Journal of Cleaner Production 305 (2021) 127229

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Journal of Cleaner Production

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

Hydrothermal assisted isolation of microcrystalline cellulose from

pepper (Piper nigrum L.) processing waste for making sustainable bio-
composite
Holilah Holilah a, Didik Prasetyoko a, *, Ratna Ediati a, Hasliza Bahruji b,
Aishah Abdul Jalil c, d, Asranudin Asranudin a, e, Susanti Dhini Anggraini f
a
Department of Chemistry, Faculty of Science and Data Analytics, Institut Teknologi Sepuluh Nopember, Sukolilo, Surabaya, Indonesia
b
Centre of Advanced Material and Energy Sciences, Universiti Brunei Darussalam, Jalan Tungku Link, BE, 1410, Brunei
c
Department of Chemical Engineering, Faculty of Chemical and Energy Engineering, Universiti Teknologi Malaysia, 81310, UTM, Skudai, Johor Bahru, Johor,
Malaysia
d
Centre of Hydrogen Energy, Institute of Future Energy, Universiti Teknologi Malaysia, 81310, UTM, Skudai, Johor Bahru, Johor, Malaysia
e
Department of Food Science and Technology, Halu Oleo University, Kendari, Indonesia
f
Department of Industrial Engineering, Universitas PGRI Ronggolawe, Tuban, Indonesia

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

Article history: Conversion of industrial pepper bio-waste (PW) to microcrystalline cellulose (MCC) was investigated as
Received 10 May 2020 reinforcement agent of natural polymer bio-composites. The investigation highlighted the importance of
Received in revised form hydrothermal assisted alkalization step for efficient separation of cellulose from lignin in PW in order to
18 March 2021
enhance the yield and the crystallinity of MCC. Cellulose was isolated from industrial pepper bio-waste
Accepted 18 April 2021
Available online 22 April 2021
via three steps i.e. alkalization, bleaching and hydrothermal assisted re-alkalization. Hydrothermal
treatment during re-alkalization step increased the release of lignin that was bound within the cellulose
Handling Editor: Prof. Jiri Jaromir Kleme
s domain via dissociation of the multiple bonds in the three-dimensional polymer and significantly
reduced the lignin concentration. Optimization of hydrolysis parameters i.e. the concentration of HCl, the
Keywords: hydrolysis temperature and time, and also the solid/liquid ratios produced 86% of MCC with high crys-
MCC tallinity index ~78.9%. The physicochemical properties of MCC isolated from PW exhibited a highly
Cellulose crystalline microcellulose structure with high thermal stability that were comparable to the physico-
Waste chemical properties of the commercial MCC. Incorporation of MCC with seaweed and chitosan natural
Pepper
polymer showed the enhancement of the tensile strength and the elongation of the bio-composites,
Bio-composites
which exhibited the potential use of MCC as reinforcement agent in bio-composites to replace tradi-
tional plastic.
© 2021 Elsevier Ltd. All rights reserved.

1. Introduction discarded as biomass waste (Direktorat Jenderal Perkebunan,

Indonesia is the second largest producer of pepper (Piper nigrum
L) in the world after Vietnam, contributes to 13% of the total global
market. Indonesia has engaged 187.545 Ha of dedicated land to
produce 90,000 tons of pepper annually (Direktorat Jenderal
Perkebunan, 2019). Pepper was processed via modern and con-
ventional methods that generally involved utilization of thresher
machine to separate the water soaked peppercorn from the peel. In
the processing of pepper, about 80% of the peel and the stalk were
* Corresponding author.
E-mail address: didikp@chem.its.ac.id (D. Prasetyoko).
2019). Since pepper is a cultivated plantation crop that can be
harvested all year round, the abundant amount of waste requires a
proper management. Disposal of the accumulated agricultural
pepper waste was carried out by open-burning causes devastating
impact to the environment. Pepper stalk contained a rich ligno-
cellulosic compound that can be used as precursor for the synthesis
of microcrystalline cellulose (MCC). MCC is defined as a type of
cellulose that undergone partial hydrolysis to form crystallites with
diameter within 1e100 mm, and the percentage of crystallinity at
55%e85% (Brinchi et al., 2013). The degree of polymerization (level-
off DP), the molecular weight and the cellulose chain length of MCC
were controlled by the duration of hydrolysis in hot acidic solution

https://doi.org/10.1016/j.jclepro.2021.127229
0959-6526/© 2021 Elsevier Ltd. All rights reserved.
H. Holilah, D. Prasetyoko, R. Ediati et al.
(Nsor-Atindana et al., 2017; Frank Ohwoavworhua et al., 2009;
Trache et al., 2016). Hydrolysis also removed amorphous compo-
sition in cellulose for the formation of high crystalline structures
(Karim et al., 2014).
MCC is a hygroscopic, non-toxic and chemically inactive mate-
rial with high surface area and high adsorption capacity that is
suitable for application in various industries. For example, MCC was
used as binder in drug delivery system (Kalita et al., 2013; Kundu
and Banerjee, 2020), advanced materials in food packaging (Merci
et al., 2019; Norain et al., 2019), and stabilizer in cosmetic in-
dustries (Ahsan et al., 2020; Buffiere et al., 2017). MCC was incor-
porated into natural polymer to enhance thermal, morphological
and mechanical properties of bio-composites (Bhatti et al., 2020;
Collazo-Bigliardi et al., 2018; Mukherjee and Kao, 2011; Norain
et al., 2019). MCC generally has high density of surface hydroxyl
group that formed hydrogen bonding with polymer matrix for the
production of bio-composites (Do Nascimento et al., 2015; Tian
et al., 2019). Boldizar et al. (1987) reported that MCC showed po-
tential as filler for polymeric material that enhanced the mechan-
ical behavior of natural polymer due to the high crystalline nature
and the short microfibrils of MCC. Since then MCC has been widely
developed as reinforcement agent in the manufacturing of bio-
composite materials (Alashwal et al., 2020; Mohan Bhasney et al.,
2020; Zhang et al., 2020).
In general, isolation of MCC from agricultural waste was carried
out via two steps treatment involving alkalization for removal of
lignin, and hydrolysis for generation of microcellulose (Collazo-
Bigliardi et al., 2018; Pachuau et al., 2019; Trache et al., 2014;
Xiang et al., 2016). Lignin is an amorphous polymer with three
dimensional structure based on phenyl propane molecule
(Abhilash and Thomas, 2017). Removal of lignin is important for the
generation of microcrystalline cellulose since lignin slowed down
the hydrolysis process (Yoon et al., 2014). Delignification reduced
the size of lignin macromolecules into small fragments that can be
easily dissolved (Chen and Wang, 2017). Delignification was often
carried out in alkaline condition such as digestion using sulfate ion
HS
and S2
, or condensation at high temperature using NaOH
(Fearon et al., 2020). High concentration of NaOH was required to
ensure efficient removal of lignin. However when NaOH was
insufficient, delignification caused further condensation of frac-
tured lignin to form larger lignin molecules (Chen and Wang, 2017).
Commercial process for removal of lignin required high pressure
oxygen condition to generate active oxygen radical for lignin
degradation. However, active oxygen radical also caused degrada-
tion of cellulose (Huang and Fu, 2013). Hydrothermal method
provided eco-friendly high pressured system that proven efficient
even without the presence of catalysts (Chen et al., 2016). Under
hydrothermal method, water was also deionized to increase the
concentration of OH
that contributed to disintegration of lignin at
minimum energy consumption (Peterson et al., 2008; Toor et al.,
2011). Delignification with NaOH under hydrothermal condition
reduced the hemicellulose and lignin content due to disintegration
of middle lamella layers of poplar wood (Shi et al., 2018). High
pressurize system in hydrothermal condition increased the surface
area of biomass, facilitated the breaking down of the cell wall
(Kro€ger et al., 2018; Tanpichai et al., 2019). Hydrothermal deligni-
fication also increased the dissociation of phenolic compound in
lignin (Arauzo et al., 2020).
Wide application of MCC in food processing, biomedical and
pharmaceutical industries has prompted research in MCC produc-
tion particularly from sustainable and natural resources. However,
to date no utilization of the industrial bio-waste from pepper in-
dustry has been reported for the formation of MCC. This study fo-
cuses on the benefit of hydrothermal assisted re-alkalization
treatment on the pepper stalk in order to efficiently remove lignin
2
Journal of Cleaner Production 305 (2021) 127229
from cellulose that subsequently enhanced the cellulose composi-
tion. The microcrystalline cellulose derived from industrial pepper
bio-waste was further investigated as potential reinforcement for
bio-composites production. To elucidate the effect of different
stages of treatment on industrial pepper bio-waste (PW), the
morphology, the structural stability and the chemical properties of
MCC were determined and finally the mechanical properties of
MCC/natural polymer bio-composites were investigated using
seaweed and chitosan with polyethylene glycol (PEG) and glycerol
as plasticizer.
2. Materials and method
Bio-waste from pepper industry was obtained from CV. Hanum
Shafira Kendari, Indonesia. The harvested pepper was soaked in
water for several days to soften the cell walls of pepper. Thresher
machine was used to separate the water-soaked peppercorn from
the stalk. The as-received stalk was treated as described in section
2.1. The following analytically graded materials were used, i.e. PEG
400, sodium hydroxide (NaOH, 99%), glycerol 85%, hydrochloric
acid (HCl, 37%) were purchased from Merck, Germany. Hydrogen
peroxide (H2O2, 30% (w/w) in H2O), chitosan (medium MW, 75e85
deacetylated) were purchased from Sigma-Aldrich, Germany and
Avicel® PH 101 (50 mm). Avicel PH 101 (a trade name of MCC) was
used as standard microcrystalline cellulose to provide comparison
on the physicochemical properties of the isolated MCC.
2.1. Preparation of industrial pepper bio-waste
Industrial pepper waste as raw material for the production of
cellulose was obtained in the form of pepper stalk. 500 g of the stalk
was immersed in deionized water at 80  C for 4 h under stirring
followed by filtration and dried at 100  C for 24 h. The stalk was
then cut into small pieces and milled to form powder labeled as PW.
Particle size analysis of PW was measured using CILAS 1190 Particle
Size Analyzer (Fig. S1 supplementary) indicated the average
diameter of PW was approximately ~143.11 mm.
2.2. Cellulose isolation from PW
Cellulose was isolated using the modified method reported by
Collazo-Bigliardi et al. (2018); Hussin et al. (2016); Prado and
Spinace  (2019). The first alkalization step involved the addition of
5% NaOH solution to the PW powder and stirred for 2 h at 90  C. The
ratio of PW sample to sodium hydroxide was kept at 1:20 (w/v). The
mixture was filtered and thoroughly washed with deionized water
to reach pH 7. The second step referred as bleaching process in
which the residual hemicelluloses and lignin were removed by
adding 1 g of the alkali treated PW powder with 40 mL of 3% (v/v)
H2O2 and 5% NaOH at solid to liquid ratio of 1:40 (w/v). The mixture
was stirred at 55  C for 90 min, cooled at 27e30  C and then
vacuum-filtered. The bleached PW was washed with deionized
water until the pH reached 6 and labeled as B-PW. The third step
was hydrothermal assisted re-alkalization, to further remove lignin
that still presence in the resulted B-PW. The re-alkalization process
was conducted using the same method with the alkalization by
modifying the conventional thermal process into hydrothermal.
The B-PW sample was mixed with 5% NaOH solution (1:20, w/v)
and the mixture was place in autoclave and heated at 90  C, 2.2 bar
for 1 h. The powder was filtered and washed with deionized water
reach pH 7, and dried at 60  C. The dried powder was denoted as H-
PW.
H. Holilah, D. Prasetyoko, R. Ediati et al.
2.3. Preparation of microcrystalline cellulose (MCC)
Cellulose that was isolated from the bleached pepper bio-waste,
B-PW and hydrothermal treated bio-waste H-PW were hydrolyzed
using hydrochloric acid to produce MCC following modification of
methods described by Hussin et al. (2016) and Zhao et al. (2018).
The B-PW and H-PW respectively were added into HCl solution
(2.5e4.5 N) at a solid/liquid ratio of 1:20, w/v. The mixture was
stirred for 60e180 min at 50e80  C. The resulted solid was filtered
and washed with distilled water until the supernatant becoming
neutral. In addition, 10-mesh sieve were used to separate the solid
residues to confirm that there were no remaining unreacted cel-
lulose in the microcrystalline cellulose powder. Lastly, the powder
was dried at 60  C for 24 h and stored for further analysis. MCC
obtained from B-PW was labeled as B-MCC, meanwhile the MCC
from H-PW was labeled as PW-MCC. Equation (1) was used to
calculate the yield of MCC.
Yield of MCCð%Þ ¼ W1=W  100 (1)
2
where W1 and W2 were the weights of MCC and PW cellulose.
2.4. Characterization
2.4.1. Composition of lignocellulosic bio-waste
Determination of lignocellulosic composition was conducted
using neutral detergent fiber (NDF), acid detergent fiber (ADF), and
acid detergent lignin (ADL) methods (Mohammed et al., 2012). The
samples were refluxed in boiled neutral detergent for 1 h to mea-
sure their NDF content. After refluxing, the solution was cooled at
29  C and was filtered. The residue from the filtration process was
washed using acetone and dried in the oven at 105  C for 6 h. The
determination of ADF content was carried out using a similar
method as NDF content determination, but acid detergent was used
instead of neutral detergent. Meanwhile, the determination of ADL
content was carried out by hydrolysis the remaining fiber from ADF
analysis with H2SO4 (72%, w/v). The determination of NDF, ADF and
ADL (%) were based on the weight differences between the pre- and
the post-treatment. The fibers were washed, dried, and burned in a
furnace at 500  C. The cellulose, hemicellulose, and lignin contents
were calculated using the following equations:
Cellulose ð%Þ ¼ ADF
ADL (2)
Hemicellulose ð%Þ ¼ NDF
ADF (3)
Lignin ð%Þ ¼ ADL (4)
2.4.2. Physicochemical properties of MCC
The moisture content was evaluated using method reported by
Jumaidin et al. (2017). 1 g of MCC was dried at 105  C with the
results were measured based on the average of three different
measurements.
.
MCð%Þ ¼ ½ðMi
Mf Þ Mi   100 (5)
where MC means the moisture content, while Mi and Mf denote the
weights before and after treatment with drying.
The ash content of the MCC (PW-MCC and C-MCC) was analyzed
by weighing the residual powder following combustion in air at
550  C to eliminate the carbon (Ohwoavworhua et al., 2009).
Pycnometer was used to measure the density of MCC with the
3
Journal of Cleaner Production 305 (2021) 127229
mass was measured with up to ±0.0001 g of accuracy, and the
volume was obtained through the displacement of Xylene. This
process was performed in triplicate to obtain the average value of
density.
The pH and the swelling index were determined by dispersion
of 5 g of MCC samples in 50 mL of deionized water. This mixture
was then centrifuged for 20 min and followed by the measurement
of pH of the supernatant. The swelling index was determined using
the initial volume (V1) and the final sedimentation volume (V2),
based on the following equation:
Swelling index ¼ V2=V (6)
1
The method reported by Fuentes-Alventosa et al. (2009) was
used to determine the water holding capacity (WHC) of MCC. The
procedure involved the homogenization of 5 g cellulose with 50 mL
of water at 25  C for 1 h, followed by centrifugation at 3000 rpm for
15 min. The supernatant was then discarded and the hydrated
cellulose accumulated and weighed. Similarly, the procedure was
also employed for the determination of oil holding capacity (OHC)
using soybean oil (density: 0.917 g/mL) and the water holding ca-
pacity and oil holding capacity were estimated using the formula
below:

g
WHC; OHCð Þ ¼ W1
Wo Wo (7)
g
where Wo and W1 denoted the weights of the dry and the swollen
samples.
2.4.3. FTIR spectroscopy
Under solid state conditions, the IR spectra were examined from
4000 to 650 cm
1 by FTIR spectrophotometer (Bruker IFS 113v)
with KBr/pellet ratio (99/1, w/w).
2.4.4. X-ray diffraction
The crystallinity of cellulose and MCC of PW was determined
using X-ray diffractometer (XRD, XDR JEOL JDX-3530) operated at
40 kV and 30 mA, with Cu Ka radiation (l ¼ 1.541 Å). The data was
recorded at 2q from 5 to 40 . The crystallinity index (CI) was
evaluated using equation (8).
CIð%Þ ¼ ½ðI002
Iam Þ = I002   100 (8)
where the maximum intensity of crystalline from the (002)
diffraction denoted as I002 (2q ¼ 22.6 ) and Iam served as the
background scatter intensity for measuring the amorphous region.
2.4.5. Morphological properties
The morphology of cellulose and microcrystalline cellulose of
PW were observed using SEM (Tescan) at 10.0 kV and FE-SEM
(JEOL). The dried sample was deposited on the double-sided cel-
lophane tape and placed on the aluminum stub. Subsequently,
Hummer Sputter Coater (Techincs EMS, Inc, VA) was used to coat
the samples at 10 milliamps, with 25 nm gold-palladium, in a ratio
of 60:40, for 3 min.
2.4.6. Thermogravimetric analysis-differential scanning calorimetry
(TGA-DSC)
TGA analysis was conducted to evaluate the decomposition
temperature and the thermal stability using Linseis STA PT-1000.
The decomposition was investigated from 28  C to 600  C (heat-
ing rate: 10  C/min, under air atmosphere). The experiment also
generated DSC thermograms through the heating process from
room temperature to 600  C.
H. Holilah, D. Prasetyoko, R. Ediati et al. Journal of Cleaner Production 305 (2021) 127229

2.4.7. FT-Raman analysis alkalization. Each isolation step plays an essential role in reducing
The microcrystalline cellulose powders (PW-MCC and C-MCC) the composition of hemicellulose and lignin. Following the first
were deposited onto glass slide and the Raman spectra were alkalization, the lignin content that was determined at 25.1% in PW
collected using SENTERRA, Bruker with wavelength Raman laser was reduced to 18.9%. In alkalization step, the hydroxide OH
ion
532 nm and maximum power 20 mW. The spectra were recorded was responsible to break the bond that formed the basic structure
from 3500 to 200 cm
1 using an operating spectral resolution of of lignin, while the Naþ ion bound into lignin to form the soluble
2 cm
1 of Raman shift, and 20 scans were averaged for each sample. sodium phenolate (Vijay et al., 2019). The dissolved lignin in the
form of sodium phenolate was filtered off as black solution and
2.5. Bio-composites preparation referred as black liquor.
Bleaching process was carried out to further reduce the
Chitosan/MCC and seaweed/MCC bio-composites were pre- remaining lignin and simultaneously improved the brightness and
pared by addition of 5 wt % of MCC to chitosan and seaweed, the purity of cellulose. Lignin concentration of B-PW was analyzed
respectively. The amount of MCC loading was chosen based on the at 12.5% and the cellulose concentration was increased to 67.6%. The
optimized loading reported by Abdul Khalil et al. (2018); Hasan hemicellulose content was also reduced from 12.2 in PW to 10.1% in
et al. (2019); Hermawan et al. (2019). PW-MCC was dispersed in B-PW. It is important to note that bleaching should preserve the
distilled water, stirred for 1 h and then the suspension was ho- cellulose content and prevent structural damage of cellulose. Per-
mogenized for 10 min (Qsonica Sonicator Q500, USA). In another hydroxyl anion (HOO
) oxidized the non-phenolic unit of lignin
part, 2.0 g of chitosan was dissolved in 2% of CH3COOH solution. through the release of one electron to form cation radical and
MCC was slowly added into the dissolved chitosan solution. Then, facilitated in the dissociation of the C-C bond (Brooks and Moore,
glycerol or polyethylene glycol (PEG) at 3.0 wt% was added, and 2000).
continuously stirred at 80  C for 20 min. Similar procedures were Lignin concentration was still significantly higher in B-PW
repeated using seaweed as natural polymer, but seaweed was only following alkalization and bleaching processes, therefore hydro-
mixed with 50 mL of distilled water. 50 mL of the suspension was thermal assisted re-alkalization was carried out to further disso-
poured on casting plate and dried in air-conditioned room for 48 h. ciate the non-phenolic unit of lignin. The hydrothermal assisted re-
The completely dry chitosan/MCC and seaweed/MCC composite alkalization method carried out in this study significantly reduced
films were stripped off from the plate. PEG and glycerol as plasti- lignin concentration to only 2.1 ± 0.5% and consequently enhanced
cizers were also added on seaweed and chitosan without the the cellulose content to 77.9%. Hydrothermal treatment in the
presence of MCC. For comparison, C-MCC was also used as filler on presence of NaOH disintegrated the macromolecular lignin struc-
seaweed and chitosan with glycerol as plasticizer. Details of the bio- tures and neutralize the organic acids produced during the process
composites film were summarized in Table 1. that was otherwise restricted due to the high stability of lignin
(Bobleter, 1994).
2.5.1. Mechanical properties of bio-composites
The tensile strength (MPa) and the elongation at break (%) of the 3.2. Optimization of hydrolysis parameters for microcrystalline
bio-composites were evaluated using universal testing machine cellulose production
(Strograph VG 10-E) at room temperature with 10 kN load ac-
cording to ASTM D-882-91. The bio-composite films were cut into The extracted cellulose (H-PW) was subsequently used as pre-
15 cm  3 cm prior to the measurement. cursor for the formation of MCC. Fig. 1a illustrated the effect of acid
concentration during hydrolysis on the yield and the crystallinity
3. Results and discussion index (CI) of PW-MCC. The variation of acid concentration was
evaluated under the following conditions (temperature: 60  C,
3.1. Composition of lignocellulosic bio-waste time: 90 min, solid/liquid ratio 1:20). The MCC yield was signifi-
cantly decreased with increasing the concentration of HCl, how-
Table 2 summarized the concentration of cellulose, hemicellu- ever, the crystallinity index improved when the HCl concentration
lose, and lignin in PW and the extracted powder following the was increased to 3.5 N. Significant reduction of MCC crystallinity
respective alkalization, bleaching and hydrothermal assisted re- was observed with further increase of HCl concentration to 4.5 N.
alkalization treatments. The cellulose content of the IPW was Hydrolysis required strong acid solution to break the glycosidic
evaluated at 39.8 ± 0.9% which was relatively higher in comparison oxygen linkage between two sugar units in cellulose (Xiang et al.,
to the cellulose content from rice husk at 33.8 ± 0.5% and coffee 2003). Proton, Hþ from acid interacted with oxygen atom in the
husk at 35.4 ± 0.9% (Collazo-Bigliardi et al., 2018). The isolation of b-1.4-glycosidic bond, while Cl
contributed to the hydrolysis
cellulose from PW was carried out following three stages of treat- process by weakening the glycoside bond (Trache et al., 2016). The
ment i.e. alkalization, bleaching and hydrothermal assisted MCC yield was reduced with increasing the concentration of acid
due to further hydrolysis of cellulose into smaller fragment (Zhao
et al., 2018). However, formation of MCC with high crystallinity
Table 1 index required hydrolysis using moderate acid concentration. Hy-
The summary of bio-composites and their compositions.
drolysis of cellulose is a heterogeneous reaction that involved
Bio-composites Natural Polymer Plasticizer Cellulose permeation of acid catalysts into the cellulose framework for
ChG Chitosan Glycerol e disintegration to form glucose and small cellulose fragment (Huang
ChP Chitosan PEG e and Fu, 2013). Kinetic analysis indicated that the initial stage of
SeG Seaweed Glycerol e hydrolysis predominantly involved degradation of amorphous
SeP Seaweed PEG e
fragment of cellulose (Stephens et al., 2008). High concentration of
ChMCCG Chitosan Glycerol MCC
ChMCCP Chitosan PEG MCC acid further disintegrated the crystalline fragments and reduced
SeMCCG Seaweed Glycerol MCC the crystallinity of MCC.
SeMCCP Seaweed PEG MCC The effect of hydrolysis time on the yield and the crystallinity of
SeCMCCG Chitosan Glycerol C-MCC MCC were presented in Fig. 1b. Hydrolysis was carried out using
ChCMCCG Seaweed Glycerol C-MCC
3.5 N of HCl with the temperature and the liquid/solid ratio was set
4
H. Holilah, D. Prasetyoko, R. Ediati et al. Journal of Cleaner Production 305 (2021) 127229

Table 2
The cellulose, hemicellulose and lignin contents (wt. percentage) of industrial pepper waste and following different treatment steps (dried PW), after alkali treatment (A-PW),
after bleaching process (B-PW) after the hydrothermal assisted re-alkalization (H-PW).

Samples Cellulosea (%) SD CV (%) Hemicellulosea (%) SD CV (%) Lignina (%) SD CV (%)

PW 39.8 0.90 2.27 12.2 0.21 1.71 25.1 0.79 3.16

A-PW 49.4 0.31 0.62 11.9 0.70 5.90 18.9 1.21 6.42
B-PW 67.6 0.69 1.02 10.1 0.10 0.99 12.5 1.33 10.62
H-PW 77.9 0.80 1.03 12.1 0.89 7.35 2.10 0.50 23.81
a
Average values from three measurement; SD: standard deviation; CV: (standard deviation (SD)/mean of data)  100.

Fig. 1. Effects of acid concentration (a), reaction time (b) and reaction temperature (c) on the yield and crystallinity index (CI) of MCC. Reaction conditions (a. 60  C, 90 min, 1:20, b.
60  C, 3.5 N, 1:20 and c. 3,5 N, 90 min, 1:20).

at 60  C and 1:20. The reaction time was varied at 60e180 min. Table 3
The physicochemical properties of the MCC isolated from pepper waste (PW-MCC)
Fig. 1b showed that the MCC yield was decreased when the hy-
and commercial MCC (Avicel PH 101).
drolysis was extended up to 150 min. However, at 90 min, the
crystallinity was significantly enhanced to 75.8% suggesting effi- Parameters PW-MCC C-MCC
cient removal of amorphous component of cellulose. Amorphous Moisture content (%) 5.30 ± 0.20 5.15 ± 0.30
cellulose consisted of disordered structures that was susceptible Ash content (%) 0.13 ± 0.03 0.15 ± 0.01
towards acid hydrolysis (Trache et al., 2016). Understanding the Density (g/cm3) 1.41 ± 0.02 1.40 ± 0.02
pH 5.90 ± 0.03 6,05 ± 0.02
mechanism of acid hydrolysis, the process occurred on the outer-
Water holding capacity 3.75 ± 0.05 2.64 ± 00.4
most surface of cellulose fiber which involved dissociation of the b- Oil holding capacity 3.10 ± 0.03 2.73 ± 0.03
1,4-glycosidic bond before further penetration into the fiber bulk Swelling index 1.63 ± 0.29 1.27 ± 0.25
structures (Kusumattaqiin and Chonkaew, 2015). Prolonged sub-
mersion of cellulose in HCl caused excessive hydrolysis of cellulose
to glucose and reduced the MCC yield. Nevertheless, increasing the The effect of variation of temperature during hydrolysis was
hydrolysis duration was also important for the removal of amor- carried out using 3.5 N of HCl concentration for 90 min at the solid/
phous components in the cellulose which subsequently increased liquid ratio of 1:20. Fig. 1c showed the yield of MCC was rapidly
the crystallinity index of MCC that represented the purity of MCC decreased with the color of MCC was changed into dark brown
(Hou et al., 2019; Zhao et al., 2018).
5
H. Holilah, D. Prasetyoko, R. Ediati et al. Journal of Cleaner Production 305 (2021) 127229

Fig. 2. FTIR spectra of PW, B-PW, H-PW, B-MCC, PW-MCC and C-MCC.

Fig. 3. X-ray diffractograms of PW, B-PW, H-PW, B-MCC, PW-MCC and C-MCC.

6
H. Holilah, D. Prasetyoko, R. Ediati et al.
when the temperatures were increased from 50 to 90  C. The
crystallinity of MCC was also enhanced when the reaction tem-
perature was increased to 70  C.
Optimization studies of acid hydrolysis of treated PW to form
MCC indicated the yield and the crystallinity index, CI of MCC were
significantly influenced by the concentration of HCl, the reaction
temperatures and the hydrolysis duration. The optimum condition
for isolation of MCC from B-PW were determined at 3.5 N of HCl
concentrations, 1:20 of solid/liquid ratio, 70  C of reaction tem-
perature and at 90 min of reaction. The MCC produced from the
optimum condition was further characterized and discussed in the
following section.
3.3. Physicochemical properties of MCC
Physicochemical properties of MCC hydrolyzed from hydro-
thermally treated PW (H-PW) were compared with commercial
MCC (Avicel PH 101) in Table 3. The moisture content of PW-MCC
was determined at 5.30 ± 0.20%, which was comparable with the
commercial MCC (5.15 ± 0.30). Moisture content determined
functionality and mechanical properties of microcrystalline cellu-
lose (Thoorens et al., 2014), with high water content generally
reducing the mechanical strength of polymer composites (Jumaidin
et al., 2017; Miao and Hamad, 2013). Visual, compaction and tensile
strength of MCC were also dependent on moisture content
(Doelker, 1993; Sun, 2008).
Ash composition in PW-MCC was analyzed at  0.15% which was
within the range of Indian Pharmacopoeia (IP) and United States
Pharmacopoeia (USP) standards and within the range reported
from MCC derived E. glaucum at approximately 0.11 ± 0.02%
(Pachuau et al. (2019) and MCC from Posidonia oceanica brown
algae at 0.75 ± 0.03% (Tarchoun et al., 2019). The pH of MCC
following immersion in water was determined at 5.90 which was
approximately similar to the commercial MCC ~6.05 ± 0.02. The
slightly acidic of MCC was presumably due to the residual acid
following the hydrolysis of cellulose.
The density of microcrystalline cellulose provided important
information during material selection and determined the perfor-
mance of bio-composites (Ilyas et al., 2018). The density of PW-MCC
was measured at 1.41 ± 0.02 g/cm3, comparable to the density of C-
MCC (1.40 ± 0.02 g/cm3). The value was also approximately similar
with the MCC that was previously reported; Avicel PH101 (1.540 g/
cm3) (Amidon and Houghton, 1995), and Avicel PH 102 (1.558 g/
cm3) (Hentzschel et al., 2012), but lower than MCC prepared from
Ensete glaucum (1.833 ± 0.47 g/cm3) (Pachuau et al., 2019).
Water holding capacity (WHC) indicated the capacity of fiber to
hold water within its domain and provided important benchmark
for the modification of viscosity and the texture of bio-composites
(Rodríguez-Ambriz et al., 2008). WHC was influenced by the crys-
talline index and the size of MCC Liu et al. (2018); Ul-Islam et al.
(2012), Zain et al. (2014). Particles size analysis of PW-MCC
(Fig. S2 supplementary) showed significantly smaller diameter
with narrow particles size distribution than C-MCC, however there
were no significant differences between the crystallinity indexes.
MCC derived from H-PW showed a higher WHC level at
3.75 ± 0.05 g/g than the commercial MCC at 2.64 ± 00.4 g/g, indi-
cating the narrow particle size distribution of PW-MCC significantly
improved the WHC level. Apart from WHC, the oil holding capacity
(OHC) of PW-MCC was also determined at 3.10 ± 0.03 g/g which
was slightly higher than the commercial MCC (2.73 ± 0.03 g/g). The
high OHC value implied the presence of high acyl group content in
cellulose. Cellulose with high OHC showed potential as food addi-
tives as it enhanced the stabilization of emulsion and high-fat food
product, and facilitated the dispersion of two unmixable liquids
(Rodríguez-Gutie rrez et al., 2014). The swelling index of PW-MCC
7
Journal of Cleaner Production 305 (2021) 127229
was also measured in water indicating that both the MCC sam-
ples from PW and commercial MCC showed relatively low swelling
index.
3.4. Fourier transform-infrared of PW and MCC from PW
Industrial pepper bio-waste mainly contained major compo-
nents of cellulose, hemicellulose and lignin, and FT-IR spectroscopy
provided information on the chemical and structural changes of
cellulose (Jonoobi et al., 2011; Mandal and Chakrabarty, 2011). The
FT-IR spectra of PW, PW after alkalization and bleaching (B-PW),
after further hydrothermal assisted re-alkalization (H-PW) and also
MCC derived from treated PW and commercial MCC were shown in
Fig. 2. In general, all samples exhibited the main cellulose molecular
absorption bands at 3400, 2905, 1640, 1430, 1060 and 897 cm
1.
The absorption band at 3400 cm
1 was assigned to the O-H group
stretching vibration that was related to the hydrophilicity of the
samples (El Achaby et al., 2019). The absorption band for aliphatic
saturated C-H stretching vibration appeared at 2901 cm
1 (Julie
Chandra et al., 2017; Singh et al., 2019). The absorption bands at
2860, 1740 and 1503 cm
1 were only observed on PW and B-PW
samples after the first alkalization and bleaching processes. The
absorption band at 2860 cm
1 was assigned to the asymmetric and
symmetric stretching vibration of the CH2 group of hemicellulose
(Abdul Razak et al., 2020). The band at 2860 cm
1 was disappeared
following further hydrothermal assisted re-alkalization (H-PW)
indicating the reduction of hemicellulose content in the samples.
The absorption band centered at 1740 cm
1 of untreated PW cor-
responded to the ketone, aldehyde or carboxylic acid groups in
hemicellulose and disappeared in H-PW (Alemdar and Sain, 2008;
Chen et al., 2009; Oun and Rhim, 2016; Trache et al., 2014). It is
interesting to note that analysis of hemicellulose content in Table 2
exhibited insignificant changes of hemicellulose concentration
following different stages of treatment. Therefore it was suggested
that the disappearances of 1740 cm
1 and 2860 cm
1 bands may be
due to the increased intensity of the adjacent 2905 cm
1 and
1640 cm
1 cellulose bands respectively due to the increase con-
centration of cellulose in the powder. The absorption band at
1503 cm
1 on PW and B-PW were assigned to C¼C aromatic skel-
etal vibration of ester linkage of lignin (Oun and Rhim, 2016; Rosa
et al., 2010). The disappearance of the ester linkage of lignin in H-
PW indicated a thorough removal of lignin from PW was achieved
following hydrothermal assisted re-alkalization step (Tarchoun
et al., 2019; Xiang et al., 2016; Zhao et al., 2018). Synthesis of
MCC from hydrothermal alkalization of PW (H-PW) showed that
the increased intensity of the absorption band at 1430 cm
1 was
corresponded to the bending vibration of symmetric CH2, known as
the “crystallinity band” (Kalita et al., 2013; Rosa et al., 2010). The
results indicated the isolation of cellulose via hydrothermal assis-
ted re-alkalization produced PW-MCC with high level of crystalline
structures. The C-O-C stretching of the b-1,4-glycosidic linkages in
PW-cellulose and C-H rocking vibration were observed at
1060 cm
1 and 897 cm
1, respectively (Singh et al., 2019; Trache
et al., 2016). In general, the results obtained from FTIR analysis
further confirmed the benefit of hydrothermal assisted re-
Table 4
Crystalline index (CI) of B-PW, H-PW, B-MCC, PW-MCC and C-MCC.
Samples Treatment Crystalinity index (%)
B-PW Bleaching, H2O2þNaOH 48.3
H-PW Hydrothermal, NaOH 51.6
B-MCC Hydrolysis of B-PW 47.8
PW-MCC Hydrolysis of H-PW 78.9
C-MCC Commercial Avicel MCC 79.2
H. Holilah, D. Prasetyoko, R. Ediati et al. Journal of Cleaner Production 305 (2021) 127229

Fig. 4. SEM micrographs of (a) PW; (b) B-PW; (c) H-PW; (d) C-MCC; (e) PW-MCC and (f) FE-SEM analysis of PW-MCC.

8
H. Holilah, D. Prasetyoko, R. Ediati et al. Journal of Cleaner Production 305 (2021) 127229

alkalization treatment to enhance lignin dissolution and further Microcrystalline cellulose was considered to have high crystalline
improved the crystallinity of MCC. structures when the crystallinity index was measured in the range
of 55e80% (Haafiz et al., 2013).
3.5. X-ray diffraction of PW and MCC from PW
3.6. Morphological properties
XRD analysis provided information of the structural changes of
The morphology of the extracted cellulose fibre from pepper
cellulose in PW following lignin removal and the transformation to
bio-waste and the microcrystalline cellulose were analyzed using
microcrystalline cellulose. X-ray diffraction pattern of all samples in
SEM and FE-SEM (Fig. 4). SEM analysis of waste pepper before
Fig. 3 showed similar diffraction peaks observed at 2q: 15e16 ,
preparation, PW showed a rough and aggregated surface area
22.6 and 34 . The industrial pepper bio-waste, PW showed a broad
consisting of hemicellulose, lignin, and other non-cellulosic sub-
diffraction peak due to the high content of hemicellulose and lignin
stances covering the untreated PW (Fig. 4a). B-PW showed the
that are naturally present in amorphous form (Owolabi et al., 2017).
corrugated texture of the cellulose fiber was disappeared following
The peaks observed at 16.3 , 22.3 , and 34.7 were corresponded to
alkali and bleaching treatments, formed a smooth and uniformed
the respective (110), (200), and (040) crystallographic planes of the
cellulose surface (Fig. 4b). H-PW obtained from further alkali-
cellulose I polymorph structure (Lu et al., 2014). No evidence of
hydrothermal treatment showed the reduction of cellulose fiber
cellulose II indicated by the absence of doublet peak at 22.6 (Rosa
rigidity that was due to the disintegration of the lignin structures
et al., 2012). Microcrystalline cellulose (B-MCC) produced from
(Fig. 4c). Following acid hydrolysis of H-PW, the small irregular
hydrolysis of B-PW showed no significant changes of the peak in-
MCC structures were produced as the result of acid penetration into
tensity suggesting the unsuccessful isolation of MCC from cellulose
the amorphous region to dissociate the b-1,4-linkage between the
following alkalization and bleaching processes. High concentration
cellulose repeating units (Fig. 4e). The morphology of PW-MCC was
of lignin ~12.5 ± 1.3% in B-PW that was not successfully removed
during first alkalization and bleaching steps affected the efficiency
of hydrolysis process for the formation of MCC. The increase of peak Table 5
intensity at 22.6 is the main indicator to confirm the formation of Thermal analysis data of B-PW, H-PW, B-MCC, PW-MCC and C-MCC.
microcrystalline cellulose with high crystallinity and structural ri- Samples Tonset(oC)a Tpeak(oC)b DH (J/g)c W residue 600  C (%)d
gidity (CIH-PW: 51.6 > CIPW-MCC: 78.9) (Rosa et al., 2012). It was
PW 300.4 332.6 1008 24.659
apparent that the crystallinity of PW-MCC obtained from hydro- B-PW 301.2 329.0 1165 1.601
thermally assisted re-alkalization of PW (H-PW) was significantly B-MCC 306.6 336.8 1240 1.059
higher than the B-PW. Low composition of lignin in H-PW allowed PW-MCC 319.8 345.6 1403 1.043
the dissociation of glycosidic bonds for the removal of amorphous C-MCC 325.3 352.2 1410 1.019
cellulose during acid hydrolysis and the formation of micro- a
Initial decomposition temperature using DSC results.
b
crystallite structures (Spagnol et al., 2012). The crystallinity index, Peak temperature of DSC.
c
CI was calculated on PW-MCC with the levels was comparable to Heat of decomposition.
d
Residual weight at 600  C.
the C-MCC index at 78.9% and 79.2% respectively (Table 4).

Fig. 5. DSC thermograms of PW, B-PW, H-PW, PW-MCC and C-MCC.

9
H. Holilah, D. Prasetyoko, R. Ediati et al.
relatively similar to the commercial MCC. However, the commercial
MCC exhibited relatively longer fiber sticks than the MCC isolated
from industrial pepper bio-waste (Fig. 4d). FE-SEM analysis of
microcrystalline cellulose, PW-MCC showed the formation of a rod
shape structures with a well-defined edges (Fig. 4f). The analysis
also revealed the formation spherical shape cellulose within
nanoscale size region presumably produced from further hydrolysis
of MCC. Particle size distribution of MCC from pepper waste was
compared with commercial MCC from Avicel (Fig. S2 supplemen-
tary). PW-MCC showed a narrow particle size distribution with the
Fig. 6. TGA (a) and DTG (b) profiles of PW, B-PW, H-PW, PW-MCC and C-MCC.
10
Journal of Cleaner Production 305 (2021) 127229
average diameter of 16.39 mm. Commercial MCC Avicel showed a
much broader distribution with the average particle size was
determined at 38.85 mm.
3.7. Thermal properties
Thermal properties of cellulose and microcrystalline cellulose
determined using DSC and TGA/DTG provided information on the
degradation pattern of the samples. DSC analysis in Fig. 5 showed
the presence of general endothermic decomposition pattern of PW.
H. Holilah, D. Prasetyoko, R. Ediati et al.
The PW-MCC showed relatively high degradation energy due to the
highly ordered molecular structures in comparison to cellulose
derived from B-PW. The enthalpy of PW-MCC and C-MCC were
determined at 1403 J/g and 1410 J/g. Table 5 summarized the
thermal analysis data from DSC that showed the onset temperature
(Tonset), the melting temperature (Tpeak) and the enthalpy of heating
(DH) of the PW-MCC were significantly higher than the rest of
cellulose derived from pepper waste, and comparable to the com-
mercial MCC, which further proved the formation of high crystal-
line PW-MCC (Azubuike and Okhamafe, 2012) (Azubuike and
Okhamafe, 2012).
TGA/DTG profiles of microcrystalline cellulose revealed two
stages of weight loss for MCC from PW (PW-MCC) that was similar
to the commercial MCC (C-MCC) (Fig. 6). However, the extracted
cellulose fibers from PW via alkali and bleaching treatments (B-
PW) and further hydrothermal assisted re-alkalization (H-PW)
indicated three stages of weight loss. The first decomposition
occurred at ~50-150  C which involved ~10% of weight loss, and was
attributed to the water evacuation from samples. The second
decomposition was observed at 250-400  C, that was due to the
degradation processes of cellulose, including dehydration, decom-
position, and depolymerization of the glycoside units. The third
thermal stage at 420e600  C, corresponded to the oxidation of
carbonaceous residues to low molecular weight gasses (Kalita et al.,
2013; Vasconcelos et al., 2017). Hemicellulose and lignin were re-
ported to have lower decomposition temperatures than cellulose
(Mora n et al., 2008). Thermal degradation of microcrystalline cel-
lulose occurred at 319  C, significantly higher than the rest of the
extracted celluloses from PW which implied high crystallinity of
MCC contributed to the thermal stability (Hussin et al., 2016;
Katakojwala and Mohan, 2019; Tarchoun et al., 2019; Trache et al.,
2014). C-MCC and PW-MCC also showed significantly lower ash
residues than the PW, B-PW and H-PW. Thermal degradation of
cellulose was described to complete at 600  C and the lower weight
of ash residue from thermal decomposition indicated the high
purity of MCC (Hou et al., 2019; Tarchoun et al., 2019).
Fig. 7. Raman spectra of MCC from PW (PW-MCC) and Avicel PH 101 as standard (C-MCC).
11
Journal of Cleaner Production 305 (2021) 127229
3.8. FT-Raman spectroscopy
Raman analysis of microcrystalline cellulose, PW-MCC was
carried out and compared with the commercial C-MCC (Fig. 7). The
results showed that the peaks observed in PW-MCC were similar
with the Raman shift of C-MCC. The characteristic peaks of
microcrystalline cellulose at 380, 897, 1095, 1120, 1380 and
1481 cm
1 were appeared with strong intensities for both com-
mercial and PW-MCC (Fechner et al., 2003). The bands observed at
380, 897, 1378, 1481 and 1121; 1095 cm
1 were assigned to the
bending vibration of CCC; HCC and HCO; HOC, HCO, and HCC; CH2
and the stretching vibration of symmetric and asymmetric of COC
in b-(1,4)-glycosidic linkage, respectively (Gierlinger et al., 2012).
The ratios between the intensity of Raman peaks at 380/1096 cm
1
(I380/I1096) and 1121/1096 cm
1 (I1121/I1096) were correlated with
the crystallinity of the fiber (Alves et al., 2016), in which the peak
intensity ratio of C-MCC and PW-MCC were calculated at 0.52; 0.49
(I380/I1096) and 0.97; 0.96 (I1121/I1096) respectively. The ratios be-
tween C-MCC and PW-MCC were relatively similar that implied the
crystallinity index of C-MCC was comparable to PW-MCC i.e. 78.9%
and 79.2%, respectively.
3.9. Bio-composites properties
MCC derived from PW (PW-MCC) was used as filler for natural
polymer chitosan and seaweed, using glycerol and polyethylene
glycol (PEG) as plasticizer. The mechanical properties of the bio-
composites i.e. the tensile strength and the elongation at break
were analyzed and presented in Fig. 8. In general, bio-composites
with glycerol as plasticizer have relatively higher tensile strength
and elongation at break than polyethylene glycol. Glycerol has
lower molecular weight and relatively higher OH- functional group
per weight than polyethylene glycol (Bourtoom, 2008; Vieira et al.,
2011). Therefore, glycerol is easy to be incorporated within the
matrix and to form strong hydrogen bond (Donhowe and Fennema,
1993; McHugh et al., 1994). Seaweed also produced bio-composites
with better mechanical properties than chitosan. Seaweed is
H. Holilah, D. Prasetyoko, R. Ediati et al. Journal of Cleaner Production 305 (2021) 127229

Fig. 8. Tensile strength (a) and elongation at break (b) of bio-composites film with glycerol and PEG as plasticizer. Chitosan (Ch) and seaweed (Se) were used as natural polymer,
meanwhile Glycerol (G) and PEG (P) were used as plasticizer.

12
H. Holilah, D. Prasetyoko, R. Ediati et al.
thoroughly dissolved in water that increased the dispersion and
distribution when incorporated with MCC (Hasan et al., 2019;
Reddy and Rhim, 2014; Rhim et al., 2013; Shankar and Rhim, 2016).
Meanwhile chitosan is insoluble in water restricting the dispersion
during bio-composites formation. The tensile strength of the chi-
tosan/MCC bio-composites showed the enhancement from
36.5 MPa to 43.4 MPa when using glycerol, and to 41.6 MPa with
PEG as plasticizer. Seaweed/MCC bio-composites also showed
enhancement of the tensile strength from 37.3 MPa to 44.3 MPa
when using PEG and 46.9 MPa with glycerol. The enhancement of
tensile strength suggested the compatibility of MCC derived from
PW as filler to transfer the applied stress from natural polymer
matrix to the MCC fiber (Abdul Khalil et al., 2018; Khan et al., 2012).
The elongation at break profile of bio-composite was evaluated to
provide information on the flexibility of the films. The addition of
MCC on seaweed when using glycerol as plasticizer showed the
increase of elongation at break from 27.0% to 38.8%. Similar
enhancement on the elongation properties of seaweed was found
when using PEG as plasticizer at 31.4%.
The tensile strength and the elongation at break of the bio-
composite films was also compared using C-MCC as filler. The
tensile strengths were measured at 40.5 MPa (chitosan) and
41.8 MPa (seaweed), meanwhile the elongation at break were
determined at 29.7% (chitosan) and 30.9% (seaweed), respectively.
PW-MCC as filler had a higher mechanical value than C-MCC pre-
sumably due to the small particle size of PW-MCC (Fig. S3 sup-
plementary). Small sized MCC occupied the sites within the natural
polymer domain to create a strong interfacial interaction with the
polymer matrix (Shankar and Rhim, 2016; Zhang et al., 2020). The
interaction between MCC and natural polymer occurred via the
formation of hydrogen bond, therefore high concentration of sur-
face hydroxyl as reported in FTIR analysis (Fig. 2) improved the
tensile strength of the bio-composites (Hasan et al., 2019). The
well-distributed and strongly adhered MCC fiber efficiently trans-
ferred the applied stress across the composite (Catto et al., 2019;
Huang et al., 2018).
SEM analysis of the top view and the cross-section of seaweed
film with PEG as plasticizer exhibited homogeneous and smooth
surface, with compact structures (Fig. S3 supplementary). However,
introduction of 5% MCC from pepper waste produced bio-
composite film with rough and uneven surfaces. The SEM analysis
was also compared with bio-composite film produced using com-
mercial Avicel MCC as filler that produced film with significantly
rougher surface than PW-MCC. The cross-section image indicated a
highly uneven surface morphology in comparison to MCC from
pepper waste that presumably due to a large particle size of C-MCC
compared to PW-MCC. SEM analysis also indicated that the MCC
fibers were successfully embedded and evenly distributed within
the seaweed bio-composite matrix. The microcrystalline cellulose
fibers isolated from industrial pepper bio-waste was compatible as
filler for seaweed and chitosan to form bio-composites in order to
strengthen the mechanical properties. The studies have demon-
strated the potential application of microcrystalline cellulose fibers
isolated from industrial pepper bio-waste to be employed as bio-
degradable composites for packaging materials in food industries.
4. Conclusion
Microcrystalline cellulose (PW-MCC) has been successfully iso-
lated from industrial pepper bio-waste via three steps of treatment
i.e. alkalization, bleaching and hydrothermal assisted re-
alkalization. The hydrothermal assisted re-alkalization step was
crucial for efficient removal of lignin from cellulose. High pressure
generated during hydrothermal treatment enhanced the dissolu-
tion of lignin that was strongly bound to the cellulose domain,
13
Journal of Cleaner Production 305 (2021) 127229
reducing lignin concentration to ~2.1 ± 0.5%. Optimum condition
for the formation of microcrystalline cellulose from PW was
determined at 3.5 N of HCl concentration, at 70  C, and for 90 min
with solid/liquid ratio of 1:20, produced 86% of MCC yield. Char-
acterization studies indicated similar physicochemical properties of
MCC derived from PW with the commercial MCC (Avicel PH 101).
FTIR and XRD analysis confirmed the formation of cellulose I
polymorph structures with high crystallinity index. The MCC dis-
played the short fiber strand morphology with well-defined edges.
Incorporation of MCC at 5% of weight loading on seaweed and
chitosan as reinforcement filler showed the improvement of the
tensile strength and the elongation at break of the bio-composites.
Finally, the investigation exhibited the potential use of industrial
pepper bio-waste to produce microcrystalline cellulose, MCC that
can be used as filler for reinforcement of bio-composites for the
production of bio-degradable food packaging.
CRediT authorship contribution statement
Holilah Holilah: Conceptualization, Methodology, Validation,
Investigation, Writing e original draft, Writing e review & editing,
Funding acquisition. Didik Prasetyoko: Supervision, Conceptuali-
zation, Validation, Writing e review & editing, Funding acquisition.
Ratna Ediati: Supervision, Conceptualization. Hasliza Bahruji:
Supervision, Conceptualization, Validation, Writing e review &
editing, Funding acquisition. Aishah Abdul Jalil: Conceptualization.
Asranudin Asranudin: Resources, Validation, Visualization. Sus-
anti Dhini Anggraini: Resources.
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.
Acknowledgement
Authors would like to acknowledge Doctoral Scholarship Fund
“BPPDN 2019” awarded to Holilah from Kemendikbud DIKTI and
doctoral research grant 860/PKS/ITS/2021 KemenristekBRIN,
Indonesia. Authors also would like to acknowledge Universiti
Brunei Darussalam Research Grant UBD/RSCH/URC/RG(b)/2019/012
for funding to H. Bahruji.
Appendix A. Supplementary data
Supplementary data to this article can be found online at
https://doi.org/10.1016/j.jclepro.2021.127229.
References
Abdul Khalil, H.P.S., Lai, T.K., Tye, Y.Y., Paridah, M.T., Fazita, M.R.N., Azniwati, A.A.,
Dungani, R., Rizal, S., 2018. Preparation and characterization of microcrystalline
cellulose from Sacred Bali bamboo as reinforcing filler in seaweed-based
composite film. Fibers Polym. 19, 423e434. https://doi.org/10.1007/s12221-
018-7672-7.
Abdul Razak, S., Mahadi, A.H., Abdullah, R., Yasin, H.M., Ja’afar, F., Abdul Rahman, N.,
Bahruji, H., 2020. Biohydrogen production from photodecomposition of various
cellulosic biomass wastes using metal-TiO2 catalysts. Biomass Convers. Biorefin.
1e12. https://doi.org/10.1007/s13399-020-01164-4.
Abhilash, M., Thomas, D., 2017. Biopolymers for Biocomposites and Chemical Sensor
Applications, Biopolymer Composites in Electronics. Elsevier Inc. https://
doi.org/10.1016/B978-0-12-809261-3.00015-2.
Ahsan, H.M., Pei, Y., Luo, X., Wang, Y., Li, Y., Li, B., Liu, S., 2020. Novel stable pickering
emulsion based solid foams efficiently stabilized by microcrystalline cellulose/
chitosan complex particles. Food Hydrocolloids 108, 106044. https://doi.org/
10.1016/j.foodhyd.2020.106044.
Alashwal, B.Y., Saad Bala, M., Gupta, A., Sharma, S., Mishra, P., 2020. Improved
properties of keratin-based bioplastic film blended with microcrystalline
H. Holilah, D. Prasetyoko, R. Ediati et al.
cellulose: a comparative analysis. J. King Saud Univ. Sci. 32, 853e857. https://
doi.org/10.1016/j.jksus.2019.03.006.
Alemdar, A., Sain, M., 2008. Isolation and characterization of nanofibers from
agricultural residues - wheat straw and soy hulls. Bioresour. Technol. 99,
1664e1671. https://doi.org/10.1016/j.biortech.2007.04.029.
Alves, A.P.P., de Oliveira, L.P.Z., Castro, A.A.N., Neumann, R., de Oliveira, L.F.C.,
Edwards, H.G.M., Sant’Ana, A.C., 2016. The structure of different cellulosic fibres
characterized by Raman spectroscopy. Vib. Spectrosc. 86, 324e330. https://
doi.org/10.1016/j.vibspec.2016.08.007.
Amidon, G.E., Houghton, M.E., 1995. The effect of moisture on the mechanical and
powder flow properties of microcrystalline cellulose. Pharm. Res. An Off. J. Am.
Assoc. Pharm. Sci. 12, 923e929. https://doi.org/10.1023/A:1016233725612.
Arauzo, P.J., Lucian, M., Du, L., Olszewski, M.P., Fiori, L., Kruse, A., 2020. Improving
the recovery of phenolic compounds from spent coffee grounds by using hy-
drothermal delignification coupled with ultrasound assisted extraction.
Biomass Bioenergy 139. https://doi.org/10.1016/j.biombioe.2020.105616.
Azubuike, C.P., Okhamafe, A.O., 2012. Physicochemical, spectroscopic and thermal
properties of microcrystalline cellulose derived from corn cobs. Int. J. Recycl.
Org. Waste Agric. 1, 1e7. https://doi.org/10.1186/2251-7715-1-9.
Bhatti, H.N., Mahmood, Z., Kausar, A., Yakout, S.M., Shair, O.H., Iqbal, M., 2020.
Biocomposites of polypyrrole, polyaniline and sodium alginate with cellulosic
biomass: adsorption-desorption, kinetics and thermodynamic studies for the
removal of 2,4-dichlorophenol. Int. J. Biol. Macromol. https://doi.org/10.1016/
j.ijbiomac.2020.02.306.
Bobleter, O., 1994. Hydrothermal degradation of polymers derived from plants.
Prog. Polym. Sci. https://doi.org/10.1016/0079-6700(94)90033-7.
Boldizar, A., Klason, C., Kuba t, J., N€ ha, P., 1987. Prehydrolyzed cellulose
aslund, P., Sa
as reinforcing filler for thermoplastics. Int. J. Polym. Mater. Polym. Biomater. 11,
229e262. https://doi.org/10.1080/00914038708078665.
Bourtoom, T., 2008. Plasticizer Effect on the Properties of Biodegradable Blend Film
from Rice Starch-Chitosan 30.
Brinchi, L., Cotana, F., Fortunati, E., Kenny, J.M., 2013. Production of nanocrystalline
cellulose from lignocellulosic biomass: technology and applications. Carbohydr.
Polym. https://doi.org/10.1016/j.carbpol.2013.01.033.
Brooks, R.E., Moore, S.B., 2000. Alkaline hydrogen peroxide bleaching of cellulose.
Cellulose 7, 263e286. https://doi.org/10.1023/A:1009273701191.
Buffiere, J., Balogh-Michels, Z., Borrega, M., Geiger, T., Zimmermann, T., Sixta, H.,
2017. The chemical-free production of nanocelluloses from microcrystalline
cellulose and their use as Pickering emulsion stabilizer. Carbohydr. Polym. 178,
48e56. https://doi.org/10.1016/j.carbpol.2017.09.028.
Catto, A.L., Dahlem Júnior, M.A., Hansen, B., Francisquetti, E.L., Borsoi, C., 2019.
Characterization of polypropylene composites using yerba mate fibers as
reinforcing filler. Compos. B Eng. 174 https://doi.org/10.1016/
j.compositesb.2019.106935.
Chen, H., Wang, L., 2017. Pretreatment strategies for biochemical conversion of
biomass. Technol. Biochem. Convers. Biomass 21e64. https://doi.org/10.1016/
b978-0-12-802417-1.00003-x.
Chen, X., Li, H., Sun, S., Cao, X., Sun, R., 2016. Effect of hydrothermal pretreatment on
the structural changes of alkaline ethanol lignin from wheat straw. Sci. Rep. 6,
1e9. https://doi.org/10.1038/srep39354.
Chen, Y., Liu, C., Chang, P.R., Cao, X., Anderson, D.P., 2009. Bionanocomposites based
on pea starch and cellulose nanowhiskers hydrolyzed from pea hull fibre: effect
of hydrolysis time. Carbohydr. Polym. 76, 607e615. https://doi.org/10.1016/
j.carbpol.2008.11.030.
Collazo-Bigliardi, S., Ortega-Toro, R., Chiralt Boix, A., 2018. Isolation and character-
isation of microcrystalline cellulose and cellulose nanocrystals from coffee husk
and comparative study with rice husk. Carbohydr. Polym. 191, 205e215. https://
doi.org/10.1016/j.carbpol.2018.03.022.
Direktorat Jenderal Perkebunan, 2019. Luas Areal Lada di Indonesia, pp. 2015e2019.
Do Nascimento, J.H.O., Luz, R.F., Galv~ ao, F.M.F., Melo, J.D.D., Oliveira, F.R.,
Ladchumananandasivam, R., Zille, A., 2015. Extraction and characterization of
cellulosic nanowhisker obtained from discarded cotton fibers. In: Materials
Today: Proceedings. Elsevier Ltd, pp. 1e7. https://doi.org/10.1016/
j.matpr.2015.04.001.
Doelker, E., 1993. Comparative compaction properties of various microcrystalline
cellulose types and generic products. Drug Dev. Ind. Pharm. 19, 2399e2471.
https://doi.org/10.3109/03639049309047196.
Donhowe, I.G., Fennema, O., 1993. The effects of plasticizers on crystallinity,
permeability, and mechanical properties of methylcellulose films. J. Food Pro-
cess. Preserv. 17, 247e257. https://doi.org/10.1111/j.1745-4549.1993.tb00729.x.
El Achaby, M., Ruesgas-Ramo  n, M., Fayoud, N.E.H., Figueroa-Espinoza, M.C.,
Trabadelo, V., Draoui, K., Ben Youcef, H., 2019. Bio-sourced porous cellulose
microfibrils from coffee pulp for wastewater treatment. Cellulose 26,
3873e3889. https://doi.org/10.1007/s10570-019-02344-w.
Fearon, O., Kuitunen, S., Ruuttunen, K., Alopaeus, V., Vuorinen, T., 2020. Detailed
modeling of kraft pulping chemistry. Delignification. Ind. Eng. Chem. Res. 59,
12977e12985. https://doi.org/10.1021/acs.iecr.0c02110.
Fechner, P.M., Wartewig, S., Füting, M., Heilmann, A., Neubert, R.H.H.,
Kleinebudde, P., 2003. Properties of microcrystalline cellulose and powder
cellulose after extrusion/spheronization as studied by Fourier transform Raman
spectroscopy and environmental scanning electron microscopy. AAPS PharmSci
5, 375e376. https://doi.org/10.1208/ps050431.
Fuentes-Alventosa, J.M., Rodríguez-Gutie rrez, G., Jaramillo-Carmona, S., Espejo-
Calvo, J.A., Rodríguez-Arcos, R., Ferna ndez-Bolan ~ os, J., Guille
n-Bejarano, R.,
Jime nez-Araujo, A., 2009. Effect of extraction method on chemical composition
14
Journal of Cleaner Production 305 (2021) 127229
and functional characteristics of high dietary fibre powders obtained from
asparagus by-products. Food Chem. 113, 665e671. https://doi.org/10.1016/
j.foodchem.2008.07.075.
Gierlinger, N., Keplinger, T., Harrington, M., 2012. Imaging of plant cell walls by
confocal Raman microscopy. Nat. Protoc. 7, 1694e1708. https://doi.org/10.1038/
nprot.2012.092.
Haafiz, M.K.M., Hassan, A., Zakaria, Z., Inuwa, I.M., Islam, M.S., Jawaid, M., 2013.
Properties of polylactic acid composites reinforced with oil palm biomass
microcrystalline cellulose. Carbohydr. Polym. 98, 139e145. https://doi.org/
10.1016/j.carbpol.2013.05.069.
Hasan, M., Lai, T.K., Gopakumar, D.A., Jawaid, M., Owolabi, F.A.T., Mistar, E.M.,
Alfatah, T., Noriman, N.Z., Haafiz, M.K.M., Abdul Khalil, H.P.S., 2019. Micro
crystalline bamboo cellulose based seaweed biodegradable composite films for
sustainable packaging. Mater. J. Polym. Environ. 27, 1602e1612. https://doi.org/
10.1007/s10924-019-01457-4.
Hentzschel, C.M., Sakmann, A., Leopold, C.S., 2012. Comparison of traditional and
novel tableting excipients: physical and compaction properties. Pharmaceut.
Dev. Technol. 17, 649e653. https://doi.org/10.3109/10837450.2011.572897.
Hermawan, D., Lai, T.K., Jafarzadeh, S., Gopakumar, D.A., Hasan, M., Owolabi, F.A.T.,
Sri Aprilia, N.A., Rizal, S., Khalil, H.P.S.A., 2019. Development of seaweed-based
bamboo microcrystalline cellulose films intended for sustainable food pack-
aging applications. BioResources 14, 3389e3410. https://doi.org/10.15376/bio-
res.14.2.3389-3410.
Hou, W., Ling, C., Shi, S., Yan, Z., 2019. Preparation and characterization of micro-
crystalline cellulose from waste cotton fabrics by using phosphotungstic acid.
Int. J. Biol. Macromol. 123, 363e368. https://doi.org/10.1016/
j.ijbiomac.2018.11.112.
Huang, X., Xie, F., Xiong, X., 2018. Surface-modified microcrystalline cellulose for
reinforcement of chitosan film. Carbohydr. Polym. 201, 367e373. https://
doi.org/10.1016/j.carbpol.2018.08.085.
Huang, Y.B., Fu, Y., 2013. Hydrolysis of cellulose to glucose by solid acid catalysts.
Green Chem. 15, 1095e1111. https://doi.org/10.1039/c3gc40136g.
Hussin, M.H., Pohan, N.A., Garba, Z.N., Kassim, M.J., Rahim, A.A., Brosse, N.,
Yemloul, M., Fazita, M.R.N., Haafiz, M.K.M., 2016. Physicochemical of micro-
crystalline cellulose from oil palm fronds as potential methylene blue adsor-
bents. Int. J. Biol. Macromol. 92, 11e19. https://doi.org/10.1016/
j.ijbiomac.2016.06.094.
Ilyas, R.A., Sapuan, S.M., Ishak, M.R., 2018. Isolation and characterization of nano-
crystalline cellulose from sugar palm fibres (Arenga Pinnata). Carbohydr. Polym.
181, 1038e1051. https://doi.org/10.1016/j.carbpol.2017.11.045.
Jonoobi, M., Khazaeian, A., Tahir, P.M., Azry, S.S., Oksman, K., 2011. Characteristics of
cellulose nanofibers isolated from rubberwood and empty fruit bunches of oil
palm using chemo-mechanical process. Cellulose 18, 1085e1095. https://
doi.org/10.1007/s10570-011-9546-7.
Julie Chandra, C.S., Bipinbal, P.K., Sunil, K.N., 2017. Viscoelastic behaviour of silica
filled natural rubber composites e correlation of shear with elongational
testing. Polym. Test. 60, 187e197. https://doi.org/10.1016/
j.polymertesting.2017.03.023.
Jumaidin, R., Sapuan, S., Jawaid, M., Ishak, M., Sahari, J., 2017. Effect of agar on
flexural, impact, and thermogravimetric properties of thermoplastic sugar palm
starch. Curr. Org. Synth. 14, 200e205. https://doi.org/10.2174/
1570179413666160921110732.
Kalita, R.D., Nath, Y., Ochubiojo, M.E., Buragohain, A.K., 2013. Extraction and char-
acterization of microcrystalline cellulose from fodder grass; Setaria glauca (L) P.
Beauv, and its potential as a drug delivery vehicle for isoniazid, a first line
antituberculosis drug. Colloids Surf. B Biointerfaces 108, 85e89. https://doi.org/
10.1016/j.colsurfb.2013.02.016.
Karim, M., Chowdhury, Z., Hamid, S., Ali, M., 2014. Statistical optimization for acid
hydrolysis of microcrystalline cellulose and its physiochemical characterization
by using metal ion catalyst. Materials 7, 6982e6999. https://doi.org/10.3390/
ma7106982.
Katakojwala, R., Mohan, S.V., 2019. Microcrystalline cellulose production from
sugarcane bagasse: sustainable process development. J. Clean. Prod., 119342
https://doi.org/10.1016/j.jclepro.2019.119342.
Khan, A., Khan, R.A., Salmieri, S., Le Tien, C., Riedl, B., Bouchard, J., Chauve, G., Tan, V.,
Kamal, M.R., Lacroix, M., 2012. Mechanical and barrier properties of nano-
crystalline cellulose reinforced chitosan based nanocomposite films. Carbohydr.
Polym. 90, 1601e1608. https://doi.org/10.1016/j.carbpol.2012.07.037.
Kro€ger, M., Klemm, M., Nelles, M., 2018. Hydrothermal disintegration and extraction
of different microalgae species. Energies 11, 1e13. https://doi.org/10.3390/
en11020450.
Kundu, D., Banerjee, T., 2020. Development of microcrystalline cellulose based
hydrogels for the in vitro delivery of Cephalexin. Heliyon 6, e03027. https://
doi.org/10.1016/j.heliyon.2019.e03027.
Kusumattaqiin, F., Chonkaew, W., 2015. Preparation and characterization of
microcrystalline cellulose (MCC) by acid hydrolysis using microwave assisted
method from cotton wool. In: Macromolecular Symposia. Wiley-VCH Verlag,
pp. 35e41. https://doi.org/10.1002/masy.201400110.
Liu, Y., Liu, A., Ibrahim, S.A., Yang, H., Huang, W., 2018. Isolation and characterization
of microcrystalline cellulose from pomelo peel. Int. J. Biol. Macromol. 111,
717e721. https://doi.org/10.1016/j.ijbiomac.2018.01.098.
Lu, Q., Lin, W., Tang, L., Wang, S., Chen, X., Huang, B., 2014. A mechanochemical
approach to manufacturing bamboo cellulose nanocrystals. J. Mater. Sci. 50,
611e619. https://doi.org/10.1007/s10853-014-8620-6.
Mandal, A., Chakrabarty, D., 2011. Isolation of nanocellulose from waste sugarcane
H. Holilah, D. Prasetyoko, R. Ediati et al.
bagasse (SCB) and its characterization. Carbohydr. Polym. 86, 1291e1299.
https://doi.org/10.1016/j.carbpol.2011.06.030.
McHugh, T.H., Aujard, J.-F., Krochta, J.M., 1994. Plasticized whey protein edible films:
water vapor permeability properties. J. Food Sci. 59, 416e419. https://doi.org/
10.1111/j.1365-2621.1994.tb06980.x.
Merci, A., Marim, R.G., Urbano, A., Mali, S., 2019. Films based on cassava starch
reinforced with soybean hulls or microcrystalline cellulose from soybean hulls.
Food Packag. Shelf Life 20, 100321. https://doi.org/10.1016/j.fpsl.2019.100321.
Miao, C., Hamad, W.Y., 2013. Cellulose reinforced polymer composites and nano-
composites: a critical review. Cellulose. https://doi.org/10.1007/s10570-013-
0007-3.
Mohammed, M.A.A., Salmiaton, A., Wan Azlina, W.A.K.G., Mohamad Amran, M.S.,
2012. Gasification of oil palm empty fruit bunches: a characterization and ki-
netic study. Bioresour. Technol. 110, 628e636. https://doi.org/10.1016/
j.biortech.2012.01.056.
Mohan Bhasney, S., Kumar, A., Katiyar, V., 2020. Microcrystalline cellulose, polylactic
acid and polypropylene biocomposites and its morphological, mechanical,
thermal and rheological properties. Compos. B Eng. 184, 107717 https://doi.org/
10.1016/j.compositesb.2019.107717.
Moran, J.I., Alvarez, V.A., Cyras, V.P., Va
zquez, A., 2008. Extraction of cellulose and
preparation of nanocellulose from sisal fibers. Cellulose 15, 149e159. https://
doi.org/10.1007/s10570-007-9145-9.
Mukherjee, T., Kao, N., 2011. PLA based biopolymer reinforced with natural fibre: a
review. J. Polym. Environ. 19, 714e725. https://doi.org/10.1007/s10924-011-
0320-6.
Norain, H.F., Kahar, A.W.M., Lilian, L., Salmah, H., Ismail, H., 2019. Enhanced prop-
erties of regenerated cellulose/corn cob biocomposite films. In: Materials
Today: Proceedings. Elsevier Ltd, pp. 2010e2015. https://doi.org/10.1016/
j.matpr.2019.06.085.
Nsor-Atindana, J., Chen, M., Goff, H.D., Zhong, F., Sharif, H.R., Li, Y., 2017. Function-
ality and nutritional aspects of microcrystalline cellulose in food. Carbohydr.
Polym. 172, 159e174. https://doi.org/10.1016/j.carbpol.2017.04.021.
Ohwoavworhua O, F., Adelakun, T., Okhamafe, A., 2009. Processing pharmaceutical
grade microcrystalline cellulose from groundnut husk: extraction methods and
characterization. Int. J. Green Pharm. 3, 97e104. https://doi.org/10.4103/0973-
8258.54895.
Ohwoavworhua, FrankO., Adelakun, T., Okhamafe, A., 2009. Processing pharma-
ceutical grade microcrystalline cellulose from groundnut husk: extraction
methods and characterization. Int. J. Green Pharm. 3, 97. https://doi.org/
10.4103/0973-8258.54895.
Oun, A.A., Rhim, J.W., 2016. Isolation of cellulose nanocrystals from grain straws and
their use for the preparation of carboxymethyl cellulose-based nanocomposite
films. Carbohydr. Polym. 150, 187e200. https://doi.org/10.1016/
j.carbpol.2016.05.020.
Owolabi, A.F., Haafiz, M.K.M., Hossain, M.S., Hussin, M.H., Fazita, M.R.N., 2017. In-
fluence of alkaline hydrogen peroxide pre-hydrolysis on the isolation of
microcrystalline cellulose from oil palm fronds. Int. J. Biol. Macromol. 95,
1228e1234. https://doi.org/10.1016/j.ijbiomac.2016.11.016.
Pachuau, L., Dutta, R.S., Hauzel, L., Devi, T.B., Deka, D., 2019. Evaluation of novel
microcrystalline cellulose from Ensete glaucum (Roxb.) Cheesman biomass as
sustainable drug delivery biomaterial. Carbohydr. Polym. 206, 336e343. https://
doi.org/10.1016/j.carbpol.2018.11.013.
Peterson, A.A., Vogel, F., Lachance, R.P., Fro €ling, M., Antal, M.J., Tester, J.W., 2008.
Thermochemical biofuel production in hydrothermal media: a review of sub-
and supercritical water technologies. Energy Environ. Sci. 1, 32e65. https://
doi.org/10.1039/b810100k.
Prado, K.S., Spinace , M.A.S., 2019. Isolation and characterization of cellulose nano-
crystals from pineapple crown waste and their potential uses. Int. J. Biol.
Macromol. 122, 410e416. https://doi.org/10.1016/j.ijbiomac.2018.10.187.
Reddy, J.P., Rhim, J.W., 2014. Characterization of bionanocomposite films prepared
with agar and paper-mulberry pulp nanocellulose. Carbohydr. Polym. 110,
480e488. https://doi.org/10.1016/j.carbpol.2014.04.056.
Rhim, J.W., Park, H.M., Ha, C.S., 2013. Bio-nanocomposites for food packaging ap-
plications. Prog. Polym. Sci. https://doi.org/10.1016/j.progpolymsci.2013.05.008.
Rodríguez-Ambriz, S.L., Islas-Herna ndez, J.J., Agama-Acevedo, E., Tovar, J., Bello-
Perez, L.A., 2008. Characterization of a fibre-rich powder prepared by lique-
faction of unripe banana flour. Food Chem. 107, 1515e1521. https://doi.org/
10.1016/j.foodchem.2007.10.007.
Rodríguez-Gutie rrez, G., Rubio-Senent, F., Lama-Mun ~ oz, A., García, A., Fern
andez-
Bolan~ os, J., 2014. Properties of lignin, cellulose, and hemicelluloses isolated
from olive cake and olive stones: binding of water, oil, bile acids, and glucose.
J. Agric. Food Chem. 62, 8973e8981. https://doi.org/10.1021/jf502062b.
Rosa, M.F., Medeiros, E.S., Malmonge, J.A., Gregorski, K.S., Wood, D.F.,
Mattoso, L.H.C., Glenn, G., Orts, W.J., Imam, S.H., 2010. Cellulose nanowhiskers
from coconut husk fibers: effect of preparation conditions on their thermal and
morphological behavior. Carbohydr. Polym. 81, 83e92. https://doi.org/10.1016/
j.carbpol.2010.01.059.
Rosa, S.M.L., Rehman, N., De Miranda, M.I.G., Nachtigall, S.M.B., Bica, C.I.D., 2012.
Chlorine-free extraction of cellulose from rice husk and whisker isolation.
Carbohydr. Polym. 87, 1131e1138. https://doi.org/10.1016/j.carbpol.2011.08.084.
15
Journal of Cleaner Production 305 (2021) 127229
Shankar, S., Rhim, J.W., 2016. Preparation of nanocellulose from micro-crystalline
cellulose: the effect on the performance and properties of agar-based com-
posite films. Carbohydr. Polym. 135, 18e26. https://doi.org/10.1016/
j.carbpol.2015.08.082.
Shi, J., Lu, Y., Zhang, Y., Cai, L., Shi, S.Q., 2018. Effect of thermal treatment with water,
H2SO4 and NaOH aqueous solution on color, cell wall and chemical structure of
poplar wood. Sci. Rep. 8, 1e9. https://doi.org/10.1038/s41598-018-36086-9.
Singh, H.K., Patil, T., Vineeth, S.K., Das, S., Pramanik, A., Mhaske, S.T., 2019. Isolation
of microcrystalline cellulose from corn stover with emphasis on its constitu-
ents: corn cover and corn cob. Mater. Today Proc. https://doi.org/10.1016/
j.matpr.2019.12.065.
Spagnol, C., Rodrigues, F.H.A., Pereira, A.G.B., Fajardo, A.R., Rubira, A.F., Muniz, E.C.,
2012. Superabsorbent hydrogel composite made of cellulose nanofibrils and
chitosan-graft-poly(acrylic acid). Carbohydr. Polym. 87, 2038e2045. https://
doi.org/10.1016/j.carbpol.2011.10.017.
Stephens, C.H., Whitmore, P.M., Morris, H.R., Bier, M.E., 2008. Hydrolysis of the
amorphous cellulose in cotton-based paper. Biomacromolecules 9, 1093e1099.
https://doi.org/10.1021/bm800049w.
Sun, C.C., 2008. Mechanism of moisture induced variations in true density and
compaction properties of microcrystalline cellulose. Int. J. Pharm. 346, 93e101.
https://doi.org/10.1016/j.ijpharm.2007.06.017.
Tanpichai, S., Witayakran, S., Boonmahitthisud, A., 2019. Study on structural and
thermal properties of cellulose microfibers isolated from pineapple leaves using
steam explosion. J. Environ. Chem. Eng. 7, 102836 https://doi.org/10.1016/
j.jece.2018.102836.
Tarchoun, A.F., Trache, D., Klapo €tke, T.M., 2019. Microcrystalline cellulose from
Posidonia oceanica brown algae: extraction and characterization. Int. J. Biol.
Macromol. 138, 837e845. https://doi.org/10.1016/j.ijbiomac.2019.07.176.
Thoorens, G., Krier, F., Leclercq, B., Carlin, B., Evrard, B., 2014. Microcrystalline cel-
lulose, a direct compression binder in a quality by design environment - a re-
view. Int. J. Pharm. https://doi.org/10.1016/j.ijpharm.2014.06.055.
Tian, Y., Zhang, K., Zhou, M., Wei, Y., Cheng, F., Lin, Y., Zhu, P., 2019. High-perfor-
mance starch films reinforced with microcrystalline cellulose made from
Eucalyptus pulp via Ball milling and mercerization. Starch - St€ arke 71, 1800218.
https://doi.org/10.1002/star.201800218.
Toor, S.S., Rosendahl, L., Rudolf, A., 2011. Hydrothermal liquefaction of biomass: a
review of subcritical water technologies. Energy 36, 2328e2342. https://
doi.org/10.1016/j.energy.2011.03.013.
Trache, D., Donnot, A., Khimeche, K., Benelmir, R., Brosse, N., 2014. Physico-chemical
properties and thermal stability of microcrystalline cellulose isolated from Alfa
fibres. Carbohydr. Polym. 104, 223e230. https://doi.org/10.1016/
j.carbpol.2014.01.058.
Trache, D., Hussin, M.H., Hui Chuin, C.T., Sabar, S., Fazita, M.R.N., Taiwo, O.F.A.,
Hassan, T.M., Haafiz, M.K.M., 2016. Microcrystalline cellulose: isolation, char-
acterization and bio-composites applicationda review. Int. J. Biol. Macromol.
https://doi.org/10.1016/j.ijbiomac.2016.09.056.
Ul-Islam, M., Khan, T., Park, J.K., 2012. Water holding and release properties of
bacterial cellulose obtained by in situ and ex situ modification. Carbohydr.
Polym. 88, 596e603. https://doi.org/10.1016/j.carbpol.2012.01.006.
Vasconcelos, N.F., Feitosa, J.P.A., da Gama, F.M.P., Morais, J.P.S., Andrade, F.K., de
Souza Filho, M. de S.M., Rosa, M. de F., 2017. Bacterial cellulose nanocrystals
produced under different hydrolysis conditions: properties and morphological
features. Carbohydr. Polym. 155, 425e431. https://doi.org/10.1016/
j.carbpol.2016.08.090.
Vieira, M.G.A., Da Silva, M.A., Dos Santos, L.O., Beppu, M.M., 2011. Natural-based
plasticizers and biopolymer films: a review. Eur. Polym. J. https://doi.org/
10.1016/j.eurpolymj.2010.12.011.
Vijay, R., Lenin Singaravelu, D., Vinod, A., Sanjay, M.R., Siengchin, S., Jawaid, M.,
Khan, A., Parameswaranpillai, J., 2019. Characterization of raw and alkali treated
new natural cellulosic fibers from Tridax procumbens. Int. J. Biol. Macromol.
125, 99e108. https://doi.org/10.1016/j.ijbiomac.2018.12.056.
Xiang, L.Y., Mohammed, M.A., Samsu Baharuddin, A., 2016. Characterisation of
microcrystalline cellulose from oil palm fibres for food applications. Carbohydr.
Polym. 148, 11e20. https://doi.org/10.1016/j.carbpol.2016.04.055.
Xiang, Q., Lee, Y.Y., Pettersson, P.O., Torget, R.W., 2003. Heterogeneous aspects of
acid hydrolysis of a-cellulose. In: Biotechnology for Fuels and Chemicals.
Humana Press, pp. 505e514. https://doi.org/10.1007/978-1-4612-0057-4_42.
Yoon, S.Y., Han, S.H., Shin, S.J., 2014. The effect of hemicelluloses and lignin on acid
hydrolysis of cellulose. Energy 77, 19e24. https://doi.org/10.1016/
j.energy.2014.01.104.
Zain, N.F.M., Yusop, S.M., Ahmad, I., 2014. Preparation and characterization of cel-
lulose and nanocellulose from pomelo (citrus grandis) Albedo. J. Nutr. Food Sci.
5, 10e13. https://doi.org/10.4172/2155-9600.1000334.
Zhang, Q., Lei, H., Cai, H., Han, X., Lin, X., Qian, M., Zhao, Y., Huo, E., Villota, E.M.,
Mateo, W., 2020. Improvement on the properties of microcrystalline cellulose/
polylactic acid composites by using activated biochar. J. Clean. Prod. 252, 119898
https://doi.org/10.1016/j.jclepro.2019.119898.
Zhao, T., Chen, Z., Lin, X., Ren, Z., Li, B., Zhang, Y., 2018. Preparation and character-
ization of microcrystalline cellulose (MCC) from tea waste. Carbohydr. Polym.
184, 164e170. https://doi.org/10.1016/j.carbpol.2017.12.024.