Polymer Testing 87 (2020) 106465

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Polymer Testing
journal homepage: http://www.elsevier.com/locate/polytest

Ocimum basilicum seed mucilage reinforced with montmorillonite for

preparation of bionanocomposite film for food packaging applications
B. Rohini a, c, S. Padma Ishwarya d, Ram Rajasekharan a, e, Arun Kumar VijayaKumar a, b, **
a
Academy of Scientific & Innovative Research (AcSIR), Ghaziabad, Uttar Pradesh, 201002, India
b
Department of Fruits and Vegetable Technology, CSIR – Central Food Technological Research Institute, Mysore, India
c
Department of Technology Scale-up, CSIR – Central Food Technological Research Institute, Mysore, India
d
Agro-processing and Technology Division, CSIR - National Institute for Interdisciplinary Science and Technology (NIIST), Thiruvananthapuram, India
e
Lipidomic Center, Department of Lipid Science, CSIR-Central Food Technological Research Institute, Mysore, India

A R T I C L E I N F O A B S T R A C T

Keywords: Use of nanocomposites is a well-established approach in enhancing the mechanical and barrier properties of
Ocimum basilicum bionanocomposite film for food packaging applications. The seed mucilage of Ocimum basilicum was employed
Seed mucilage for the preparation of bionanocomposite films with montmorillonite (MMT) as nanofiller. The films were pre­
Montmorillonite
pared by solvent-casting method at varied solution pH (1, 3, 5 and 9) and MMT loading (1%, 3%, 5%, 10%, 15%
Exfoliated nanocomposite
and 20%). The films were characterized for physical, mechanical and barrier properties in addition to micro­
Bionanocomposite film
Tensile strength structure and X-ray diffraction pattern. XRD analysis revealed the exfoliated dispersion of MMT at pH 9, con­
Water vapour permeability firming its effective interaction with the bionanocomposite film. Maximum film tensile strength was achieved at
a lower MMT load of 5%. Water vapour permeability reduced with increase in MMT loading up to 5%, followed
by an increase at higher MMT loadings. Film formed at pH 9 showed tensile strength of 17.3 � 0.33 MPa and
reduced water vapour permeability (WVP) of 0.21 g mm.m 2.hr 1.kPa 1.

1. Introduction mucilage is comparable to that of commercial food hydrocolloids in

A shift in trend towards the biodegradable food packaging materials
has been observed in the recent times. An increased awareness on the
non-biodegradable nature of packaging materials based on petrochem­
ical products is the driving force underlying the identification of novel
biopolymers. The advantages of biodegradable films with respect to food
packaging applications include barrier to transfer of moisture, oxygen
and flavours and the consequent increase in shelf life of food products
[1]. Plant polysaccharides and proteins have received substantial
attention as raw materials for the preparation of bionanocomposite
films, of which the former is known for their abundant availability at
low cost [2]. However, the major limitation of plant polysaccharide
based biopolymers is the poor mechanical properties in comparison with
their synthetic counterparts [3,4]. The above constraint is known to be
combated by the approach of bionanocomposites, owing to the presence
of nanoscale fillers in the polymer matrix that impart improved me­
chanical and barrier properties [5,6].
The seed mucilage of basil (Ocimum basilicum) is an interesting
candidate for the biodegradable film preparation. The basil seed
terms of its functional properties [7] and ability to produce films with
good appearance and satisfactory mechanical properties [8]. On hy­
dration of the basil seeds, a swollen mucilaginous mass is obtained from
the outer pericarp layer [9]. The swelling on hydration has been
attributed to the chemical composition of basil seed mucilage [10],
which is mainly constituted by polysaccharides, more specifically, glu­
comannan and xylan [11,12]; 1975; [13]. The applications of basil seed
mucilage have been recognized as pharmaceutical excipients [14], for
the biosorption of heavy metals such as cesium-137 and strontium-90
[15] and as natural coagulants for wastewater treatment in the textile
industries [16]. A study by Khazaei et al. [8] has indicated the appli­
cability of basil seed mucilage for the preparation of biodegradable
edible film by varying different concentrations of glycerol as plasticizer
(25–50%), for the improvement of mechanical and barrier properties of
the biodegradable film. Amini et al.(2015) studied influence of different
plasticizers and fatty acids on properties of basil seed gum edible film
and observed that polyols when used as plasticizers improved the me­
chanical and barrier properties of the edible film. A recent study by
Thessrimuang et al. [17] has developed modified basil seed mucilage

* Corresponding author. Department of Fruits and Vegetable Technology, CSIR – Central Food Technological, Research Institute, Mysore, India.
E-mail address: arun@cftri.res.in (A.K. VijayaKumar).

https://doi.org/10.1016/j.polymertesting.2020.106465
Received 27 December 2019; Accepted 29 February 2020
Available online 30 March 2020
0142-9418/© 2020 Elsevier Ltd. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
B. Rohini et al.
film by using di-carboxylic acid as crosslinkers to improve its physical
and thermal properties. Thus, the present focus is on incorporating
montmorillonite as nanofiller with basil seed mucilage in preparation of
bionanocomposite film using glycerol as plasticizer.
The advent of nanotechnology in the field of food packaging is a
promising sign to resolve the concerns of poor mechanical and barrier
properties of natural polymers. Bionanocomposites are formed when the
biopolymer matrix is reinforced with nanomaterials such as layered
silicate (Kumar et al., 2010), wherein, nanomaterials are a class of
materials with at least one of their dimensions in the range of nano­
meter. Bionanocomposites are environment-friendly, non-toxic, biode­
gradable and eventually a good replacement for the conventional
petroleum-based packaging materials. Montmorillonite (MMT) or
nanoclay is one of the widely exploited nanomaterials for the prepara­
tion of bionanocomposites [18]. MMT consists of an aluminum layer
sandwiched between two layers of tetrahedral silicon sheets in the ratio
of 2:1. These clay platelets with a high surface area of 750 m2/g and
aspect ratios in the order of 100–500 are separated by thin layers of
water known as gallery [19]. The aforesaid features render it an excel­
lent filler material that adds up strength and barrier properties to the
polymeric material. MMT has been found efficient in improving the
barrier properties of many biopolymers such as agar, chitosan, starch,
whey protein, soy protein, gelatin, tilapia skin gelatin, fish gelatine and
hydroxypropyl methyl cellulose (Nagarajan et al., 2015 [20,21]; Mondal
et al., 2013; [5,22–25]. The effectiveness of bionanocomposites depends
on the mode of dispersion of nanomaterials in the polymeric matrix and
the demonstration of functionality at the lowest loading or filler content,
i.e. less than 5% by weight [26]. In this context, the pH of film forming
solution is known to have a vital influence on the mode of dispersion of
MMT and the biopolymer-MMT interaction (Mc Atlee, 1969).
The present study is aimed at the development of an improved bio­
nanocomposite film based on basil seed mucilage and to evaluate the
physical, mechanical, barrier and microstructural properties of the
resulting film as a function of MMT loading, after identifying the pH of
film forming solution which facilitates the most favourable mode of
dispersion and interaction with the polymeric matrix.
2
Polymer Testing 87 (2020) 106465
2. Materials and methods
2.1. Materials
Genotype RR-25 (O. basilicum) with high mucilage yield was iden­
tified after thorough screening of a number of Basil accessions. The seed
material of RR-25 was obtained from the crop cultivation and seed
production in the Research Farm of CSIR-CFTRI, Mysore, India. Mont­
morillonite, K10 was purchased from Sigma Chemicals, Bangalore,
Glycerol was purchased from Merck, Bangalore. Sodium hydroxide
(NaOH) and hydrochloric acid (HCl) were obtained from Merck (Ban­
galore) and Hi-media (Bangalore) respectively.
2.2. Extraction of mucilage from O. basilicum seeds
The seeds were carefully cleaned to remove stones, dirt material and
were packed in a polythene bag stored in dry and cool place until
extraction of mucilage. The seeds were soaked in distilled water for
about 2.5 h of duration, during which very thick mucilage was formed
around the seed coat. The seed to water ratio was fixed to 1:65.98 (w/v)
(Razavi et al., 2009). The hydrated seed containing mucilage was stirred
using propeller; the solution was further subjected to centrifugation to
separate mucilage from seeds by Westfalia separator. Mucilage solution
was dried in the hot air oven at 56 � C, and dried mucilage was ground to
powder form.
2.3. Preparation of MMT solution
The MMT solution was prepared by dispersing 5% of MMT with
respect to the weight of mucilage (1% w/v in deionized water) and
stirred for about 1 h. The solution was centrifuged at 2500 rpm for 10
min to separate the unsuspended MMT and discarded from the sus­
pended one. About 20 ml of an aliquot was dried in a hot air oven to
know the amount of MMT present in the suspension.
2.4. Preparation of bionanocomposite film from basil seed mucilage
Initially, the films were prepared as a function of varying pH of the
film forming solution at 5% MMT loading (as described in Section 2.3) to
study the effect of pH on film properties. pH of the mucilage solution was
adjusted to 1, 3, 5 and 9 (�0.5) using 1 N NaOH or 1 M HCl solution. The
B. Rohini et al. Polymer Testing 87 (2020) 106465

solution pH which demonstrated improved physical, mechanical and
barrier properties observed was set constant for the further preparation
of bionanocomposite films at varied MMT loadings, i.e. 1%, 3%, 5%,
10%, 15% and 20%. Solvent casting method was adopted for the prep­
aration of bionanocomposite films. The detailed procedure is discussed
below and represented with the schematic diagram.
Schematic representation of experimental procedure.
The required amount of MMT was added to the solution of adjusted
pH. The solution was stirred for about 2 h until all the mucilage added
have been dispersed completely without any formation of lumps. To the
film-forming solution, 50% glycerol (1% w/v of powdered mucilage)
was added with continuous stirring for 30 min. Air bubbles formed if any
were removed completely by centrifugation at 2000 rpm for 5 min. The
resultant film-forming solution was poured onto a Teflon coated glass
plate (25 cm � 35 cm), spread evenly and dried at 46 � C overnight. The
film was prepared in duplicates for each pH solution and control was
maintained without the addition of MMT and change in pH. Later, the
film was peeled off and stored in desiccator until further use.
2.5. Sample characterization
2.5.1. Determination of physical properties of films
ried out according to ASTM E96/E96M 14 standards [48]. The
desiccant method was followed in which calcium chloride (CaCl2) was
filled to 1/4 inch of cups. Above the cup, film was cut and sealed using
melted paraffin. The cups were placed in humidity chamber of 98% RH
and 38 � C. The weight of the cups was measured to nearest of 0.001 for
every hour and the difference in the weight was calculated and plotted
as function of time. The slope of each line was calculated by linear
regression (R2 > 0.99). Water vapour transmission rate (WVTR) and
water vapour permeability (WVP, g.mm.kPa 1 h 1 m 2) were calcu­
lated using Eqs. (6) and (7) respectively.
Slope
WVTR ¼ (6)
Area
WVTR X Δx
WVP ¼ (7)
Δp
where, Δx is the thickness of the film (mm) and Δp is the vapour pres­
sure difference across the test specimen. The values were measured in
duplicates.
2.5.4. XRD analysis
X-ray diffraction spectra were obtained using X-ray diffractometer
(Rigaku, Japan), using Cu-kα radiation at a wavelength of 1.54 Å. The
diffractogram was collected in reflection mode by scanning the 2θ range
from 1 to 60� , with a step size of 1� min 1 at room temperature.

2.5.1.1. Film thickness and density measurements. The thickness of the

2.6. Statistical analysis
film was measured using digital micrometer, M120 (United Kingdom) to
the nearest value of 0.001 mm. About six readings were taken in 6
The data obtained from continuously randomized design was
random positions for each film, and the average thickness was calcu­
analyzed statistically with One-way ANOVA using MINITAB version 17
lated, and these values were used for calculating the tensile strength and
software. Tukey’s test was used to find the differences in the mean
water vapour permeability. The density of the film was calculated for
values of properties of the film with a confidence level of 95%.
three replicates using eq. (1)
Film weightðgÞ 3. Results and discussion
Film density ¼ (1)
Film areaðcm2 Þx thicknessðcmÞ
3.1. Screening for high mucilage yielding basil accession
2.5.1.2. Moisture content. The % moisture was measured using digital
Moisture Analyzer (MB-45, OHAUS, Switzerland) at 105 � C. The anal­ 15 indigenous and exotic O. basilicum accessions were evaluated for
ysis was conducted in triplicates for each film and the mean value was the yield of seed mucilage, of which, 9 promising accessions with
calculated. moderate to high mucilage yield were identified (Table 1). Among the 9
accessions, genotype RR-25, a collection from the research farm of CSIR-
2.5.1.3. Film microstructure. The film surface was evaluated using the CFTRI campus, Mysore, Karnataka, India resulted in the highest muci­
scanning electron microscopy (Leo 435 VP, Leo Electronic Systems, lage yield of 21.9%, and thus identified as a potential source for the
Cambridge, UK). The films were mounted on specimen holder and preparation of bionanocomposite film. The photograph of RR-25 field
sputter-coated with gold and observed at 15 kV and vacuum of 9.75 � view is shown in Fig. 1 and Fig. 2(a) illustrates the formation of mucilage
10 5 Torr. around the seed coat on hydration of the O. basilicum seeds. Bio­
nanocomposite film was prepared using MMT and O. basilicum muci­
2.5.2. Mechanical properties lage is shown in Fig. 2(b).
The mechanical properties of films were determined using Food
Texture measuring instrument LLOYD-LR-5K (United Kingdom). The 3.2. Effect of solution pH on the X-ray diffractogram of bionanocomposite
tensile strength and percentage elongation was calculated by the film
maximum load and extension at break of the film according to ASTM
D882 standards [47]. Prior to test, the films were cut in the rectangular X-ray diffraction study provides clear evidence on the mode of
size of 2.5 cm � 10 cm. During analysis, test conditions such as the inner
distance of 50 mm and speed limit of 50 mm min 1 was maintained. Table 1
Tensile strength and % elongation for films of varying pH was calculated Mucilage content (%) of Ocimum basilicum accessions.
using Eq. (4) and Eq. (5) mentioned below. Accessions Mucilage content (%)
LoadðNÞ RR- 6 10.6 � 0.88
Tensile strength ¼ (4) RR- 8 12.1 � 0.71
Areaðmm2 Þ
RR-14 11.0 � 0.45
RR- 17 13.9 � 0.59
Extension at breakðmmÞ
%Elongation ¼ x100 (5) RR- 20 12.5 � 0.32
inner grip distanceðmmÞ RR- 21 13.7 � 0.21
RR- 25 21.9 � 0.98
2.5.3. Water vapour permeability RR- 26 14.0 � 0.82
RR- 27 20.1 � 0.71
The water vapour transmission rate and permeability test were car­

3
B. Rohini et al. Polymer Testing 87 (2020) 106465

The exfoliated dispersions of clay platelets are the best-suited for­

mation for maximum use of its properties [19]. Lagaley et al. (2003)
reported that the solution pH has a significant influence on the disper­
sion of MMT, where acidic pH of solution results in intercalated MMT

Fig. 1. Field view of RR-25 at CFTRI campus.

dispersion of MMT layers in the polymeric matrix. Fig. 3(a)-(b) show the
diffractograms for MMT nanoclay and that of RR-25 control film and RR-
25 bionanocomposite films formed with 5% MMT at varying pH of film-
forming solution. Fig. 3(a) shows the peak at a 2θ angle of 19.5� cor­
responding to MMT with clay inter-layer spacing value (d-spacing) of
4.46 Å. Muthuvel, Krishnakumar & Swaminathan [27] reported similar
observations on the MMT diffractogram. Peak was observed at 22.2� for
the RR-25 control film. Films formed at pH1 and 3 depicted peaks at 2θ
angle of 17.33� (d-spacing ¼ 5.11 Å) and 16.60� (d-spacing ¼ 5.33 Å)
respectively. For the film formed at pH5, a peak shift was observed at 2θ
angle of 19.12� (d-spacing ¼ 4.63 Å), corresponding to MMT. On the
other hand, the peak at 19.5� was absent for the film formed at pH 9. The
shift in diffraction peak to lower angle occurs after the formation of
nanocomposites [5,28].
The shift in peak for films formed at pH1 and pH3 was lower than
that for MMT, in addition to the increasing value of d-spacing. This can
be explained by the fact that d-spacing value increases when there is
intercalation of biopolymer within the interlayer region of clay, which
causes a shift in the diffraction peak as observed in case of starch and
agar films reinforced with Na-MMT (Slavutsky, Bertuzzi, Armada, Gar­
cía & Ochoa, 2012; [5]. In this ‘intercalated’ mode of dispersion, the clay Fig. 3. X-ray diffractogram of, (a) MMT; (b) RR-25 film and bionanocomposite
particles are known to be stacked or aggregated in the polymeric matrix films prepared under different pH of film forming solution.
at acidic pH, where the polymer gets attached to the interlayer region of
clay. These intercalated structures lead to moderate interaction with the
polymer (Mc Atee et al., 1969; [19]. However, the negligible shift in Table 2
peak value for film formed at pH5 can be due to the non-interactive Physical properties of RR-25 bionanocomposite films at varying pH of film
forming solution.
effect between RR-25 and MMT. This non-interactive mode of disper­
sion of the nanoclay has been termed as ‘tactoid’, which is characteristic Sample Thickness (mm) Moisture content (%) Density (g/ml)
of undispersed clay layers and poor interaction with the polymer [29]. pH1 0.0267 � 0.000758a 13.02 � 0.0200b 2.85 � 0.00755ab
The disappearance of peak at an angle of 19.5� for the film formed at pH3 0.024 � 0.001720ab 12.25 � 0.0100c 2.3 � 0.189b
pH9 can be due to the complete dispersion of MMT platelets into a single pH5 0.025 � 0.002381a 13.42 � 0.0208d 2.7 � 0.01143c
pH9 0.027 � 0.000675a 13.85 � 0.0208e 3.4 � 0.01338d
layer, corresponding to the formation of an ‘exfoliated’ structure at basic
Control 0.021 � 0.00412b 14.36 � 0.0153a 3.54 � 0.1384a
pH (Mc Atee et al., 1969). The development of exfoliated nano­
composites films at low level of MMT addition (5%) has been reported Data is presented as Mean � Standard deviation (n ¼ 3). Values in each column
by Alboofetileh, Razei, Hosseini & Abdollahi [21]. with same letters are not significantly different (p < 0.05).

Fig. 2. (a) Formation of mucilage from RR-25 seeds on hydration; (b) Bionanocomposite film using O. basilicum mucilage and MMT.

4
B. Rohini et al.
platelets or flocculant type dispersion whereas basic pH results in
exfoliation of MMT layers. Also, the neutral state was observed to be in
the pH range of 5–6.5. Thus, the inferences obtained in this study are in
accordance with the above observations in the literature.
3.3. Effect of solution pH on the bionanocomposite film properties
3.3.1. Physical properties
Table 2 shows the effect of varying pH on the film thickness. There
was a significant increase (p < 0.05) in the thickness of test films casted
at varying pH of the film forming solution when compared to that of the
control film. However, there was not much variation in the mean values
of thickness between the films formed at varied pH of the solution.
Therefore, the presence of MMT had a significant effect on the
improvement of film thickness rather than the pH of film forming so­
lution. The density of test films was lesser than that of the control film;
but increased significantly with increase in the pH of film forming so­
lution. The above had a direct correlation with the increasing moisture
content of the films with increase in solution pH, as would be explained
below.
Variation in moisture content of films (Table 2) may be attributed to
the variations in interaction between the biopolymer, MMT and glycerol
content under different pH conditions. When there is an effective
interaction between the biopolymer, glycerol and MMT, there is a less
exposure of hydroxyl groups of glycerol to interact with water mole­
cules. It was evident from XRD analysis that intercalated dispersion
occurs in film formed at pH1 and pH3 that corresponds to moderate
interaction between biopolymer, MMT and glycerol, leading to lower
moisture content in the film. Film formed at pH5 had comparatively
higher moisture content than those at pH 1 and 3, owing to the poor
dispersion of MMT that enhances the water holding property of glycerol
Fig. 4. Effect of change in pH of the film forming solution on (a) Tensile
strength of bionanocomposite film (b) % Elongation of bionanocomposite film
(Different alphabets indicate the statistical difference, P < 0.05).
5
Polymer Testing 87 (2020) 106465
Fig. 5. Difference in WVP with change in pH of film forming solution (Different
alphabets indicate the statistical difference, P < 0.05).
molecule to hold much of the water molecules leading to increase in
moisture content (Khazaei et al., 2014). Highest moisture content of the
film formed at pH9 may be due to the release of bound water molecules
during exfoliation. Theng [30] reported release of bound water mole­
cules present in the interlayer region of MMT during the interaction
between biopolymer and MMT; in turn, these bound water molecules
will be released in bulk solution as free water molecules.
3.3.2. Mechanical properties
The mechanical properties of the bionanocomposite film formed at
varied pH conditions were evaluated by the determination of tensile
strength and percent elongation (Fig. 4(a) and (b)). The tensile strength
of film improved at pH9 to 17.30 � 0.3265 MPa, compared to the
control film which had a tensile strength of 10.47 � 0.491 MPa. There
was significant difference observed in the tensile values at different pH
(p < 0.05), attributed to the difference in the interaction between MMT
and RR-25 biopolymer at varied pH. The intercalated mode of dispersion
of MMT in the RR-25 biopolymers at pH1 and 3 resulted in moderate
tensile strength, corroborating with the moderate interaction as
explained above.
At pH9, the tensile strength improved due to exfoliated dispersion of
MMT platelets resulting in a very strong interaction, as indicated by the
X-ray diffractogram. However, poor tensile strength was observed at
pH5 due to the undispersed MMT platelets and the consequent weak
interaction with RR-25 biopolymer in addition to the neutral behaviour
of MMT. Similar relationship was observed between % elongation and
pH with pH9 resulting in the improved elongation compared to other
films. The enhanced tensile strength along with improved elongation is
the pre-requisite for a packaging material and this was observed in the
bionanocomposite film formed at pH9.
Fig. 6. X-ray diffractogram of RR-25 bionanocomposite films prepared with
different MMT loadings.
B. Rohini et al. Polymer Testing 87 (2020) 106465

Table 3 WVP is caused by the lengthening of tortuous pathway for the diffusion
Physical properties of RR-25 bionanocomposite films at varying MMT load. of water vapour molecules between the impermeable layers of nanoclay
%MMT Thickness (mm) Moisture content (%) Density (g/ml) [31].
e e
1 0.021 � 0.0004 15.18 � 0.01 3.85 � 0.01a
3 0.024 � 0.00815de 15.4 � 0.02d 3.3 � 0.01c 3.4. Effect of MMT loading on bionanocomposite film properties
5 0.0264 � 0.0007d 13.82 � 0.01f 3.4 � 0.0b
10 0.033 � 0.0013c 19.7 � 0.01a 3.04 � 0.01d As explained above, pH9 facilitated exfoliated dispersion of MMT
15 0.039 � 0.0016b 18.8 � 0.01b 2.95 � 0.01e platelets leading to improved mechanical properties and reduced water
20 0.048 � 0.0010a 18.5 � 0.01c 2.5 � 0.01f
vapour permeability. Therefore, further experiments on the effect of
Data is presented as Mean � Standard deviation (n ¼ 3). Values in each column MMT loading on the film properties were carried out at pH9 of the film
with same letters are not significantly different (p < 0.05). forming solution.

3.3.3. Barrier property 3.4.1. X-ray diffraction

The WVP for RR-25 bionanocomposite films under different pH of
film forming solution is shown in Fig. 5. The WVP was improved when
compared to control film with the addition of 5% MMT and also there
was a significant (p < 0.05) improvement in WVP values under different
pH conditions.
The control film had permeability values as 0.25 g mm.hr 1.m 2.
kPa 1 and film formed at pH1 and 3 showed similar values of 0.24 g mm.
hr 1.m 2.kPa 1. But at pH5, a WVP value of 0.3 g mm.hr 1.m 2.kPa 1
was observed, which was greater than that of the control film. Exfoliated
dispersion facilitates exposure of individual platelets or biopolymer to
interact and creates the slow movement of water vapour into the film
thereby reducing permeability. Hence, film formed at pH9 showed lesser
permeability value of 0.21 g mm.hr 1.m 2.kPa 1. Fahranaky et al. [20]
explained the decrease in WVP with increase in clay content. Nanoclay
particles with their large surface-to-volume ratio function as physical
barriers against the movement of water molecules. The reduction of
The diffractograms of films formed at pH9 at varying MMT load (1%–
20%) are shown in Fig. 6. MMT loading was found to have a greater
influence on the intensity of diffraction peak rather than the peak angle.
Increase in MMT content enhanced the peak intensity at an angle of
19.5� . Similar relationship between nanoclay content and peak intensity
has been observed in starch based nanocomposite films formed with
Cloisite Naþ [18], MMT and hectorite [32] and the agar based coun­
terparts with Cloisite Naþ [5]. Also, it was observed that the ‘exfoliated’
dispersion of MMT was not sustained at higher MMT load, in spite the
pH of film forming solution was maintained at 9. The formation of
intercalated and exfoliated dispersion of MMT has been observed to
depend on the type and amount of MMT used [33].
3.5. Effect of MMT loading on bionanocomposite film properties
3.5.1. Physical properties
Thickness of the films formed at different MMT loading is given in
Table 3. Film thickness increased significantly (p < 0.05) with increase
in the MMT loading, attributed to the increase in solid content [34].
Similar relationship between film thickness and nanoclay content has
been observed in starch-based films [35] and alginate films [21].
Films containing 1% and 3% MMT showed similar values for mois­
ture content. However, at 5% MMT, there was a significant drop in the
moisture content, followed by an increase at MMT content >5%
(Table 3). The decrease in moisture content with the increase in MMT
concentration has been ascribed to the presence of clay in the films [21].
Density values of the bionanocomposite films exhibited a direct corre­
lation with the moisture content and hence a similar trend of change was
observed with increase in the MMT load (Table 3).

3.5.2. Mechanical properties

The tensile strength (TS) and percentage elongation (%E) are shown
in Fig. 7(a) and (b), respectively. It was observed that the TS for 1%
MMT was 14.6 � 1.3 MPa, which increased at 5% MMT (17.3 � 0.5
MPa), but decreased with further increase in MMT load (7.84 � 0.5 MPa,

Fig. 7. Effect of varying MMT load on (a) Tensile strength of bionanocomposite

film and (b) %Elongation of bionanocomposite film (Different alphabets indi­ Fig. 8. Difference in WVP with change in MMT loading (Different alphabets
cate the statistical difference, p < 0.05). indicate the statistical difference, P < 0.05).

6
B. Rohini et al.
at 20% MMT). Thus, a significant difference (p < 0.05) was observed,
except at 1% and 3% MMT. Increase in TS with increase in the clay
content have been observed in many other biopolymer-based nano­
composite films [18,22,36]. The improved mechanical property has
been attributed to the various reasons such as the resistance to defor­
mation force exerted by the clay with its high surface area (about 750
m2/g), high aspect ratio (50–1000) and very high elastic modulus (178
GPa) [26,29,37,38] and by the stronger interfacial interaction between
polymer matrix and intercalated silicate layer through hydrogen or ionic
bonds [18]. The decrease in TS beyond a MMT load >5% might be owed
to the formation of stacked clays due to incomplete dispersion of MMT in
the polymer matrix at high levels of clay concentration. TS value of the
basil seed mucilage based nanocomposite film obtained in this study is
higher than that of a synthetic LDPE film and basil seed mucilage film
using succinic acid as plasticizer which is 13 MPa [39] and 15.4 � 1.02
MPa (Thessrimuang & Prachayawarakorn,2019) respectively.
The change in % elongation values with increase in MMT content
followed the same trend as the tensile strength. The %E value obtained
in this study at 5% MMT loading is much lesser than that obtained by
Khazaei et al. [8] with basil seed mucilage film incorporated with 50%
Fig. 9. Effect of MMT loading on the microstructure of bionanocomposite films: (a) MMT nanoclay (b) Control (c) bionanocomposite film with 5% MMT (d) bio­
nanocomposite film with 10% MMT (e) bionanocomposite film with 15% MMT.
7
Polymer Testing 87 (2020) 106465
glycerol (39.37 � 2.79%) and Slavutsky et al. [40] with brea gum/MMT
film at 5% MMT content (9.31 � 1.31%).
3.5.3. Water vapour permeability (WVP)
The water vapour permeability for films with varying MMT load at
pH9 of the film forming solution is shown in Fig. 8.
WVP is dependent on the solubility and diffusivity of water in the
biopolymer matrix, which in turn is a function of the interaction be­
tween nanoclay and the polymeric matrix [40]. WVP values for films
with 1% and 3% MMT were almost equal to 0.32 g mm.hr 1.m 2.kPa 1.
Film formed with 5% MMT showed reduced WVP values of 0.21 g mm.
hr 1.m 2.kPa 1. With further increase in the MMT load, the films
exhibited poor water vapour barrier property. These observations can be
corroborated with the absence of ‘exfoliated’ dispersion and the conse­
quent ineffective interaction with the biopolymer matrix, in the nano­
composite films formed at high MMT load. This might have led to the
increase in WVP due to the presence of more voids in the film that causes
the water vapour to easily permeate across it. On the other hand, the
decrease in WVP of nanocomposite films at 5% MMT can be due to the
dispersion of silicate layers with large aspect ratios in the polymer
B. Rohini et al.
matrix that renders the nanocomposite film impermeable to water
vapour (Rhim & Ng, 2007). In addition, the layered structure of MMT
hinders and/or delays the transmission of water vapour through the film
matrix due to the tortuosity of the pathway [41,42], by increasing the
effective path length for diffusion [43]. Similar observations on the
improvement of WVP with MMT addition have been reported by several
authors (Agar-based nanocomposite films: [5]; Starch-based nano­
composite films: [18]; chitosan-based nanocomposite films: [25,36]; soy
protein [22]: and whey protein-based nanocomposite films: [44].
Thus, as observed in the case of mechanical properties, 5% MMT load
was found optimal for the preparation of nanocomposite films, from the
purview of the reduction in WVP. The obtained WVP values of basil seed
mucilage based nanocomposite films at 5% MMT in this study (of 0.21 g
mm.hr 1.sqm 1.kPa 1) is much lesser than that of starch/MMT film
(1.06 � 0.09 g mm/kPa.h.m2; at 6% MMT [24]; starch/poly vinyl
alcohol/sodium montmorillonite nanocomposite films (0.718–1.430 g
mm/kPa.h.m2, at 1–20% loading of MMT [45]; and soy protein iso­
late/MMT films (2.96 � 0.10 g mm/kPa.h.m2, at 5% MMT loading;
Kumar et al., 2010). In general, films with low WVP are compatible with
food products with low or intermediate moisture content, which in­
cludes but not limited to dried fruits, nuts, flours, jams and jellies,
confectionery, biscuits and chips [46].
3.6. Film microstructure
The microstructure of films bears a significant relationship with their
mechanical and barrier properties. Fig. 9 shows the SEM micrographs of
the outer surface of the films formed at varied MMT loading.
The dispersed spherical to irregular shaped structures in the SEM
images correspond to MMT platelets. SEM micrographs of the bio­
nanocomposite film showed smooth and uniform surface morphology
without pores or cracks in contrast to the control RR-25 film without
MMT addition. At a MMT content of 5%, MMT platelets were well
dispersed in the bio-nanocomposite film (Fig. 9(b)). This confirmed the
exfoliation of MMT in the bio-nanocomposite film at a MMT content of
5% and pH9. The film surface turned rougher with increase in the MMT
content. Some aggregates of MMT were found at 10% & 15% loadings
(Fig. 9(d) & (e)), which might be due to the poor dispersion of MMT in
the polymeric matrix, as explained above.
4. Conclusion
This study demonstrated the preparation of bionanocomposite films
from basil seed mucilage and montmorillonite nanoclay. The pH of the
mucilage solution had a significant influence on the dispersion mode of
MMT in polymeric matrix, as revealed by the XRD studies. The ‘exfoli­
ated’ dispersion of MMT platelets in the biopolymer matrix formed at
pH9 was advantageous in terms of strong interaction, improved tensile
strength and reduced water vapour permeability. Thus, at 5% MMT
loading and pH9 of the film forming solution, the basil seed mucilage
based bionanocomposite films had the lowest values for WVP and
highest tensile strength.
Authors contribution
Conceived and designed the experiments: AKV RR. Performed the
experiments: RB PIS. Analyzed the data: AKV RR RB. Contributed re­
agents/analysis tools: AKV RR. Wrote the paper: AKV RR RB.
Acknowledgment
The authors acknowledge Dr. C. Anantharamakrishnan, Director,
IIFPT, Tanjavur, India, and Dr. Prasanna Anjenaya Reddy for their
support in executing the project. The authors would like to acknowledge
the Central Instrumentation Facility and Services at CSIR-CFTRI for
providing experimental facilities. We gratefully acknowledge the
8
Polymer Testing 87 (2020) 106465
financial support from Council of Scientific and Industrial Research,
New Delhi, India for carrying out this research.
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