miscible solvent
was analytical grade ethyl acetate (Fermont, Mexico), which was
selected for preparing the nanocapsules because of its low tox-
®
icity (ICH class: 3). The distilled water was of Milli-Q quality
(Millipore, USA, Bedford, MD). Xanthan gum was purchased from
®
Sigma-Aldrich (Germany) and used as a continuous matrix for the
coatings. Propylene glycol was used as the plasticizer (99%) (Sigma-
Aldrich, USA). All other reagents were of analytical grade and were
used with no further purification.
Melons (Cucumis melo, var. cantaloupe) were purchased at a
local wholesale market in Cuautitlan Izcalli, Mexico, and selected
according to size, uniformity of shape and external skin color. Dam-
aged or defective melons were discarded. The fruits selected were
randomly divided into 10 kg batches and kept in refrigerated con-
ditions at 4 ◦ C for 24 h prior to use.
2.2. Nanoparticle preparation
The nanocapsules (Ncs) and nanospheres (Nsp) were pre-
pared using the emulsification-diffusion method, considering
the optimization methodology proposed by Zambrano-Zaragoza,
Mercado-Silva, Gutiérrez-Cortez, Castaño-Tostado, and Quintanar-
Guerrero (2011). Briefly, ethyl acetate 100 mL and water 180 mL
1875
were mixed to equilibrium (saturation of both solvents in order, to
avoid solvent transference during the process). Then, 80 mL, of the
organic phase (ethyl acetate saturated with water) and 160 mL of
the aqueous phase (water saturated with ethyl acetate) were used
to prepare the Ncs. PCL (∼500 mg) and ␤-carotene (300 mg) previ-
ously dissolved in sunflower oil (700 mg), were then dissolved in
®
the organic phase, while Pluronic F-127 (4.8 g/L) was dissolved in
the aqueous phase. These two components were mixed and emul-
®
sified at 4000 rpm for 10 min (Ultra-Turrax T18, IKA, Germany),
followed by the addition of excess water (720 mL) to promote dif-
fusion of the solvent into the aqueous phase and Ncs formation.
Finally, the excess solvent was removed under reduced pressure
®
at 70 mmHg and 30 ◦ C (RV10, IKA Labotechnik, Germany) and the
dispersion obtained was diluted to 1 L with water. The Nsp were
prepared in the same way as the Ncs, but without ␤-carotene. Thus,
the final solid concentrations of materials in the Ncs dispersions
were 0.3 g/L of ␤-carotene, 0.7 g/L of sunflower oil, 0.5 g/L of PCL,
®
and 4.8 g/L of Pluronic F-127.
2.3. Particle size (PS), polydispersion index (PDI) and zeta
potential (␨)
PS distribution and the PDI were determined by the laser light-
scattering technique at 25 ◦ C (He/Ne: 633 nm, 10 mW) and a fixed
angle of 173◦ for 60 s using a Z-sizer 4 (Zetasizer Nano Series
Malvern Ltd., UK). The scattering intensity data were analyzed by
digital correlation under the unimodal analysis mode. Dispersions
®
were diluted with Milli Q until the appropriate particle concen-
tration was reached, as indicated by the particle counts per second
within the sensitivity range. The zeta potential (␨), was evaluated by
electrophoretic motion using standard polystyrene dispersions as
the references (␨ = −55 mV). This parameter measures the degree
of repulsion between adjacent particles. All measurements were
performed in triplicate at 25 ◦ C.
2.4. Coating formulations and their application on fresh-cut
melon
The batches tested in the study were as follows: 1) nanocap-
sules (Ncs). 500 mL of the dispersion were diluted with water to
1 L, 2) nanocapsules with xanthan gum (Ncs/XG): 500 mL of the
dispersion were mixed with 3 g of xanthan gum and diluted to 1 L;
3) nanospheres (Nsp): 500 mL of the dispersion were diluted with
water to 1 L; 4) nanospheres with xanthan gum (Nsph/XG): 500 mL
of the dispersion were mixed with 3 g of xanthan gum and diluted
to 1 L with water; and, 5) xanthan gum (XG): 3 g of xanthan gum
were dispersed in 1 L of water. Finally, one batch with no coating
of any kind was set aside as a control.
The whole melons were sanitized by immersion in a chlo-
rine solution at 90 ppm for 5 min. All cutting boards, utensils and
containers were also sanitized to minimize contamination due to
microorganisms. The melons were peeled and cut into spheres
using small scoops. After cutting, the melon pieces were placed
in a 1% CaCl2 solution and then immersed by batches into the cor-
responding nanodispersion coating. All samples were drained at
4 ◦ C to remove any excess coating. Once coated, the spheres were
packed in 250-mL polypropylene containers with ≈90 g of fruit,
and then placed in refrigerated storage at 4 ± 1 ◦ C and 85% relative
humidity (RH) for 21 days.
2.5. ˇ-Carotene encapsulation efficiency (EE)
The loading efficiency of the ␤-carotene was evaluated follow-
ing the method described by Tan et al. (2014), with modifications.
Briefly, a known amount of the ␤-carotene Ncs were dispersed in a
1876 M.L. Zambrano-Zaragoza et al. / Carbohydrate Polymers 157 (2017) 1874–1882
constant volume of distilled water, then 1 mL of this sample was put
in contact with 5 mL of cyclohexane to extract the ␤-carotene. The
sample was placed in a magnetic stirrer at 250 rpm, and aliquots
®
were withdrawn after 24 h, then filtered in Millipore vacuum
equipment using filter paper (sieve = 0.2 ␮m). The ␤-carotene con-
tent was determined spectrophotometrically at ␭ = 480 nm versus a
calibration curve prepared under the same conditions. The loading
efficiency was determined in triplicate using Eq. (1).
mass of ˇ − carotene (mg)
EE% = × 100 (1)
Total mass of the NP (mg)
2.6. Morphological studies
The morphological characterization of Ncs was carried out by
scanning electron microscopy (SEM) using the methodology pro-
posed by Zambrano-Zaragoza et al. (2011). The samples were
coated with gold using a JFC-1100 Sputter coater (JEOL, Tokyo,
®
Japan) (∼20 nm) and observed under a JSM 5600 LV-SEM LV
microscope (JEOL, Tokyo, Japan) at a resolution of 5 nm and 28 kV
with a chamber pressure of 12–20 Pa.
The morphological characterization of the surfaces of the mel-
ons with the different coatings was performed by longitudinal
tissue section after the spheres were dipped into the corresponding
coating system, followed by coating with gold, as described above.
The voltage used when taking the micrographs was 20 kV.
2.7. Oxygen consumption rate
The effects of the different coatings and the modification of
the O2 consumption rate in relation to the modified atmosphere
generated by treatment were determined using the static method
(Iqbal, Rodrigues, Mahajan, & Kerry, 2009). Briefly, melon spheres
of known weight were placed inside 170-mL glass bottles which
were immediately sealed. Samples of head space gas were drawn
off through a needle inserted into a septum placed in the top of
® The ␤-carotene release profile was evaluated every third day
each bottle and connected to an oxygen analyzer (Quantek Instru-
ments model 905, USA) in order to obtain the oxygen concentration
expressed as a fraction of the volume inside the container. Changes
in O2 content were monitored for 21 days. The O2 consumption
rate (RO2 ) was measured by the difference in O2 concentrations at
different time intervals, as expressed in Eq. (2).
(yi O2 − yO2 ) Vf
RO2 = ∗ (2)
(t − ti ) W
where: RO2 is the respiration rate, yiO2 and yO2 are the O2 con-
centration in the gas mixture at time ti -any time except time zero
expressed in hours- respectively; W is the weight of the apples (kg),
and Vf is the free volume inside the container.
2.8. Physicochemical changes
Juice leakage from the melon spheres was measured using the
method proposed by Zhang, Sampundo, Pothakos, Sürengil, and
Devlieghere (2013). Briefly, packages were tilted at a 20◦ angle
for 5 min, after which the accumulated liquid was recovered with
a 5 mL syringe. Results were reported as the amount of liquid
volume (mL) recovered per 100 g of fresh-cut melon in each con-
tainer. The juice from approximately 90 g of sample was extracted
from the polypropylene containers for each sample and used to
measure total soluble solids (TSS) using an ABBE refractometer
(Spectronic, Inc., USA). Titratable acidity (TA) was verified by titra-
tion with 0.1 N NaOH at pH 8.1 and expressed as citric acid (mg)
(Zambrano-Zaragoza et al., 2014). Finally, pH was measured with
a potentiometer (Philips Harris model E3039018G/K, Shenstone
England) equipped with an electrode (model P43-120). All mea-
surements were performed in triplicate.
2.9. Color evaluation
The effect of the different nanosystem coatings on the color
of fresh-cut melon spheres was determined by measuring their
surface color with a Minolta CR-300 colorimeter (Konica Minolta,
Tokyo, Japan) every 3 days during the 21 days of storage. Readings
were obtained on a CIELAB scale (L*, a*, b*). Three spheres from
each treatment were tested. Numerical values for the L*, a* and b*
parameters were used to calculate chroma (C*), ◦ Hue and white-
ness index (WI) using Eqs. (3), (4) and (5), respectively, following
Ferrari, Sarantópoulos, Carmello-Guerrero, and Hubinger (2013).
 1/2
C ∗ = a∗2 + b∗2 (3)
◦
 ∗ ∗

Hue = arc tang b /a (4)
 1⁄2
WI = 100 − (100 − L∗ )2 + a∗2 + b∗2 (5)
2.10. Firmness
The firmness of the fresh-cut melon spheres was measured
every third day up to day 18 of storage. The spheres were cut
in half, and 5 wedges from each batch were evaluated using an
INSTRON 4411 with a 50-N load cell, equipped with a fixed, 6 mm-
thick punch (Instron, Mass., USA). Each sample wedge was fractured
by a downward motion using a velocity of 2.5 mm s−1 to determine
the maximum force (highest N value) required to penetrate 5 mm
into the wedge. This measurement was used as the firmness index.
These measurements were made in quintuplicate (Silveira, Aguayo,
Chisari, & Artés, 2011).
2.11. Kinetic release of ˇ-carotene
during the 21 days of storage at 4 ◦ C. Each melon sphere (2.5 cm)
was placed in 40 mL of cyclohexane under stirring for 90 min (Ther-
molyne Cimarez, USA), then the samples were filtered through
®
a 0.2 ␮m membrane using Millipore vacuum equipment. The
amount of ␤-carotene was determined spectrophotometrically at
␭ = 480 nm versus a calibration curve (0–200 ␮g/mL) using cyclo-
hexane as the solvent. These measurements were performed in
triplicate at 25 ◦ C. In order, to determine the ␤-carotene in the
melon samples, as well as the effect of xanthan gum on these
batches, changes in ␤-carotene content were also determined.
The ␤-carotene profiles were analyzed to determine the release
type and mechanism by fitting the data to different mathematical
models; namely, Peppas’ semi-empirical (Eq. (6)), and first-order
(Eq. (7)) models, the Higuchi model for matrix systems (Eq. (8)), and
models that evaluate the values of the coefficient (R2 ) to describe
the quality of the fit.
Mt
= Kt n (6)
M∞
LnQt = LnQ0 ± k1 t (7)
1
Qt = kH t ⁄2 (8)
where Mt /M∞ is the fraction of ␤-carotene released at time t, K is
the Korsmeyer-Peppas kinetic constant, and n is the release expo-
nent that determines the ␤-carotene release mechanism (Zandi
et al., 2014). Qt is the amount of ␤-carotene released at time t, Q0 is
the amount of ␤-carotene released at time zero, and k1 and kH are
the release rate constants for the first-order model, and Higuchi’s
model, respectively.
 M.L. Zambrano-Zaragoza et al. / Carbohydrate Polymers 157 (2017) 1874–1882
Fig. 1. Micrographs of the structure of ␤-carotene Ncs and ␤-carotene Ncs on the surface of fresh-cut melon: a) ␤-carotene Ncs; b) surface of fresh-cut melon; c) surface of
fresh-cut melon with ␤-carotene Ncs; and d) surface of fresh-cut with ␤-carotene Ncs/XG.
2.12. Statistical analysis
®
Results were analyzed using MINITAB 14 to perform an anal-
ysis of variance (ANOVA) that compared the effectiveness of the
different coatings on the conservation of the melon spheres. The
mean values obtained from the ANOVA were subjected to a Tukey
test for difference between means. Treatments were considered
significantly different at a level of 5%.
3. Results and discussion
3.1. Particle size, PDI and zeta potential (␨)
The results of the analyses performed to determine particle size,
the polydispersion index (PDI), and the ␨ of the nanoparticle sys-
tems employed in preparing the coatings determined that mean
particle size was between 122 and 220 nm, indicating that incor-
porating xanthan gum increased particle size only slightly. The PDI
was below 0.35 in all cases, suggesting a narrow size distribution.
No statistical difference (p > 0.05) was found among treatments
in this regard. The particle size distribution showed monomodal
behavior in all cases, including the samples with xanthan gum.
Similar results were obtained with the nanoparticles that contained
␤-cyclodextrin (Memişoğlu, Bochot, Sen, Duchêne, & Hincal, 2003).
The ␨ was negative and the values obtained – from −38.5 to
41.3 mV – established that there was good repulsion between par-
ticles that limited aggregation and improved the functionality of
1877
the coatings. These values were used to determine the repulsion
between the nanoparticles in the aqueous medium that provided
the best stability in the Ncs (Alishahi et al., 2011; Badea, Lăcătuşu,
Badea, Ott, & Meghea, 2015).
3.2. Encapsulation efficiency and morphological studies
The encapsulation efficiency of the ␤-carotene loaded into poly-
␧-caprolactone Ncs was 65%. The rest of the active ingredient was
lost in the continuous phase. Even so, the efficiency determined was
higher than that reported by Singh, Shakil, Kumar, Walia, and Kar
(2015), who found that the highest efficiency in their nano micelles
was 28%. Therefore, adding poly-␧-caprolactone to the formation
of the Ncs substantially increased the encapsulation efficiency of
the ␤-carotene.
Fig. 1a–d shows typical micrographs of the ␤-carotene Ncs. A
dense halo is visible around the spheres, suggesting the presence of
a smooth-shelled PCL membrane around the oily nanodroplets that
contain ␤-carotene (Fig. 1a). The size of the nanoparticles corre-
sponds to that reported by light scattering. Fig. 1b shows the surface
structure of the untreated fresh-cut melon spheres. Similar struc-
tures were shown by Beaulieu, Bruce, Ingber, and Lea (2011). Fig. 1c
and d shows post-treatment images in which capsular entities are
seen to be distributed uniformly on the surface. When xanthan gum
was used, plaques with embedded Ncs were observed (Fig. 1d).
1878 M.L. Zambrano-Zaragoza et al. / Carbohydrate Polymers 157 (2017) 1874–1882
Fig. 2. Oxygen consumption for the coatings applied on fresh-cut melon
with respect to control samples. Control; XG = xanthan gum;
Ncs = nanocapsules; Ncs/XG = nanocapsules/xanthan gum;
Nsp = nanospheres; Nsp/xanthan gum.
3.3. Oxygen consumption rate
Fig. 2 shows the changes associated with the oxygen consump-
tion of the fresh-cut melon. It is important to note that applying
coatings contributed greatly to decreasing the rate of oxygen con-
sumption in all samples, but that the most significant decrease
occurred on day 6 of storage at 4 ◦ C. The samples coated with ␤-
carotene Ncs mixed with xanthan gum showed the best results,
as the rate of oxygen consumption decreased by 49% compared
to controls; while the melon coated only with xanthan gum pro-
duced the lowest reduction, at just 20%. The ANOVA (p > 0.05)
showed no statistically-significant difference at 12 days of stor-
age among the samples coated with Ncs, Nsp and Nsp/XG, a result
likely attributable to the nanosystems that were infiltrated into the
fruit. The samples without ␤-carotene (Nsp) achieved only a low
control of the browning index and were less effective in reducing
oxygen consumption than those coated with Ncs. The high oxygen
consumption observed in the control samples can be attributed to
changes associated with stress due to the cutting and treatment of
the fruit during preparation, even though these were minimal (Li &
Thomas, 2014). It is important to point out that the use of coatings
with different components causes a selective modification of the
fruit’s permeability to gases, thus resulting in an increased con-
centration of CO2 and a decrease in the amount of O2 (Olivas &
Barbosa-Cánovas, 2005). In the present study, ␤-carotene Ncs inte-
grated into a polysaccharide matrix helped reduce the respiration
rate of the fruit because, while XG alone decreased the respiration
rate by 16% compared to controls, the addition of Ncs produced a
reduction in this activity of 47%. These results suggest that particle
size and the composition of the Ncs had a direct effect on how the
different coatings were formed.
3.4. Physicochemical changes
Table 1 shows the results for juice leakage, TSS, pH, and the
titratable acidity of the fresh-cut melon spheres. Controls had the
highest juice leakage from 9 days of storage on, at 2.18%, followed
by the fruit coated with xanthan gum only, in which the loss
was above 2.94% after 15 days of storage at 4 ◦ C, though this dif-
ference was not statistically significant (p > 0.05) with respect to
controls at the end of the storage period. In contrast, the use of
xanthan gum combined with Nsp did not generate any statistically-
significant difference among means at the end of storage (␣ = 0.05).
The time of storage there is a statistically significant difference
and according to Tukey test the C and XG show a similar behav-
ior. In addition, the use of coatings with Nsp and Ncs with and
without XG contributes to decrease juice leakage. According to
Rojas-Graü, Soliva-Fortuny, and Martín-Belloso (2008), the use of
polysaccharides alone does not contribute to reducing weight loss
per se, but incorporating a coating based on Ncs/XG does help con-
trol juice leakage through the fruit surface. This behavior can be
attributed to the sealing capacity of the poly-␧-caprolactone wall
that formed in the samples treated with Ncs combined with XG.
This treatment also decreased the oxygen consumption associated
with the antioxidant effect of the ␤-carotene. On the other hand,
edible coatings create a semi-permeable barrier that can be mod-
ified by incorporating lipid structures (Duan, Jiang, & Zhao, 2011;
Maqbool, Alderson, Mohamed, Siddiqui, & Zahid, 2011). When the
Ncs/XG coating was applied, there was a significant reduction of
juice leakage at 21 days of storage at 4 ◦ C, which was lower than that
reported by Manzocco, Da Pieve, and Maifreni (2011). Moreira et al.
(2014), meanwhile, studied the effect of trans-cinnamaldehyde
microencapsulated in cyclodextrins and incorporated into pectin-
chitosan-CaCl2 . They reported juice losses of 0.8–1.0% at 15 days,
indicating that the effect on reducing juice leakage was related to
a cross-linking between the polymer wall and the polysaccharide
matrix wall that contributed to form a modified atmosphere. In
the present study, the use of Ncs incorporated into a xanthan gum
matrix (Ncs/XG) succeeded in reducing the loss of juiciness to below
1% after 15 days of storage.
Table 1 also presents the SSC, which ranged from 8.51–7.28◦ Brix
for controls and samples coated with XG, respectively. This slight
change may be related to the higher rate of juice loss and inherent
enzymatic activity. The two-way ANOVA (␣ = 0.05), the samples
showed a statistical difference between the means of the treat-
ments as a function of time and with respect to the composition of
the coating. The Tukey test also revealed that the C, XG and Ncs/XG
coatings had no statistically-significant differences in relation to
the SSC, whereas the samples coated with Ncs and Nsp showed only
minor variations in their SSC content. In their study of cantaloupe
using edible antimicrobial coatings based on pectin and chitosan,
Martiñon, Moreira, Castell-Perez, and Gomes (2014) also reported
minor variations in SSC (of 8.5–10◦ Brix) compared to the initial
concentration. The changes in pH and acidity are summarized in
Table 1, which demonstrates that the samples with Nsp and Ncs
plus ␤-carotene, with or without xanthan gum, had lower pH after
9 days of storage. This is attributable to the molecular dispersion
and fracturing of the PCL, which allows water to attack the bonds
of the polymer chains, thus shortening their length to produce chain
fragments with carboxylic groups that are solubilized in the fruit
and degraded to ␧-hydroxycaproic acid. It is common for aque-
ous media to generate system acidification (Bölgen, Menceloǧlu,
Acatay, Vargel, & Pişkin, 2005; Lacoulonche et al., 1999).
 M.L. Zambrano-Zaragoza et al. / Carbohydrate Polymers 157 (2017) 1874–1882 1879

Table 1
Physicochemical and color changes during storage of fresh-cut melon at 4 ◦ C as a function of treatment (mean ± standard deviation).
◦
Treatment Days Juice leakage (%) TSS pH TA (mg) L* WI Hue C*

C 0 0.00 ± 0.00 8.17 ± 0.15 6.37 ± 0.10 0.10 ± 0.00 69.00 ± 0.00 60.87 ± 0.31 70.50 ± 0.32 23.87 ± 0.56
9 2.18 ± 0.21 7.83 ± 0.15 6.17 ± 0.49 0.10 ± 0.03 57.67 ± 1.54 52.76 ± 0.77 67.35 ± 1.55 20.95 ± 0.99
15 3.20 ± 0.10 7.47 ± 0.10 5.60 ± 0.59 0.11 ± 0.05 55.00 ± 1.73 50.67 ± 0.92 62.04 ± 1.49 20.20 ± 0.94
21 3.43 ± 0.23 7.28 ± 0.14 5.57 ± 0.43 0.11 ± 0.02 52.33 ± 1.15 48.35 ± 1.12 59.02 ± 1.76 19.88 ± 1.12
XG 0 0.06 ± 0.00 8.51 ± 0.09 6.60 ± 0.78 0.10 ± 0.06 66.00 ± 1.83 57.65 ± 0.61 69.76 ± 0.48 25.24 ± 0.46
9 0.53 ± 0.02 8.40 ± 0.21 6.70 ± 0.34 0.10 ± 0.04 62.00 ± 1.54 55.46 ± 1.27 67.29 ± 1.15 23.24 ± 0.46
15 2.94 ± 0.15 7.48 ± 0.06 5.63 ± 0.46 0.12 ± 0.02 53.67 ± 1.77 48.03 ± 1.31 64.16 ± 1.47 23.52 ± 1.17
21 3.16 ± 0.17 7.34 ± 0.19 5.65 ± 0.15 0.12 ± 0.03 51.00 ± 1.02 45.65 ± 1.97 62.63 ± 1.76 23.50 ± 0.85
Nsp 0 0.19 ± 0.00 8.40 ± 0.10 6.20 ± 0.58 0.09 ± 0.02 67.00 ± 1.00 57.92 ± 0.73 69.83 ± 0.15 26.10 ± 0.19
9 1.27 ± 0.10 8.00 ± 0.27 5.60 ± 0.27 0.11 ± 0.05 55.00 ± 1.42 49.39 ± 0.82 69.34 ± 0.18 23.15 ± 0.73
15 1.45 ± 0.08 8.17 ± 0.16 5.33 ± 0.61 0.16 ± 0.06 55.00 ± 1.98 48.94 ± 2.02 62.97 ± 0.43 24.14 ± 0.46
21 1.64 ± 0.28 8.15 ± 0.27 5.52 ± 0.54 0.15 ± 0.04 55.00 ± 1.42 48.83 ± 1.14 61.82 ± 0.51 24.35 ± 0.53
Nsp/XG 0 0.00 ± 0.00 8.17 ± 0.15 6.57 ± 0.10 0.10 ± 0.00 68.00 ± 1.00 60.53 ± 0.53 69.28 ± 0.60 23.09 ± 0.49
9 1.14 ± 0.10 8.33 ± 0.10 6.53 ± 0.28 0.10 ± 0.05 58.33 ± 2.52 51.90 ± 1.06 70.09 ± 0.89 24.00 ± 0.49
15 1.43 ± 0.05 7.47 ± 0.10 6.30 ± 0.39 0.10 ± 0.02 60.33 ± 2.81 53.26 ± 2.54 66.68 ± 0.76 24.68 ± 0.65
21 1.56 ± 0.08 7.58 ± 0.14 6.42 ± 0.27 0.10 ± 0.03 57.67 ± 3.05 51.34 ± 1.33 64.04 ± 078 23.99 ± 0.56
Ncs 0 0.38 ± 0.17 8.40 ± 0.10 6.50 ± 0.21 0.10 ± 0.01 69.33 ± 0.57 59.20 ± 0.78 72.03 ± 0.41 26.91 ± 0.67
9 0.51 ± 0.06 8.23 ± 0.21 6.27 ± 0.47 0.09 ± 0.01 66.33 ± 1.00 58.49 ± 0.92 70.02 ± 0.43 24.26 ± 0.60
15 1.30 ± 0.15 7.85 ± 0.69 5.67 ± 0.52 0.12 ± 0.06 57.33 ± 1.52 51.31 ± 0.83 66.66 ± 0.72 23.45 ± 0.42
21 1.38 ± 0.46 8.12 ± 1.13 5.30 ± 0.32 0.12 ± 0.05 57.33 ± 1.15 51.27 ± 0.59 66.01 ± 0.46 23.53 ± 0.52
Ncs/XG 0 0.03 ± 0.00 8.47 ± 0.35 6.30 ± 0.27 0.10 ± 0.02 69.67 ± 1.15 59.40 ± 0.72 71.37 ± 0.61 26.98 ± 0.26
9 0.45 ± 0.05 8.27 ± 0.46 6.27 ± 0.26 0.09 ± 0.03 67.00 ± 1.00 58.43 ± 0.89 71.05 ± 0.54 25.27 ± 0.27
15 0.98 ± 0.06 8.35 ± 0.39 5.67 ± 0.49 0.12 ± 0.02 67.00 ± 1.00 59.06 ± 0.76 69.37 ± 0.61 24.22 ± 0.31
21 1.21 ± 0.17 7.89 ± 0.47 5.07 ± 0.14 0.11 ± 0.42 62.00 ± 1.00 54.84 ± 0.98 69.61 ± 0.98 24.40 ± 0.28

C = Control, XG = xanthan gum, Ncs = Nanocapsules, Ncs/XG = nanocapsules/xanthan gum, Nsp = nanospheres, Nsp/XG = nanospheres/xanthan gum. C* = Chroma; WI = whiten
index.

3.5. Color changes
Table 1 shows the changes in the factors of WI, C* and ◦ Hue
for controls and the samples coated and stored at 4 ◦ C for 21 days.
The control, Ncs and XG melons all showed significant decreases in
WI in excess of 25% with respect to the initial condition. The most
effective coating in this regard was Ncs/XG, as WI remained sta-
ble during storage with no statistically-significant difference as a
function of storage time (p < 0.05) up to 21 days. A similar behavior
was observed for the samples coated with xanthan gum, as their
WI remained basically constant up to 9 days, with only a slight
decrease of 3.3%. In contrast, the melon pieces coated with Nsp
failed to maintain their WI during the storage period, showing that
the Ncs had an influence on sustaining the fruit’s original color due
to their antioxidant capacity. Ferrari et al. (2013) reported a loss
of whiteness of 15% for control samples and 3.1% when melons
were coated with CaCl2 and pectin. This decrease in the WI sug-
gests an effect of transparency or translucency; a change that may
be associated with a process of deterioration.
The decrease in C* is associated with the intensity of the fruit’s
color (Table 1). The systems with nanoparticles had the best reten-
tion of C* in the fresh-cut melon during storage, with the pieces
coated in Ncs/XG achieving the minimum loss of color intensity of
only 4.03%. In terms of effectiveness, this coating was followed by
the Nsp/XG composition. It is apparent that particle size had an
important effect on the changes associated with the intensity and
brightness of the fruit, because a smaller size modified the light
refraction index. Also, poly-␧-caprolactone enhanced the conser-
vation of color intensity. Amaro et al. (2013) obtained a reduction
of C* of up 19.5% when fresh-cut cantaloupe was treated with
1-methylcyclopropane, while Moreira et al. (2014) evaluation of
incorporating microcapsules made of cinnamaldehyde into differ-
ent types of packaging and multi-layer coatings with essential oil
microencapsulated in cyclodextrins found that C* decreased by
approximately 15% during storage.
Finally, Table 1 also shows the changes associated with hue
angle, a measure of the color saturation of fresh-cut melon and
its evolution during the storage period. The largest decrease was
observed in the control samples, which developed a red pigment
as a result of browning. In contrast, the samples with XG, Nsp and
Nsp/XG showed no statistically-significant differences (p > 0.05), as
the change of just 11.4% suggested only slight browning. The melon
coated with Ncs/XG had the lowest change in hue angle (7%), likely
due to the protective effect of the ␤-carotene nanoencapsulated in
the polysaccharide matrix.
3.6. Firmness
Fig. 3 shows the changes of firmness in the fresh-cut melon.
The statistical analysis showed a statistically-significant difference
between means of treatment, as it showed that at 3 days of stor-
age the mean firmness of the melon pieces did not differ among
samples except for controls. Later, all samples of fresh-cut melon
suffered a loss of firmness, except those coated only with Ncs. Here,
once again, the control samples showed the most pronounced loss
of firmness; a finding which confirms that the use of a coating with
nanoparticles contributes significantly to maintaining fruit texture.
The melon pieces coated with XG had a decrease of 21.45% at 6 days
compared to controls, which showed a loss of 39% in the same
period. The application of Ncs with ␤-carotene helped conserve
firmness, as the decrease in this case was only 9.9% for the melon
coated with Ncs, and just 2% for the spheres immersed in Ncs/XG.
These findings suggest that the xanthan gum and ␤-carotene Ncs
exerted a synergistic effect. Poverenov et al. (2014) used an electro-
static layer-by-layer technique to apply chitosan-alginate coatings
on melon, and reported a texture decrease of 2.45–2.3 N at 7 days. In
another study, Dilmaçünal, Erbaş, Koyuncu, Onursal, and Kuleaşan
(2014) used hazelnut oil with vitamin E as the antioxidant. They
found that this treatment was the one that best controlled the tex-
ture of Kikagac melon, with a maximum loss of 39.8% at 12 days of
storage. These results allow us to affirm that incorporating Ncs into
a polysaccharide matrix helps maintain the firmness of fresh-cut
melon for longer time periods.
3.7. Release of ˇ-carotene
The adequate release of an active ingredient by means of a poly-
mer matrix depends on many factors, including active solubility
1880 M.L. Zambrano-Zaragoza et al. / Carbohydrate Polymers 157 (2017) 1874–1882

Fig. 3. Changes in the firmness of fresh-cut melon. Control; XG = xanthan gum; Ncs = nanocapsules; Ncs/XG = nanocapsules/xanthan gum; Nsp = nanospheres;
Nsp/XG = nanospheres/xanthan gum.

and loading, particle size, the release medium, the presence of other
components in the formulation, the type of encapsulation, and the
architecture of the system, among others (Quirós-Sauceda, Ayala-
Zavala, Olivas, & González-Aguilar, 2014; Zandi et al., 2014). Hence,
the analysis of release profiles is a complex task that entails con-
sidering all these factors in order to accurately assess the release
mechanism and time-dependence. Given that fruits themselves
contain a certain amount of antioxidant – called “native” – it was
first necessary to determine this quantity using the control sam-
ples at the different evaluation intervals, and then subtract it from
the total amount in order to correctly compile the release profiles.
Fig. 4 shows the behavior of the release of ␤-carotene from the
melon spheres used in this experiment under the different sys-
tems. Controls and the XG treatment showed an average “native”
␤-carotene content of 2.3 ± 0.23 ␮g/g of fruit, an amount similar
to that obtained by Amaro et al. (2013) for cantaloupe. In con-
trast, the systems containing Ncs alone and Ncs/XG had a peculiar
profile characterized by a first step in which controlled release of ␤-
carotene reached maximum amounts of 25.5 and 39.7 ␮g/g of fruit
for Ncs and Ncs/XG, respectively, up to the ninth day. An immedi-
ate release (tburst ) was evident for both systems: 3.6 and 9.7 ␮g/g
for Ncs and Ncs/XG, respectively. This controlled release can be
explained by the diffusion of ␤-carotene and/or the opening of the
Ncs on the fruit. The difference in the amount released between
these two systems may be attributable to their respective retention
capacities, such that xanthan gum helps retain more Ncs on the sur-
face of the fruit by forming a thin film. The data from this first step
were analyzed using Korsmeyer-Peppas’ semi-empirical equation
(Mt /M˛ = Ktn ), according to this equation and the results analyzed
it is suggested that the transport mechanism and release form is
Fickian Type (Kaunisto, Marucci, Borgquist, & Axelsson, 2011).
The logarithmic form of this equation was used to obtain
Fig. 4. Release of ␤-carotene; C = Control, XG = xanthan gum;
the exponential “n” for each system, generating values of 0.32 Ncs = nanocapsules; Ncs/XG = nanocapsules/xanthan gum.
(r2 = 0.989) and 0.30 (r2 = 0.997) for Ncs and Ncs/XG, respectively.
Thus, both systems have a time-dependent Fickian release (t∼0.3 ).
Considering the distribution of the Ncs over the fruit, we can sur- by Higuchi to explain the release of pharmaceutical ointments
mise that the behavior of these systems resembles that of a matrix (Higuchi, 1961). When our data were adjusted in accordance with
system in which the nanoparticles could remain “undissolved” or Higuchi’s model (Mt versus t0.5 ), results showed good correlations
penetrate regions with high ␤-carotene concentrations. In addition, of r2 = 0.996 and r2 = 0.945 for Ncs and Ncs/XG, respectively. These
there may be an outer region where all ␤-carotene is dissolved calculations suggest that the systems do indeed have a matrix-
entirely (i.e., leaving no particles), but follows a gradient gov- like behavior, which could be explained by a pseudo-steady-state
erned by a process of diffusion, which can be explained by Fick’s approach in which the ␤-carotene is easily available for transfer
laws. These are only assumptions, but they suggest those proposed from the Ncs to the fruit. It is important to mention that a cross-
current process occurs during storage, as exudation “washes” the
 M.L. Zambrano-Zaragoza et al. / Carbohydrate Polymers 157 (2017) 1874–1882
Fig. 5. General aspect of fresh-cut melon during storage at 4 ◦ C.
Ncs into the fruit while simultaneously limiting the amount of ␤-
carotene that enters the pulp. This process would prevail from day
9 through day 22, and may well explain the second curve of the
slope.
Finally, Fig. 5 shows the general aspect of the fresh-cut melon
during storage. Only the lower part of the packaging is shown in
order to facilitate the visualization of juice leakage and changes in
product appearance. Observe that samples coated with Ncs show
the best behavior.
4. Conclusions
Based on these findings, it appears that the inclusion of xanthan
gum contributes to retaining the Ncs and this, in turn, increases
␤-carotene content. Thus, this coating contributes significantly to
maintaining the quality of fresh-cut melon (Cucumis melo, var.
cantaloupe). Apparently, this mechanism operates because the
xanthan gum forms a film that helps retain ␤-carotene; a behavior
that correlates with lower indices for measures of the modifications
of the physical properties of fresh-cut melon during storage.
The release profile of ␤-carotene though the Ncs and Ncs/XG
coatings showed two characteristic slopes. The initial phase was
marked by the controlled release of ␤-carotene (days 0–9) with
tburst. The Korsmery-Peppas model indicated a release pattern that
can be explained by time-dependent Fickian diffusion. The first
stage was found to fit quite well with Higuchi’s model, thus sug-
gesting a matrix-like behavior in which the nanocapsules take the
place of “undissolved” particles with high ␤-carotene concentra-
tion. The second slope, in contrast, was marked by a steady decline
in the ␤-carotene content (day 9–22) that may be explained by the
continuous exudation of the fruit that would carry away some of
the ␤-carotene.
1881
Acknowledgements
The authors acknowledge and appreciate the technical support
provided by Dr. María de los Angeles Cornejo Villegas, and the finan-
cial support for projects PAPIIT IT200814, PAPIME PE206614 from
the DGAPA and CONACYT INFRA-251940.
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