Scientia Horticulturae 240 (2018) 496–508

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Scientia Horticulturae
journal homepage: www.elsevier.com/locate/scihorti

Shelf life extension of strawberry by temperatures conditioning, chitosan T

coating, modified atmosphere, and clay and silica nanocomposite packaging
⁎
Hossein Barikloo, Ebrahim Ahmadi
Department of Biosystem Engineering, Faculty of Agriculture, Bu-Ali Sina University, Hamedan, Iran

A R T I C LE I N FO A B S T R A C T

Keywords: In order to investigate the changes of gas inside the packaging as well as qualitative and mechanical properties of
Modified atmosphere packaging coated strawberry under modified atmosphere conditions, during the storage period, a study was conducted.
Nanoclay First, the C0S0, C1S0.75 and C2S0.75 nanocomposite films were prepared using a molten mixture method (S and C
Nanosilica represent nanosilica and nanoclay, respectively). Then, the strawberry was packed under modified atmosphere
Oxygen
conditions (10% oxygen, 15% carbon dioxide and 75% nitrogen). Treatments included of two different tem-
Temperature
perature (4 °C and 25 °C), three levels the packaging (C0S0, C1S0.75 and C2S0.75) and two levels the coating
(chitosan-coated and uncoated). The measurement parameters including chemical properties (pH, soluble solids
content), physical properties (weight loss) and mechanical properties (firmness, elasticity modulus), as well as
changes in the proportions of gases inside the package were evaluated and analyzed based on the Completely
Randomized Multivariate Factorial (CRF) design. The results of variance analysis of weight loss, pH, soluble
solids content, firmness, modulus of elasticity and changes in oxygen and carbon dioxide indicated that the main
and interaction effects were significant at (p < 0.05). According to the SEM images, distribution of nanoparticles
is almost uniform and there is no accumulation in various areas of the fracture surface. During the storage
period, the variations of the dependent parameters at 4℃ were less than ones at 25℃. Moreover, at two tem-
peratures of 4 °C and 25 °C, the trend of variations of oxygen gas in the coated samples was different from the
non-coated sample, so that it was slowing down until the second day and then increased. Finally, the results
indicated that the coated sample packed with the C1S0.75 nanocomposite film had the most optimal mode in
terms of the changes in physical, chemical and mechanical properties during the storage period.

1. Introduction
The revolution in the standard of living, eating habits, and in-
creasing awareness of nutrition has pushed consumers from an energy
providing diet to the diet with a balanced nutrient profile along with
metabolic, physiological, health and functional benefits (Panghal et al.,
2018). So fruits and vegetables often placed on the list of consumer’s
priorities. Strawberry (Fragaria ananassa) contains a diverse range of
vitamins and minerals and it has a general acceptance because of its
nutritional value. However, strawberry due to its relatively high me-
tabolic activity, is highly perishable and subjected to rapid post-harvest
losses, which, in turn, poses a serious challenge to its marketing. Since
Strawberry is a luxury high price fruit, so it has to be preserved and may
be converted in different types of processed products and marketed
scientifically to fetch foreign exchange and to conserve the national
economy. (Panghal et al., 2009). Various studies have shown that the
methods of coating and proper packaging of agricultural products and
combining these procedures are important ways to maintaining
strawberry fruit quality, assisting more appropriate storage, and de-
creasing the damages in storage.
Edible coatings have the advantage of controlling oxygen, carbon
dioxide, and moisture transfer (Jayakumar et al., 2007; Lazaridou and
Biliaderis, 2002). Improving the mechanical properties of the products
is also an advantage of these coatings (Han, 2007). Chitosan is a natural
polymer and a non-toxic, eco-friendly and biodegradable compound
that also has antimicrobial properties (Tripathi et al., 2008). The ben-
eficial effects of chitosan on titratable acidity, tissue firmness and re-
duction of rot in strawberry (Miranda-Castro, 2016), and raspberry
(Jaqueline et al., 2014) were reported. According to studies conducted
by Lazaridou and Biliaderis (2002), chitosan-based coatings have the
potential for improving fresh fruit storage duration, decreasing oxygen
content, decreasing ethylene production, and increasing internal carbon
dioxide.
The polyolefin elastomer (POE) film is very suitable for resistance to
moisture penetration, but their resistance against penetration of gases
and aromatic materials are low (Du et al., 2012). The use of mineral

⁎
Corresponding author.
E-mail address: eahmadi@basu.ac.ir (E. Ahmadi).

https://doi.org/10.1016/j.scienta.2018.06.012
Received 16 February 2018; Received in revised form 19 May 2018; Accepted 6 June 2018
Available online 23 June 2018
0304-4238/ © 2018 Elsevier B.V. All rights reserved.
H. Barikloo, E. Ahmadi
nano-fillers in packaging materials has several advantages including:
the improving maintenance, improving aroma, taste, color, texture and
stability during storage and transportation, reducing rot, preventing the
entry and exit of gases and increasing thermal stability of the polymers
(Othman, 2014). Yang et al (2010) indicated that content of acid be-
tween nano-containers and containers of conventional polymers to
preserve strawberry over storage life has a significant difference. Ad-
ditionally, strawberries available nano containers had more market-
ability, compared to ones contained in polymer containers. Other stu-
dies also demonstrated that nano packaging had favorable impacts on
the organic, physical, chemical and physiological quality of fresh
strawberry compared to polyethylene packaging (Zhong et al., 2007).
The packaging method with modified atmosphere conditions is also
one of the most important ways to increase the shelf life of strawberry
fruit (Gholami et al., 2018). The maintenance with the controlled at-
mosphere leads to a decrease in the rate of chemical and biochemical
reactions, as well as the reduction of growth of pathogenic micro-
organisms which, in turn, cause maintaining quality and extending
storage life of products (Gholami et al., 2018). In a research done,
strawberries were packaged in a MAP containing 15% carbon dioxide
and 8% oxygen and sachets of ethylene absorbent and moisture (Seckin
and Caner, 2011). The results of qualitative properties such as firmness,
soluble solids content and color indicated that these characteristics
retained in the optimum conditions for 14 days and in addition, the
fungal growth, under these storage conditions, significantly decreased
(Seckin and Caner, 2011). In a study by Nielson and Leufven (2008),
two cultivars of Corona and Honeoye strawberries for seven days of
storage, were examined. The effect of controlled atmosphere (9–12%
carbon dioxide and 11–14% oxygen) on strawberries were evaluated
and then a decrease in respiratory and growth of microbial, were re-
ported.
As above-mentioned Strawberry is prone to mechanical damage,
drying and physiological impairment during storage period. In addi-
tion, due to the having general acceptance and the nutritional value of
strawberries, packaging, storage and transportation of this fruits are
important. In this study, in order to maintain the quality of this fruit
during the storage period, we combined the methods of the chitosan-
based coating and packaging of nanocomposites under modified at-
mosphere conditions. Then, we examined the qualitative, mechanical
characteristics and changes in the gas inside the strawberry package
with using clay-silica PE/POE/PA6 nanocomposites film and coating
chitosan under modified atmosphere conditions during storage period.
2. Materials and methods
Strawberry cultivar Gavita (Sphericity coefficient: 0.897 ± 0.051,
Diameter: 27.25 ± 2.05, Mass: 13.85 ± 0.35), was provided from a
hydroponic based greenhouse, Hamedan, Iran. Experiment period data:
August 11, 2017–September 28, 2017. All samples were harvested ar-
ranged in rows at 16 × 13 cm2 dimensions boxes. Samples were kept at
ambient (25℃ and 30% humidity) and cold storage conditions (4℃ and
95% humidity). In order to conduct the experiments, fruits were ex-
posed to temperatures of 4℃ and 25℃ and kept at three packaging type
including C0S0, C1S0.75, C2S0.75; C and S denotes nano-clay and nano-
silica, respectively.
2.1. Coating strawberry
Strawberry fruits were completely evaluated in terms of physical
damages, fungal infections, and size homogeneity after harvesting from
hydroponic culturing greenhouse. Then, the fruits were divided into
coated and un-coated fruits. Fruits to be coated were prepared with
chitosan emulsion, submerged for 2 min, and kept at 20℃ for 1 h for
drying the surface coating through airflow. Chitosan emulsion was
prepared by addition of 5 g r chitosan (SIGMA) into 1 l acetic acid 1% to
making 0.5% chitosan concentrations. After complete dissolution of
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Scientia Horticulturae 240 (2018) 496–508
Table 1
Combined mass of materials for producing the films of PE/POE/PA6 nano-
composites contains clay and silica nanoparticles.
Variable E-g-MA (%) S (%) C (%) POE (%) LDPE (%) PA6a (%)
C0S0 5 0 0 0 85.5 9.5
C1S0.75 5 0.75 1 14 71.4 7.9
C2S0.75 5 0.75 2 27 58.62 6.2
a
Polyamide 6.
chitosan, pH was adjusted at 5 via 1 N NaOH and eventually 2 ml tween
80 was added to the solution (Petriccione et al., 2015).
2.2. Nanocomposite film production and its structure analysis
Melt mixing of clay nanocomposite was done by using a co-rotating
twin-screw extruder (Coperion-Germany) with L/D = 62, D = 75, and
speed = 500 rpm. The temperature at various areas of the extruder
from feeding portion to exit parts was established at 125, 145, 155, 170,
185, 195, and 200 °C. The pressure of the melt and the temperature at
dying were equal to 5 bar and 200 °C, respectively. After assuring the
cleanness of the route and establishing above-mentioned conditions,
polyethylene, Polyolefin Elastomers (POE), Polyamide 6 (PA6), soil clay
nano-particles and organically modified silica at specified percents
(Table 1) weighed (including C0S0, C1S0.75, C2S0.75; C and S denotes
nano-clay and nano-silica, respectively), mixed, and entered the ex-
truder case via loss in weight feeder. Finally nano composite compound
cut into granule by strand pelletizer. For producing the film, a single
screw extruder and casting roll (Brabender-Germany) was used. At this
stage, granules were placed in the machine via feeding section after
melting and primary film shaping, melted film were dispersed on the
cooling roller as a thin film. Finally, during the cooling, they were
pulled via several cold rollers and finally rotated around a tube.
Scanning electron microscopy (SEM) was conducted for observing
the failure surface of nano-composites. In order to achieving the op-
timum failure surface, samples put in liquid nitrogen. After complete
fractionation, samples fractures and their fracture section area coated
by a thin gold layer via coating instrument and evaluated by scanning
electron microscopy (MIRA3 SEM-FEG, Italy) at 3 kV under neutral
atmosphere at 35 kx (Zhong et al., 2007).
2.3. Strawberry fruit packaging and analyzing their different properties
In this step, 1080 strawberries samples were divided into three
groups of 360 samples for three types of nanocomposite packaging
containing C0S0, C1S0.75 and C2S0.75. Also, each 360-strawberries group
was divided into two coated and uncoated groups, each containing 180
strawberries. These steps were replicated for 25℃. Totally, 2160
strawberries were selected (180 (number of fruits for each level) × 2
(coating levels) × 3 (packaging film levels) × 2 (temperature levels
including 4℃ and 25℃). 10 fruits were put in each packaging and packs
were closed using thermal binding. Next, the samples were kept at 4
and 25℃ and physical (weight loss), chemical (pH and soluble solids
content), and mechanical (firmness and elasticity modulus) traits of
strawberries were evaluated. The present study was conducted as fac-
torial based on completely randomized design with three replications
including two coating levels, three film levels, and two temperature
levels to evaluate the effect of selected variables on the shelf life of
strawberry fruits. For this purpose, nanocomposite films were prepared,
their nanostructure was evaluated, and then fruits were packed with
these films. Afterward, various physical, chemical, and mechanical
traits were measured during the storage. Data were analyzed via SPSS
v20 software and the results were discussed.
Weight loss was determined by weighing the strawberry before and
during the storage period and the percentage of weight Loss was
H. Barikloo, E. Ahmadi
calculated according to the following Eq. (1):
W0 − Wt
Weight Loss (%) = × 100
W0 (1)
Where W0 is the initial weight of the strawberry and Wt is the weight at
time t (with t = 2, 4, 6, 8, and 10 days).
In order to determine the percentage soluble solids from each
sample, a refractometer (Model: 3820 (PAL-2), Resolution: ± 0.1%)
was utilized and pH value using and pH amount of each sample was
determined using a pH-meter (744 Metrohn, Made in china), (OECD
standard, 1998). Mechanical traits and fruit firmness were measured
through penetration test using materials test machine Zwick/Roell (bt1-
fr0. 5th. d14. Xforce hp, Made in Germany) having 500 N load cell
(Germany) with small diameter with concave probe of 6.4 mm, pene-
tration depth of 3 mm, and crosshead speed was set at of 10 mm/s.
Mechanical properties including firmness and elasticity modulus were
calculated from the force-deformation curve. Elasticity modulus was
obtained from Bosonic theory and Eq. (4). Stress and strain were ob-
tained respectively from Eqs. (2) and (3) respectively.
F chemical and mechanical properties of strawberry fruit
σ= , A = πr 2 (r = 3.2 mm)
A (2)
L0 − L1
ε=
L0 (3)
3/2
0.338 F (1 − ν 2) 1 11/3 1 1 1/3
E= ⎡K1 ( R1 + R′1 ) + K2 ( R2 + R′2 ) ⎤
D9/2⎣ ⎦
R1 = R′1 = R2 = R′2 and K1 = K2 = K (4)
Where E is elasticity modulus (MPa), F is the force (N), υ is a Poisson
ratio, and D (mm) is the deformation. K is a constant extracted from
standards (ASABE, 2008) and R (mm) is the radius of curvature in
contact points calculated from apparent dimensions (Barikloo and
Ahmadi, 2013).
MAP Henkelman 200a machine (made in Germany) was used for
packaging of strawberries. The percentages of oxygen, carbon dioxide
and nitrogen gases were selected 10%, 15% and 75%, respectively
(Soltani et al., 2015). Moreover, atmosphere changes inside the package
were measured using (WIT-GASETECHNIK Oxybaby 6i O2/CO2 Made in
Germany) device.
2.4. Statistical analysis
Data analyses were performed using Statistical Package for the
Social Sciences (SPSS v20). The analysis of variance (ANOVA) of the
data was conducted in the form of randomized multivariate factorial
(2 × 3 × 2) design. Main and interaction effects were evaluated.
3. Results and discussion
3.1. The SEM image of packaging film
SEM images represent a uniform distribution and a complete se-
paration of clay plates from each other. In addition, they show that the
nanoparticles have been well enclosed by a polymer substrate, which
indicates good compatibility with nanoparticles and polymers. The
obtained results are similar to those reported in other studies (Casariego
et al., 2009). As shown in Fig. 1(B, C), the distribution of clay and silica
nanoparticles is almost appropriate and aggregation at various areas in
failure surface, is not apparently observed. Surface view of nano-
composites is also intensive, and the presence of clay and silica nano-
particles in the images is completely clear. At higher concentrations, the
accumulation of nanoclay and nanosilica in films of C1S0.75 and C2S0.75
has increased compared to C0S0 film (without nanoparticles). However,
the increase in aggregation did not appear to be non-uniform or as parts
of high density. It seems that, the addition of nanoclay and nanosilica to
PE/POE/PA6 films leads to a compact and folded structure which can
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Scientia Horticulturae 240 (2018) 496–508
be well prevent from release of water vapor. During the storage period
in a cold environment such as a refrigerator, if moisture is created in-
side the packages, the nanosilica particles will absorb it and thus pre-
vent from spoilage of fruit in the package (Duncan, 2011).
According to the optical microscope images, it was observed that
nanosilica particles act as a nuclear matter in the growth of crystals.
Hence, the entry of these particles increases the number of crystals.
Clay silicate layers in nanocomposites leads to a significant reduction in
the permeability of gases from the polymer matrix compared to pure
polypropylene. Additionally, improving clay content in the nano-
composite reduces the permeability rate (Marsh and Bugusu, 2007;
Duncan, 2011). The most important reason for using these nano-
composites is the improvement of their mechanical, thermal properties
and impermeability compared to pure polymer or conventional com-
posites (Leslie and Ann, 2009). These nanocomposites have desirable
properties such as modulus and high resistance (Malwela and Sinha,
2011).
3.2. Effect of temperature, type of packing and coating on physical,
The results of the variables variance analysis for physical, chemical
and mechanical properties have been summarized in Table 2. The main
effects of temperature, day, film type of packaging and coating, and
their interaction effects on most of the measured characteristics were
significant.
3.2.1. The effect of temperature, type of the packing film and coating on
strawberry weight loss during the storage period
According to Table 2, the data related to the percentage weight loss
in all main effects and interactions effects including (tempera-
ture × packaging film), (temperature × coating), (temperature × day),
and (coating × day) had a significant difference at p < 0.05. Ac-
cording to the results obtained at two temperatures of 4 and 25 °C,
weight loss in the chitosan-coated samples that have been packed with
C0S0, C1S0.75 and C2S0.75 nanocomposite films under modified atmo-
sphere conditions, were lower than similar samples without chitosan
coating (Fig. 2). Based on the mean comparison (Fig. 2), it can be
concluded that weight loss percentage at 4℃ in all treatments was less
than that at 25℃. At 25 °C, weight loss in the C2S0.75 nanocomposite
film without coating and the C1S0.75 nanocomposite film having chit-
osan coating were the highest and lowest, respectively, while at 4 °C, in
C0S0 nanocomposite film without coating and C2S0.75 nanocomposite
film with coating were the highest and lowest, respectively.
At 25 °C, changes in the weight loss of fruit packed with C0S0,
C1S0.75 and C2S0.75 nanocomposite films without coating compared to
first day, were equal to 1.22, 1.35 and 1.64 respectively, while it in the
same samples, but with chitosan coating, were equal to 1.16, 1.13 and
1.34, respectively. A review of fruit weight loss percentage at 4 °C
showed that changes in the weight loss of fruit in the uncoated samples
that packed with C0S0, C1S0.75 and C2S0.75 nanocomposite films were
0.68, 0.65 and 0.51 (%) and in the samples having coating were 0.38,
0.26, and 0.3 (%) compared to the first day, respectively. Regarding the
percentages of nanoclay and nanosilica, the changes in weight loss were
not regular, but in general, they have been less in the C1S0.75 nano-
composite film compared to the C0S0 and C2S0.75 nanocomposite films.
The surface coating of strawberry fruit leads to reduction of surface
pores diameter, respiration and the water loss (Khalifa et al., 2016). The
effect of chitosan coating on the delaying in weight loss of strawberry
fruit in comparison with other cellulose derivatives, has been reported
by Riberio et al (2007). Park et al. (2005) stated that chitosan coating
solely decreases the weight of fresh strawberries by about 15%, com-
pared to uncoated samples. The use of clay nanosilicate packaging has
been reduced the weight loss percentage in strawberry (Zandi et al.,
2013; Neeta et al., 2013). The Nano-packaging, not only has kept
moisture in the fruit of the apple, but also it has prevented its weight
H. Barikloo, E. Ahmadi Scientia Horticulturae 240 (2018) 496–508

Fig. 1. SEM image PE/POE/PA6 nanocomposites films with different percentages of clay and silica nanoparticles.

loss (Zhou et al., 2011). Reducing respiration rate causes down the
percentage of product weight loss (Lu and Toivonen, 2000). On the
other hand, a higher percentage of weight loss in uncoated samples that
packed with C0S0, C2S0.75 films, at 4 °C and 25 °C, can be attributed to
more suitable ventilation and more favorable conditions for the re-
spiratory process, that is consistent with the results already obtained
(Artes-Hernandez et al., 2006). Also, attention to how to placement of
clay and silica nanoparticles in between the polymer, coating of chit-
osan on fruit, and modified atmosphere with optimal gas composition
conditions has led to lower weight loss in samples packed with C1S0.75,
C2S0.75 nanocomposite films with coating at temperatures of 25 °C and
4 °C, respectively. Losing water of fruit depends on the force caused by
the water vapor pressure deficit between the fruit tissue and the sur-
rounding air as well as tissue resistance against this force (Soltani et al.,

Table 2
Analysis of variance (mean square) for effects of temperature (TE), package type (PA), coating (CO) and day (D) on the characteristic weight loss, pH, Soluble solids
content (%), Firmness (N) and Young modulus (N/mm2) strawberry fruit.
Variable df Mean Square (M.S)

Weight loss (g) pH SSC (%) Firmness (N) Young modulus (N/mm2)

TE 1 10.434a .026a 0.410ns 56.348a 0.002a

PA 2 0.290a 0.090a 0.362ns 16.856a 0.005a
CO 1 1.226a 0.065a 1.883a 0.032a 55.651a
D 5 20.952a 1.039a 2.096a 0.011a 31.177a
TE × PA 2 0.851a 0.002ns 2.262a 0.434a 0.003ns
TE × CO 1 0.085a 0.030a 0.488ns 0.273ns 0.002a
TE × D 5 5.618a 0.137a 1.332ns 2.888a 0.002a
PA × CO 2 0.091ns 0.046a 10.656 14.172a 0.010a
PA × D 2 0.099ns 0.186a 2.509ns 3.652ns 0.001ns
CO × D 5 0.564a 0.060a 4.605a 3.972a 0.002a
TE × PA × CO 2 .047ns 0.318a 1.141a 8.427a 0.004a
TE × PA × D 10 0.316ns 0.066a 2.856a 3.354ns 0.001ns
TE × CO × D 5 0.062ns 0.259a 5.117a 0.519ns 0.002a
PA × CO × D 10 0.189ns 0.302a 4.637a 3.484ns 0.002ns
TE × PA × CO × D 10 0.162ns 0.098a 2.353ns 3.936ns 0.001ns
Error 144 3.094 0.367 20.563 41.034 0.021
CV (%) – 3.391342 0.1619 0.90296 2.586652 4.582718

ns: No significant difference.

a
Significant difference at 5%.

499
H. Barikloo, E. Ahmadi
Fig. 2. Effects of temperature, coating, and packaging film on changes of strawberry weight loss during storage and modified atmosphere conditions.
2015). The pressure deficit rises with increasing temperature and de-
creasing moisture. Given that the strawberry fruit has a thin skin and it
quickly loses its water, therefore, its packaging with the modified at-
mosphere can maintain the vapor pressure around of the fruits at an
upper limit which, in turn, prevents water loss and weight loss of fruits.
3.2.2. Effect of temperature, packaging and coating on the chemical and
mechanical properties of strawberry
The results of variance analysis of variables for the measured che-
mical properties, have been presented in the Table 2. As can be seen,
about pH, all the main effects and the interaction effects, except for the
interaction effect (temperature × packing), were significant. Moreover,
the results of analysis of variance for percentage of SSC showed that
except for the effects of temperature and packing as well as the inter-
action effects (temperature × coating), (temperature × day),
(packing × day) and (temperature × coating × day × packing), all the
main and interaction effects were significant.
According to Fig. 3, it can be concluded that in all treatments,
changes of pH at 4 °C are lower than 25 °C. Regarding to the results
obtained at two temperatures of 4 and 25 °C, it is clear that changes of
pH in the chitosan-coated samples that have been packed with C0S0,
C1S0.75 and C2S0.75 nanocomposite films under modified atmosphere
conditions, were lower than similar samples without chitosan coating
(Fig. 3). This is due to reduction of respiration rate of strawberries
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Scientia Horticulturae 240 (2018) 496–508
packed in optimum atmosphere and appropriate coating conditions.
Because of the reducing respiratory rate of the fruit at the refrigerator
temperature, the chemical changes in the fruit tissue occur at a lower
rate. The favorable effect of nanocomposite film and chitosan coating
with low permeability plays a remarkable role in decreasing oxygen
content and consequently decreasing the pH change (Gholami et al.,
2018). Moreover, cultivar type and storage conditions affect the in-
ternal process and post-harvest behaviors. Generally, changes in the pH
of the fruit during storage can be due to metabolic activity such as
respiration, which, in turn, causes the consumption of sugars and or-
ganic acids of the fruit (Yun et al., 2017). Considering to Fig. 3, under
ambient conditions, pH value in all three types of packaging had an
increasing trend over time until 8th day and was decreasing trend
afterwards. This irregular pattern may be related to the decomposition
of cell wall until eighth day (Kerry and Butler, 2008). Generally, during
the storage period, the changes in pH at 25 °C in the C0S0 nano-
composite film without coating and in C1S0.75 nanocomposite film
having coating were highest and lowest values, respectively. However,
changes of pH at 4 °C were irregular, so that in C0S0, C1S0.75 and C2S0.75
nanocomposite films with coating had an incremental trend until 8th
day, then had a downward trend. The incremental changes in pH value,
in 25 °C, in samples packed with C0S0, C1S0.75 and C2S0.75 nano-
composite films with coating compared to the first day, were equal to
0.55, 0.16 and 0.27 and for uncoated samples, this changes were equal
H. Barikloo, E. Ahmadi
Fig. 3. Effects of temperature, coating, and packaging film on changes of strawberry pH during storage and modified atmosphere conditions.
to 0.1, 0.06 and 0.25, respectively. At 4 °C, the trend of pH changes in
samples packed with C0S0, C1S0.75 and C2S0.75 nanocomposite films
without coating compared to the first day, were equal to 0.1, 0.006 and
0.023, and in coated samples were equal to 0.07, 0.19 and 0.44 re-
spectively. Ayala-Zavala et al (2007) reported that pH value in straw-
berry fruit increases during the storage. Kader and Watkins (2000)
stated that the difference between pH in the researchers' findings may
depend on the different effects of increased oxygen on the respiration
rate in the product. With carbon dioxide gas treatment in the straw-
berry fruit store, Cordenunsi et al. (2003) concluded that the pH in-
creased and acidity decreased simultaneously. This change was de-
pendent on the concentration of carbon dioxide. In a study, during the
28 days of storage, the acidity of packed Kiwi fruits with nanocomposite
and pure polyethylene decreased. During this period, there was no
significant difference in amount of acidity in none of packaging film,
but after a 42-days storage, the acidity of the samples in the pure
polyethylene were less than ones in the nanocomposite film (Hu et al.,
2011).
As can be seen in Fig. 4, there is no considerable difference in
changes in soluble solids content under ambient conditions up to day 2
and the increasing was the same. From days 2 to 8 of storage, the lowest
amount of decreasing changes for soluble solids were observed for
C1S0.75 nanocomposite film with coating and highest amount was ob-
served for C2S0.75 nanocomposite film without coating; however, from
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Scientia Horticulturae 240 (2018) 496–508
day 8 to day 10, a decreasing trend was observed for soluble solids
changes. An increase in Brix percentage and pH during storage might be
associated with the transformation of pectic substances, starch hydro-
lysis, and dehydration of fruits (Goncalves et al., 2000). Moreover, at
25 °C, changes of soluble solids in samples packaged with C0S0, C1S0.75
and C2S0.75 nanocomposite films with coating under modified atmo-
sphere until the 6th day had an increasing trend and then had a de-
creasing trend, so that the minimum (0.37) and maximum (1) changes
were observed in C1S0.75 and C2S0.75 respectively. The changes in the
soluble solids content in samples packed with C0S0 and C1S0.75 nano-
composite films without coating and in samples packed with C0S0,
C1S0.75 and C2S0.75 nanocomposite films having coating under atmo-
sphere conditions at 4 °C were an increasing trend until the fourth day,
and after that, these changes had a decreasing trend. However, the
highest (0.85) and lowest (0.45) changes of soluble solids content were
observed in films of C0S0 and C1S0.75 having coating, respectively. Also,
in samples packed with C2S0.75 nanocomposite film without coating, the
process of changes in the soluble solids content was reduced until 4th
day and then it increased. Generally, the lowest changes in soluble
solids content between nanocomposite films were observed in the
C1S0.75 film having coating under atmosphere conditions. Increasing
soluble solids during storage could be due to the conversion of carbo-
hydrates to sugars and other soluble compounds that it was occurred
through metabolic processes and decrease in fruit water, which in turn
H. Barikloo, E. Ahmadi
Fig. 4. Effects of temperature, coating, and packaging film on changes of strawberry SSC (%) during storage and modified atmosphere conditions.
causes an increase in soluble solids concentration (Yun et al., 2017).
Moreover, as shown in Fig. 4, during storage period, the soluble solids
content has been increased, and in samples that were packed in C1S0.75
film and coated with chitosan compared to other samples, increasing
the soluble solids content is lower, in other words, the speed of fruit
ripening in them are less. This can also be attributed simultaneously to
reduction of respiration (Gholami et al., 2018) and absorption of
ethylene emitted from fruit by nanoclay particles in nanocomposites
(Kerry and Butler, 2008). The coatings reduce the respiration rate of
fruit through delaying in use of organic acids in respiratory enzymatic
reactions. Additionally, they prevent from the increasing in soluble
solids content by slowing down the speed of fruit ripening (Hernandez-
Munoz et al., 2008; Gholami et al., 2018). In the other words, nano-
composite films have prevented a sharp increase in soluble solids
compared to other samples in reducing the amount of metabolism and
reducing the respiration of fruit. In another investigation, in the 28 days
since the beginning of storage, the soluble solids content of kiwi fruit
was increased in both the polyethylene and nanocomposite packages,
and the improving speed of this property, for fruit that were packed
with pure polyethylene was more (Hu et al., 2011). Moreover, Nano
packaging has significantly maintained the soluble solids content in
cherry cv. Syaahe Mashhad compared to conventional polyethylene
containers, and prevented its excessive increase in 45 days of storage
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Scientia Horticulturae 240 (2018) 496–508
(Zandi et al., 2013). At the end of the storage period, the coated
treatments had a higher sugar content than the control sample. On the
other hand, the coating action creates a protective layer on the surface
of the fruit, and the oxygen penetration in it is done very slowly
(Javanmard, 2008). With using this packaging and coating system, the
respiratory rate can be reduced. Reducing the rate of respiration leads
to the retaining the desired taste of the product, reducing the changes of
organic acids, maintaining the optimal pH and preventing the loss of
sugar, which ultimately leads to increment of storage time of straw-
berry.
One of the important factors in marketability of fruit is its tissue
firmness, which is effective on its quality. The results of variance ana-
lysis of variables for mechanical properties are summarized in Table 2.
The results of analysis of variance related to the main effects including
temperature, type of packaging film, type of coating and day on the
amount of firmness and elasticity modulus were significant at
P < 0.05. In addition, the interaction effects such as tempera-
ture × film packing, temperature × day, coating × film packing,
coating × storage period, temperature × film packing × coating on the
amount of firmness was significant at P < 0.05 (Table 2).
According to the obtained results (Fig. 5), the variation of the
firmness and the elasticity modulus during the storage period at 4 °C
was less than 25 °C. The produced ethylene amount, the fruit ripening
H. Barikloo, E. Ahmadi
Fig. 5. Effects of temperature, coating, and packaging film on changes of strawberry firmness during storage and modified atmosphere conditions.
and respiration intensity are effective factors in tissue alteration (Pretel
et al., 1999). Depending on the storage conditions, the fruit ripening,
respiration and ethylene production at high temperature is more than
low temperature. For this reason, firmness loss due to soften of the
texture in the normal environments is happen faster compared to re-
frigerator (Hernández-Munoz et al., 2006). At both temperatures (4 °C
and 25 °C), the changes of the firmness and the elasticity modulus in
samples packed with C0S0, C1S0.75 and C2S0.75 nanocomposite films
under modified atmosphere conditions having chitosan coatings were
less than samples without coating (Figs. 5 and 6). Generally, during the
storage period and at 25 °C, the maximum and minimum of changes in
firmness (1.62 and 0.88) were observed in C0S0 nanocomposite film
without coating and C1S0.75 nanocomposite film with coating, respec-
tively. The trend of these changes was reducing until the eighth day,
then, with modest changes, it was increasing. Also, at temperature of
4 °C, variations trend of firmness was decreasing until eighth day, and
then increased, so that the highest (2.02) and lowest (0.38) changes
were observed in C0S0 without coating and C2S0.75 with coating, re-
spectively. In general, the C1S0.75 film with chitosan coating had the
best conditions in the case of firmness control, under atmosphere con-
ditions. Hernández-Munoz et al (2006) founded that after 4 days of
storage, firmness values for strawberries coated with chitosan were
higher than those for uncoated samples. The activity of enzymes of
polygalacturonase, β-galactosidase and pectin-methyl esterase are as
most important factors in softening of fruit texture and any factor that
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Scientia Horticulturae 240 (2018) 496–508
inhibits the activity of these enzymes will result in better tissue firmness
(Yun et al., 2017). Nano packs have led to maintain of strawberry fruit
strength, in addition, they have prevented from too much softening of
fruit during the 18 days of storage (Sogvar et al., 2016). The employing
of coating, Nano-packaging and proper temperature are effective in
achieving this goal (Sogvar et al., 2016). Moreover, maintenance of
strawberries in an optimal modified atmosphere conditions has a fa-
vorable effect on decreasing the amount of firmness during storage
period. Regarding to the presence of clay and silica nanoparticles in the
polymer matrix, the film's permeability has been reduced which, in
turn, caused the decreasing the gas exchange in the nanocomposite
film. Also, the combined employing of nanocomposite coatings and
films leads to improving carbon dioxide, absorbing of the ethylene,
maintaining of moisture inside the package (due to the presence of clay
and silica nanoparticles in nanocomposites), as well as delaying the
process of softening the fruit during storage (Kerry and Butler, 2008).
ANOVA results of main effects of temperature, packaging film type,
coating type, and storage period on elasticity modulus showed that the
mentioned factors had a significant impact on elasticity modulus and
interaction effects of (temperature × coating), (temperature × day),
(coating × film packing), (coating × day), (temperature × film
Packaging × coating) and (temperature × coating × day) on elasticity
modulus were statistically significant (P < 0.05; Table 2). But other
interaction effects of factors on elasticity modulus were not statistically
significant. According to the obtained results (Fig. 6), the variation of
H. Barikloo, E. Ahmadi
Fig. 6. Effects of temperature, coating, and packaging film on changes of strawberry elasticity modulus during storage and modified atmosphere conditions.
the elasticity modulus during the storage period at 4 °C was less than
one at 25 °C. The reason for this phenomenon is the respiration of fruit
and the production of ethylene, which leads to increment of fruit ri-
pening as well as directly effects on the softness of tissue (Perkins-
Veazie, 2010).
By investigation of the elasticity modulus, it was found that its
changes at 25 °C was more than at 4 °C and it also in sample with
coating was less than sample without coating. In general, during the
storage period, the changes of elasticity modulus at 25 °C in the C2S0.75
nanocomposite film without coating and in the C1S0.75 nanocomposite
film with coating, were the highest (0.055) and lowest (0.0028)
amount, respectively. The process of these changes was shrinking until
the eighth day, and then with modest changes, it increased. At 4 °C, the
elasticity modulus changes were decreasing until the eighth day and
then it was increasing, so that the highest (0.036) and the lowest
(0.00066) amount of changes were related to the C0S0 film without
coating and the C2S0.75 film with coating, respectively. But generally, in
the case of controlling changes of elasticity modulus under modified
atmosphere conditions, the most optimal of conditions were observed
in the C2S0.75 film with chitosan coating. The better performance of the
film with chitosan coating may be due to the reduction of strawberries
respiration activity, which leads to delaying the ripening and softening
of strawberry fruit (Sogvar et al., 2016). The investigation of the de-
creasing trend related to changes of elasticity modulus at 4 °C indicated
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Scientia Horticulturae 240 (2018) 496–508
that the elasticity modulus of fruit in samples packed with nano-
composite films containing C0S0, C1S0.75 and C2S0.75 without coating
were equal to 0.0362, 0.025 and 0.029 compared to the first day, and in
samples having coating, this changes were equal to 0.025, 0.01 and
0.00066, respectively. Furthermore, at 25 °C, the variations in elasticity
modulus of fruit in samples packed with C0S0, C1S0.75 and C2S0.75 na-
nocomposite film without coating were equal to 0.0449, 0.0324 and
0.055, and in the samples with coating were equal to 0.0103, 0.0028,
and 0.0142, respectively. Factors affecting fruit elasticity modulus and
firmness include cell wall strength, cell to cell contact, and cellular
turgor pressure, as well as degradation of the middle lamella of the cell
wall in cortical parenchyma cells during ripening (Perkins-Veazie,
2010). The elasticity modulus and firmness of fresh strawberries of all
groups showed a decreasing trend during the storage, which it was si-
milar to the results reported in (Fan et al., 2009). The loss in elasticity
modulus during storage of pear fruits occurred due to breaking down of
enzymes, loss of water, and degradation of pectic substances present in
the fruit (Nath et al., 2012).
3.2.3. Evaluation of the effect of temperature, type of packaging and coating
on oxygen and carbon dioxide changes inside the package
Results obtained from variance analysis of variables for oxygen and
carbon dioxide changes in three types of C0S0, C1S0.75 and C2S0.75 na-
nocomposite films under modified atmosphere are presented in Table 3.
H. Barikloo, E. Ahmadi Scientia Horticulturae 240 (2018) 496–508

Table 3 The results of variance analysis related to the main effects of tem-
Analysis of variance (mean square) for effects of temperature (TE), package perature, type of packaging film, type of coating and day on the changes
type (PA), coating (CO) and day (D) on the characteristic O2 (%) and CO2 (%) in oxygen and carbon dioxide gases were significant at P < 0.05. Apart
strawberry fruit. from the interaction effect of (temperature × packaging
Variable df Mean Square (M.S) film × coating × day), all of the interaction effects on oxygen changes
and interaction effect (temperature × packaging film), (tempera-
O2 (%) CO2 (%) ture × packaging film × coating) and (temperature × packaging
TE 1 536.642 a
94.010a
film × coating × day) on carbon dioxide gas changes, were significant
PA 2 211.624a 95.764a at P < 0.05, (Table 3). By examining the results in Fig. 7, it is observed
CO 1 3564.539a 1127.510a that changes of oxygen gas at 25 °C are more than the temperature of
D 5 352.514a 1086.470a 4 °C, additionally, these changes in chitosan-coated sample are less
TE × PA 2 11.598a 0.554ns
compared to the uncoated sample.
TE × CO 1 864.150a 219.615a
TE × D 5 118.258a 32.321a At 25 °C, the trend of oxygen changes in the samples of without
PA × CO 2 148.543a 66.914a coating which packed with the C0S0, C1S0.75 and C2S0.75 nanocomposite
PA × D 2 73.260a 32.072a films were reducing until the tenth day of storage, in which the C1S0.75
CO × D 5 1146.707a 410.036a
and C2S0.75 samples have the most and the least variations, respectively.
TE × PA × CO 2 13.050a 0.601ns
TE × PA × D 10 33.911a 9.015a
Moreover, the numerical values of oxygen changes relative to the first
TE × CO × D 5 55.630a 23.587a day, at 25 °C, in the samples of without coating which packed with the
PA × CO × D 10 55.630a 23.587a C0S0, C1S0.75 and C2S0.75 nanocomposite films are equal to 10.4, 18.65
TE × PA × CO × D 10 11.858ns 5.203 ns and 14.85, respectively. Oxygen changes in chitosan-coated samples
Error 144 236.845 34.650
which packed with C0S0, C1S0.75 and C2S0.75 nanocomposite films were
CV (%) – 1.18596 1.039543
decreasing trend until the second day of storage and then they had an
ns: No significant difference. incremental and almost the same trend, so that the lowest and the most
a
Significant difference at 5%. amount of this changes were observed in the C0S0 and C1S0.75 samples.

Fig. 7. Effects of temperature, coating, and packaging film on changes of strawberry O2 during storage and modified atmosphere conditions.

505
H. Barikloo, E. Ahmadi
Fig. 8. Effects of temperature, coating, and packaging film on changes of strawberry CO2 during storage and modified atmosphere conditions.
Also, at 4 °C, the oxygen values in the uncoated samples which packed
with C0S0, C1S0.75 and C2S0.75 nanocomposite films were decreasing
until the tenth day, while the maximum and minimum changes of
oxygen were related to C0S0 and C2S0.75 samples. As can be seen in
Fig. 8, variations of carbon dioxide gas at a temperature of 25 °C are
higher than 4 °C, while the amount of this variations in chitosan-coated
sample is less than the uncoated sample. At 25 °C, the carbon dioxide
values increased in the uncoated samples, which packed with C0S0,
C1S0.75 and C2S0.75 nanocomposite films, until the tenth day of storage,
and the highest and lowest amount of it's were obtained for the un-
coated sample packed with C0S0 film and the chitosan-coated sample
packed with C1S0.75 film. However, the trend of carbon dioxide changes
in the chitosan-coated samples packed with C0S0, C1S0.75 and C2S0.75
films were decreasing and its almost were identical.
Also, at 4 °C, the carbon dioxide values, in samples packed with
C0S0, C1S0.75 and C2S0.75 nanocomposite films, were decreasing until
the tenth day of storage, and the highest and lowest amount of it's were
observed in the uncoated sample packed with C0S0 film and the chit-
osan-coated sample packed with C2S0.75 film. The investigation of
carbon dioxide changes at 4 °C indicated that these changes in the un-
coated samples packed with nanocomposite films containing C0S0,
C1S0.75 and C2S0.75, compared to the first day, were equal to 11.4, 10.45
and 7.2 and in the chitosan-coated samples, these values were equal to
0.4, 0.3 and 0.55, respectively. In packaging system with modified
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Scientia Horticulturae 240 (2018) 496–508
atmosphere, the selection of suitable films to create the primary at-
mosphere and its preservation in long-term, is very essential (Mangaraj
et al., 2009). If the permeability of packaging film relative to oxygen
and carbon dioxide matches with the respiration of products, balance of
modified atmosphere will be created in the package and the shelf-life of
the product will be also increased (Pretel et al., 2000). In order to
packaging of fresh products under MAP conditions, it is possible to use
from films with high, medium and low permeability. The amount of
changes in oxygen and carbon dioxide gases in 25 °C compared to the
4 °C, were more. This is due to fact that at higher temperatures, the
amount of strawberry respiration is higher which, in turn, leads to more
oxygen consumption and more carbon dioxide production (Sandhya,
2010). If the temperature in the warehouse increases, the packaging
effect with the modified atmosphere method, will be reduced. Because
in this case, the amount of solubility of CO2 decreases and the re-
spiration rate increases. In fact, with the increasing temperature,
oxygen consumption in fresh product is raised, as a result, the shelf-life
of the product is declined (Sandhya, 2010). In this regards, Lopez
Briones et al. (1993) stated that the two factors as the respiration of
product and permeability of the packaging film, which depend on
temperature, change the atmosphere inside the package in which con-
taining living productions, so that those cause carbon dioxide removal
and oxygen entry. However, the effect of temperature on respiration
rate is higher. In permeable packaging, doing respiration, as the oxygen
H. Barikloo, E. Ahmadi
in the pack is consumed, a constant change occurs in composition of the
package atmosphere, so that, first the carbon dioxide concentration
increases against oxygen, since the film is always more sensitive to
carbon dioxide than oxygen, carbon dioxide is released faster than
input oxygen, but since the film is always more permeable to carbon
dioxide than oxygen so, the outlet of carbon dioxide will occur faster
than the entry of oxygen (Aday et al., 2011). With the consumption of
oxygen and production of carbon dioxide, the gradient of gas will be
created inside the package (Aday et al., 2011). This gradient removes
the carbon dioxide and it leads to the entry of oxygen. Initially, the
propulsion is low, but it will ultimately create a balance (Smith and
Stow, 1984). Nano fillers can be considered as impervious barriers to
the movement of oxygen molecules. Additionally, silicate and clay
layers can also be effective as an impervious barrier against the
movement of oxygen molecules (Zeppa et al., 2009). In this regards,
Moraru et al. (2003) indicated that the uniform distribution of nano-
particles inside the plastic makes the layer extremely impenetrable, in
the other hand, the penetration and exchanging of carbon dioxide,
oxygen and vapor between the two outer and inner spaces is declined.
Generally, in the packaging of products, oxygen is considered as an
undesirable element. In fact, warehouses having the changed and
controlled atmosphere the amount of respiration via limiting the ex-
change of oxygen and carbon dioxide, decrease and as a result they
increase products life (Yun et al., 2017). In this way, by consumption
oxygen and producing of carbon dioxide, the concentration of oxygen in
the packaging environment is reduced and the carbon dioxide con-
centration is increased. This results in a reduction the amount of re-
spiration and other oxidative reactions which can lead to enzymatic
browning, thus the quality and shelf life of the product is increased
(Serrano et al., 2006).
4. Conclusion
In the present research, SEM images represent a uniform distribu-
tion and a complete separation of clay plates from each other. On the
other hand, these images show that nanoparticles have been well en-
closed by a polymer substrate, which indicates good compatibility with
nanoparticles and polymers. In general, during the storage period,
strawberries coated in a PE/POE/PA6 nanocomposites film with a
percentage of the C1S0.75 mixture preserved well mechanical and qua-
litative properties in ambient and refrigerated conditions. The changes
in parameters such as weight loss, pH, soluble solids, firmness, oxygen,
carbon dioxide and elasticity modulus at 4 °C were less than 25 °C
during the storage period. The weight loss in chitosan-coated straw-
berry at 25 °C and 4 °C had the lowest amounts in films of C1S0.75 and
C2S0.75, respectively, that it can be attributed to the favorable effect of
chitosan coatings and clay and silica nanoparticles. The pH variations in
the refrigerated conditions had an irregular trend, however, these
changes had the lowest amount in ambient temperature and in the
chitosan-coated sample that packed with C1S0.75 film. The minimum
and maximum of changes the soluble solids, at 25 °C and 4 °C, were
observed in C1S0.75 and C0S0 films, respectively. About the controlling
of the penetration force in modified atmosphere conditions, in general,
the C1S0.75 film with coating had the most optimal conditions, however,
at temperature of 4 °C, the coated sample packed with C2S0.75 film had
less variations of penetration force. Based on obtained results, the
changes in elasticity modulus, at 25 °C, were more compared to 4 °C,
and also, these changes in the chitosan-coated sample were less than the
uncoated sample. Oxygen changes in coated samples, until the second
day, had a decreasing trend and then, they increased. Also, these
changes in the sample without coating had a decreasing trend, in ad-
dition at ambient temperature was higher than refrigerator tempera-
ture, so that these variations in the C2S0.75 film were the lowest amount.
Carbon dioxide changes had a diminishing trend in both ambient and
refrigerated conditions. Also, changes in carbon dioxide in samples
coated with chitosan at ambient and refrigerator temperatures, in the
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Scientia Horticulturae 240 (2018) 496–508
C1S0.75 and C2S0.75 films were the lowest value, respectively. The results
of this investigation indicated that the employing of clay-silica PE/
POE/PA6 nanocomposites film with chitosan coating under atmosphere
conditions is useful for maintaining the qualitative and mechanical
properties of fruit and improving the shelf life, which is important in
terms of marketability and exporting.
Appendix A. Supplementary data
Supplementary material related to this article can be found, in the
online version, at doi:https://doi.org/10.1016/j.scienta.2018.06.012.
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