Fibers and Polymers 2022, Vol.23, No.

10, 2885-2896 ISSN 1229-9197 (print version)

DOI 10.1007/s12221-022-4006-6 ISSN 1875-0052 (electronic version)

Evaluation of Reinforced and Green Bioplastic from

Carrageenan Seaweed with Nanocellulose
Fatmawati Adam1,2*, Nor Amira Othman1, Nur Hidayah Mat Yasin1,
Chin Kui Cheng3, and Nurul Aini Mohd Azman1
1
Faculty of Chemical and Process Engineering Technology, Universiti Malaysia Pahang, 26300 Gambang,
Kuantan, Pahang, Malaysia
2
Center of Advanced Research in Fluid Flow, Universiti Malaysia Pahang, 26300 Gambang,
Kuantan, Pahang, Malaysia
3
Department of Chemical Engineering, College of Engineering, Khalifa University, Abu Dhabi 127788,
United Arab Emirates
(Received November 13, 2021; Revised February 28, 2022; Accepted March 5, 2022)

Abstract: In this work, bioplastic was formulated through the incorporation of cellulose into refined carrageenan. Three
different sizes of cellulose were used microcrystalline cellulose (MCC), cellulose nanowhisker (CNW) and nanofibrillated
cellulose (NFC). The CNW was mechanically nano-grinded and NFC was ultrasonicated to produce nano particle for MCC.
The CNW has the smallest particle size distribution and an average size of 54 nm. The abundance of hydroxyl groups in the
cellulose structure forms intermolecular interactions with the carrageenan and increases the viscosity (293.90 mPas), shear
stress (27.90 Pa) and tensile strength (19.87 MPa) of Carra-CNW bioplastic. This revealed increasing intensity and a shift of
1
H-NMR due to hydrogen becoming closer to an electronegative atom in both micro and nano size cellulose. At a larger
cellulose size, Carra-MCC is thermally stable and the activation energy (Ea) is 73.81 kJ/mol higher than Carra-CNW Ea at
47.14 kJ/mol. Cellulose incorporated into a carrageenan matrix is a potential eco-material that could replace conventional
plastic where the sample decomposes up to 65 % of the initial weight in biodegradability and compostability test. The water
vapour permeability value is in the range of 3.43 to 4.09×10-14 g s-1 m-1 Pa-1. In conclusion, carrageenan incorporated with
cellulose potentially suitable for uses such as food packaging applications based on the results of the mechanical strength,
thermal stability and decomposition tests conducted.
Keywords: Bioplastic, Green technology, Carrageenan, Nanocellulose, Polysaccharide seaweed

Introduction Bioplastic or bio-based plastics are usually derived from

renewable biological resources to reduce pollution and
Plastics are extremely important because of their usage in promote a sustainable environment. Bioplastic was introduced
package goods. Plastics are cheap and permit single-use in as an alternative to replace petroleum-based plastics [9].
many applications including medical tools and surgery [1]. There are several resources used to produce bioplastic
This has resulted in global plastic pollution of about ⁓1.4 to including plants such as starch, wheat, corn and barley [10].
3.7 million tons per year [2]. In certain cases, heavy metals Other sources of bioplastic include bacterial based
and organometallic compounds are also added into the polyhydroxyalkanoates (PHA), polyhydroxybutyrate (PHB),
polymer matrix of plastics to optimize properties such as algel-based microalgae and seaweeds [11]. Compared with
being lightweight, hygenic and strong as well as to reduce conventional plastic, bioplastics are less dependent on fossil
production costs. These additives can gradually leach into fuels, non-toxic, easier to be disposed of, renewable and eco-
the environment through the life cycle of plastics known as friendly [9-12]. The application of bioplastic is broad, such
microplastics (MPs) [2-4]. MPs are defined as small as for food packaging, medical care, agriculture and
particles of plastic that integrate at high densities into electronics [13,14]. Despite this, there are several drawbacks
millimetre to sub-millimetre size [5,6]. The size of MPs is of bioplastic such as poor heat ability, thermal instability and
less than 5 nm and they are usually derived from personal brittleness which need to be improved. It is difficult to
care products or cosmetics; secondary MPs are fragments of produce a low-cost bioplastic that has a mechanical properties
larger plastics [5,7]. MPs have become a global concern due comparable to conventional plastics [13].
to their effects on environment and human health particularly Seaweed offers advantages compared to terrestrial plants,
aquatic environments. Aquatic species are affected by the such as low cost and being abundantly available in nature
presences of the MPs due to the shape, colour and dimension [15]. Polysaccharides from seaweeds have been getting
of the particles, as well as additive types and the chemical attention as a main component in the production of bioplastic.
contaminants [8]. Carrageenan is a natural polysaccharide derived from
seaweed. It is widely used in the food industry as a coagulant
*Corresponding author: [email protected] or an emulsifier [16]. Several studies have applied carrageenan

2885
2886 Fibers and Polymers 2022, Vol.23, No.10 Fatmawati Adam et al.

to the production of bioplastic film. The addition of plant oil particles size. Alberola et al. (2007) highlighted that the
and polyethylene glycol 400 (PEG-400) into carrageenan interphase thickness of the biopolymer has no effect on the
biocomposites has reduced the tensile strength of the film to size of the nano-filler particles. Nevertheless, adding
6.65 MPa and 17.96 MPa, respectively [17,18]. Bioplastic different sizes, dimensions and morphologies of nanofillers
film produced from kappa-carrageenan at 3 % w/v shows will affect the matrix behaviour of the biopolymer [34]. The
enhanced physical and mechanical properties with the addition of 5 % wt/v microfibrillated cellulose can improve
addition of PEG300 in the range 9.12-9.32 MPa [19]. A the tensile strength of polymeric films by up to 40 % [35].
study was conducted on semi-refined carrageenan incorporated There is insufficient work on the biopolymer characteristics
with arabic gum for hard capsule applications. The addition produced by varying the load transfer between the filler and
of 67 % w/w carrageenan-arabic gum mix produced a tensile the matrix using various sizes of cellulose filler such as
strength 36.21 MPa [20]. However, several limitations exist nanowhiskers (CNW). Hence, this study aims to investigate
particularly in term of water vapour permeability and water the effect of cellulose sizes towards the mechanical, physical,
resistance since carrageenan is naturally hydrophilic and the chemical, thermal stability and decomposition properties of
bioplastic film formulated is brittle [15,21]. Hence, it carrageenan bioplastic films.
requires additives such as crosslinker, filler and toughening
agent to improve the bioplastic properties. Glyoxilic acid Experimental
crosslinker was incorporated with the carrageenan and the
film disintegrated when in contact with water rather than Preparation of Cellulose Nanowhiskers (CNW)
swelling [22]. There are two types of crosslinking; chemical Microcrystalline cellulose (MCC) was purchased from
and physical. Chemical crosslinking increases the swelling Sigma-Aldrich (USA) at a size of ⁓50 µm (ground using
ability of the film and physical crosslinking promote the nanogrinder machine (Fritsch, USA) equipped with P7
disintegration and degradation of the film in the environment Premium Line). Approximately 5 g of MCC was dissolved
[23]. in 20 ml of deionized water to produce wet MCC before the
Cellulose is one of the most abundant natural polymers grinding process. Wet MCC was placed in grinder well with
which can be found as a structural component in the cell 5 mm grinding balls. The wet MCC was ground for
wall of plants and algae and is also a derivative from several 5 minutes and then rested for 3 minutes rest over an hour’s
types of bacteria [24]. It contains β-1,4-D anhydro glucopyranose duration at a 700 rpm stirring rate. Then, the particle size of
which displays a regular network of inter- and intramolecular ground MCC was measured using a nano-particle analyser.
hydrogen bonds [25,26]. Pure cellulose from plants or algae
can be treated with chemicals to produce several types of Preparation of Nanofibrillated Cellulose (NFC)
derivatives such as hydroxyethylcellulose, ethylcellulose, Nanofibrillated cellulose (NFC) was prepared by dissolving
hydroxypropyl cellulose, cellulose acetate, cellulose nitrate, approximately 5 g of MCC in 500 ml of deionized water.
carboxymethyl cellulose and others. The addition of cellulose The solution was then stirred at 200 rpm overnight. The
into the bioplastic formulation can enhance the mechanical sonication process was carried out using a Qsonica (Q700,
properties, biodegradability and thermal properties of the USA) instrument for 1 hour at an amplitude of 20 %. The
films [27,28]. Microcrystalline cellulose (MCC) offers a solution was kept stationary until three layers of the solution
great potential to enhance the properties of the bioplastic were obtained. The bottom layer was removed and the top
blend [29,30]. Nanocellulose has also gained considerable two layers were kept in a stationary position overnight to
interest due to its unique properties. Nanocellulose is either obtain two separated layers. The upper layer sample was
derived from native cellulose fibres or is based on the degree collected for measurement to ensure NFC was successfully
of fibrillation. The fibrillated cellulose is usually known as produced.
cellulose nanowhiskers (CNW) and nanofibrillated cellulose
(NFC). Both can be produced by mechanical methods such Preparation of Carrageenan-Cellulose Biocomposite Film
as grinding and high-intensity ultrasonication [31,32]. Nano- Three grams of carrageenan were dissolved in 200 ml of
grinding is a mechanically rapid process which can rupture deionized water at 60 °C for 1 hour. In another beaker, 2 g of
the hydrogen bonding interaction and the cell walls to obtain microcrystalline cellulose (MCC) and 1 g of hydroxypropyl-
ultra-thin cellulose [30]. An addition of nanocellulose into methylcellulose (HPMC) was dissolved in 50 ml of deionized
the formulation of bioplastic, can result in transparent films water. After a homogenous solution of carrageenan was
with good resistance against oxygen. Consequently, it is obtained, MCC was added into the solution and continuously
suitable for food packaging [33,34]. stirred for 30 minutes. Polyethylene glycol (PEG) and
Factors which influence the effects of nanoparticles on HPMC were added into the solution and then stirred for
biopolymer processing include the mechanical properties of another 30 minutes at 60 °C. Finally, glyoxylic acid was
inorganic nanofillers, uniformity nanofiller dispersion within added into the solution as a crosslinker and stirred continuously
the polymer matrix and the volume percentage of the filler for 30 minutes. Then the solution was cast and left to dry at
Reinforcement of Nanocellulose in Carrageenan Matrix Fibers and Polymers 2022, Vol.23, No.10 2887

room temperature (37 °C). The above steps were repeated with TexturePro CT V1-8 Build 3.1. The bioplastic film was
for CNW and NFC samples. cut into a dumb-bell shape (2 cm×10 cm) and fixed to the
tensile machine at both ends prior to the mechanical testing
Viscometer Analysis with 50 kg load cell. The method followed ASTM D882-12
Hydrocolloid properties involve the observation of the and a crosshead speed of 50 mm/min was employed. The
viscosity, shear stress and shear rate of the bioplastic elongation at break (EAB) and tensile strength of the
solution using a Viscometer (Brookfield, Rheo 3000, USA) bioplastic film was calculated using the following equations
equipped with LCT 25. Approximately 16.5 ml of carra- [37,38]:
cellulose solution was programmed at a speed of 300 rpm
 load at break 
with 100 MPoints at a constant temperature of 40 °C. Tensile strength = ------------------------------------------------------------------------------------- (3)
 original width   original thickness 
Opacity Analysis and Water Contact Angle
Film opacity analysis was conducted by cutting the film Elongation at break (%) = elongation at rupture-  100
------------------------------------------------- (4)
initial length
according to the test cell and directly inserting it. The film
was measured at 600 nm using a UV-vis Specrophotometer SEM Morphology
(Shimadzu, Japan). The film opacity was calculated using The surface morphology of the film sample was observed
the following equation: using a HITACHI Tabletop Microscope (Benchtop SEM)
TM4000/TM3030Plus Scanning Electron Microscope and
Abs600
T = ------------------ (1) viewed under 5000X magnification. A 1 cm×1 cm bioplastic
x
film was placed on the specimen stub and coated with 10 nm
where T is the transparency of the films, Abs600 is the value thick gold before observation. Gold acts as an electrically
of the absorbance at 600 nm and x is the thickness of the conductive surface to prevent the accumulation of electrostatic
films. charges on the samples.
Meanwhile for water contact angle analysis, the sessile FTIR and 1H NMR Analysis
drop method via a Contact Angle Analyzer (KSV CAM 101; The FTIR spectra of the bioplastic were analysed using a
KSV Instruments Ltd., Helsinki, Finland) at room temperature Perkin-Elmer ATR-FTIR Spectrometer. FTIR analysis was
was used to measure the interfacial tensions of the solid- used to determine the presence of the functional group of
liquid (wetting film properties). A syringe with 5 µl of water bioplastic film. The FTIR spectra were analysed and
was released to the film surface. Images were shot after 10 identified according to the FTIR library. The spectra were
seconds of contact and documented after the water was collected from 4000 cm-1 to 500 cm-1 frequency range with
released onto the film surface. The angle between the 16 times number of scan to reduce the noise.
tangent and the baseline at the drop boundary was measured. Meanwhile, 1H NMR analysis was recorded using NMR
spectrometer (Bruker Ultra Shield Plus, Germany) at
Water Vapour Permeability Analysis 500 MHz and at room temperature (27 °C). Samples were
Crucible was filled with approximately 6 ml of distilled prepared in DMSO-d6 and tertramethylsilane (TMS) as the
water and a film sample was securely covered over the rim. internal standard. The chemical shift were measured respected
The crucibles were placed inside a desiccator (50±5 % RH to the remaining proton resonance of DMSO-d6.
and 23±2 °C) containing silica beads. The weights were Moisture Content
monitored at 1 h interval over 8 h periods and WVP was The moisture content of the bioplastic film was measured
calculated using the following equation: using moisture meter (MS-70 with SHS inside, Japan).
Approximately 0.1 g of the film was placed onto the plate of
w  l
WVP = --------------- (2) the moisture meter and the film was heated at 105 °C. The
AtP
heating stopped automatically once the constant moisture
where Δw is the weight difference (g), l is the film thickness content value was obtained [39].
(m), A is the exposed area of the film (m2), t is the time Thermal Properties
elapsed, and P is the partial pressure difference of the water Thermal properties were determined from Differential
vapour across the film (Pa). The method was adapted from Scanning Calorimetry (DSC) and Thermo Gravimetric
[36]. Analysis (TGA). For DSC, the analysis was carried out
using a TA-Instrument DSC/Q 1000. 0.6 mg of bioplastic
Biocomposite Film Characterisation film was heated at 10 °C/min from 0 °C until 250 °C. The
Tensile Strength glass transition temperature (Tg), melting temperature and
Characterisation of the mechanical properties of tensile enthalpy (Tm and ΔHm), crystallization temperature and
strength was carried out using a textile analyser testing enthalpy (Tc and ΔHc) were recorded. From the enthalpy
machine (CT3-Texuture Analyzer, Broolfield, USA) equipped value, the crystallinity index can be calculated from the
2888 Fibers and Polymers 2022, Vol.23, No.10 Fatmawati Adam et al.

following equation:
 Hm – Hc 
Crystallinity index (%) = -----------------------------
-  100% (5)
H m
TGA analysis was run on a Mettler Toledo thermal
analyser. The sample was heated from 0 °C up to 800 °C (at
10 °C/min) in airflow. The mass loss and differential thermal
analysis profiles were recorded. The activation energy was
calculated from the result obtained from TGA thermal
analysis based on Arrhenius theory and using the Broido’s
model as per the following equation [40]:
E
ln ln  --- = –  -----a  --1- – C
1
(6)
 y  R   T Figure 1. Particle size distribution of CNW and NFC nanocelluloses.

where Ea is the activation energy, R is the gas constant

(8.314 J mol-1 K-1) and C is the constant value of the slope.
Broido’s model was selected because it is the simplest
method to calculate the activation energy of polysaccharide
materials [40].
Degradability Test and Compostability Test
Degradation tests were conducted in formic acid at 30 °C.
200 mg of bioplastic film was placed in 20 ml humic acid in
polypropylene tubes for 30 minutes within 5 hours. Then,
the bioplastic film was rinsed with deionized water and dried
out in a hot oven at 60 °C for four days. The amount of
degraded bioplastic film was obtained by measuring the
difference between the initial weight and the final weight of
the bioplastic film. Commercial paper and conventional
petroleum-based plastic samples were taken for the
compostability test in comparison with the bioplastic samples.
Samples were weighed separately and buried approximately
10 cm deep in soil for 1 month at room temperature (37 °C).
The mass of each sample was weighted and the appearances
were observed on alternate days (i.e. days 0, 5, 10, 15, 20,
25, and 30) [41].

Results and Discussion

Cellulose Fibrillations
The particle size distribution of both CNW and NFC
fibrillated celluloses are shown in Figure 1. The particle size
distribution of CNW was distributed in the range 5.62 to
190.10 nm with an average diameter of 54.30 nm. For NFC,
the particle size distribution was in the range 68.06 to
1990.00 nm with an average diameter of 474.50 nm. CNW
has a much smaller particle size compared to NFC. The Figure 2. Morphology of particle size distribution of FESEM (a)
original particle size of MCC was 50 µm. The micro particles CNW and (b) NFC; and TEM images of (c) NFC.
of MCC were broken into smaller sizes after grinding to
obtain nano size CNW. The MCC was sonicated to reduce distribution of the nano-size particles. CNW particles had
the size of MCC from 50 µm to 474.50 nm of NFC. The smaller size of 54.30 nm and the highest intensity (8.2 %).
largest particle size of CNW is the smallest particle size of The sharp and narrow peak of NFC indicates that the particle
NFC obtained. Thus, the grinding method successfully reduced size of NFC was homogenous with most of the particle sizes
a smaller nano-size particle compared to the sonication of NFC being of the order 474.50 nm. Nanoparticle size was
method. The broad shape of the CNW graph shows the in the range 1 to 100 nm [42]. Hence, NFC represents the
Reinforcement of Nanocellulose in Carrageenan Matrix Fibers and Polymers 2022, Vol.23, No.10 2889

fine particles of MCC since the average size of NFC was MCC film. Visual observations found that, Carra-CNW and
above 100 nm. Carra-NFC are more flexible compared to Carra-MCC film
The narrow particle size distribution of NFC particles which is more rigid. Carra-MCC is more rigid and opaque
shows that the particle tend to agglomerate [43]. This is in due to the greater cellulose size used. A similar effect was
agreement with Figure 2 where under the observation of reported in a study, where due to the insolubility of cellulose
FESEM, the NFC particles agglomerate compared to the and the solution of the film Carra-MCC was cloudier at high
CNW particles. The TEM images (Figure 2(c)) also show concentrations of MCC [44]. Based on Table 1, Carra-MCC
the agglomeration of the NFC particles as per highlighted in has the highest opacity (12.89) compared to 5.82 and 3.63
the circled area. for Carra-NFC and Carra-CNW, respectively. MCC cellulose
has greater in particle size compared to the CNW and NFC
Carrageenan-Cellulose Biocomposite Film cellulose which demonstrates that the particle size of the
The appearance of each carrageenan-cellulose film is cellulose affects the appearance of the biocomposite film
different according to the types of the cellulose used. In obtained. The contact angle was measured for each bioplastic
Figure 3, Carra-CNW and Carra-NFC have a smoother sample. The values in Table 1 reflect that Carra-CNW shows
surface and are more transparent compared to the Carra- the highest hydrophobicity ability compared to the control
Carra, Carra-MCC and Carra-NFC. Carra-CNW has the
smallest (54.30 nm) cellulose nanoparticles and produced a
70.31 ° water contact angle value suggesting a good
dispersion of cellulose particles in the formulation solution.
Water vapour permeability (WVP) for Carra-MCC, Carra-
CNW and Carra-NFC are 3.58×10-14 g s-1 m-1 Pa-1, 3.43×
10-14 g s-1 m-1 Pa-1, and 4.09×10-14 g s-1 m-1 Pa-1, respectively.
WVP values show a minor difference between the cellulose
sizes which were used in the carrageenan formulation
matrix. In comparison to WVP value of carrageenan
plasticized with glycerol film, the value is lower at 3.26×10-8
g s-1 m-1 Pa-1. The addition of pomegranate seed oil into the
carrageenan film produced 2.35×10-8 to 8.47×10-9 g s-1 m-1
Pa-1 WVP film [45]. Using chitosan with graphene oxide at
different mixing temperatures produced WVP in the range
of 11.91-9.56×10-8 g s-1 m-1 Pa-1 [36]. In this work the film
samples exhibit a good water vapour barrier which is
suitable as a future food packaging materials.

Biocomposite Film Characterisation

Figure 3. Appearances of (a) Carra-MCC, (b) Carra-CNW, and (c) Hydrocolloid and Tensile Properties
Carra-NFC samples. Table 2 compares the viscosity, tensile strength and
elongation at break (EAB) of the Carra-cellulose samples.
Viscosity represents the intermolecular interaction in the
Table 1. Opacity measurement and water contact angle of Carra- formulation solution between carrageenan, cellulose, crosslinker,
MCC, Carra-CNW and Carra-NFC thickener agent, plasticiser and water. Viscosity can also be
Type of cellulose Opacity Water contact angle (°) correlated with the homogeneity and uniformity of the
Control Carra 4.71±0.37 46.81±1.32 solution and will affect the tensile strength of the film.
Viscosity reflects the intermolecular interaction in the
Carra-MCC 3.63±0.10 49.04±0.25
formulation solution such as the Van der Waals and hydrogen
Carra-CNW 12.89±0.46 70.31±0.74
bonding [23]. κ-carrageenan has established a hydrogen
Carra-NFC 5.82±0.12 57.73±1.86 bonding interaction between glyoxylic acid through physical

Table 2. Tensile strength, EAB and viscosity of Carra-MCC, Carra-CNW and Carra-NFC
Type of bioplastic Shear stress (Pa) Viscosity (mPas) Tensile strength (MPa) EAB (%)
Carra-MCC 96.11±2.02 264.63±6.83 11.15±0.40 44.29±1.32
Carra-CNW 127.90±1.17 293.90±2.11 19.87±0.53 39.16±0.01
Carra-NFC 102.36±0.52 262.78±1.89 16.39±0.33 37.06±0.65
2890 Fibers and Polymers 2022, Vol.23, No.10 Fatmawati Adam et al.

crosslinking. The oxygen and sulphur atoms in κ-carrageenan

demonstrated the most negative electrostatic potential.
Meanwhile, the positive regions are localised at the hydrogen
functional group in glyoxylic acid [23]. In another work, the
addition of 1.6 wt./v% of CNC crosslinked with alginic acid
into semi refined carrageenan produced 33.08 MPa tensile
strength and a 1.17 mPas of viscocity for hard capsule
application [46].
The incorporation of carrageenan with different sizes of
cellulose (MCC, CNW and NFC) shows the different values
of shear stress of each film. Carra-NCW has the highest
value of shear stress (127.90 Pa) compared to Carra-NFC
and Carra-MCC which are 102.36 Pa and 96.11 Pa,
respectively. Viscosity can be defined as the coefficient of
friction that was obtained when shear stress (friction force)
was applied to the fluid or solution. The increasing value of
shear stress between the layers of the solution would
increase the viscosity of the solution tremendously [47].
Hence, Carra-CNW has a higher value of viscosity (293.90
mPas) compared to Carra-MCC and Carra-NFC values
(264.63 mPas and 262.78 mPas, respectively).
The viscosity can be correlated to the tensile strength of
the film samples [48]. Tensile strength shows that Carra-
CNW has the highest value (19.87 MPa) followed by Carra-
NFC (16.39 MPa) and Carra-MCC (11.15 MPa). The
increment of tensile strength is correlated with the increasing
degree of substitution value of OH bonds in CNW and NFC.
The substituition of carboxymethyl group in fibrillated MCC
leads to an increase in intermolecular interaction between
the composite chains [48]. Additives such as xanthan gum
and glyoxal most likely promoted the strong intermolecular
interaction under alkaline conditions which increased the
wheat gluten-plasticiser compatibility for bioplastic production
[49]. PLA was incorporated with starch and natural cellulose
fibre (with added glycerol) to produce a tensile strength of Figure 4. SEM surface morphology of (a) Carra-MCC, (b) Carra-
approximately 20 MPa with the increased concentration of CNW, and (c) Carra-NFC.
natural cellulose fibres [50]. The reduction of tensile
strength in Carra-MCC might be due to the agglomeration of
MCC in the biocomposite matrix [51]. Besides this, the SEM Morphology
lower value of tensile strength also due to the lower Ther surface morphology of the biocomposite films of
intermolecular interaction of the hydroxyl group of Carra-MCC, Carra-CNW, and Carra-NFC are shown in
carrageenan with MCC [52]. High values of elongation at Figure 4 under 5000x magnification. In Figure 4(a), Carra-
break (EAB) leads to material with high toughness. Carra- MCC has a rough, jagged surface. This is because MCC did
MCC has an EAB value of 44.29 %, higher than Carra- not dissolve and dispersed well in water. MCC can only be
CNW (39.16 % of EAB) and Carra-NFC (37.06 % of EAB), dissolved in NaOH at a concentration of 25-30 % [53]. The
respectively. EAB of the biofilm is usually manipulated by surface morphology of both Carra-CNW and Carra-NFC is
the presence of the plasticiser. The changes in the value of smoother and less jagged compared to Carra-MCC. CNW
EAB might be due to the insufficient amount of plasticiser in and NFC are able to disperse well in water compared to
the matrix to increase the flexibility and the continuation of MCC. This can be attributed to the particle size of CNW and
the matrix chains. The condition might be due to the NFC which are smaller and more easily diffuse into the
substituent carboxymethyl group in both CNW and NFC solution matrix compared to MCC.
cellulose which increased the intermolecular interaction and FTIR and 1H NMR of Biocomposite
require a higher concentration of plasticiser in the matrix to The FTIR spectra of carrageenan, Carra-MCC, Carra-
obtain high EAB values. CNW and Carra-NFC are shown in Figure 5, to determine
Reinforcement of Nanocellulose in Carrageenan Matrix Fibers and Polymers 2022, Vol.23, No.10 2891

Figure 5. FTIR spectra of carrageenan, Carra-MCC, Carra-CNW

and Carra-NFC.

the presence of the functional groups of biofilm sample. The

band at 1223 cm-1 is due to the -SO of sulphate esters and
1034 cm-1 indicates the glycosidic linkage of carrageenan
that usually appears in the 1010-1080 cm-1 region [54]. Band
841 cm-1 belongs to the D-galactose-4-sulfate as it appears in
840-850 cm-1 range [54]. The 925-935 cm-1 region indicates
the presence of 3,6-anhydro-D-galactose of carrageenan
which,, in this work appears at 923 cm-1.
The wide band of 3381-3393 cm-1 in Carra-MCC, Carra-
CNW and Carra-NFC indicates the stretching vibration of
-OH groups of the cellulose which appears in the region
between 3200 and 3500 cm-1 [55,56]. The band that appears
at 2875 cm-1 in Carra-MCC and 2874 cm-1 in both Carra-
CNW and Carra-NFC, belongs to the asymmetrically
stretching vibration of the -CH group in the pyranoid ring of
cellulose appeared in 2905 cm-1 region [57]. The peak at
1647 cm-1 in Carra-MCC, 1637 cm-1 in Carra-CNW and
1647 cm-1 in Carra-NFC shows the -OH adsorbed water
vibration [57] and -H intermolecular interaction occurs,
increasing the intensity of the peak. Finally, the peak in the
1062-1063 cm-1 region in all Carra-cellulose spectra can be
attributed to the -CO group of cellulose which appeared at
Figure 6. (a) 1H-NMR spectrum of refined carrageenan, (b) 1H-
1059 cm-1. NMR spectra of (i) MCC, (ii) CNW, and (c) 1H-NMR spectrum of
Spectra of Carra-MCC, Carra-CNW and Carra-NFC shows Carra-MCC bioplatic film.
the combination of several peaks from both carrageenan and
cellulose used in the matrix. It is expected that the hydroxyl
group (-OH) of cellulose plays an important role with not alter any of the functional groups from MCC structure.
carrageenan through interaction with the sulphate group The increasing intensity and shifting of the band indicates
forming hydrogen bonding between the network formation the intermolecular interaction that occurs between carrageenan
[39]. The interaction of the intermolecular bonds is shown in and cellulose in the biocomposite matrix.
the intensity and shifted peak at 1223 cm-1 in carrageenan Figure 6(a) shows 1H-NMR of the refined carrageenan
spectrum to 1247 cm-1carra-cellulose spectrum. It can also spectrum which was divided into two main units. G-unit
be seen that the entire spectrum has an identical peak. refers to alternating 3-linked β-d-galactopyranose and 4-
Hence, the method used to obtain both CNW and NFC did linked 3,6-anhydro-α-D-galactopyranose refers to DA-units.
2892 Fibers and Polymers 2022, Vol.23, No.10 Fatmawati Adam et al.

The trend is similar to kappa-carrageenan 1H-NMR in the

previous literature [58-60]. The spectrum in Figure 6(a),
demonstrated that G-unit has two points of chemical shift:
G-1 (1H; 2.79 ppm) and G-4 (1H; 2.68 ppm). Meanwhile for
the DA-unit, there are four points of chemical shift: DA-1
(1H; 2.71 ppm), DA-3 (O-1H; 2.41 ppm), DA-4 (O-1H; 2.47
ppm) and DA-5 (1H; 2.51 ppm). In comparison to the
previous work [60-62], the chemical shift obtained is
different due to the different solvents used to dissolve
carrageenan prior to the analysis. DMSO-d6 caused the
peaks obtained in this study to be lower in comparison to the
chemical peak obtained using deuterated water (D2O) and
deuterated chloroform solvents.
In Figure 6(b), the 1H-NMR spectra of MCC and CNW
fibrillated cellulose are identical. This is in agreement with
the FTIR spectrum (Figure 5) of both MCC and CNW which
are also identical due to the similarity of the chemical Figure 7. DSC thermogram trends of Carra-MCC, Carra-CNW
structure. Figure 6(b)(i) shows four main points of chemical and Carra-NFC.
shift for MCC namely 1H triplet (2.40-3.50 ppm), 1H
quartet (2.40-3.50 ppm), 1H doublet (3.00-3.53 ppm), O-1H properties of the plastic, reducing the tensile strength and
(3.70 ppm) which are similar to Hameed et al. (2018) for making it more brittle [66].
cellulose fibrillated proton NMR [63]. Meanwhile, Figure Thermal Properties
6(c) represents the 1H-NMR spectrum of Carra-MCC Figure 7 represents the differential scanning calorimetry
bioplastics where interaction occurs at DA-unit of refined (DSC) analysis for each carra-cellulose bioplastic film. An
carrageenan and O-1H of MCC. The chemical shift endothermic profile for each carra-cellulose films begins at
increased the hydrogen became closer to the electronegative 180 °C due to the heat absorption for dehydration of water
atom. In the NMR spectrum, increments of electronegativity and the decomposition of the biofilm [40]. The decomposition
can be observed as the peak becomes more downfield, of the biofilm is attributed to the occurrence of the
shifting to the left. heterolysis and thermolysis of glycosidic bonds along the
Moisture Content ordered chains. The higher onset temperature is often
The moisture content results of bioplastic samples associated with higher thermal stability. This kind of behaviour
demonstrated that the addition of cellulose used in the is usually related to the high degree of crystallinity of
carrageenan matrix increased the water content. Cellulose is cellulose [27,63,64]. The glass transition (Tg), crystallisation
capable of increasing the water content of the bicomposites temperature (Tc), crystallization enthalpy (ΔHm), melting
or biofilm produced [64]. The control Carra film had temperature (Tm), melting enthalpy (ΔHm) and crytallinity
approximate moisture content about 13.90 %. The moisture percentage (XcH) values are listed in Table 4.
content of Carra-MCC, Carra-CNW and Carra-NFC were Tg values for all bioplastic samples are indifferent,
15.75 %, 11.29 % and 13.02 %, respectively (as shown in however, there is a slight difference in Tm of Carra-NFC
Table 3). The value of moisture content was not really compared to Carra-CNW and Carra-MCC. Tg for κ-
affected much by the type of cellulose. The hydrophilic carrageenan is at 118 °C [67]. The addition of cellulose acts
property of the carrageenan has increased the affinity as a nano-size reinforcement filler because it is able to fill
between plasticizer and water, hence increasing the water and diffuse into carrageenan matrices. This then strengthens
absorption [65]. This resulted in unsuitable bioplastic for the matrix-chains in the film samples. All sample peaks are
most applications because the water absorption alters the observed before the Tm value may suggest an incomplete
homogenous structure of the film samples. Carra-MCC
shows many small peaks compared to Carra-CNW and
Table 3. Moisture content of Carra-MCC, Carra-CNW and Carra- Carra-NFC due to the non-homogeneous film samples of
NFC bioplastic film Carra-MCC. Tm is determined by the transition of the
Type of cellulose Moisture content (%) crystalline region. Tc is the transition process from amorphous
Control Carra 13.90±0.52
to crystalline states. Carra-NFC has the highest values of Tm
and Tc and this suggests that a strong intermolecular
Carra-MCC 15.75±0.42
interaction formed in this sample. The higher enthalpy of
Carra-CNW 11.29±0.25 melting demonstrated the cross-linking ability of carrageenan
Carra-NFC 13.02±0.25 and cellulose by glyoxylic acid which exhibits a narrow
Reinforcement of Nanocellulose in Carrageenan Matrix Fibers and Polymers 2022, Vol.23, No.10 2893

Table 4. Melting temperature and melting enthalpy of Carra-MCC, Carra-CNW and Carra-NFC biofilm
Melting Melting Crystallisation Crystallisation Crytallinity
Type of Glass transition
temperature enthalpy temperature enthalpy percentage
cellulose (Tg, °C)
(Tm, °C) (ΔHm, J/g) (Tc, °C) (ΔHc, J/g) (XcH)
Carra-MCC 150.73 171.40 23.69 180.87 432.17 94.40
Carra-CNW 151.59 174.25 27.69 184.63 449.84 93.84
Carra-NFC 152.80 177.75 32.98 186.85 506.44 93.49

Figure 8. (a) DTG and (b) weight loss plot of Carra-MCC, Carra-CNW and Carra-NFC.

crystallisation peak implying excellent crystallite size ash mass is approximately 8.21 %, 9.06 % and 10.69 % for
distribution. Stronger hydrogen bonding formed in Carra- the Carra-MCC, Carra-CNW and Carra-NFC samples,
NFC. The crystallinity percentage in all bioplastic samples respectively.
are unchanged and are not affected by the cellulose sizes. All Carra-cellulose bioplastic have almost identical stages
Thermogravimetric analysis (TGA) was carried out to of thermal decomposition. There is not much difference in
determine the thermal stability and decomposition of carra- the total weight loss of each carra-cellulose sample film. The
cellulose bioplastic films. In Figure 8(a), DTG represents the highest weight lost is 64.86 % for Carra-CNW. The weight
peak at which temperature caused the maximum weight loss loss for Carra-MCC is 63.50 % and Carra-NFC is 60.30 %.
during film decomposition. The maximum values for The activation energy of the film was calculated according
decomposition temperature for Carra-MCC, Carra-CNW to Arrhenius theory using Broido’s model derived from the
and Carra-NFC are 247.6 °C, 250.6 °C and 231.6 °C, highest peak of DTG. Figure 9(a), (b) and (c) show the
respectively. In another work [68], the addition of glycerol Broido’s plot of Carra-MCC, Carra-CNW and Carra-NFC.
and tocopherol improved the thermal stability of the semi- The slope represents the value of activation energy (Ea).
refined carrageenan films from 229.5 °C to 266-277 °C. In Carra-MCC has an activation energy value of 73.81 kJ/mol,
this work, the different sizes of cellulose increased the Carra-NFC is 59.14 kJ/mol and Carra-CNW is 47.14 kJ/mol.
decomposition temperature to 231-250 °C. The weight loss Higher values of Ea, indicate that the film is more thermally
(see Figure 8(b)) shows the decomposition stage which stable. The changes of the Ea values are due to the
occurs on the bioplastic film samples. In Figure 8(b), all the compatibility of the cellulose size which interacts with
samples experienced several stages of decomposition during carrageenan. Ea for control carra is 27.88 kJ/mol which is
the thermal degradation process. The main decomposition lower than carra-cellulose bioplastic film. Carra-MCC is
stage occurs from 150-360 °C which indicates bioplastic more thermally stable compared to Carra-CNW and Carra-
depolymerisation [69]. A temperature of 120-260 °C indicates NFC. High thermal stability causes the sample to take a
the decomposition of the plasticiser, PEG [70] and at 250- longer time to decompose [40]. Micron size cellulose
360 °C, indicates the thermal decomposition of the reinforced into carrageenan bioplastic exhibited the most
polysaccharide backbone in the carrageenan [71]. The final stability compared to nano-sized samples. This suggests that
stage occurs at 600-800 °C represented by the horizontal line cellulose size is an important component for bioplastics.
and indicates that all samples decomposed. The residue or High energy was required to decompose Carra-MCC. This is
2894 Fibers and Polymers 2022, Vol.23, No.10 Fatmawati Adam et al.

Figure 9. Broido’s plot of (a) control Carra, (b) Carra-MCC, (c) Carra-CNW, and (d) Carra-NFC.

in agreement with the morphological observations in Figure

3(a) in which Carra-MCC has a rough surface compared to
Carra-CNW and Carra-NFC. The degradation routes in both
DSC and TGA plots are the same due to the identical
functional group for all types of cellulose used since both
CNW and NFC maintain the cellulose structure from MCC.
A lower Ea value usually exhibits a faster degradation for a
Carra-CNW sample.
Degradability Test and Compostability Test
Degradability tests were carried out to evaluate the
bioplastic biodegradability. Figure 10 shows the percentage
of the weight loss of carra-cellulose film in humic acid. Both
Carra-CNW and Carra-NFC dissolved completely after 200 Figure 10. Weight loss of Carra-cellulose films with the time of
minutes of immersion time. Carra-MCC dissolved completely exposure in humic acid.
at 270 minutes. In comparison with the degradation time of
carra-cellulose films, petroleum-based plastic and conventional
paper was also considered as a reference. The graph shows
that petroleum-based plastic was only able to degrade up on
to 80 %. Conventional paper degraded after 270 minutes of
immersion with 90 % weight loss.
Compostability tests were carried out to evaluate the
capability of the bioplastic films to be degraded in soil. The
observations were conducted for 30 days. Figure 11 shows
the comparison of the weight loss between conventional
paper, petroleum-based plastic and all type of bioplastic
samples. The weight loss of both conventional paper and
petroleum-based plastic did not change from the start of the
test until day 30. All types of bioplastic samples show the
weight loss from the beginning until day 30. The weight loss Figure 11. Weight loss of paper, conventional plastic, Carra-MCC,
of Carra-MCC, Carra-CNW and Carra-NFC at day 30 was Carra-CNW and Carra-NFC in 30 days.
Reinforcement of Nanocellulose in Carrageenan Matrix Fibers and Polymers 2022, Vol.23, No.10 2895

49 %, 65 % and 58 %, respectively. In a study, the degradation 4. X. Cheng, H. Shi, C. D. Adams, and Y. Ma, Environ. Sci.
of the film produced by the incorporation of starch and Pollut. Res., 17, 1323 (2010).
microorganism such as fungi and bacteria increases the mass 5. M. Whitt, K. Vorst, W. Brown, S. Baker, and L. Gorman, J.
loss up to 87.6 % compared to the control film at 55.1 % Plast. Film Sheeting, 29, 163 (2013).
within 15 days of burial [41]. This suggests that the chemical 6. G. F. Schirinzi, I. Pérez-Pomeda, J. Sanchís, C. Rossini, M.
components used in the formulation of bioplastics are able to Farré, and D. Barceló, Environ. Res., 159, 579 (2017).
influence the degradation rate of the bioplastic. According to 7. A. A. Horton, A. Walton, D. J. Spurgeon, E. Lahive, and C.
European Standard EN1343, the requirement for the Svendsen, Sci. Total Environ., 586, 127 (2017).
biodegradable plastic to be considered biodegradable and 8. S. Rainieri, N. Conlledo, B. K. Larsen, K. Granby, and A.
compostable is the ability to break down completely under Barranco, Environ. Res., 162, 135 (2018).
the right conditions into water, carbon dioxide (CO2) and 9. F. Gironi and V. Piemonte, Energy Sources, Part A
biomass after six month without leaving any toxic residue Recover. Util. Environ. Eff., 33, 1949 (2011).
[41]. Based on the weight loss graph, the bioplastic film’s 10. S. R. Vijayan, P. Santhiyagu, R. Ramasamy, P. Arivalagan,
on-going degradation has not yet plateaued. This suggests G. Kumar, K. Ethiraj, and B. R. Ramaswamy, Enzyme
that the Carra-films degrade quickly and are able to fulfil the Microb. Technol., 95, 45 (2016).
requirement to be considered as biodegradable plastic. 11. N. Rajendran, S. Puppala, M. Sneha Raj, B. Ruth
Angeeleena, and C. Rajam, J. Pharm. Res., 5, 1476 (2012).
Conclusion 12. S. Brockhaus, M. Petersen, and W. Kersten, J. Clean.
Prod., 127, 84 (2016).
In conclusion, the incorporation of micro and nano sizes of 13. F. Bilo, S. Pandini, L. Sartore, L.E. Depero, G. Gargiulo,
cellulose leads to different bioplastic properties. Carra-MCC A. Bonassi, S. Federici, and E. Bontempi, J. Clean. Prod.,
is more stable with the highest Ea based on the thermal 200, 357 (2018).
effect. Incorporation of 54.30 nm of cellulose improved the 14. S. Papong, P. Malakul, R. Trungkavashirakun, P. Wenunun,
surface smoothness, hydrophobicity and lower opacity, and T. Chom-In, M. Nithitanakul, and E. Sarobol, J. Clean.
resulted in the highest tensile strength and EAB. CNW Prod., 65, 539 (2014).
particles are able to increase the intermolecular interaction 15. B. B. Sedayu, M. J. Cran, and S. W. Bigger, Carbohydr.
such as hydrogen bonding and Van der Waals due to the Polym., 216, 287 (2019).
highest viscosity and shear stress value in the formulation 16. Y. Qin, “Bioactive Seaweeds for Food Applications:
solution. 1H-NMR spectra reflect the formulation of hydrogen Natural Ingredients for Healthy Diets”, 1st ed., pp.135-
bonding in Carra-MCC, Carra-CNW and Carra-NFC. The 152, Academic Press, 2018.
NMR analysis should also be evaluated for the possibility of 17. R. Nur Fatin Nazurah and Z. A. Nur Hanani, Carbohydr.
Van der Waals intermolecular interaction between carrageenan Polym., 157, 1479 (2017).
and cellulose at different cellulose sizes. 18. G. Sun, T. Liang, W. Tan, and L. Wang, Food Hydrocoll.,
85, 61 (2018).
Acknowledgements 19. M. P. Sudhakar, D. Magesh Peter, and G. Dharani, Environ.
Sci. Pollut. Res., 28, 33899 (2021).
The authors would like to acknowledge Universiti Malaysia 20. F. Adam, J. Jamaludin, S. H. Abu Bakar, R. Abdul Rasid,
Pahang (UMP) for the project funding (RDU1803187), and Z. Hassan, Cogent Eng., 7, 1765682 (2020).
UMP student research funding (PGRS200353) and for 21. S. Shojaee-Aliabadi, H. Hosseini, M. A. Mohammadifar,
providing facilities in undertaking this work. A. Mohammadi, M. Ghasemlou, S. M. Hosseini, and R.
Khaksar, Carbohydr. Polym., 101, 582 (2014).
Conflicts of Interest 22. F. Adam, M. A. Hamdan, S. H. Abu Bakar, M. M. Yusoff,
and R. Jose, Chem. Eng. Commun., 208, 741 (2021).
The authors declare that they have no competing interests 23. S. H. Abu Bakar and F. Adam, Malaysian J. Anal. Sci., 21,
979 (2017).
References 24. C. Bishop, “Vacuum Deposition onto Webs, Films and
Foils”, 2nd ed., William Andrew, USA, 2011.
1. J. J. Klemeš, Y. Van Fan, and P. Jiang, Energy Sources, 25. M. El Achaby, N. El Miri, H. Hannache, S. Gmouh, H.
Part A Recover. Util. Environ. Eff., 43, 1549 (2021). Ben Youcef, and A. Aboulkas, Int. J. Biol. Macromol., 117,
2. J. R. Jambeck, R. Geyer, C. Wilcox, T. R.Siegler, M. 592 (2018).
Perryman, A. Andrady, R. Narayan, and K. L. Law, 26. D. Trache, M. H. Hussin, C. T. Hui Chuin, S. Sabar, M. R.
Science, 347, 6223 (2015). N. Fazita, O. F. A. Taiwo, T. M. Hassan, and M. K. M.
3. N. Ahmad, M. Nasibullah, F. Hassan, and M. Rahman, Int. Hasfiz, Int. J. Biol. Macromol., 93, 789 (2016).
Res. J. Environ. Sci., 1, 32 (2012). 27. N. Herrera, A. P. Mathew, and K. Oksman, Compos. Sci.
2896 Fibers and Polymers 2022, Vol.23, No.10 Fatmawati Adam et al.

Technol., 106, 149 (2015). 50. F. Masmoudi, A. Bessadok, M. Dammak, M. Jaziri, and E.
28. V. S. Karande, A. K. Bharimalla, N. Vigneshwaran, P. G. Ammar, Environ. Sci. Pollut. Res., 23, 20904 (2016).
Kadam, and S. T. Mhaske, Iran. Polym. J., 23, 869 (2014). 51. H. Kargarzadeh, M. Mariano, J. Huang, N. Lin, I. Ahmad,
29. S. Suvachittanont and P. Ratanapan, J. Chem. Chem. Eng., A. Dufresne, and S. Thomas, Polymer, 132, 368 (2017).
7, 1136 (2013). 52. M. A. Hamdan, N. A. Ramli, N. A. Othman, K. N. Mohd
30. C. Borsoi, H. L. Ornaghi, L. C. Scienza, A. J. Zattera, and Amin, and F. Adam, Mater. Today, 42, 56 (2021).
C. A. Ferreira, Polym. Polym. Compos., 25, 563 (2017). 53. W. Neng, D. Enyong, and C. Rongshi, Langmuir, 24, 5
31. V. Kumar, R. Bollström, A. Yang, Q. Chen, G. Chen, P. (2008).
Salminen, D. Bousfield, and M. Toivakka, Cellulose, 21, 54. V. Webber, S. M. de Carvalho, P. J. Ogliari, L. Hayashi,
3443 (2014). and P. L. M. Barreto, Food Sci. Technol., 32, 812 (2012).
32. F. Li, P. Biagioni, M. Bollani, A. Maccagnan, and L. 55. M. M. Rahimi Kord Sofla, R. J. Brown, T. Tsuzuki, and T.
Piergiovanni, Cellulose, 20, 2491 (2013). J. Rainey, Adv. Nat. Sci. Nanosci. Nanotechnol., 7, 035004
33. M. Österberg, J. Vartiainen, J. Lucenius, U. Hippi, J. (2016).
Seppälä, R. Serimaa, and J. Laine, ACS Appl. Mater. 56. J. Li, Y. Wang, X. Wei, F. Wang, D. Han, Q. Wang, and L.
Interfaces, 5, 4640 (2013). Kong, Carbohydr. Polym., 113, 388 (2014).
34. D. Brown, V. Marcadon, P. Mélé, and N. D. Albérola, 57. N. Jia, S. M. Li, M. G. Ma, J. F. Zhu, and R. C. Sun,
Macromolecules, 41, 1499 (2008). BioResources, 6, 1186 (2011).
35. F. Lin, R. Zheng, J. Chen, W. Su, B. Dong, C. Lin, B. 58. E. M. Vilén, L. C. E. Lundqvist, D. Jouanneau, W. Helbert,
Huang, and B. Lu, Carbohydr. Polym., 205, 244 (2019). and C. Sandström, Biomacromolecules, 11, 3487 (2010).
36. H. L. F., C. P. Tan, R. M. Zawawi, and N. H. Za, Food 59. F. Van De Velde, L. Pereira, and H. S. Rollema, Carbohydr.
Packag. Shelf Life, 28, 100663 (2021). Res., 339, 2309 (2004).
37. M. Lubis, A. Gana, S. Maysarah, M. H. S. Ginting, and M. 60. L. Youssouf, L. Lallemand, P. Giraud, F. Soulé, A. Bhaw-
B. Harahap, IOP Conference Series: Mater. Sci. Eng., 309, Luximon, O. Meilhac, C.L. D'Hellencourt, D. Jhurry, and
012100 (2018). J. Couprie, Carbohydr. Polym., 166, 55 (2017).
38. Maulida, M. Siagian, and P. Tarigan, J. Phys. Conf. Ser., 61. W. A. K. Mahmood, M. M. R. Khan, and T. C. Yee, J.
710, 012012 (2016). Phys. Sci., 25, 123 (2014).
39. M. A. Hamdan, F. Adam, and K. N. Mohd Amin, Int. J. 62. N. G. Voron’ko, S. R. Derkach, M. A. Vovk, and P. M.
Innov. Sci. Res. Technol., 3, 457 (2018). Tolstoy, Carbohydr. Polym., 151, 1152 (2016).
40. J. Jamaludin, F. Adam, R. A. Rasid, and Z. Hassan, Chem. 63. T. M. Hameed, A. H. Dawood, I. S. Arif, and Y. D.
Eng. Res. Bull., 19, 80 (2017). Saihood, J. Pharm. Sci. Res., 10, 2768 (2018).
41. M. Carissimi, S. H. Flôres, and R. Rech, Algal Res., 32, 64. M. A. S. Azizi Samir, F. Alloin, and A. Dufresne,
201 (2018). Biomacromolecules, 6, 612 (2005).
42. I. Khan, K. Saeed, and I. Khan, Arabian J. Chem., 12, 908 65. N. Wahyuningtyas and H. Suryanto, J. Mech. Eng. Sci.
(2019). Technol., 1, 24 (2017).
43. V. Baheti, R. Abbasi, and J. Militky, World J. Eng., 9, 45 66. A. Jones, M. A. Zeller, and S. Sharma, Prog. Biomater., 2,
(2012). 12 (2013).
44. N. A. Othman, F. Adam, and N. H. Mat Yasin, Mater. 67. N. Khairuddin, I. I. Muhamad, W. A. W. Abd Rahman, and
Today, 41, 77 (2021). B. M. Siddique, Sains Malaysiana, 49, 323 (2020).
45. E. Sogut, A. M. Ili Balqis, Z. A. Nur Hanani, and A. C. 68. K. H. Abd Hamid, W. A. Wan Yahaya, N. A. Z. Mohd
Seydim, Polym. Test., 77, 105886 (2019). Saupy, M. P. Almajano, and N. A. Mohd Azman, J. Food
46. M. A. Hamdan, M. A. Khairatun Najwa, R. Jose, D. Process. Preserv., 43, 3937 (2019).
Martin, and F. Adam, Food Hydrocoll. Heal., 1, 100023 69. H. Suryanto, F. A. Kharismawan, A. W. Solichin Rahmawan,
(2021). R. T. Sahana, M. Muhajir, and U. Yanuhar, IOP Conference
47. S. L. Hii, J. Y. Lim, W. T. Ong, and C. L. Wong, J. Eng. Series: Mater. Sci. Eng., 494, 012077 (2019).
Sci. Technol., 11, 1 (2016). 70. R. Colussi, V. Pinto, S. L. M. El Halal, B. Biduski, L.
48. P. Rachtanapun and N. Rattanapanone, J. Appl. Polym. Prietto, D. D. Castilhos, E. Zavareze, and A. R. G. Dias,
Sci., 122, 3218 (2011). Food Chem., 221, 1614 (2017).
49. M. Jiménez-Rosado, L. S. Zarate-Ramírez, A. Romero, C. 71. J. T. Martins, M. A. Cerqueira, and A. A. Vicente, Food
Bengoechea, P. Partal, and A. Guerrero, J. Clean. Prod., Hydrocoll., 27, 220 (2012).
239, 117994 (2019).