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International Journal of Biological Macromolecules xxx (2017) xxxxxx

Contents lists available at ScienceDirect

International Journal of Biological Macromolecules

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

Physicochemical and microstructural properties of a novel edible lm

synthesized from Balangu seed mucilage
Atina Sadeghi-Varkani, Zahra Emam-Djomeh , Gholamreza Askari
Transfer Research center for controlled release, Phenomena Laboratory (TPL), Department of Food Science, Technology and Engineering,College of
Agriculture and Natural resources, University of Tehran

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

Article history: This paper reports the synthesis of a novel edible lm from Balangu seed mucilage (BSM) as a new carbo-
Received 10 June 2017 hydrate source. Optimal formulation of the proposed edible lm was found through fabricating several
Received in revised form 5 November 2017 distinct lms with different concentrations of BSM and glycerol. The effect of these formulation vari-
Accepted 6 November 2017
ables on the physical, mechanical, thermal, barrier, and microstructural properties of the manufactured
Available online xxx
lms was then investigated. Optimal formulation of the BSM edible lm was then determined based
on the measured mechanical and barrier characteristics. These characteristics were found to deteriorate
Keywords:
with an excessive use of glycerol which caused non-homogeneity of the lms as observed through scan-
Biodegradable lm
Balangu seed mucilage
ning electron micrographs. In-depth analysis of the optimal BSM lm properties was performed through
Film optimization investigating its oxygen permeability, Fourier transform infrared spectroscopy, atomic force microscopy,
X-ray diffraction, and water sorption isotherm. The superior mechanical and barrier characteristics of the
obtained optimal BSM edible lm make it a potential candidate for packaging that aim at an extended
shelf-life.
2017 Elsevier B.V. All rights reserved.

1. Introduction erties, plasticisers such as glycerol [7,911], glycol [12], sorbitol

Widespread application of synthetic petroleum packaging in
the food industry has negatively impacted the environment over
the past century [1]. Bio-based packaging materials have there-
fore been introduced as a green alternative in the past decades,
among which, edible lms have gained more attention due to their
environmentally-friendly characteristics, vast variety and avail-
ability, non-toxicity, and low-cost [2]. Edible lms are commonly
produced using lipids, carbohydrates, and proteins; however, car-
bohydrates are more common due to their abundant sources [3].
Carbohydrates such as starch [4], cellulose [5], chitosan [6],
gum, and mucilage [7] were the subject of many studies for
biodegradable coatings and lms. Water-solubility and gel-forming
characteristics of carbohydrates have enabled manufacturing of a
new class of edible lms that are good barriers against oxygen and
can be used for increasing the food shelf-life by preventing from
microbial spoilage [8]. In order to improve carbohydrate lm prop-
This research did not receive any specic grant from funding agencies in the
public, commercial, or not-for-prot sectors.
Corresponding author at: Center of Excellence in Biothermodynamics, University
of Tehran, Tehran, Iran.
E-mail address: ememj@ut.ac.ir (Z. Emam-Djomeh).
[9,12,13], xylitol [12], and sucrose [9] are usually added to the lm
solution in the production stage. This weakens the biolm molec-
ular interactions leading to an improved exibility and toughness
as well as reduced glass transition temperature. On the downside,
the hydrophilic nature of glycerol causes poor water vapour barrier
characteristics and increased water solubility in carbohydrate lms
[14].
This study uses mucilage of Balangu plant seed for the rst time
as a new source of carbohydrate for fabricating novel edible lms
for food packaging. Balangu plant (Lallemantia iberica F. & C. M.),
classied in the mint family, is locally known as iberica dragons
head or lions head, and is famous for use in traditional medicines.
This plant, originally from the Caucasus and the Middle East, is
spread throughout European and eastern countries [15]. Balangu
seed mucilage (BSM) features many advantages over most of its
polymer counterparts, including a lower production cost, higher
efciency, and better medicinal properties.
BSM polymer, in its pure form, is not exible enough for packag-
ing, thus glycerol was used in this study as a plasticizer in the lm
formulation. To this aim, several concentration levels of glycerol
and BSM were used in fabricating edible lm samples of different
characteristics. These characteristics, including thickness, mois-
ture content, water solubility, water vapour permeability, oxygen
permeability, colour and transparency, and mechanical properties

https://doi.org/10.1016/j.ijbiomac.2017.11.029
0141-8130/ 2017 Elsevier B.V. All rights reserved.

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were experimentally measured with respect to the glycerol and
BSM concentration levels in lm formulation. Scanning electron
microscopy images of the fabricated lm samples were used to
investigate their distinct characteristics. Ultimately, the optimal
formulation of the proposed BSM edible lm was determined based
on the obtained experimental results to have an improved mechan-
ical and barrier characteristics. Further investigations were then
carried out on the optimal BSM lm properties by obtaining their
Fourier transform infrared spectroscopy, atomic force microscopy,
X-ray diffraction, and water sorption isotherm. The results of this
study indicate that the developed optimal BSM edible lm features
promising properties for food packaging applications aiming at an
extend shelf-life.
2. Materials and methods
2.1. Materials
Balangu seeds used in this study were native to Iran and
obtained through local suppliers. Ethanol of 96% purity was pur-
chased from Ghadir Industries (Tehran, Iran). Glycerol with%95
purity was from SigmaAldrich (Aldrich, MO, USA). Other materi-
als used in the experiments, including magnesium nitrate, sodium,
and chloride, were obtained from Merck Corporation (Readington
Township, NJ, USA).
2.2. Extraction of BSM
Balangu seeds were stored in vacuum packages at 25 C before
commencing the experiments. In order to purify the seeds, they
were sieved and soaked in ethanol (96% w/v) for 30 min to remove
dust and dirt. The seeds were then dried in a conventional oven at
30 C for 12 h, after which, the seed mucilage was extracted through
hydration process. To this aim, the seeds were soaked in distilled
water and mechanically stirred at 40 C for 3 h. The seed to water
ratio was chosen to be 1:35 based on preliminary experiments.
The seed-water compound was then ltered through a cheese-
cloth to remove the seeds from the BSM solution. In the nal step,
the obtained BSM solution was centrifuged at 3000 rpm for 15 min
to remove any impurity. The resultant puried BSM solution was
freeze-dried and vacuum-packed to preserve its characteristics for
use in the next stages of this work.
2.3. Synthesis of the proposed edible BSM lms
Four concentration levels of deried BSM measuring at 0.4%,
0.8%, 1.2% and 1.6% (w/v) were used with three concentration lev-
els of glycerol equaling 10%, 35%, and 60% (w/w) with respect to
the mucilage weight, resulting in twelve distinct edible lms. For
each concentration of dried BSM, an aqueous mucilage solution
was prepared by dissolving the respective amount of dried BSM,
obtained in Section 2.2, in distilled water. A homogeneous solution
was achieved by stirring the resultant compound using a magnetic
stirrer at a constant speed of 500 rpm at 25 C for 4 h (RCT-basic,
IKA, Staufen, Germany). Different lm-forming solutions were then
obtained by adding different concentrations of glycerol to the BSM
solution. pH of each solution was maintained at 8.01. The solu-
tions were homogenised using a high-speed dispersing instrument
at 25 C and the speed of 10,000 rpm for 20 min (Ultra-Turrax T25,
IKA, Staufen, Germany). To enhance the edible lm characteristics,
air bubbles and insoluble solids were removed from the lm-
forming solution by centrifuging at the speed of 1000 rpm at 25 C
for 10 min. BSM edible lms were fabricated by casting a specic
amount of the solution on Teon plates followed by a dehydra-
tion process using an air convection oven at 40 C and 40% relative
humidity (RH) for 24 h. In the nal step, the lms were peeled off
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the plates and conditioned for a minimum of 20 h at ambient tem-
perature in a desiccator lled with saturated magnesium nitrate
(52% RH).
2.4. Physical properties of the BSM lms
Thicknesses of the manufactured lms were measured using a
digital micrometer with a precision of 0.01 mm. Each lm sample
was gauged at ve random regions, and the mean value was taken
as the associated thickness.
In order to measure the moisture content of the lms, samples
with 2 cm diameter were dehydrated in an oven at 105 C for 24 h,
after which, the sample weights remained constant. The measured
difference in the sample weight after this period is referred to as the
moisture content of the sample. This experiment was performed
three times on each lm sample, and the mean value was taken as
the moisture content the associated BSM lm.
In order to obtain water solubility (WS) of the lms, a sample of
each lm was heated in the oven at 105 C for 24 h to reach a sta-
ble moisture level. The weight of the sample was then measured
(Wi ), after which, it was immersed in 30 ml of distilled water. The
solution was stirred gradually at 25 C for 24 h and then ltered
through a paper lter. The paper lter which contained the undis-
solved portion of the lm was dehydrated in the oven at 105 C for
24 h. Subsequently, the amount of the undissolved substance was
measured (Wf ). Water solubility was then calculated by:
Wi Wf
WS(%) = 100 (1)
Wi
The method for measuring water vapour permeability (WVP)
for the BSM lms was adopted from the standard ASTM E96-
95. Each lm sample was sealed using parafn wax over a glass
cup to isolate the cup inner-atmosphere from the outside (inner
diameter = 3.8 cm, height = 4.3 cm). The cup was initially lled with
anhydrous calcium chloride to ensure 0% RH. The cup was put inside
a desiccator with saturated sodium chloride solution to maintain
75% RH. The lower RH of the cup inner-atmosphere absorbs humid-
ity from the desiccator and into the anhydrous calcium chloride
resulting in an increase in the cups weight. This experiment was
performed for 48 h with the cup weights being measured every 6 h.
Water vapour permeability of the lm sample was then determined
by:
GL
WVP = (2)
A t P
where G is the increase in the weight (g), L is the thickness of the
edible lm (mm), A is the effective cross-sectional area (mm2 ), t is
the time duration of the permeation (h) and P is the difference
between the pressure of the cup atmosphere and the desiccator
(kPa). The reported WVP value is the mean value of ve separate
measurements.
Oxygen permeability (O2 ) for a lm sample was measured
according to the standard ASTM D3985. This experimental test
was performed at 23 C and 50 1% RH. The BSM lm sample was
masked with aluminium foils on both sides leaving a circular uncov-
ered area of 5 cm2 on the surface. The masked lms were then
placed into a test cell that isolates both sides of the lm, with pure
oxygen owing on one side and a ow of neutral gasses comprised
of 98% nitrogen and 2% hydrogen on the other side. The system was
left to reach equilibrium for 10 h, after which, oxygen permeability
was calculated according to:
Oxygen transmission rate average lm thickness
O2 = (3)
Difference in partial pressure between both sides of the lm
Where oxygen transmission rate is in cm3 /m2 , average lm thick-
ness is in m, and pressure is measured in kPa.
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The method for measuring transparency of the manufactured
lms is in accordance with ASTM D1746-09. Accordingly, the
manufactured lm samples were cut into rectangular shapes and
conditioned at 53% RH and 25 C for 24 h. A spectrophotometer (CE
2502, CECIL, Cambridge, U.K.) was then used to measure the trans-
parency of the lm samples. The instrument was rst calibrated by
an empty cell at 560 nm wavelength. Transparency of this empty
cell was taken as reference for this experiment. The lm samples
were then separately placed in the cell, and the readings from the
instrument were recorded.
A colorimeter was used to determine the colour of the lms
based on a*, b* and L* values which represent the levels of
greenness-redness, blueness-yellowness, and lightness of the lms,
respectively. Each lm sample was cut into a rectangular piece and
put in a standard plate with known values of as , bs and Ls . Sub-
traction of these standard values from the colorimeter readings, a*,
b* and L*, gives the actual colour factors of the lm sample. Total
colour difference factor, E, yellow index (YI), and white index (WI)
of the samples were then calculated by:
 was attached to an aluminium surface. The equipment was set to AC
2
E = (a as )2 + (b bs ) + (L Ls )2
142.86 (b bs )
YI =
(L Ls )
 roughness in several parts of the illustration.
WI = 100 (100 (L Ls ))2 + (a as )2 + (b bs )
Five consecutive readings were performed for each factor, and
their mean value were taken as the nal value.
Mechanical properties of the manufactured lms including their
tensile strength (TS) and elongation at break (E) were measured
based on the ASTM D882-12C2012 standard. The equipment used
for this measurement was DMTA (Testometric Co. Ltd., Rochdale,
U.K). Each lm was cut into a strip with a 90 mm length and 100 mm
width that was conditioned at 25 C and 52% RH for 48 h. The thick-
ness of each lm strip was measured at ve regions, and their mean
was taken as the sample thickness. Each lm strip was clamped
between two grips with an initial distance of 60 mm. The lm strip
was then subject to an extension (at a rate of 10 mm/min) and the
associated force and distance were recorded. The obtained data was
then used to calculate TS and E of the lm samples according to:
Load on the lm causing it to break(N)
TS =
Crosssection of the lm (mm2 )
Final gauge length Initial gauge length
E=
Initial gauge length
Five consecutive readings were performed for each lm sample,
and their mean was taken as the nal value.
2.5. Structural characterization of the BSM lm
Morphology of the BSM lms was observed by scanning elec-
tron microscopy (SEM) (Philips XL30, Amsterdam, Netherlands).
To this aim, each lm sample was cut into a 2 2 mm sheet and
dehydrated at 45 C for 12 h. They were then covered by a thin
layer of goldpalladium in a high vacuum coating system using a
sputter coater (Q150R, Sussex, U.K.). SEM images from each sample
cross-section were then taken at an acceleration voltage of 20 kV
to investigate the effect of glycerol and dried BSM concentration
levels on the lm structure.
Thermal properties of the manufactured lms were measured
by a differential scanning calorimetry (DSC) (Mettler Toledo,
Switzerland). 10 mg of each sample lm was cut into small pieces
and placed into the equipments aluminium crucible pan. An empty
pan of the same characteristics was taken as a reference. The pan
temperature was varied from 120 to 140 C at a rate of 10 C/min.
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3
Nitrogen was used as the purge gas in this process at a ow rate
of 20 ml/min. Glass transition temperature (Tg ) and melting point
temperature (Tm ) of the sample lms were then measured by the
equipment. The measurements were performed thrice, and the
mean value was reported.
Fourier transform infrared (FTIR) spectra of the BSM lm sam-
ples was obtained at room temperature using a spectrometer
(Bruker FRA 106/S FT-Raman, Germany). A horizontal attenuated
total reectance (ATR) equipped with a ZnSe crystal plate was used
to obtain the spectra in the range of 4004000 cm1 with a resolu-
tion of 2 cm1 .
X-ray diffraction (XRD) analysis was performed for the BSM
lms using an X-ray diffractometer (SIEMENS D5000, Germany).
The diffraction angle was set in a range of 4 80 at the rate of
1 /min while the current and voltage were equal to 40 kV and 40 mA
respectively. Copper was used as an anode.
Atomic force microscopy (AFM) (Dualscope/Rasterscope C26,
DME, Denmark) was used to investigate the surface morphology
of the optimal BSM lm sample in ambient condition. The sample
(4) non-contact mode. A tapping mode cantilever was used by a reso-
nance frequency of 150190 kHz and force constant of 2560 N/m.
The resulting data was used to generate a 3D image of the sample
(5)
surface. DME-SPM (2.1.1.2) software was then used to measure the
2
(6)
2.6. Water sorption isotherm
The method for obtaining water sorption isotherm of the opti-
mal BSM edible lm was adopted from [16]. Six straps of the optimal
BSM lm (3 3 cm) were put over silica gel in a desiccator (0% RH)
for one week to remove their moisture content. The dehydrated lm
samples were then put in six individual desiccators with 22.51%,
43.16%, 53.1%, 69.6%, 74.5%, and 85.1% RH. The experiment tem-
perature was kept at 25 C. The weights of the lm samples were
measured every 12 h until they reached equilibrium. Water sorp-
tion at a given RH was then calculated from the mass increase of
the dried lm sample after equilibration.
The experimentally measured water sorption behavior was
modelled by GuggenheimAndersondeBoer (GAB) and Brunauer,
Emmett and Teller (BET) equations, respectively expressed by:
(7)
M0 C k aw
M= (9)
(8) (1 k aw )(1 k aw + C k aw )
M0 C aw
M= (10)
(1 aw )(1 aw + C aw )
Where M and M0 are the equilibrium and monolayer moisture con-
tents (g water/g dry solid), respectively, aw is the water activity, C
is a constant, and k is a factor correcting the multilayer molecule
properties corresponding to the bulk liquid.
2.7. Statistical analysis
Properties of the fabricated BSM edible lms with respect to
their glycerol and mucilage concentration levels were investigated
by repeating each experiment for a minimum of three times. The
mean value of the measurements was then reported as the nal
value. Microsoft Excel 2013 and SPSS 23.0 software were used to
analyse the results. Analysis of variance (ANOVA) was used to anal-
yse the obtained experimental data. Duncans multiple range test
was then used to establish differences between treatments at a 5%
level.
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Table 1
Effect of different dried BSM and glycerol concentrations on thickness, moisture content, and solubility of the fabricated BSM edible lms.

Dried BSM Glycerol Concentration Thickness (mm) Moisture Content (%) Water Solubility (%) WVP (g/Pa s m 10 11 ) Transparency (%T cm)
Content (%w/v) (%w/w)

0.4 10 0.042 0.002 k 10.4 0.24 i 28.33 0.16 f 7.483 0.391 j 53.16 0.40 a
35 0.043 0.002 k 11.33 0.27 h 37.02 0.30 e 15.57 0.350 g 52.73 0.87 a
60 0.044 0.004 j 11.72 0.28 h 46.06 0.26 d 23.43 0.272 d 52.96 0.80 a

0.8 10 0.048 0.007 i 14.49 0.34 g 15.74 0.34 h 6.363 0.284 k 45.16 0.45 b
35 0.049 0.002 h 15.64 0.18 f 17.58 0.23 g 12.593 0.261 h 44.70 0.78 b
60 0.050 0.002 g 17.15 0.26 e 20.26 0.13 g 20.383 0.346 e 44.80 0.55 b

1.2 10 0.058 0.002 f 18.46 0.35 d 9.48 0.26 k 4.363 0.284 l 38.20 0.90 c
35 0.059 0.004 e 19.86 0.27 c 10.36 0.22 j 9.593 0.261 i 37.60 0.60 c
60 0.060 0.003 d 20.88 0.37 b 11.54 0.27 i 18.383 0.346 f 37.63 0.72 c

1.6 10 0.065 0.002 c 23.68 0.24 a 50.44 0.72 c 34.696 0.489 c 24.94 0.73 d
35 0.070 0.002 b 23.41 0.35 a 55.44 0.48 b 41.540 0.363 b 23.86 0.80 de
60 0.074 0.002 a 23.66 0.29 a 58.37 0.43 a 56.526 0.270 a 23.07 0.86 f

3. Results and discussion 3.1.2. Moisture content

Experimental tests as described in Section 2 were performed on
the fabricated BSM lm samples with different concentrations of
glycerol and dried BSM. These results and their analysis are ulti-
mately used in this section to determine the formulation of the
optimal BSM edible lm with desirable barrier and mechanical
characteristics.
3.1. Physical properties of the fabricated BSM lms
Preliminary experiments were performed to determine the
proper range of mucilage and plasticiser concentrations in the lm-
forming solution. It was found that lms with 0.8%1.6% (w/v) dried
BSM content had a clear and homogeneous appearance. Moreover,
lms with a minimum of 0.4% (w/v) dried BSM content were easily
peeled off the casting plates. However, lms with dried BSM con-
tent of more than 1.6% (w/v) were viscous with gummy properties.
It was also revealed that using glycerol as a plasticiser in the
lm-forming solution led to lms with more uniform surfaces that
are lighter and more exible. This is attributed to the polymer-
plasticiser bonds caused by the hydroxyl groups of glycerol [25].
According to the experiments, BSM lms with a glycerol concen-
tration of lower than 10% (w/w) of the dried BSM content were
brittle and inexible. On the contrary, overuse of glycerol in the
lm-forming solution caused the resultant BSM lms to become
sticky and hard to peel off the casting plates. From the performed
preliminary experiments, the maximum acceptable glycerol con-
centration in the BSM lm-forming solution was found to be 60%
(w/w) of its dried mucilage content.
3.1.1. Thickness
Thicknesses of the fabricated BSM lms were measured to be
within 0.042 nm0.068 nm as shown in Table 1. According to the
measurements, at a xed dried BSM content, an increase in the
glycerol concentration caused the lm thickness to increase sig-
nicantly (p < 0.05). This is caused by penetration of the glycerol
molecules in the mucilage polymer chains, breaking their molecu-
lar interactions especially their hydrogen bonds. Thus, more water
vapour is absorbed into the lm structure causing an increase in
the lm thickness.
According to Table 1, at a xed glycerol level, increasing the
mucilage content caused a signicant increase in the lm thickness
(p < 0.05). This complies with the previous study for Ghatti gum
edible lms [11].
The measured moisture content of the fabricated BSM lms with
respect to the dried BSM and glycerol concentrations are shown in
Table 1. Evidently, an increase in the dried mucilage content has sig-
nicantly increased the moisture content of the manufactured lms
(p < 0.05). This is explained by the hydroxyl groups of the mucilage
polymer which form hydrogen bonds with water molecules, result-
ing in a slight increase in the moisture content of the lm.
Similarly, from Table 1 it is observed that an increase in the
glycerol concentration of the lm-forming solution results in an
increased moisture content of the BSM lms. The addition of
glycerol to the lm-forming solution causes polymerpolymer
and polymer-water bonds to break, leading to formation of
glycerol-polymer and glycerol-water connections [17]. This causes
absorption of the water molecules in the resultant biopolymer
structure, leading to a signicant increase in the moisture content
of the edible lm as seen in Table 1.
It is concluded from the experiments that moisture content
of the BSM edible lms is directly proportional to their mucilage
and glycerol concentration levels. Similar results were reported in
the literature on Ghatti gum and cress seed mucilage edible lms
[11,18,19].
3.1.3. Water solubility
Edible lms are generally soluble in water. This feature, if con-
trolled, can be used in many drug delivery applications in medicine.
In such applications, the drug is encapsulated within an edible lm
which is dissolved after a certain time in the water, releasing the
drug in the body.
Table 1 lists the water solubility of the fabricated BSM edible
lms with respect to their mucilage and glycerol concentration lev-
els. Evidently, increasing the glycerol concentration from 10%60%
(w/w) causes an increase in the water solubility of the edible lms
(p < 0.05). It is also observed from Table 1 that the rate of increase
in the water solubility of the fabricated lms with respect to the
glycerol level is inversely related to their dried mucilage content. In
particular, higher water solubility of the fabricated lms is observed
at lower dried mucilage contents. Similar results were reported in
the literature on edible lms based on chia seed and Ghutti gum
[11,20].
Water solubility of polymers is affected by their cross-linked
network structure and crystallization between their polymer
chains. As explained earlier, polymer-glycerol bonds in glycerol-
infused lms cause an increase in hydrophilic feature of the edible
lm resulting in an increased water solubility [21].

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3.1.4. Water vapour permeability

Water vapour permeability (WVP) of the manufactured lms
were measured as listed in Table 1 with respect to the mucilage and
glycerol concentration levels. It is observed that adding glycerol in
the lm forming solution increases WVP of the resultant lms. The
measured WVPs with respect to the glycerol level is lowest in the
lms with 1.2% dry mucilage content where increasing the glycerol
concentration from 10% to 60% caused a signicant increase in the
WVP from 4.36 to 18.38% (p < 0.05).
WVP of edible lms are dependent on their water solubility.
When in contact with water vapour, the hydrophilic structure of
the edible lm absorbs the water vapour causing it to condense
and dissolve on the lm surface. The water molecules then gradu-
ally move through the lm, evaporating from its other side. From
the previous subsection, the solubility of the manufactured lms
increased with the glycerol concentration level due to the disrup-
tion caused by the low weight glycerol molecules in the polymeric
chains. This fact causes the porosity of the edible lm to increase
in microscopic levels, increasing its permeability to water vapour
with an increased glycerol concentration level.
From Table 1, at a xed glycerol concentration level, increas-
ing the dried BSM content of the fabricated lms from 0.4% to 1.2%
resulted in a decrease in their WVP. As the mucilage content of
the edible lms is increased, the polymeric chains within the lm
align in a more orderly manner, making the lm a better barrier
against water vapour. On the contrary, increasing the mucilage
content beyond a certain level causes gaps between the polymeric Fig. 1. Mechanical properties of the manufactured lms with respect to their
chains, increasing their permeability to water vapour (as seen in mucilage and glycerol concentration levels. (a) Tensile strength. (b) Elongation at
break.
the lm with a mucilage concentration of 1.6% in Table 1). These
results are conrmed through SEM images of the fabricated lms
as will be seen in the next subsections (Fig. 2). Similar behavior yellowness. No statistical differences were observed for a* with
was observed on edible lms synthesized from Ghatti gum [11]. respect to the mucilage and glycerol concentration levels (p < 0.05).
The reported WVP of the proposed BSM edible lms in this section
are lower than many edible lms in the literature, including acetyl 3.1.7. Mechanical properties
acylglycerol [22], sodium caseinate mixed with acetylated [23] and Mechanical properties such as tensile strength and elongation
whey protein isolate mixed with palmitic acid, stearyl alcohol, or at break are of prime importance in packaging applications. These
beeswax [24]. characteristics are measured for the fabricated BSM lms as shown
in Fig. 1 with respect to the mucilage and glycerol concentration
levels. From Fig. 1(a) and (b), increasing the glycerol concentration
3.1.5. Light transmittance (Transparency)
of the BSM lms leads to a signicant increase in the elongation
All the fabricated lms regardless of their mucilage and glyc-
at break and a decreased tensile strength, regardless of the dried
erol concentration levels were transparent enough to be used as
BSM content. Evidently, the addition of glycerol is improving the
see-through lms for packaging applications. Transparencies of the
functional properties of the fabricated biodegradable lms. This is
fabricated lms are reported in Table 1. It is observed that with
caused by the penetration of glycerol molecules between the poly-
an increase in the glycerol concentration level, transparency was
mer chains, reducing their intermolecular interactions which leads
not signicantly changed. Moreover, an increase in the dried BSM
to an increase in the mobility between the molecular layers [25].
content caused a signicant reduction in the transparency of the
According to Fig. 1(a) and (b), tensile strength and elongation
fabricated edible lms. This behavior is attributed to the increased
at break for the fabricated lms are directly proportional to their
intermolecular interactions and reduced mobility of the polymer
dried BSM content up to 1.2% w/v. However, at high mucilage levels
chains with an increased dried BSM content, leading to increased
(1.6% w/v) tensile strength and elongation at break were signi-
opacity for the edible lm.
cantly reduced despite of the glycerol concentration. In particular,
increased mucilage content leads to an increasing molecular inter-
3.1.6. Color measurement action between the polymer chains, strengthening the edible lms.
Colour of the BSM lms were evaluated in terms of a*, b* and L* However, beyond a certain threshold of mucilage content (1.2% w/v
factors using a colorimeter and their YI and WI were found as listed for the fabricated BSM lms), tensile strength and elongation at
in Table 2. It is observed that L* and WI increase with an increased break are reduced due to the resultant toughness, weakening of the
glycerol level while a*, b*, and polymer chains, and non-homogeneity in the polymer structure.
YI exhibit an insignicant variation (p < 0.05) indicating a whiter These ndings are conrmed through SEM images shown in the
lm structure with an increased plasticiser concentration. next section (Fig. 2). Similar behavior to those obtained in this study
On the other hand, an increase in the mucilage content has have been reported in the literature for edible lms synthesized
caused a signicant decrease in L* which indicates the lm light- from Psyllium seed, Salep seed, and Ghatti gum [7,10,11].
ness. Therefore, darkness of a BSM lm is directly proportional to The obtained experimental results of Fig. 1 indicate that optimal
its mucilage content. Moreover, WI which represents the whiteness tensile strength of 15.3 MPa and elongation at break of 90%, can be
of the lm has reduced with increased mucilage level. Additionally, reached at 1.2% w/v of dried BSM content and 35% glycerol con-
b* and YI are signicantly increased with the increasing mucilage centration. These results demonstrate an improvement over other
content which shows the tendency of the fabricated lms toward edible lms synthesized from bitter vetch, Chia seed and Opuntia

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Table 2
Effect of different mucilage and glycerol concentrations on color compounds of the fabricated BSM edible lms.

Dried BSM Glycerol concentration L* a* b* WI YI

content (%w/v) (%w/w)

0.4 10 34.48 0.39 c 4.34 0.25 a 5.71 0.43 c 34.09 0.38 c 23.66 1.72 g
35 38.44 0.21 b 4.73 0.19 a 5.59 0.32 c 38.00 0.17 b 20.77 1.13 g
60 41.48 0.43 a 4.44 0.59 a 5.43 0.31 c 41.06 0.41 a 18.71 1.21 g

0.8 10 24.44 0.23 f 4.44 0.60 a 8.56 0.39 b 23.82 0.15 f 50.01 1.85 e
35 26.98 0.18 e 5.01 0.28 a 8.59 0.45 b 26.30 0.25 e 45.53 2.71 ef
60 28.73 0.27 d 4.58 0.33 a 8.53 0.51 b 28.07 0.23 d 42.40 2.33 f

1.2 10 18.32 0.41 i 4.52 0.19 a 15.61 0.52 a 16.72 0.42 i 121.77 4.81 b
35 18.89 0.26 h 4.50 0.22 a 15.35 0.46 a 17.33 0.34 h 116.11 4.88 c
60 20.43 0.23 g 4.56 0.40 a 15.80 0.45 a 18.74 0.30 g 110.56 4.39 d

1.6 10 15.28 0.42 j 4.73 0.52 a 15.56 0.40 a 13.73 0.42 j 145.59 6.09 a
35 14.95 0.77 j 4.58 0.41 a 15.90 0.46 a 13.35 0.73 k 152.17 8.10 a
60 14.81 0.74 j 4.95 0.42a 15.63 0.37 a 13.25 0.78 k 151.08 10.68 a

Fig. 2. SEM micrographs for cross-sections of the fabricated BSM edible lms. (a) Fabricated lm with 0.4% dried BSM content and 35% glycerol. (b) Fabricated lm with 1.2%
dried BSM content and 35% glycerol. (c) Fabricated lm with 1.6% dried BSM content and 35% glycerol.

Ficus-indica mucilage, where tensile strength was measured at 5,

13.2, and 0.9 MPa, respectively [20,26,27]. The tensile strength of
the optimum lm was also lower than some bio-polymer such as
high amylose starch-composited gelatin lms [28].

3.1.8. Scanning electron microscopy

SEM micrographs for BSM lm samples with 0.4%, 1.2%, and 1.6%
dried BSM content and 35% glycerol concentration are shown in
Fig. 2. It is observed that the fabricated lm with 1.2% BSM content is
smoother with a more homogeneous structure compared with the
other lms of 0.4% and 1.6% mucilage content in which fractures
and holes were observed. SEM images also revealed small pores
and cracks in the lm structures that are attributed to insoluble
materials in the lm-forming solution. These were minimal in the Fig. 3. DSC thermograms of BSM lms containing 1.6 dry content with 60% glycerol,
lm with 1.2% mucilage content. 1.6 dry content with 0% glycerol, 0.4 dry content with 60% glycerol, and 0.4 dry
content with 0% glycerol.

3.1.9. Thermal properties

Four lm samples were fabricated with their dried BSM con- Table 3
Thermal properties of BSM lms with respect to their dried BSM content and glycerol
tent and glycerol concentration set to their respective higher and
concentration.
lower boundaries as shown in Table 1. Glass transition tempera-
ture (Tg ) and melting point temperature (Tm ) of the fabricated BSM Dried BSM Content (%) Glycerol Concentration (%) Tg ( C) Tm ( C)

lms were then measured using a DSC unit. These temperatures 0.4 0 43.56 112.89
are the phases at which the substance structure undergoes changes 60 46.7 104.06
from amorphous to viscose rubbery and from solid to liquid states, 1.6 0 47.15 105.35
respectively. The measured DSC curves for the BSM lms are plot- 60 71.95 100.88
ted in Fig. 3 and the obtained Tg and Tm are listed in Table 3. These
results imply that Tg and Tm of the fabricated lms are reduced by
increasing the glycerol concentration. This behavior can be justi- hydroxyl groups and hydrogen bonds with an increasing glycerol
ed by the free volume theory that describes a reduction in the concentration. This phenomenon weakens the biopolymer struc-

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Table 4 results in the previous sections where an increase in the glycerol

Physical, mechanical, and barrier properties of the optimal BSM edible lm with
concentration led to an increase in WVP, solubility and moisture
1.2% dried BSM content and 35% glycerol concentration.
content of the lms.
Physical properties Barrier and mechanical properties The broad band observed in the range of 30003600 cm1 in
Thickness (mm) 0.059 0.004 O2 (cm3 m/m2 kPa) 6.25 Fig. 4 is attributed to the stretch of bonded hydroxyl (O H), and
Moisture content (%) 19.86 0.27 WVP(g/Pa s m 1011 ) 9.593 0.261 the band in the range of 27003000 cm1 refers to the C H bonds
Water solubility (%) 10.36 0.22 E (%) 90.27 1.33 stretching. The FTIR spectra of both samples had undergone notable
Transparency (%T cm) 37.60 0.60 TS (MPa) 15.37 0.29
changes in these regions. A comparison between the two spectra
Color
shows that the peaks in the O H stretching region have shifted
L* 18.89 0.26
from 3338 cm1 for the pure BSM to 3301 cm-1 in the optimal
a* 4.50 0.22 BSM lm. This is attributed to the addition of glycerol to the edi-
b* 15.35 0.46 ble lm formulation, resulting in development of new hydrogen
WI 17.33 0.34 (O H) bonds. These new O H bonds could be attributed to elec-
YI 116.11 4.88
trostatic attractions between polymerpolymer, glycerolglycerol,
and polymer-glycerol. Moreover, free hydroxyl groups of glycerol
could change the wavenumber of the peak 3338 cm1 . On the other
hand, a hydrogen bond reaction causes a reduction in the C H
bonds, leading to lower wavenumbers for the peak in the C H
stretching band of the FTIR spectrum of the optimal BSM lm as
seen in Fig. 4. In particular, the associated peak to the C H bonds
is shifted from 2923 cm1 for the pure BSM to 2919 cm1 in the
optimal BSM lm.
In an FTIR spectrum, peaks at 819, 894, and 1020 cm1 are
related to C O bond stretching [32]. Therefore, in Fig. 4, the peaks
around 894 and 1020 cm1 could be related to O C stretching.
Moreover, peak appearing in the region of 1596 cm1 are reported
as carboxyl ion stretching band (COO ) [33]. These peaks are
observed in the spectra of Fig. 4 for BSM lms and have increased
by the addition of glycerol in the lm structure.
Fig. 4. FTIR spectra of the optimal BSM-based edible lm in comparison with that
of the pure BSM.
3.2.2. X-ray diffraction (XRD)
XRD patterns of pure BSM and the optimal BSM edible lm
ture, causing an increase in the molecular movements, leading to with 1.2% dried BSM content and 35% glycerol concentration were
an increased intermolecular free space that yields a reduction in Tg obtained as shown in Fig. 5. These patterns are used to deter-
and Tm . mine the structure characterization of the samples. According to
From Fig. 3 and Table 3, all lm samples have a Tg between 43 Fig. 5, pure BSM exhibited a sharp peak at 2 = 5.56 indicating
to 72 C which is higher than that of lms manufactured based its crystalline structure. This peak was disappeared by addition of
on high-density polyethylene (125 C) [29], gellan (84 C), and plasticizer into the lm forming solution, showing a high compati-
pure glycerol liquid (93 C) [30]. This indicates a good potential bility and a high interaction between the BSM polymer and glycerol
for BSM as a carbohydrate source for packaging applications. [25]. Moreover, a broad peak was observed at 14.78 and 22.64 in
the XRD pattern of pure BSM, demonstrating its amorphous struc-
3.2. Characterization of the optimal BSM lm ture. The intensity of these peaks were increased by the addition of
glycerol.
The measured properties of the fabricated BSM lms with dif-
ferent dried BSM content and glycerol concentration, as discussed 3.2.3. Oxygen permeability
in Section 3.1, revealed that barrier and mechanical properties, as Polysaccharide-based edible lms have desirable barrier char-
decisive factors in packaging applications, are at their optimum acteristics against non-polar molecules such as oxygen and aroma
value at 1.2% dried BSM content and 35% glycerol concentration. compounds. This is mainly due to the hydrophilic groups and
Therefore, this mucilage/glycerol combination is chosen as the opti- hydrogen bonds of such edible lms. Oxygen permeability is an
mal formulation of the BSM edible lm. Physical, mechanical, and important factor in packaging which directly affects the shelf life
barrier properties of the optimal BSM edible lm are listed in of a product.
Table 4. Oxygen permeability of the optimal BSM lm was obtained
according to the method described in Section 2 and was measured
3.2.1. Fourier transform infrared (FTIR) at 6.25 cm3 m/m2 KPa at 50% relative humidity for 24 h. This value
FTIR spectra of pure BSM and the optimal BSM edible lm were is higher than lms that are based on synthetic polymers such as
measured as shown in Fig. 4. The most considerable differences polyethylene terephthalate [34] and polyvinylidene chloride [35],
between the two FTIR spectra in the range of 4004000 cm1 was and biopolymer-based edible lms such as those from quince seed
seen in 1408, 1597, 1735, 2919, and 3301 cm1 . mucilage [17]. Evidently, the proposed optimal BSM edible lm
The specic peak of glycerol in its FTIR spectrum is expected at has promising characteristics for packaging applications compared
1456 cm1 [31]. From Fig. 4, in the optimal BSM lm this peak is with its synthetic polymer and biopolymer counterparts.
shifted to 1408 cm1 . This is due to the interaction between glyc-
erol and biopolymer in the lm structure. A comparison between 3.2.4. Atomic force microscopy (AFM)
the two spectra of Fig. 4 indicates that addition of glycerol slightly AFM images of the optimal BSM lm are shown in Fig. 6. The
increases the peak intensity at 1733 cm1 which is attributed to results show that roughness of the optimal BSM lm is 8.38 nm,
water. This is because of the interaction between glycerol and water indicating a smooth surface. Moreover, a uniform distribution of
molecules in the biolm structure. This conrms the obtained the lm-forming material is observed leading to formation of a

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Fig. 5. XRD pattern the optimal BSM edible lm in comparison with that of pure BSM.

Fig. 6. AFM images of the optimal BSM lm.

Table 5 water vapour sorption of the lm sample. The k-factor obtained in

Parameters of GAB and BET water sorption isotherm models.
Model constants C K
GAB 0.571 0.944
BET 0.254
homogeneous lm structure. This indicates the high compatibility
of BSM and glycerol due to their hydroxyl nature. SEM micro-
graphs conrms the AFM images. Roughness of fabricated optimal
BSM lm is considerably lower than similar lms based on konjac
glucomannan-ethyl cellulose (51 nm) [36].
AFM images of the optimal BSM lm as shown in Fig. 6 also
contain some large rugged areas which it could be attributed to the
micro or nanoscale insoluble particles in the biopolymer matrix
[37].
3.2.5. Water sorption isotherm
Water sorption isotherm of the optimal BSM lm describes
the relationship between the equilibrium moisture content and
the water activity of the lm sample at a specied temperature.
This also reects valuable details of the lm microstructure. The
isotherm model of the optimal BSM lm is essential to predicting
its stability of packaging and storage. Moisture adsorption isotherm
of the optimal BSM lm in the range of 22%85% is measured, on
which, GAB and BET models are t as shown in Fig. 7(a) and (b),
respectively. The constants of these models and their R2 are shown
in Table 5. From this table, a good tting with the experimental data
is observed by the GAB model (R2 = 99.607) in comparison with the
BET model (R2 = 93.916). In the GAB model, the constant k is less
than unity. A reduction in the k-factor indicates a reduction in the
this study (0.944) was less than SPI lm [38], and cassava starch
R2 [39], indicating good properties of the manufactured optimal BSM
99.607 lm for packaging in different humidities.
93.916 According to Table 6, the water sorption isotherm of the opti-
mal BSM lm is of sigmoid-shape. A slight increase in the slope
of the isotherm is observed at low aw regions, while at high aw
region the slope increases signicantly. This is due to swelling of
the hydrophilic structure in the bio-polymer matrix.
4. Conclusion
This paper investigated the feasibility of producing polysaccha-
ride edible lms from BSM as a new carbohydrate source. Glycerol
was used in the lm formulation as a plasticizer for increasing the
exibility and elongation of the fabricated lms. Different dried
BSM contents (0.4, 0.8, 1.2, and 1.6% (w/v)) and glycerol concentra-
tions (10, 35, and 60% (w/w)) were used to optimize the fabricated
BSM lm characteristics. Experimental tests were performed to
measure physical, mechanical, thermal, barrier, and microstruc-
tural properties of the fabricated lms with respect to their BSM
and glycerol concentration levels.
The obtained results indicated that increasing the glycerol con-
centration from 10 to 60 (%w/w) led to a signicant increase in
the lm thickness from 0.042 to 0.044 mm when a low dried BSM
content (0.4 (%w/v)) was used. However, the lm thickness varied
from 0.065 to 0.074 mm at a high dried BSM content (1.6 (%w/v)).
The increase in the glycerol concentration level led to increased
intermolecular spacing in the fabricated lms, making it easier for
the water vapour molecules to transfer through the lm, increasing

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Fig. 7. Water sorption isotherm of the optimal BSM lm at 25 C. (a) GAB model. (b) BET model.
the solubility (in the range of 9.48%58.37%), water vapour perme-
ability (in the range of 4.3656.53 g/Pa s m 10 11 ), and moisture
content (in the range of 10.4%23.68%) for the fabricated lms. The
increased intermolecular spacing also led to a decreased tensile
strength, regardless of the dried BSM content. Elongation at break of
the fabricated lms was also increased in the range of 32.25%94.5%
with the increasing glycerol concentration. The variation trends of
the two latter properties were observed up to the threshold of 1.2
(%w/v) BSM content, after which, the effect was reversed. This was
due to the aggregation of the excessive mucilage content in the lm
matrix, weakening the structural integrity which leads to a lower
resistance against fracture.
Transparency of the manufactured lms was not signicantly
affected by the glycerol concentration level. However, it was
decreased with an increasing dried BSM content, measured at
53.16%24.94% in the lms with a low glycerol concentration (10%),
and from 52.96% to 23.07% in the lms with a higher glycerol con-
centration (60%). The increasing dried BSM content also decreased
white index from 34.09 to 13.73 at a lower glycerol concentration
(10%) and from 41.06 to 13.25 at a higher glycerol concentration
(60%), ultimately leading to darker lms.
Optimal formulation of the proposed BSM edible lm for pack-
aging applications with 1.2% dried BSM content and 35% glycerol
concentration was determined based on the highest barrier and
mechanical characteristics. FTIR, XRD, and AFM analysis were per-
formed on the optimal BSM edible lm. It was shown that the
optimal BSM edible lm exhibits mechanical, water vapour perme-
ability, and oxygen permeability properties that were comparable
to those of other popular polysaccharide edible lms, making it a
potential candidate for packaging applications.
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Please cite this article in press as: A. Sadeghi-Varkani, et al., Int. J. Biol. Macromol. (2017), https://doi.org/10.1016/j.ijbiomac.2017.11.029