International Journal of Biological Macromolecules 260 (2024) 129637

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International Journal of Biological Macromolecules

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

Extraction and characterization of cellulose nanoparticles from palm kernel

meal for potential application in active food packaging
Warakagoda Widanalage Don Rumali Chamathka a, Tsun-Thai Chai b, Eng Tong Phuah c,
Jun Xian Wong d, Sook Ngoh Chen e, Ali Yassoralipour a, *
a
Department of Agricultural and Food Science, Faculty of Science, Universiti Tunku Abdul Rahman (UTAR), Jalan Universiti, Bandar Barat, 31900 Kampar, Perak,
Malaysia
b
Department of Chemical Science, Faculty of Science, Universiti Tunku Abdul Rahman (UTAR), Jalan Universiti, Bandar Barat, 31900 Kampar, Perak, Malaysia
c
Food Science and Technology, School of Applied Sciences and Mathematics, Universiti Teknologi Brunei, Jalan Tungku Link, Mukim Gadong A BE1410, Brunei
Darussalam
d
Department of Food Science, Faculty of Food Science and Technology, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia
e
Department of Allied Health Science, Faculty of Science, Universiti Tunku Abdul Rahman (UTAR), Jalan Universiti, Bandar Barat, 31900 Kampar, Perak, Malaysia

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

Keywords: The research aimed to explore the potential of palm kernel meal (PKM) as a sustainable source of cellulose
Cellulose nanoparticles nanoparticles (CNPs) for active food packaging. The CNPs were isolated using a combination of chemical
Palm kernel meal techniques, such as alkaline treatment, bleaching, and acid hydrolysis. The characterization of the CNPs was
Chemical isolation methods
analysed using various techniques, including scanning electron microscopy, Fourier-transform infrared spec­
Morphological features
Active food packaging
troscopy, X-ray diffraction, and UV–visible spectroscopy. The findings revealed that chemical processing effec­
tively removed lignin and hemicellulose from PKM. The SEM morphology confirmed the separation of the CNPs,
resulting in the production of 40–100 nm spherical cellulose nanoparticles, while XRD and FTIR analyses
confirmed their purity and composition. Moreover, the UV–visible spectroscopy exhibited high transmittance
rates, indicating the potential of CNPs as reinforcing agents for polymer matrices. The significance of utilising
PKM as a valuable fibre source for extracting CNPs can be recommended for developing active food packaging.

1. Introduction
Oil palm (Elaeis guineensis), which comes from West Africa, is one of
the most important agricultural crops for Malaysia’s economic growth.
Malaysia is the world’s second-largest palm oil producer and the main
agricultural crop in Malaysia [1,30]. Because oil palm plantations are
taking over more and more land, there is a lot of waste left behind at oil
palm plantations. According to estimates, the oil palm industry produces
around 55 t of total dry matter (TDM) annually per hectare of land [18].
This biomass comes in the form of palm kernel shell (PKS), oil palm
fronds (OPF), oil palm empty fruit bunches (OPEFB), oil palm trunks
(OPT), oil palm leaves (OPL), and oil palm mesocarp fibres (OPMF)
[22,23,37].
However, the biomass residue left over after the palm oil extraction
from oil palm fruits is known as palm kernel meal (PKM), which consists
of 39.5 % of cellulose weight [8]. In most cases, it will be discarded as
waste in the palm oil mill. However, it is used to generate electricity by
acting as fuel for boilers to produce steam. These oil palm wastes have
caused environmental pollution by producing millions of tonnes of
waste every year and destroying the ecosystem around them by piling up
waste from open combustion on the ground [1,10,29].
In addition to palm oil, the palm oil sector produces around 169.72
million metric tonnes of solid waste after the extraction of palm oil,
accounting for 85.5 % of the total biomass waste generated in Malaysia
[50]. Palm kernel meal, which is the main by-product after the extrac­
tion of palm oil, which is a plentiful and reusable resource, is a great way
to make packaging that is also environmentally sound. However, this
agricultural waste material has not been used in many industries. Waste
valorisation turns palm kernel meal, which is a byproduct of the palm oil
industry, into cellulose nanoparticles that help create a circular econ­
omy by lowering waste and increasing resource efficiency [29]. The fact
that cellulose, which is the main ingredient in palm kernel meal, natu­
rally breaks down in nature is not harmful to the environment. In
addition to being environmentally friendly, the CNP in PKM in the food

* Corresponding author.
E-mail address: aliyas@utar.edu.my (A. Yassoralipour).

https://doi.org/10.1016/j.ijbiomac.2024.129637
Received 11 October 2023; Received in revised form 3 December 2023; Accepted 30 December 2023
Available online 21 January 2024
0141-8130/© 2024 Elsevier B.V. All rights reserved.
W.W.D.R. Chamathka et al.
packaging polymer matrix expectantly acts as a reinforcing material,
which keeps food fresh longer and increases its shelf life. PKM is a
readily available raw material that makes cellulose-based active food
packaging possible.
In response to the need for innovative materials derived from readily
available and recyclable resources, extensive research is being con­
ducted on agro-industrial waste, particularly cellulose-based waste,
which holds promise as a solution for environmental pollution [48].
These materials can be used to produce nanocomposite materials with
potential applications in various industries such as construction, elec­
tronics, the cosmetics industry, packaging, and even biomedicine
[22,47]. Since cellulose is a nanostructured natural polymer and has
unique properties, including low density, mechanical strength, and a
larger surface area, as well as the potential for structural and chemical
modification, non-toxicity, and biodegradability, it is the most suitable
among various organic materials for a variety of types of applications
involving nanomaterials. As an example, it has been demonstrated that
the incorporation of CNPs as reinforcing agents in composites can lead to
the development of materials that exhibit enhanced mechanical prop­
erties [13,16,17]. One innovative approach is to separate cellulose
nanoparticles (CNP) from renewable resources and use them in active
packaging as a replacement for synthetic fibres to provide better phys­
ical, mechanical, and barrier properties [38]. Therefore, composite
materials can be strengthened by incorporating nanocellulose as rein­
forcing agents, resulting in enhanced mechanical characteristics [28].
Even though PKM is a better source of cellulose, no research has been
done on extracting nanocellulose from palm kernel meal as its potential
application in packaging material.
Nanocellulose is defined as cellulosic particles with nanoscale
structural dimensions. In recent years, there has been a growth in in­
terest in the synthesis of nanocellulose due to its non-toxic and biode­
gradable characteristics, as well as its potential due to its impact on the
nanoscale [6,14]. Cellulose can be made through a number of chemical,
physical, physiochemical, and biological processes, such as high-
pressure homogenization, ultrasonication, and acid hydrolysis [12].
This study aimed to extract and isolate CNP from PKM by a chemical
process including alkaline treatment to remove alkali-soluble compo­
nents, in essence, lignin and hemicelluloses that dissolve in the solution,
bleaching treatment and acid hydrolysis process to remove the amor­
phous regions of the cellulose and release cellulose nanocrystals from
the cellulose substrate [7,9]. Then the characterization of CNP to see
whether it has promising characteristics for use as a major component of
the polymer matrix in developing active packaging film. Starting with
the raw fibres, making CNPs requires three main steps: early processing
of the fibres, separation of the CNPs, and then processing of the hy­
drolyzed celluloses. The processing of fibres includes bleaching and al­
kali treatments. In contrast, CNP isolation often involves acid hydrolysis.
In the final process, purification and sonication are done for the
hydrolysed cellulose [5]. Scanning electron microscopy (SEM), UV–vi­
sible spectroscopy, and Fourier transform infrared spectroscopy (FTIR)
assessed isolated nanoparticles’ morphological and structural proper­
ties. X-ray diffraction (XRD) analysis was used to describe the charac­
teristics of the cellulose nanoparticles isolated from palm kernel meal.
Also, thermogravimetric analysis (TGA) was used for determining heat
stability in cellulose nanoparticles.
2. Materials and methods
Cellulose nanoparticles (CNP) from palm kernel meal (PKM) using
alkaline, bleaching, and acid hydrolysis processes. The cellulose nano­
particles were characterized using optical transmittance, X-ray diffrac­
tion (XRD) analysis and scanning electron microscopy (SEM). The
crystallinity index (Crl) value was calculated from the XRD patterns, and
SEM was used to examine how different treatments affected the mate­
rials’ morphology. Fourier transform infrared (FTIR) spectroscopy to
study the changes in functional groups generated by CNP and various
2
International Journal of Biological Macromolecules 260 (2024) 129637
treatments.
2.1. Materials
Palm kernel meal (PKM) samples were collected from KLK Sdn. Bhd.
Palm Oil Mill and stored in a refrigerator until use. Sodium hydroxide
(NaOH), sodium chlorite (NaClO2), glacial acetic acid, sulphuric acid
(H2SO4), potassium bromide (KBr), and distilled water were of analyt­
ical or reagent grades and used without further purification.
2.2. Preparation of cellulose nanoparticles using palm kernel meal
PKM waste was dried at 50 ◦ C for around 12 h, ground into a fine
powder, and sieved through a 0.25 mm sieve. The CNP extraction was
performed with slight modifications to the method demonstrated in
[7,9].
2.2.1. Alkaline process
Ground and sieved PKM powder was subjected to an alkaline process
using 4 % NaOH (w/v) at 50 ◦ C for 180 min, with the procedure repeated
3–6 times to improve the crystalline properties. This process breaks
down the hemicellulose and depolymerizes the lignin, leading to the
breakdown of covalent bonds between the lignocellulose components.
The sample was washed and filtered as described by Chieng et al. [8].
2.2.2. Bleaching process
The alkali-treated fibres were subjected to around 4–6 rounds of
bleaching at 80 ◦ C for 4 h using a mixture of equal parts of aqueous
NaClO2 (1.7 %, w/v), distilled water, and acetate buffer (a mixture of
2.7 g of NaOH and 7.5 mL of glacial acetic acid per 100 mL of distilled
water). The process was repeated 6–8 times to obtain white-colour
cellulose, and the fibres were filtered, rinsed with distilled water, and
air-dried as described by Chieng et al. [8].
2.2.3. Acid hydrolysis process
The resulting cellulose fibres were subjected to acid hydrolysis using
52 % (w/w) of H2SO4 at 45 ◦ C for 45 min. It is to eliminate the cellulose’s
amorphous areas and liberate the nanocrystals from the cellulose sub­
strate. The hydrolysed cellulose sample was then diluted with cold
distilled water to stop the process and centrifuged three times at 10,000
rpm at 10 ◦ C for 10 min to remove any extra acid and water-soluble
pieces [32]. The acid hydrolysis was stopped by adding ice-cold
distilled water, and the cellulose was rinsed with distilled water 6–8
rounds to achieve a pH of 7. The nanocrystals were then dispersed using
an ultrasonic processor (sonicator – Cole Parmer CPX 130, United
States), with the resultant suspension freeze-dried and ground to elim­
inate any clumps before characterization, as described by Abral [2] with
some modifications.
2.3. Characterization of cellulose nanoparticles (CNP)
The characterization of CNP extracted using PKM was done by four
instruments, namely UV–visible spectroscopy, Scanning Electron Mi­
croscopy (SEM), X-ray diffraction (XRD), and Fourier-transform infrared
spectroscopy (FTIR).
2.3.1. Optical transmittance
A UV–visible spectrophotometer (Thermo Scientific, GENESYS 10S,
United States) was used to measure the transmittance rate of a con­
centration of 0.1 % nanocellulose suspensions at wavelengths ranging
from 200 to 800 nm.
2.3.2. X-ray diffraction (XRD) analysis
The crystallinity index of the CNCs was ascertained from the XRD
patterns. The samples were scanned using an X-ray diffractometer
(Shimadzu XRD 6000, Kyoto, Japan) at a rate of 2 per minute and an
W.W.D.R. Chamathka et al.
angle range of 10◦ to 40◦ . In order to measure the crystallinity of the
samples, the crystallinity index value was calculated. The formula for
the crystallinity index (CIr) is as follows:
(I002 − Iam)
CIr (%) = × 100
I002
where Iam is the peak intensity of the amorphous region and I002 is the
peak intensity corresponding to the crystalline region [44].
2.3.3. Scanning electron microscopy (SEM)
SEM was used to examine how different treatments affected the
materials’ morphology. Alkaline-treated PKM, bleached cellulose, and
CNP made from PKM were all observed using a JEOL scanning electron
microscope (JEOL JSM-6701F Tokyo, Japan) at a 4 kV accelerating
voltage. In order to ensure proper conductivity before analysis, the
samples were prepared by positioning them on a metal holder and
coating them with a platinum coating system at 10 mA for 55 s.
2.3.4. Fourier transform infrared (FTIR) spectroscopy
To study the changes in functional groups generated by raw PKM,
various treatments, such as alkaline, bleached, and cellulose nano­
particles of the FTIR spectra were recorded on a Fourier transform
infrared Spectrometer. Being an optically transparent material for light
in the infrared region of analysis, KBr was used as a carrier for the IR
spectrum sample. The 400–4000 cm− 1 range was used for the FTIR
spectrum analysis.
2.3.5. Thermogravimetric analysis (TGA)
The thermal stability of the materials was determined using ther­
mogravimetric analyses (TGA) performed on a Mettler Toledo (TGA/
DSC 3+) thermogravimetric analyser. Thermograms were taken from
25 ◦ C to 700 ◦ C at the heating rate of 10 ◦ C/min and a nitrogen flow rate
of 20 mL/min.
2.3.6. Development of the food packaging film
Polyvinyl alcohol (PVA) (6 % wt) was gradually added to 94 mL of
hot distilled water and continuously stirred at 90 ◦ C for 45–60 min until
the solution was completely dissolved. 6 % PVA solution (10 mL) and
mixed with 1 % of PKM CNP mixed in distilled water (10 mL). The so­
lution (PKM CNP) container was put in a water bath sonicator for 20 min
before being mixed with PVA. The same protocol was used for 2 % and 3
% PKM CNP solutions. The optimum ratio for the experiments was
selected as PVA 3 % wt. Then the solution was poured into plastic petri
dishes (9 cm) greased with coconut oil and evaporated at room tem­
perature for 48 h (Fig. 7).
3. Results and discussions
3.1. Extraction of cellulose nanoparticles (CNP) using palm kernel meal
(PKM)
Table 1 demonstrates the extraction of CNP from PKM was success­
fully extracted by using the chemical extraction method in the current
study and the percentage of the yields of cellulose nanoparticles ob­
tained from various sources of agricultural crop residues.
Table 1
Percentage of the yield of various sources of agricultural crop residues.
Cellulose source Percentage of the yield (%) Reference
Palm kernel meal 20.08 ± 0.96 % Current study
Coir 23.5 % [9]
banana rachis 28.6 % [9]
Kapok 33.7 % [9]
Sisal 38.8 % [9]
Giant reed 34.50 ± 0.34 % [45]
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International Journal of Biological Macromolecules 260 (2024) 129637
Fig. 1 illustrates the physical appearances of four types of PKM:
untreated, finely ground raw PKM, PKM treated with alkali, bleached
PKM, and PKM-CNP after acid hydrolysis. The treatments resulted in
changes in the colour of the fibres, as demonstrated in Fig. 1a. Finely
ground raw PKM (Fig. 1a) typically appears as a coarse powder with a
light brownish colour. The texture became even smoother after a longer
grinding process and sieving using a 0.25 mm sieve. The application of
alkali treatment had an impact on the colour, as alkaline-treated PKM
turned a darker colour than the original PKM during the treatment
(Fig. 1b) due to the removal of alkali-soluble components, in essence,
lignin and hemicelluloses that dissolve in the solution. Therefore, the
outcome appeared dark brown during the process and even when dried.
The bleaching treatment resulted in a relatively white appearance of the
fibres, as shown in Fig. 1c. After alkali treatment, the cellulose material
undergoes the bleaching process to further improve its purity and colour
[47]. Acidified NaClO3 delignification has been the most frequently used
method for removing lignin as an early step in cellulose extraction [45].
Bleaching agents break off phenolic compounds present in lignin and
residual pigments and remove the non-cellulosic matter [44]. Thus,
whitening the cellulose fibres and resulting in a whiter appearance. The
bleached cellulose of PKM may have a more refined texture and a white
or slightly off-white colour. After the acid hydrolysis, the obtained CNP
had a whitish colour, as shown in Fig. 1d. The resulting CNP extracted
from PKM appeared as a whitish, translucent material with a fine and
smooth texture.
3.2. Characterization of cellulose nanoparticles (CNP)
The characterization of CNP extracted using PKM was done by four
instruments, namely UV–visible spectroscopy, Scanning Electron Mi­
croscopy (SEM), X-ray diffraction (XRD), and Fourier-transform infrared
spectroscopy (FTIR).
3.2.1. UV visible spectroscopy analysis
Fig. 2 illustrates the optical transmittance of CNP amphiphilic sus­
pension of PKM in the light range of 200–800 nm.
The CNP was the result of the removal of lignin, and hemicellulose
during the alkaline, bleaching and acid hydrolysis process and the break
of amorphous regions during the ultrasonication process. A CNP sus­
pension (0.1 %) was used for the analysis process. Findings of the CNP
shows a very high (> 90 %) transmittance rate at a visible light range of
400–600 nm (Fig. 2). The results revealed that the cellulose fibres
containing a higher transmission of light are effectively degraded into
uniformly forming CNPs, with only a trace amount of remaining
aggregated cellulose nanoparticles. Transmittance can be measured
directly using a spectrophotometer and the process involves trans­
mitting light through. It involves passing light through the sample and
quantifying the amount of light that passes through, providing infor­
mation about the transparency and transmittance of the CNP and the
dispersion of the particles without agglomeration. It is crucial to observe
before incorporating nanocellulose into the polymer matrix before the
development of the packaging film. Studies showed the use of CNPs as
the optimum calibration standard for UV–visible spectrometry and
found that a concentration of 0.1 % was ideal for measuring the
absorbance of light at specific wavelengths [34,39].
The studies found that using a higher concentration of CNP led to
increased light scattering and reduced the accuracy of the absorbance
measurements. Moreover, the study done by Sirviö et al. [35] revealed
that the nanocelluloses were utilised as lightweight fillers to enhance the
strength of a polyvinyl alcohol (PVA) matrix. These fillers hold excellent
UV-absorption properties when combined with the PVA material [35].
The option to utilise a low concentration, specifically 0.1 w/w% of
cellulose nanoparticles (CNP), in UV–vis spectrometer examination to
assess optical qualities prior to their incorporation into the packaging
matrix. One reason can be the sensitivity of a UV–vis spectrometer,
which is frequently employed for the analysis of light absorption and
W.W.D.R. Chamathka et al. International Journal of Biological Macromolecules 260 (2024) 129637

Fig. 1. Progressive changes in the physical appearances of palm kernel meal (PKM) following different treatments. (a) finely ground raw PKM, (b) alkali-treated
cellulose of PKM, (c) bleached cellulose of PKM, (d) cellulose nanoparticles (CNP) extracted using PKM.

Fig. 2. Optical transmittance of PKM CNP suspensions at 0.1 %, 1 % and 2 %.

transmission in various materials. When the concentration of a sub­
stance increases, the transmittance rate decreases, as shown in Fig. 2.
Therefore, the use of a reduced concentration of CNP (0.1 w/w%) en­
ables the uniform and homogeneous nature of the spectrometer to be
precise and sensitive in observing the optical properties of the nano­
material [35].
Moreover, prior to conducting UV–vis analysis, it is important to
confirm that the sample exhibits homogeneity and proper dispersion
[39]. Elevated levels of CNP might lead to greater aggregation or
condensation. Therefore, there are difficulties in achieving sample ho­
mogeneity. The utilisation of a reduced concentration facilitates the
homogeneous dispersion of CNP within the solvent or matrix. It enables
more accurate and reliable measurements that are representative of the
system.

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W.W.D.R. Chamathka et al. International Journal of Biological Macromolecules 260 (2024) 129637

Similar results were found by Sirviö et al. [34] and Visanko et al. [39]
on amphiphilic cellulose nanocrystals (CNCs) suspensions (0.1 %) at
visible light wavelengths ranging from 350 to 800 nm and the presence (a)
of nano-sized particles were further confirmed by optical transmittance.
The highest transmittance in the visible light range of 400–700 nm may
be attributed to the degradation of cellulose fibre [34,39].
Optical transmittance is crucial for cellulose nanoparticles used in
polymer matrix composites for transparent applications like packaging
films. CNP’s size, shape, lightweight, surface chemistry, concentration,
and compatibility with the polymer matrix affect their transmittance.
Well-dispersed CNPs with high aspect ratios can reinforce the polymer
and enhance its mechanical properties without impacting transparency
[4,24,27]. A composite film made with nanocellulose fibrils with 80 %
light transmittance produced from maize husks was reported to be
highly visible and have increased tensile strength when nanocellulose
was incorporated in the polymer matrix [40]. Similarly, a study by
Silvério et al. [42] found that the optical transmittance of CNC extracted
from corncob-reinforced polyvinyl alcohol (PVA) films decreased with
increasing CNC content and showed a high transmittance percentage
even at high CNC loadings. Optical transmittance measurements have
confirmed the presence of these components in the films, and their (b)
(b)
distinctive optical characteristics are dependent on their size, shape,
concentration, aggregation state, and refractive index [42].

3.2.2. Scanning electron microscopy (SEM) analysis

Scanning electron microscopy (SEM) was used to examine the sur­
face morphological features, composition, and orientation of the PKM
CNPs and fibres treated with various treatments. The different ways the
fibre is treated change its shape and expose its surface by removing non-
cellulosic macromolecular components like hemicelluloses, lignin,
pectin, oils, and other impurities that cause fibrillation [8].
Fig. 3 shows the outcomes of the morphology study of CNPs in palm
kernel meal. The formation of cellulose nanoparticles is a result of acid
hydrolysis’s removal of lignin and hemicellulose. During the acid hy­
drolysis process, hydronium ions (H3O+) target the available amor­
phous regions of cellulose and facilitate the hydrolytic breaking of
glycosidic linkages [45]. Then, cellulose particles go through the soni­
cation process to separate the coagulated cellulose particles. The for­ Fig. 3. SEM images of Cellulose Nanoparticles of PKM under the magnifications
mation of cellulose nanoparticles with a spherical shape leads to the of (a) X 5000 and (b) X 50,000.
elimination of lignin and hemicellulose during acid hydrolysis. Thus, the
CNP in Fig. 3 illustrates the topographical images of the palm kernel diffractometer. Fig. 4 shows the CNP produced from PKM (data range
meal-derived cellulose nanoparticles. The particle analysis covered a between 10◦ and 40◦ ). The elasticity, thermal stability, absorptive ca­
scale range of 48.1–108.7 nm. Most of the particles in the covered area pacity, and other physical characteristics of the fibre that are significant
were homogeneous and spherical. It was feasible to determine the for industrial uses have been determined using cellulose crystallinity. An
morphology and elementary size of the particles of PKM CNP using SEM
analysis.
The morphology of the cellulose nanoparticles obtained from PKM
was more similar to cotton linters’ morphological features. It was
revealed in the study by Ram and Chauhan [31] that the spherical cel­
lulose nanocrystals obtained from cotton linter powder were 45–75 nm,
which is smaller than the size of the CNP obtained in the current study.
A study by Kargarzadeh et al. [21] showed that spherical CNPs
significantly enhanced the mechanical and barrier characteristics of the
film, including greater tensile strength and reduced water vapour
permeability in active packaging films for food applications.
Similarly, a study by Zaini et al. [14] observed the various aspect
ratio-induced nucleation/reinforcing and barrier effects of
nanocellulose-incorporated food packaging revealed that the CNP ob­
tained from lyocell fibres were under the size of 30 nm. It suggests that
the selection of the starting raw material also plays a role in particle size.
The type of fibre can influence the morphological formation of the iso­
lated nanocellulose after the sonication process [14].

3.2.3. X-ray diffraction (XRD) analysis

The X-ray diffraction (XRD) pattern was used to assess the crystalline
index and purity of the sample of the PKM CNP using an X-ray Fig. 4. X-ray diffraction (XRD) of CNP extracted using PKM.

5
W.W.D.R. Chamathka et al.
increase in crystallinity is anticipated to improve tensile strength and
rigidity, providing stronger resistance to breaks. Thus, it enables the
development of nanocomposites with better mechanical and physical
properties [19].
The nanocellulose samples present four diffraction peaks at 2θ = 15◦ ,
16.7◦ , 20.6◦ , 22.4◦ , and 34.5◦ , corresponding to (1− 10), (1–10), (110),
(200), and (004), respectively. These prominent diffraction peaks in the
pattern of palm kernel meal CNP correspond to the typical crystalline
cellulose structure. Those peaks represent the crystallographic planes of
the monoclinic cellulose I (Iβ lattice) where it is more stable [6,11,44].
The CNC extracted from PKM had their crystalline index determined
using the XRD patterns. The proportion of cellulose’s crystalline to
amorphous regions is known as the crystallinity index (CrI), and the CrI
% of CNP extracted using PKM was 78.8 %. A study of the character­
ization of cellulose nanocrystals from oil palm mesocarp fibre done by
Chieng et al. [8] obtained 77.80 % of the crystallinity index in cellulose
nanocrystals isolated by oil palm mesocarp, which was lower than the
current study. Compared to the current study, relatively lower crystal­
linity indices for iso-, n-, and tert-butylamino-functionalized cellulose
nanocrystals were determined as 52.7 %, 56.4 %, and 57.0 %, respec­
tively, in the study of Visanko et al. [39]. The physical characterization,
including the crystallinity index, exhibited the incorporation of CNP in
the PVA matrix to develop an active packaging film that contains sig­
nificant mechanical (tensile strength and elongation) properties, ther­
mal stability, and a lower oxygen transfer rate [32].
A higher crystallinity index of Crl% is due to the lignin and hemi­
celluloses being removed from the cellulose fibres during the repeated
bleaching process. During the acid hydrolysis process, the sulphuric acid
permeates the amorphous region of the cellulose during the acid hy­
drolysis process. It enables the hydrolytic fragmentation of glycosidic
bonds and subsequently releases individual nanostructures [15,20].
The crystallinity index (Crl) is the ratio of crystalline cellulose to
amorphous cellulose. The crystallinity of cellulose has been used to test
Fig. 5. Fourier transform infrared (FTIR) spectra of Alkaline treated raw PKM, bleached PKM and cellulose nanoparticles of PKM.
6
International Journal of Biological Macromolecules 260 (2024) 129637
the fibre’s elasticity, thermal stability, and ability to absorb water,
which indicates important physical properties for industrial uses. An
increase in crystallinity is expected to make the material stronger and
more rigid. Therefore, it shows that it might be possible to make
nanocomposites with better mechanical properties using these nano­
structures [8,15].
An observed reduction in the CrI of CNP indicates that the extraction
duration of 45 min was excessively harsh given the hydrolysis conditions
applied [44,45]. Consequently, the acid treatment of 45 min not only
eliminated the amorphous component of cellulose but also partially
disrupted its crystalline nature [44]. Previous studies have also reported
a comparable outcome resulting from an extended hydrolysis duration
[44,45].
3.2.4. Fourier transforms infrared spectroscopy (FTIR)
FTIR analysis is an effective technique for comprehending alterations
in structure, chemical composition, physical state, and even minor
changes in the structure of cellulose during its different treatment stages.
The study of the functional groups presented in PKM and an analysis of
the changes caused by various treatments was achieved using the FTIR
analysis. Fig. 5 illustrates the spectra of alkaline-treated raw PKM,
bleached PKM, and acid hydrolysed PKM (CNP). The peaks at 3421
cm− 1 in all of the spectra correspond to O–H stretching and reflect the
tendency to be hydrophilic [22]. When treated with an alkaline sub­
stance, the intensity of the peak increases due to the concentration of OH
ions increase. This occurred due to the alkaline substance reducing the
hydrogen bonding within the hydroxyl groups and non-cellulose matter
[36,48]. The decrease of the peaks at 2896 cm− 1, which shown in the
spectrums correlates to C–H stretching might also be attributed to this
reaction. This indicates the disappearance of the typical peaks of lignin
and hemicellulose in PKM after the alkaline and bleaching processes
[46]. A peak intensity at 1732 cm− 1 that is unique to the spectra of
OPMF is caused by the C– –O stretching of carbonyl functional groups
W.W.D.R. Chamathka et al.
from the hemicellulose and lignin components [43].
After alkaline treatment, the C––O stretching is mitigated as a result
of the elimination of carboxylic groups, which may be remaining fatty
acid residues from the fibre surface. The peak at 1234 cm− 1 in the OPMF
spectrum, which corresponds to the syringyl ring and C–O stretching of
lignin and xylan, exhibits a reduction in intensity before diminishing.
Mazlita et al. [26] explained this occurred due to the lignin and a small
amount of hemicellulose have been removed from the PKM during the
chemical treatments.
The CNP spectrum’s peak at 1163 cm− 1 indicated the existence of a
sulphated group, possibly as a result of the cellulose’s sulfonation during
the process of acid hydrolysis. The CNP spectrum has 1423 cm− 1, 1324
cm− 1, 1045 cm− 1, and 896 cm− 1 peaks, which correspond to typical
cellulose absorption peaks. Each peak can be assorted as CH2 bending,
CH2 rocking, C–O stretching, or C–H or CH2 bending, respectively [8].
Furthermore, the peak at 896 cm− 1 can also be attributed to the typical
structure of cellulose due to the glycosidic connections of the glucose
ring of the cellulose chain [22]. The existence of glycosidic bonds serves
as proof of the formation of polysaccharides through the bonding of the
Fig. 6. (a) Thermal Gravimetric analysis (TGA) and (b) Derivative Thermogravimetric analyses (DTG) curves of Bleached PKM cellulose and PKM CNP.
7
International Journal of Biological Macromolecules 260 (2024) 129637
anomeric carbon atom of saccharides, indicating the presence of a cel­
lulose structure [8].
The removal of hemicellulose and lignin from the cellulose during
the extraction process leads to a reduction in peaks in the FTIR spectra of
bleached cellulose and acid-hydrolysed nanocellulose particles [3].
Furthermore, FTIR analysis is vital to examine the qualitative changes in
crystallinity for both raw materials before and after the synthesis pro­
cedure. The FTIR analysis used to assess changes in the crystalline
structure of cellulose using light wavelengths, which can be a useful tool
in evaluating the effects of certain processes, such as dissolving and
reforming, on cellulose [33]. Thus, FTIR can provide valuable infor­
mation about the chemical composition and structure of nanocellulose-
based active packaging films and can help researchers optimize the
properties of these materials for various applications.
3.2.5. Thermal gravimetric analysis (TGA)
The analysis of the thermal stability of fibres is a crucial factor in
assuring the enhancement of the characteristics of composites shown in
Fig. 6. The thermogravimetric analysis (TGA) graph of cellulose
W.W.D.R. Chamathka et al.
Fig. 7. Demonstrates the development of the packaging film and its physical appearance when incorporating CNP extracted using PKM along with PVA. (a) PVA film,
(b) PVA film containing 1 % CNP, (c) PVA film containing 2 % CNP and (d) PVA film containing 3 % CNP.
nanoparticles generally exhibits the reduction in mass of the nano­
particles in relation to changes in temperature.
TGA was carried out to assess the compatibility of the samples for
bio-composite processing. The thermal decomposition parameters were
obtained using the TGA curves, as shown in Fig. 6. The initial observed
decrease in weight, occurring at temperatures around 90 ◦ C - 100 ◦ C was
ascribed to the process of evaporation of water in the CNP sample
[45,48]. The weight loss was observed at around 4 %.
Onset Temperature (T5%) which is the temperature at which the
sample begins to exhibit a significant weight loss at 207.5 ◦ C and
261.18 ◦ C in the bleached cellulose sample and CNP respectively. The
T10% signifies the temperature at which a substantial portion (10 %) of
the bleached cellulose sample at 240 ◦ C and CNP at 302.67 ◦ C sample’s
weight has been lost. The next stage encompassing the temperature
range of 180–300 ◦ C is associated with the breakdown of hemicellulose.
The third stage, occurring within the temperature range of 300–400 ◦ C,
is attributed to the disintegration of cellulose. On the other hand, the
fourth stage is associated with the degradation of lignin, which takes
place gradually and is accompanied by a significant deterioration from
400 ◦ C to the end temperature (800 ◦ C) [48].
The distinct processes are documented in both Bleached and CNP
thermograms. On the other hand, the TGA curve of bleached and CNP
progressed to higher temperatures and the weight loss percentages of
hemicellulose and lignin dropped, suggesting a decrease in their
respective quantities. Those observations show the higher thermal sta­
bility is largely attributed to the chemical treatment when hemicellulose
and lignin are eliminated [48,49]. A smaller residue is also reported for
CNP, which verifies the elimination of non-cellulosic materials. The
residue yield percentage of bleached cellulose obtained was 42.3 % and
8
International Journal of Biological Macromolecules 260 (2024) 129637
CNP was 8 %. It is noteworthy to mention that CNP demonstrates a
steady behaviour at 160 ◦ C -180 ◦ C, which corresponds to the composite
curing temperature.
4. Conclusion
The comprehensive chemical methods employed in this study,
namely alkaline treatment, bleaching, and acid hydrolysis, have suc­
cessfully enabled the isolation of cellulose nanoparticles from PKM. The
morphological analysis conducted using SEM has provided definitive
evidence regarding the precise size of the extracted nanostructures,
further confirming the successful extraction of cellulose nanoparticles.
Moreover, the higher transmittance observed in UV–visible spectros­
copy indicates efficient disintegration of cellulose fibres into uniformly
distributed CNPs. The XRD analysis has shed light on the purity, crys­
tallinity index, and structural parameters of the extracted CNPs, while
the FTIR analysis has revealed valuable insights into the material’s
composition and structural modifications. Collectively, the findings of
this study suggest that PKM can serve as a preferable fibre source in the
production of CNPs. Furthermore, the crystallinity index, morphological
features, surface topography, and particle size of the CNPs derived from
PKM, as elucidated in this research, highlight their potential for uti­
lisation in the polymer matrix of active packaging films to enhance the
film’s properties. These findings contribute to the expanding knowledge
base in the field of cellulose nanoparticle extraction and present prom­
ising avenues for further research and practical applications in active
packaging materials.
W.W.D.R. Chamathka et al.
CRediT authorship contribution statement
Warakagoda Widanalage Don Rumali Chamathka: Conceptuali­
zation, Data curation, Methodology, Project administration, Visualiza­
tion, Writing – original draft. Tsun-Thai Chai: Resources, Writing –
review & editing. Eng Tong Phuah: Resources, Writing – review &
editing. Jun Xian Wong: Data curation, Methodology, Visualization,
Writing – review & editing. Sook Ngoh Chen: Data curation, Method­
ology, Visualization, Writing – review & editing. Ali Yassoralipour:
Conceptualization, Data curation, Methodology, Project administration,
Resources, Supervision, Writing – review & editing.
Declaration of competing interest
All authors have participated in (a) conception and design, or anal­
ysis and interpretation of the data; (b) drafting the article or revising it
critically for important intellectual content; and (c) approval of the final
version.
This manuscript has not been submitted to, nor is under review at,
another journal or other publishing venue.
The authors have no affiliation with any organization with a direct or
indirect financial interest in the subject matter discussed in the
manuscript
Data availability
Data will be made available on request.
Acknowledgement
This study was supported by the research funding under Universiti
Tunku Abdul Rahman Research Funding (Project Number: IPSR/RMC/
UTARRF/2021-C1/A01) at the Faculty of Science, Universiti Tunku
Abdul Rahman, Kampar, Perak, Malaysia.
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