International Journal of Biological Macromolecules 240 (2023) 124412

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International Journal of Biological Macromolecules

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

Development of water-resistant paper using chitosan and plant-based wax

extracted from banana leaves
Anyaporn Boonmahitthisud a, b, c, Chontirat Booranapunpong a, Chalunthon Pattaradechakul a,
Supachok Tanpichai d, e, *
a
Department of Materials Science, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand
b
Center of Excellence in Green Materials for Industrial Application, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand
c
Center of Excellence on Petrochemical and Materials Technology, Chulalongkorn University, Bangkok 10330, Thailand
d
Learning Institute, King Mongkut's University of Technology Thonburi, Bangkok 10140, Thailand
e
Cellulose and Bio-based Nanomaterials Research Group, King Mongkut's University of Technology Thonburi, Bangkok 10140, Thailand

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

Keywords: On being exposed to water, cellulose paper swells and its mechanical properties become weak. In this study,
Hydrophobicity natural wax with an average particle size of 12.3 μm extracted from banana leaves was mixed with chitosan to
Oleophilicity prepare coatings applied on paper surfaces. Chitosan efficiently dispersed banana leaf-extracted wax on paper
Packaging
surfaces. The mixed chitosan and wax coatings considerably influenced paper properties, including yellowness,
Oil sorption
Water resistance
whiteness, thickness, wettability, water absorption, oil sorption, and mechanical properties. The coating induced
Mechanical properties hydrophobicity in the paper, resulting in a significant increase in the water contact angle from 65.1 ± 7.7◦
Banana leaves (uncoated paper) to 123.2 ± 2.1◦ , and a decrease in water absorption by ⁓64 % to 52.6 ± 1.9 %. The coated
Wax paper demonstrated an oil sorption capacity of 212.2 ± 2.8 %, which was ⁓43 % greater than that of the un­
Waterproof coated paper (148.2 ± 5.5 %), and the tensile strength of the coated paper improved under wet conditions
compared to the uncoated paper. Additionally, a separation of oil in water was observed for the chitosan/wax
coated paper. Based on these promising results, the paper coated with chitosan and wax could be used for direct-
contact packaging applications.

1. Introduction cellulose, is sensitive to moisture and water owing to its hydrophilic

The global food packaging market could reach up to 464 billion USD
by 2027 owing to its importance in food distribution and supply chains
observed during the COVID-19 pandemic [1,2]. Biodegradable pack­
aging is seen as an alternative to petroleum-based and nonbiodegradable
materials owing to the environmental issues associated with circular
economy campaigns, regulations, and standards [3]. Plastics, glass,
paper, and aluminum are typically used in food packaging; however,
papers and paper boards are the most important materials, accounting
for the maximum share in the global packaging market sector [4,5]. The
global market for papers and paper boards in 2018 (⁓425 million tons)
almost doubled in quantity compared to that in 1992 [6,7]. Moreover,
paper utilization in the packaging sector in Europe in 2018 was ⁓50 %,
which was 10 % higher than that of petroleum-based polymer packaging
[2].
Paper, a biodegradable and sustainable material that is composed of
characteristics and porous structure, limiting its use [3,8]. Our previous
work [9] has shown that the tensile strength of cellulose paper consid­
erably weakened by ⁓90 % when in contact with water owing to
swelling caused by the loss of interaction between cellulose fibers.
Improving the physical and mechanical properties of papers when
exposed to moisture and water is challenging. To overcome this
obstacle, coating the surface of the paper would be promising. Wax, a
long-chain hydrophobic substance, is typically used as a coating mate­
rial for fruit preservation and food packaging owing to its high water
resistance [10]. Depending on the source, waxes are classified as pe­
troleum or natural waxes. Natural waxes are an interesting renewable
and ecofriendly wax owing to their abundance and biodegradability.
Bananas are one of the most economically important crops; they
account for 16 % of the global fruit cultivation. The plant is cultivated in
tropical and subtropical regions in 130 countries, including Brazil, India,
China, and Indonesia [11–15]. Banana plants can produce only one

* Corresponding author at: Learning Institute, King Mongkut's University of Technology Thonburi, Bangkok 10140, Thailand.
E-mail address: supachok.tan@kmutt.ac.th (S. Tanpichai).

https://doi.org/10.1016/j.ijbiomac.2023.124412
Received 5 January 2023; Received in revised form 30 March 2023; Accepted 7 April 2023
Available online 11 April 2023
0141-8130/© 2023 Elsevier B.V. All rights reserved.
A. Boonmahitthisud et al.
bunch of bananas containing 3–20 hands (a hand of bananas comprises
up to 20 bananas) [15]. After harvesting, considerable agricultural
wastes, including banana pseudo-stems and leaves, are generated
[11–13,16]. The waste generally ends up in landfills and emits green­
house gases, a source of air pollution [12,13,15]. Numerous attempts
have been made to use banana pseudo-stem residues as a raw material in
many applications, such as bioethanol, biodegradable polymers, paper,
and fibers [7,14–17]. However, only a few researchers have studied the
potential use of banana leaves. The surfaces of banana leaves are
covered by wax that comprises long-chain fatty acids and alcohols
[12,13]. Yanagida et al. [12] and Chrumanee et al. [13] have found that
wax extracted from banana leaves possessed similar characteristics,
including chemical composition and physicochemical properties
(melting point, crystal structure, and acid and saponification numbers),
to carnauba wax.
Although wax has been successfully extracted from banana leaves,
studies on the use of wax extracted from these banana leaves are limited.
The objective of this work was to use banana leaf-extracted wax to
develop a biodegradable waterproof contact material. Various concen­
trations of wax extracted from banana leaves were dispersed in chitosan
solutions to prepare suspensions as chitosan could act as an emulsifier to
stabilize wax via electrostatic interaction between positive charges of
chitosan and negative charges of wax [18,19]. These mixed suspensions
were applied on one side and both the upper and lower layers of paper.
The properties of the coated paper, including color properties,
morphology, wettability, water absorption, oil sorption, and mechanical
properties under dry and wet conditions, were studied. This work
highlights the potential of chitosan and banana leaf-extracted wax
coatings in water-resistant paper development.
2. Experimental
2.1. Materials
Fresh banana leaves (Musa sapientum L.) for wax extraction were
purchased from a local market in Bangkok, Thailand. The leaves were
washed with tap water, oven-dried at 65 ◦ C for 2 h, and cut into small
pieces (~3 × 3 cm2). High-molecular-weight chitosan (2100 kDa) was
supplied by Marine Bio Resource Co., Ltd. (Samutsakhon, Thailand).
Kraft paper and vinegar with 5 % acetic acid were provided by TS
Papertube Co., Ltd. (Bangkok, Thailand) and Kewpie (Thailand) Co.,
Fig. 1. Photograph of the chitosan and banana leaf-extracted wax suspensions.
2
International Journal of Biological Macromolecules 240 (2023) 124412
Ltd. (Bangkok, Thailand), respectively. Ethanol and n-hexane were ob­
tained from Chemical Co., Ltd. (Bangkok, Thailand).
2.2. Extraction of wax from banana leaves
Banana leaves (100 g) were dispersed in 600 mL of n-hexane in a
batch reactor at 90 ◦ C for 5 h to extract wax. The extracted solution was
cooled down to room temperature and filtered to remove the solvent
using a rotary evaporator. The extracted wax was decolorized by stirring
for 10 min in ethanol, followed by centrifugation. The banana leaf-
extracted wax was oven-dried overnight at 60 ◦ C.
2.3. Preparation of chitosan and banana leaf-extracted wax coatings
Five different weight fractions of chitosan and banana leaf-extracted
wax coatings (10:0, 9:1, 7:3, 5:5, and 3:7) were prepared, as shown in
Fig. 1. Chitosan was dissolved initially in a 5 % acetic acid solution at
65 ◦ C for 4 h. The banana leaf-extracted wax was then added to the
chitosan solution, and a suspension with a total solid content of 1 wt%
was continuously stirred for 24 h. The homogenously mixed suspensions
of chitosan/banana leaf-extracted wax are referred to as “XW,” in which
X is the weight ratio of the banana leaf-extracted wax in the suspension.
Notably, no phase separation of these suspensions was observed after
leaving at room temperature for 7 days.
2.4. Preparation of papers coated with chitosan and banana leaf-
extracted wax
Suspensions of chitosan/banana leaf-extracted wax (⁓5 mL) were
applied on a 20 × 20 cm2 Kraft paper using a No. 16 coating rod. The
coated paper samples were oven-dried at 110 ◦ C for 10 min and then
kept in a desiccator at temperature of 30 ± 3 ◦ C for further character­
ization. Paper samples coated with chitosan and banana leaf-extracted
wax are referred to as XWY, while X is the weight ratio of the banana
leaf-extracted wax in the suspension, and Y is the number of the coated
paper side (1 for one-sided coating and 2 for both upper- and lower-
sided coating). For example, 0W2 is a sample coated with only chito­
san on both the upper and lower sides of the paper.
A. Boonmahitthisud et al.
2.5. Color properties and thickness
The colors of paper surfaces with or without coatings were measured
using a CIE-L*a*b* system with a Color-Eye 7000A spectrophotometer
(X-rite, Inc., USA). The L*− (0 (darkness) to 100 (lightness)), a*− (−
indicates greenness and + indicates redness), and b*− values (− in­
dicates blueness and + indicates yellowness) were determined. The
whiteness index (WI) and yellowness index (YI) were determined ac­
cording to ASTM E313-20 [20]. Moreover, the total color difference
(ΔE) between the coated and uncoated paper was calculated using the
following equation [21,22].
√̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅
( * )2 ( )2̅
ΔE = L0 − L* + (a*0 − a* )2 + b*0 − b*
where L*0 , a*0 , and b*0 are the color parameters of the uncoated paper, and
L*, a*, and b* are the color parameters of the coated paper.
The thicknesses of the uncoated and coated paper samples were
measured from five randomly selected points using a micrometer (0.01
mm, Mitutoyo, Japan).
2.6. Morphology
The surface morphologies and fracture surfaces of the uncoated
paper and paper coated with chitosan and banana leaf-extracted wax
were analyzed using scanning electron microscopy (SEM) with a JEOL
JSM-6400 (Japan) scanning electron microscope (5 kV accelerating
voltage). The paper samples were sputter-coated with a thin layer of
gold under vacuum to avoid charging during examination [23,24].
2.7. Chemical structure
The chemical structures of chitosan, banana leaf-extracted wax, and
paper samples coated with chitosan/banana leaf-extracted wax were
characterized using Fourier-transform infrared (FTIR) spectroscopy
(Nicolet 6700, USA). The chitosan and banana leaf-extracted wax were
mixed with KBr, and the mixture was compressed into a disc. All the
samples were scanned over a frequency range of 4000–400 cm− 1 in the
attenuated total reflectance (ATR) mode [9,24].
2.8. Wettability
Water and soybean oil wettability were analyzed by measuring the
contact angles of uncoated and coated paper samples using a contact
angle instrument (Kyowa, DM-CE1, Japan). A liquid droplet was
deposited on the surface of a sample, and the contact angles of the
droplet were recorded at holding times between 0 and 120 s [9].
2.9. Water absorption and oil sorption
The uncoated and coated paper samples (3 × 3 cm2) were oven-dried
at 50 ◦ C for 24 h and then weighed to record their initial weights (Wi).
The samples were then immersed in 10 mL of either distilled water or oil
at room temperature (30 ± 3 ◦ C) for 24 h. Liquid accumulated on the
sample surfaces was gently wiped off before determining their final
weights (Wf). Liquid absorption (L) was calculated using the following
equation [25].
(( ))
Wf − Wi
%L = × 100 (2)
Wi
2.10. Mechanical properties
The mechanical properties of the uncoated and coated paper speci­
mens were characterized using a universal testing machine (Tinius
Olsen, 5ST, USA) with a 500-N load cell and a cross speed of 15 mm
min− 1. Specimens (10 × 50 mm2) were prepared with a gauge length of
International Journal of Biological Macromolecules 240 (2023) 124412
25 mm. Before testing in the dry state, specimens were kept in a desic­
cator at room temperature (30 ± 3 ◦ C) for 24 h, while the specimens
were immersed in distilled water for 6 h for the wet conditions [9].
2.11. Statistical analysis
A one-way analysis of variance (ANOVA) was used to determine
significant differences between datasets. For datasets with a statistically
significant difference in the mean (p < 0.05), Duncan's multiple range
test was performed using SPSS Statistics 29.0.
3. Results and discussion
(1)
3.1. Appearance and thickness
A photograph of the coated and uncoated cellulose papers is shown
in Fig. 2, and Table 1 presents the color parameters (L*, a*, and b*), WI
and YI of the paper coated with chitosan and banana leaf-extracted wax,
and ΔE between the coated and uncoated paper samples. The applica­
tion of chitosan and banana leave-extracted wax coatings on paper
resulted in similar L*-values (darkness–lightness) for both the coated
and uncoated paper. However, a decrease in the a*-value (green­
ness–redness) and an increase in the b*-value (blueness–yellowness)
were observed for the coated paper. These color changes could be
attributed to chitosan. The 0W1 samples exhibited lower WI and higher
YI values compared to the uncoated paper. This effect is likely due to the
filling of most pores in the paper with chitosan, which can cause yel­
lowing owing to the reaction of functional groups in the chitosan
[9,26,27]. The addition of wax to the coatings resulted in a decrease in
WI for the coated paper, but an increase in YI. With increasing wax
loading in the coating, no change in WI and YI was observed. Addi­
tionally, there was no significant difference (p ≥ 0.05) in WI and YI
values between the single-side coating and doubled-side coating of
paper. Notably, the application of chitosan and wax coatings is not ex­
pected to have an effect on the visual appearance of the paper, as
indicated by the ΔE value of < 2.0 [28].
The thicknesses of paper samples coated with chitosan and banana
leaf-extracted wax are presented in Fig. 3. The uncoated samples had an
average thickness of 183.6 ± 0.5 μm. An increased thickness that varied
with banana leaf-extracted wax content was observed after coating. For
example, the 1W1 paper had a thickness of 187.4 ± 4.6 μm, while 7W1
had a thickness of 191.6 ± 2.9 μm. An increase in the thickness of the
coated paper agreed with the findings of Akhtar et al. [29]. In their
study, the increased film thickness was associated with an increased
beeswax content. However, coating both sides of paper with chitosan
(0W2) did not result in any significant increase (p ≥ 0.05) in thickness
compared to that of the 0W1 sample. It has been reported that the chi­
tosan filling in cavities between fibers mainly dominated after coating
for one or two layers, and the formation of the continuous chitosan film
on the paper surface was observed when the paper was coated with three
layers of chitosan [9,24]. On the other hand, after coating both sides of
the paper, an increased thickness was more pronounced for all the pa­
pers coated with chitosan and banana leaf-extracted wax. A significant
increase in thickness (p < 0.05) to 196.1 ± 3.0 μm was observed for
1W2, while 7W2 was 200.1 μm-thick. The increase in thickness of the
coated papers could be mainly attributed to the solid wax on their sur­
face rather than pore filling and film formation of chitosan.
3.2. Morphology
Fig. 4 shows the surface morphologies of the paper coated with
chitosan and banana leaf-extracted wax. Similar surface morphologies of
one-sided and both-sided coated paper samples were observed. Paper is
a porous material in which cavities are formed among cellulose fibers;
however, these cavities mostly disappeared after chitosan application
(0W). This disappearance was attributed to the cavity filling by chitosan
3
A. Boonmahitthisud et al. International Journal of Biological Macromolecules 240 (2023) 124412

Fig. 2. Photograph showing the appearance of papers coated with chitosan and banana leaf-extracted wax on one side and both upper and lower sides.

Table 1
Color parameters (L*, a*, and b*), whiteness index (WI), and yellowness index
(YI) of coated and uncoated paper and ΔE between coated and uncoated sam­
ples. Superscript letters are used to indicate significant differences (p < 0.05)
among the samples tested under the same condition.
Material L* a* b* WI YI ΔE

Uncoated 92.51 ± − 0.33 ± 1.48 ± 74.99 2.63 ± –

0.13ab 0.02a 0.05a ± 0.30a 0.09a
0W1 92.49 ± − 0.45 ± 1.91 ± 72.92 3.39 ± 0.47
0.08ab 0.01b 0.02b ± 0.31b 0.05b ±
0.09a
1W1 92.45 ± − 0.58 ± 2.72 ± 67.79 4.86 ± 1.28
0.10abc 0.04cd 0.41de ± 0.06de 0.79f ±
0.47c
3W1 92.36 ± − 0.58 ± 2.66 ± 69.16 4.73 ± 1.21
0.19bcd 0.04cd 0.12de ± 0.61de 0.21def ±
0.09c
5W1 92.18 ± − 0.53 ± 2.62 ± 68.88 4.70 ± 1.38
0.17d 0.04c 0.21d ± 1.37d 0.38def ±
0.01c
7W1 92.59 ± − 0.58 ± 2.69 ± 70.3 ± 4.71 ± 1.22
0.13a 0.03cd 0.01de 1.60d 0.11cd ±
0.06c
0W2 92.24 ± − 0.54 ± 2.24 ± 71.35 3.89 ± 0.77 Fig. 3. Thicknesses of papers coated with chitosan and banana leaf-extracted
0.01cd 0.01cd 0.04c ± 0.12c 0.10c ± wax on one side and both upper and lower sides. Bars and error bars present
0.02b the averages and standard deviations, respectively. Letters are used to indicate
1W2 92.19 ± − 0.68 ± 2.76 ± 69.38 4.93 ± 1.35
significant differences (p < 0.05) among the samples tested under the
0.06d 0.04f 0.10de ± 0.93de 0.20de ±
same condition.
0.15c
3W2 92.41 ± − 0.64 ± 2.79 ± 68.59 4.98 ± 1.34
0.03abc 0.04ef 0.02de ± 0.08de 0.03ef ± aggregates were observed from coatings with high wax concentration
0.03c owing to a limited chitosan content to disperse the wax. Moreover, after
5W2 92.34 ± − 0.59 ± 2.75 ± 70.22 5.10 ± 1.38
0.12bcd 0.01de 0.19de ± 0.09de 0.07cd ±
the coating application, the coated paper was dried at a temperature of
0.10c 110 ◦ C, which was higher than the melting temperature of banana leaf-
7W2 92.24 ± − 0.59 ± 2.96 ± 70.04 5.30 ± 1.54 extracted wax (74–82 ◦ C) [12,13]. This may lead to the formation of wax
0.02cd 0.01cde 0.04e ± 0.99e 0.07cde ± aggregates on the paper surfaces. The SEM images of the fracture sur­
0.04c
faces of the paper coated with chitosan and banana leaf-extracted wax
are shown in Fig. 5. Chitosan and wax were mainly deposited on the top
from the interaction between negative charges from the cellulose and surface of the paper. Zhang et al. [32] have observed high permeation of
the positive charges from chitosan [30]. This mechanism is consistent chitosan through pores between fibers when the chitosan concentration
with previous works [9,24,31]. It has been reported that chitosan in the paper coating was <1.0 wt%.
initially filled the pores and formed a thin layer afterward [9,24]. After
the application of chitosan and banana leaf-extracted wax coatings, wax
3.3. Chemical structure
particles were observed on all the paper surfaces. Particularly, the
number of wax particles increased with an increased wax concentration.
The FTIR spectra of the coated and uncoated paper samples are
The wax particles measured by a particle size analyzer (Mastersizer
shown in Fig. 6. The banana leaf-extracted wax presented absorption
2000, Malvern Panalytical Ltd., UK) at 25 ◦ C had a mean size of 12.3 μm.
bands at 2954, 2915, 2846, 1735, 1465, 1163, and 720 cm− 1. The ab­
Notably, although the paper was coated with mixed chitosan and banana
sorption peaks located at 2915 and 2846 cm− 1 correspond to the
leaf-extracted wax, pores in the paper were observed when the chitosan
stretching vibrations of C–H in fatty acid chains, while that at 1735
fraction was lower than that of the wax. The pores existed because
cm− 1 is owing to the stretching vibrations of ester carbonyls. Hydro­
insufficient chitosan filled the cavities in the paper. Further, wax
carbon vibrations correspond to the bands located at 2954 and 1465

4
A. Boonmahitthisud et al.
Fig. 4. Scanning electron microscopy (SEM) images of the surfaces of the cellulose paper samples coated with chitosan and banana leaf-extracted wax. An arrow is
used to indicate the location of the wax of the surface.
cm− 1. The bands at 1163 and 720 cm− 1 are attributed to amide groups
[33,34]. These characteristic peaks are similar to those of beeswax
[33–35]. On the other hand, the FTIR spectrum of chitosan exhibited
typical characteristic peaks at 2880, 1640, 1553, 1250, 1150, and 1080
cm− 1. The broad peak in the range of 3000–3600 cm− 1 corresponds to
O–H and N–H stretching, while the duplet bands at 2880 cm− 1 are
assigned to the asymmetric and symmetric C–H stretching of chitosan
[9,24,36]. The N-acetylglucosamine structure of chitosan is verified
from C– – O stretching (amide I) located at 1640 cm− 1, N–H bending
(amide II) located at 1553 cm− 1, and N–H cobending (amide III)
located at 1250 cm− 1 [37,38]. The typical peaks at 1150 and 1080 cm− 1
are owing to the symmetric C–O–C stretching of the pyranose unit
[9,24,36].
The uncoated paper sample presented an FTIR spectrum with char­
acteristic peaks from cellulose: 1430 cm− 1 (bending vibration of CH2),
1160 cm− 1 (C–O–C stretching of the β-1,4-glycosidic linkages), and
5
International Journal of Biological Macromolecules 240 (2023) 124412
1060 cm− 1 (C–O–C stretching vibration of the pyranose rings)
[39–41]. The board peak located between 3000 and 3600 cm− 1 is owing
to O–H stretching vibration, while the peak at 2920 cm− 1 corresponds
to C–H stretching in the cellulose structure [39–41]. The typical peaks
representing chitosan and wax were observed with the introduction of
chitosan and banana leaf-extracted wax. The intensity of these peaks
depended on the amount of chitosan and wax on the surfaces of the
paper samples. The intensity of the absorption bands at 2915, 2846,
1735, 1465, and 720 cm− 1 that are associated with wax was consider­
able in the 3W, 5W, and 7W samples, indicating the incorporation of
wax on the surfaces. Both one-sided and both-sided coated paper sam­
ples exhibited similar FTIR spectra.
3.4. Wettability
Water contact angle measurements are employed to differentiate the
A. Boonmahitthisud et al.
Fig. 5. SEM images of the fracture surfaces of the cellulose paper samples coated with chitosan and banana leaf-extracted wax.
Fig. 6. Fourier-transform infrared (FTIR) spectra of the chitosan, banana
leaf–extracted wax, uncoated paper, and paper coated with chitosan and ba­
nana leaf-extracted wax.
hydrophobicity and hydrophilicity of the materials. When the contact
angle of water droplets is > 90◦ , a surface is considered hydrophobic
[42]. Fig. 7(a) shows the water contact angles of the paper coated with
chitosan and banana leaf-extracted wax. The uncoated paper had a
contact angle of 65.1 ± 7.7◦ , and the water droplet was immediately
absorbed by cellulose. However, with chitosan coatings, the paper (0W)
had a contact angle of 104.3 ± 2.9◦ . Chitosan has been reported to
induce hydrophobicity during film solidification owing to the functional
group rearrangement of chitosan molecules, including hydroxyl and
amine groups, parallel to film surfaces [43–45]. By incorporating hy­
drophobic wax, the contact angles of the coated paper samples
increased. The sample 1W had a contact angle of 108.2 ± 3.7◦ . The
hydrophobicity of the coated papers increased with an increased wax
concentration. The water contact angles of 3W and 5W were 116.5 ±
3.9◦ and 123.2 ± 2.1◦ , respectively. No increment in the contact angle
was observed for 7W. This might be owing to wax aggregates. The dis­
tribution and concentration of wax on paper surfaces influenced surface
hydrophobicity [37]. Controlling the wax distribution on paper surfaces
to obtain the superhydrophobic paper is a subject of future work.
6
International Journal of Biological Macromolecules 240 (2023) 124412
Notably, no difference in contact angles between papers coated on one
or both sides was observed. This finding was owing to water contact
angles being measured between the water droplet and the surface.
Additionally, the changes in contact angles of the coated paper with
holding times were monitored. The immediate disappearance of the
water droplet on the uncoated paper was observed owing to the cellulose
absorption. The contact angle of 0W considerably decreased from 104.3
± 2.9◦ (holding time of 0 s) to 87.9 ± 1.7◦ (holding time of 120 s). This
reduction could be owing to water absorption by chitosan and cellulose
[37]. After incorporating wax, no significant change (p ≥ 0.05) in the
water contact angles between the holding time of 0 s and 120 s was
observed for the 1W, 3W, and 5W samples. This indicates that wax could
prolong the water–cellulose contact. However, the significant reduction
(p < 0.05) in water contact angles between the holding time of 0 s and
120 s occurred for 7W. This reduction might be attributed to the wax
aggregates and less chitosan on the paper surface, which allowed water
molecules to interact with cellulose within the paper.
Oil contact angles on the uncoated and coated paper samples were
also measured to study the effect of wax distributed on paper surfaces. A
surface is considered oleophilic when an oil contact angle is < 90◦ , and
when the oil contact angle is < 10◦ the surface is superoleophilic
[42,46]. Fig. 7(b) presents oil contact angles of the paper coated with
chitosan and banana leaf-extracted wax as a function of holding times,
and Fig. 7(c) shows the wetting behavior of water and oil droplets on the
surface of the paper samples (Uncoated and 7W). All uncoated and
coated paper samples were found to be oleophilic. The oil contact angle
was 29.0 ± 2.6◦ for the uncoated paper surface, and the contact angle
decreased to 12.4 ± 1.2◦ after 120 s. This reduction could be attributed
to the oil uptake by cellulose fibers [47]. With chitosan on the surface of
the paper (0W), the oil contact angle slightly increased to 32.9 ± 3.2◦ .
This increase might be owing to chitosan hindering oil uptake. After
incorporating banana leaf-extracted wax, the 1W, 3W, 5W, and 7W
samples exhibited similar oil contact angles at the holding time of 0 s.
However, the oil contact angle of the coated paper decreased signifi­
cantly with increasing holding time. The 0W sample exhibited a
reduction in the oil contact angle from 32.9 ± 3.2◦ to 27.5 ± 3.6◦ at a
holding time of 120 s, whereas those of 1W and 3W decreased from 32.2
± 3.8◦ to 21.5 ± 3.5◦ and from 33.1 ± 1.7◦ to 18.3 ± 1.4◦ , respectively.
A decrease in the oil contact angle at 120 s-holding time was more
noticeable with an increase in wax concentration in the chitosan/wax
coating. 5W and 7W exhibited oil contact angles of 9.8 ± 3.3◦ and 8.1 ±
3.0◦ at 120 s-holding time, respectively. This could be owing to oil
sorption caused by wax [48]. Soy, soy-bee, and carnauba waxes have
been reported to be superoleophilic with an oil contact angle of 0 ◦ C
A. Boonmahitthisud et al. International Journal of Biological Macromolecules 240 (2023) 124412

Fig. 7. (a) Water and (b) oil contact angles of papers coated with chitosan and banana leaf-extracted wax between 0 and 120 s holding times. (c) Oil (blue dye) and
water (red dye) droplets on the uncoated and coated (7W) paper samples. (For interpretation of the references to color in this figure legend, the reader is referred to
the web version of this article.)

[48]. were 84.3 ± 4.0 %, 74.4 ± 2.5 %, and 67.5 ± 2.0 %, respectively. These
reduced values suggest that banana leaf-extracted wax limited water
absorption on the papers. A high reduction in water absorption has been
3.5. Water absorption and oil sorption
reported when utilizing a high concentration of beeswax in composites
[37]. Moreover, a noticeable decrease in water absorption was observed
Water absorption is one of the important material features for ma­
when both sides of the paper were coated. The water absorption for 0W2
terials. Fig. 8(a) presents water absorption of the paper coated with
decreased by almost 35 % to 78.2 ± 4.2 % in comparison with that of
chitosan and banana leaf-extracted wax. The uncoated paper showed a
0W1. Reductions in water absorption to 54.2 ± 3.1 % and to 52.6 ± 1.9
high water absorption capacity of 147.9 ± 8.8 %, which was attributed
% were observed for 5W2 and 7W2, respectively. Although a solidified
to numerous hydroxyl groups in cellulose molecules [49]. With chitosan
wax concentration of > 50 % was used in the coatings, the coated paper
coating, the water absorption of the 0W1 sample decreased to 130.0 ±
still absorbed water. This finding could be attributed to several factors
1.8 %. This reduction could be attributed to chitosan bonding with
such as water penetrating through the paper via uncoated parts, chito­
cellulose molecules, allowing few available functional groups in cellu­
san adsorption, and incomplete coverage of the paper surfaces by wax
lose and chitosan to interact with water. A similar reduction in water
due to its morphology and aggregates.
absorption by incorporating chitosan in paper has been reported [49].
Oil sorption was monitored to study the effect of chitosan and ba­
Coating paper with 0.5 % chitosan solution reduced water absorption by
nana leaf-extracted wax coatings on the papers. Fig. 8(b) presents oil
33.7 %, and with increasing chitosan concentration to 2 %, the water
sorption of the uncoated paper and paper coated with chitosan and
absorption of the paper considerably decreased by 58.7 %. Water mol­
banana leaf-extracted wax. The oil uptake of the uncoated paper was
ecules on the coated paper required a longer time to penetrate than on
148.2 ± 5.5 %. The oil uptake by cellulose fibers is owing to combined
the uncoated paper [49]. According to our previous studies [9,24], when
mechanisms, including adsorption, absorption, and interfiber capillary
the chitosan coated paper samples were exposed to water under various
sorption within fibers [47,50]. Fine cellulose fibers with a high surface
conditions (5 ◦ C for 10 days, 40 ◦ C for 10 days, 70 ◦ C for 2 h, and 100 ◦ C
area have been reported to absorb higher amounts of oil than coarse
for 30 min), no significant differences (p ≥ 0.05) in overall migration
fibers owing to larger spots for surface adsorption and interfiber capil­
between the uncoated and chitosan coated paper samples were found.
lary uptake [47]. When the paper was coated with chitosan, the oil
The occurrence of overall migration might be owing to leaching of the
sorption was 135.1 ± 5.4 % for 0W1 and 97.7 ± 6.9 % for 0W2. Chi­
fragmented fibers from sides of the paper rather than chitosan. The
tosan acted as a protective layer to delay cellulose molecules from
water absorption of the paper decreased with the introduction of wax.
interacting with oil, causing a decrease in oil sorption. With introduction
1W1 had a water absorption of 120.7 ± 6.5 %. The water absorption
of wax, a considerable increase in oil sorption capacity was observed.
decreased significantly (p < 0.05) with increased wax concentrations.
The 1W1 sample demonstrated an oil sorption capacity of 101.3 ± 4.1
The values of water absorption for the 3W1, 5W1, and 7W1 samples

7
A. Boonmahitthisud et al. International Journal of Biological Macromolecules 240 (2023) 124412

Fig. 8. (a) Water absorption and (b) oil sorption of the papers coated with chitosan and banana leaf-extracted wax on one side and on the upper and lower sides.
Letters are used to indicate significant differences (p < 0.05) among the samples tested under the same condition. Sorption of high-density oil (dyed red) under water
by (c) uncoated paper and (d) paper coated with chitosan and banana leaf-extracted wax on both sides (7W2), and (e) oil sorption of the 7W2 sample after soaking in
water for 10 min. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

%. With increased wax concentrations (corresponding to decreased
chitosan contents) in the coatings, the oil sorption capacity of the coated
paper increased significantly. The 3W1, 5W1, and 7W1 samples
demonstrated an oil sorption capacity of 125.8 ± 7.9 %, 161.0 ± 1.3 %,
and 183.8 ± 5.1 %, respectively. However, coating the upper and lower
layers of the paper with chitosan and banana leaf-extracted wax pro­
vided higher oil sorption than coating only one side of the paper. Oil
sorption values of the 5W2 and 7W2 samples were 169.3 ± 5.9 % and
212.2 ± 2.8 %, respectively, which represented a 73 % and 117 %
improvement over 0W2 and a 14 % and 43 % improvement over the
uncoated paper. This increment in oil sorption could be due to high
contents of wax on paper surfaces, which could adsorb oil [48]. It has
been reported that the natural wax microparticles, with the average size
of ⁓120 μm, exhibited the high sorption capacity up to 5 times their dry
weight for various types of oils including crude oil, mineral oil, and
hexadecane [48].
Fig. 8(c–e) shows a separation of high-density oil (dyed red) in water
for the uncoated paper and the paper coated on both sides with chitosan
and banana leaf-extracted wax (7W2). Although the oil uptake was
148.2 ± 5.5 % for the uncoated paper, no oil sorption was observed. This
was due to the fact that when the paper is immersed in water, cellulose
molecules immediately absorb water and the paper is completely wet,
causing ineffective removal of oil precipitation in water [25]. Mean­
while, the 7W2 sample exhibited oil sorption (shown by red marks on its
surface) and oil film formation on its surface [25]. These phenomena
were attributed to the wax acting as a protective layer that hindered the
interaction between cellulose and water. Yuqing et al. [25] have applied
beeswax-coated lignin particles on cotton surfaces and found no water
absorption of this material when immersed in water. The blockage of
water penetration into the material was due to the formation of an air
layer between beeswax-coated lignin particles on cotton surfaces. The
layer was formed from the combined mechanisms of the low surface
energy of beeswax and lignin and the micro/nanoscale structures of the
particles. To investigate the potential of the protective layer generated
by wax under longer immersion times, the 7W2 sample was immersed in
distilled water for 10 min before oil sorption measurement. Oil sorption
of the 7W2 sample was still observed (Fig. 8(e)). Notably, although the
paper coated with chitosan/banana leaf-extracted wax showed better oil
sorption capacity than the uncoated paper, the oil sorption capacity of
the coated paper is still far below the requirement for oil sorbent ap­
plications. Porosity and pore size of oil absorbents are essential for oil
sorption [51]. Cellulose aerogels are porous-structured materials with
ultrahigh porosity that exhibit oil sorption efficiency of > 150 times
their own weight [51,52]. Based on our findings, chitosan/banana leaf-
extracted wax coated paper could be more beneficial for food packaging
rather than as an oil absorbent material. Although the coated paper
showed the high oil absorption properties, its application may be limited
in some cases where it could affect the sensory quality of fatty foods.

8
A. Boonmahitthisud et al.
Absorption of oil or fat by the coated paper from these foods may could
reduce their taste or texture. However, the coated paper prepared in this
work could be useful for reducing excess oil from deep-fried foods such
as chicken, French fries, and cookies.
3.6. Mechanical properties
The mechanical properties of paper are commonly investigated from
tensile strength and elongation at break [53]. The paper samples were
characterized under two different conditions (dry and wet states). The
dry mechanical properties, including tensile strength and elongation at
break, were measured when the paper samples were in a dry state; on the
other hand, the wet mechanical properties were investigated when the
paper samples were completely wet. The dry tensile strength and elon­
gation at break of the coated and uncoated papers are shown in Fig. 9.
The uncoated samples had the dry tensile strength of 21.7 ± 1.4 MPa,
and with a coating on one side, the dry tensile strength the paper coated
with chitosan (0W1) increased to 23.7 ± 1.1 MPa. This increase could be
attributed to an enhanced stress-transfer process in the paper from the
formation of interfacial adhesion between chitosan and cellulose [24].
Papers coated with high-molecular-weight chitosan have exhibited
higher tensile strength than those coated with low-molecular-weight
chitosan [24]. The difference could be owing to the many cavities fil­
led by chitosan in the paper. 1W1 had similar tensile strength to 0W1.
The 3W1 sample exhibited a tensile strength of 25.7 ± 1.7 MPa, which
was higher than those of 0W1 (23.7 ± 1.1 MPa) and 1W1 (23.8 ± 0.9
MPa). Moreover, 7W1 showed similar tensile strength to that of the
uncoated paper. These results suggest that chitosan played a vital role in
Fig. 9. Tensile strength and elongation at break of the uncoated and coated papers measured under dry and wet conditions. Letters are used to indicate significant
differences (p < 0.05) among the samples tested under the same condition.
9
International Journal of Biological Macromolecules 240 (2023) 124412
improving the mechanical properties of the paper, whereas wax had no
effect on mechanical properties because wax only deposited on the
surface of the papers. Furthermore, the coated paper exhibited consid­
erably higher elongation at break than the uncoated paper, but all the
coated paper samples had similar dry elongation at break. This might be
owing to the stress relaxation of cellulose fibers within the paper when
in contact with water during coating [9,53]. Moreover, the mechanical
properties of the paper coated on one and both sides were comparable
due to an increased thickness of the paper coated on both sides, as shown
in Fig. 3.
Additionally, the mechanical properties of the coated papers soaked
in water for 6 h were characterized. As expected, when the paper is in
contact with water, hydrogen bonding between cellulose molecules is
broken, affording poor mechanical properties [9]. The tensile strength of
the uncoated paper soaked in water was ⁓86 % (3.0 ± 0.5 MPa) lower
than its dry tensile strength (21.7 ± 1.4 MPa). A significant decrease (p
< 0.05) in tensile strength measured under wet conditions was observed
for all the paper samples. When chitosan and banana leaf-extracted wax
were introduced on one side of the paper, no significant improvement
was observed in the wet tensile strength, indicating that coating only on
one side of the paper was not sufficient in protecting the paper from
interacting with water. However, the wet tensile strength improved
significantly (p < 0.05) when the upper and lower layers of the paper
were coated. The tensile strength of the 0W2 sample (3.9 ± 0.2 MPa)
was ⁓24 % higher than that of the 0W1 (3.1 ± 0.2 MPa) and ⁓32 %
higher than that of the uncoated paper, indicating that chitosan acted as
an adhesive to bond cellulose fibers and prevented paper swelling [9].
The 1W2 sample showed the wet tensile strength of 3.8 ± 0.3 MPa,
A. Boonmahitthisud et al.
representing an improvement of ⁓22 % and ⁓30 % compared to the
1W1 sample and uncoated paper, respectively, while the wet tensile
strength of 3W2 (4.2 ± 0.4 MPa) was higher than that of the 3W1 sample
and uncoated paper, showing a ⁓16 % and ⁓43 % improvement,
respectively. However, less difference in tensile strength between papers
with one-sided and two-sided coatings was found when the chitosan
concentration was equal to or less than the wax content. Furthermore,
values of the wet elongation at break of 0W1 (9.1 ± 1.0 %), 1W1 (8.9 ±
1.2 %), 3W1 (9.4 ± 1.7 %) and 5W1 (8.1 ± 1.4 %) were higher than that
of the uncoated paper (7.5 ± 1.0 %). The improvement in the wet
elongation at break could be attributed to the flexibility of chitosan
[9,53]. However, when the wax content in the coating was greater than
the chitosan concentration, 7W1 exhibited the wet elongation at break
of 7.1 ± 0.6 %, which was similar to that of the uncoated paper. A
pronounced improvement in the wet elongation at break was observed
when a two-sided coating was applied. Elongation at break values of the
0W2, 1W2, and 3W2 samples were 10.7 ± 1.6 %, 10.7 ± 1.6 %, 10.1 ±
1.1 %, respectively.
It has been reported that some acetic acid was volatized from chi­
tosan films during drying, but the presence of residue acetic acid was
still found in the dried chitosan films. The residue acetic acid weakened
the ordered structure of chitosan molecules, leading to a decrease in
mechanical and barrier properties of the chitosan films [54]. Never­
theless, the application of a single layer of chitosan on paper resulted in
a minimal increase in the grammage of paper (1.95 g m− 2 or 0.000195 g
cm− 2) [9]. This small content of residue acetic acid left in chitosan is
unlikely to obstruct the use of the paper coated with chitosan and wax
for direct-contact packaging applications.
4. Conclusions
Wax was successfully extracted from banana leaves and mixed with
chitosan to prepare paper coatings. The effects of chitosan and banana
leaf-extracted wax coatings on the properties of paper were investigated.
Chitosan was applied to disperse wax particles on the paper surfaces;
however, wax aggregates were found when a chitosan concentration
was lower than a wax content in the coatings. Although chitosan and
banana leaf-extracted wax affected the whiteness and yellowness of the
paper, this was difficult to notice by eyes, which is desirable for appli­
cations particular to paper appearance. Chitosan and banana leaf-
extracted wax coatings induced hydrophobicity in the papers. The un­
coated paper had a water contact angle of 65.1 ± 7.7◦ , and the cellulose
fibers in the paper immediately absorbed water. Conversely, the paper
coated with chitosan and banana leaf-extracted wax had a water contact
angle of 123.2 ± 2.1◦ , which was maintained for at least 120 s. More­
over, after coating with chitosan and banana leaf-extracted wax, the
coated paper exhibited a decrease of ⁓64 % in water absorption from
147.9 ± 8.8 % to 52.6 ± 1.9 %. Further, an increased oil sorption to
212.2 ± 2.8 % was detected for the coated paper in comparison to the
uncoated paper (148.2 ± 5.5 %). Owing to the hydrophobicity induced
by the coating, the paper had less swelling under water. This resulted in
an improvement in the wet tensile strength of the coated paper by ⁓43
% compared to that of the uncoated paper. These promising results
suggest that the water-resistant paper could be used for packaging ap­
plications. Additionally, the paper could be beneficial for absorbing
excess oil from deep-fried foods such as chicken, French fries, and
cookies.
CRediT authorship contribution statement
Anyaporn Boonmahitthisud: Conceptualization; Data curation;
Formal analysis; Funding; Investigation; Methodology; Project admin­
istration; Resources; Writing - original draft; Writing - review & editing.
Chontirat Booranapunpong: Conceptualization; Data curation; Formal
analysis; Investigation; Methodology. Chalunthon Pattaradechakul:
Conceptualization; Data curation; Formal analysis; Investigation;
10
International Journal of Biological Macromolecules 240 (2023) 124412
Methodology. Supachok Tanpichai: Conceptualization; Data curation;
Formal analysis; Funding; Investigation; Methodology; Project admin­
istration; Resources; Writing - original draft; Writing - review & editing.
Declaration of competing interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influence
the work reported in this paper.
Data availability
Data will be made available on request.
Acknowledgments
This project is funded by National Research Council of Thailand
(NRCT) (Grant No. N41A640090). The instrumental facilities were
supported by the Department of Materials Science, Faculty of Science,
Chulalongkorn University. S. Tanpichai would like to express heartfelt
admiration for his wonderful spouse for being a great mom to his two
enthusiastic sons and thank his parents for their blessings and contin­
uous support.
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