Received: 10 March 2021 Revised: 1 July 2021 Accepted: 13 July 2021

DOI: 10.1002/pc.26232

RESEARCH ARTICLE

Extraction of nanocellulose from sugarcane bagasse and its

characterization for potential applications

R. K. Gond1 | M. K. Gupta1 | Mohammad Jawaid2

1
Department of Mechanical Engineering,
Motilal Nehru National Institute of
Abstract
Technology Allahabad, Prayagraj, Uttar The development of food packaging films from nanocellulose has received
Pradesh, 211004, India
attention from researchers. In this study, the chemo-mechanical method has
2
Laboratory of Biocomposite Technology,
been used to obtain nanocellulose from sugarcane bagasse. The shape and size
Institute of Tropical Forestry and Forest
Products (INTROP), Universiti Putra of the nanocellulose were examined by field emission scanning electron
Malaysia (UPM), Serdang, Selangor, microscopy (FE-SEM) using ImageJ software, and the result was found to be
Malaysia
in good agreement with that of dynamic light scattering (DLS) analysis.
Correspondence Various characterizations such as X-ray diffraction (XRD), Fourier transform
M. K. Gupta, Department of Mechanical infrared (FTIR), thermo-gravimetric analysis (TGA), and ultraviolet visible
Engineering, Motilal Nehru National
Institute of Technology Allahabad, (UV–Vis) spectrometer for the analysis of crystallinity, chemical structure,
Prayagraj 211004, Uttar Pradesh, India. thermal stability, and optical properties were performed for fibers and
Email: [email protected], mnnit.
nanocelluloses. Moreover, antibacterial evaluation was also carried out using
[email protected]
the disc diffusion method with Bacillus and Escherichia coli bacteria. The
removal of noncellulosic elements from fibers after treatment and grinding
was revealed by FTIR analysis. Furthermore, nanocellulose exhibited not only
higher thermal stability and crystallinity but also acceptable antibacterial prop-
erties. Based on the results of this study, polymeric films and composites of the
presented nanocellulose might be utilized in food packaging, plastic
packaging, and container fabrication.

KEYWORDS
antibacterial analysis, characterization, nanocellulose, sugarcane bagasse

1 | INTRODUCTION fibrous residue that can be obtained by crushing the stem

of sugarcane in the mill for the production of juice. Major
In this era, one of the main sources of sugar is sugarcane portion (60%–70%) of bagasse is used as fuel to generate
plant, which account for 60% global sugar production. steam and electricity in power stations, and the
Among all the sugarcane-growing countries, Brazil comes remaining (40%–30%) is used in paper products and alco-
at the first position for maximum production (386.2 mil- hol industries.2,3 Every 10 tons of sugarcane produces
lion ton) with an area of 5.343 million ha, followed by nearly 3 tons of bagasse. Around 54 million tons of dry
India with (289.6 million ton) an area of 4.608 million ha bagasse is produced per year in the world.4 The
and China (92.3 million ton) with an area of 1.328 mil- dry bagasse normally contains a large amount of cellulose
lion ha.1 In India, Uttar Pradesh has the largest area contents, which can be used to produce cellulose-derived
(around 50%) for sugarcane production, followed by products.
Tamil Nadu, Maharashtra, Andhra Pradesh, Karnataka, Cellulose is the most abundant, biodegradable, natu-
Gujarat, Haryana, Bihar, and Punjab. Bagasse is pulpy ral, and renewable polymer existing in hemp, sugarcane

Polymer Composites. 2021;1–13. wileyonlinelibrary.com/journal/pc © 2021 Society of Plastics Engineers. 1

2 GOND ET AL.

bagasse, wood, rice husk, areca nut, bamboo, cotton, and reported by Fourier transform infrared (FTIR) analysis.31
other plant-based materials.5 Recently, nanocellulose Boufi and Chaker32 used a conventional high-speed
from plant fibers have been the center of attraction blender to obtain the nanocellulose of corn stalk. The
among researchers owing to their excellent characteris- nanoscale of nanofibrils with 3 nm width was confirmed
tics such as high specific area, low thermal expansion by atomic force microscopy AFM analysis. Moreover, the
coefficient, low density, and good optical and mechanical enhancement in tensile modulus and strength of its 10 wt
properties.6 These nanocellulosees have been utilized in % reinforced composite was 125 and 10 times higher than
biomedical applications such as tissue engineering, drug that of the polymer matrix. The cellulose nanocrystals
delivery, wound dressings, and medical implants, and (10–25 nm diameter) of grain straw were extracted by
several other applications such as films, paper transistors, acid hydrolysis, and a very high aspect ratio, high crystal-
display devices, e-papers, head phone diaphragms, sensors, linity index, and high thermal stability were reported for
water purifiers, EMI shielding, and flexible light-emitting this.33
diodes.7–16 In packaging applications, the use of non- Ifuku et al.34 obtained the nanofibers with a width of
renewable and nonbiodegradable materials responsible for 10–20 nm using the grinding method under acidic condi-
environmental imbalance is very common. Therefore, there tions. Deep et al.35 isolated banana nanocellulose by
is a great need for renewable and biodegradable materials using the steam explosion technique, and its morphology,
(i.e., biomass/natural fibers) for packaging applications. chemical composition, and thermal properties were char-
Many researchers have proposed the applications of bio- acterized. In another study, the nanocellulose of banana,
mass materials for packaging applications .17–20 Recently, jute, and pineapple was extracted using the steam explo-
the use of polymer (polycaprolactone, polylactic acid, poly- sion technique and was proposed for advanced nanotech-
glycolic acid, polybutylene succinate, polyvinyl alcohol, and nological applications.36 Alemdar and Sain37 obtained
chitosan)-based films and composites of nanocellulose has nanofibers, with a diameter in the range of 10–80 nm,
noticeably been increased in packaging applications from wheat straw using the chemo-mechanical technique.
because of their biodegradability, eco-friendliness, huge Maiti et al.38 used three different materials—China cotton,
availability, and low cost.21–25 Li et al.26 presented a critical South African cotton, and waste tissue papers—to produce
review and suggested that nanocellulose has the potential nanocellulose using the acid hydrolysis technique. They
to be used in packaging applications. In another review, the examined that the length and diameter of the extracted
application of nanocellulose-based films and composites nanocelluloses range from 50 to 200 nm and from 10 to
was proposed for packaging by Ferrer et al..25 Paakko 90 nm, respectively, with higher thermal stability and
et al.27 suggested the applications of microfibrillated cellu- crystallinity. Sodium chlorite, acetic acid, and nitric acid,
lose in packaging, filtration, and high-performance biode- and formic acid methods were carried out to extract the
gradable nanocomposites. jackfruit nanocrystals where the sodium chlorite method
Some researchers have successfully extracted the was found to be the most suitable method for the isolation
nanocellulose/nanocrystals of the various natural fibers of nanocrystals as it produced a maximum yield of cellu-
using the different methods such as chemo-mechanical, lose.39 Abraham et al.40 used a novel eco-friendly method
cryocrushing, and high-speed grinding.28 Mandal and (i.e., steam pretreatment) to extract nanocellulose from
Chakrabarty2 obtained the nanocellulose of bagasse by coir fibers. The diameters of the extracted nanocellulose
acid hydrolysis subjected to characterization by various were found in the range of 5 to 50 nm. Similar extraction
techniques. Thermal stability and the crystalline behavior method was used by Chirayil et al. to obtain the
were found to improve for nanocellulose as compared to nanocellulose from isora fibers .41 This method consisted
other fibers. The same method was also used to isolate of alkali treatment, bleaching, acidic steam treatment, and
the nanocrystalline cellulose of bamboo, and it was found homogenization. In another work, the same method was
that the crystallinity index of nanocrystalline cellulose used to isolate the nanocellulose from pine apple leaf
(71.98%) is higher than that of other non-wood mate- fibers.42
rials.29 Chandra et al.30 achieved the nanocellulose of Oliveira et al.15 used the electrospinning method to
areca nut husk with an average diameter less than 3– develop the bio-tissue based on nanocrystalline cellulose
5 nm and a good aspect ratio ranging 120–150 using the from bamboo fibers. The maximum resistance to traction
chemo-mechanical method. In this work, highly ther- and modulus was shown by the bio-tissue with 5% of cel-
mally stable and crystalline nanofibrils were obtained. lulose. The greater thermal, mechanical, and optical
The cellulose nanocrystals of rice husk with a diameter properties of films based on cellulose nanofibers from
in the range of 10 to 15 nm were obtained by acid hydro- rice straw were reported by Sharma et al..16 Hamawand
lysis, and its crystallinity was found to increase. Further- et al.43 in their review discussed technologies for particle
more, the removal of noncellulosic elements was size reduction and their potential applications. Although
GOND ET AL. 3

many studies have reported on the isolation and charac-

terization of nanocellulose from different natural fibers, a
few works have been carried out on the characterization
of nanocellulose for packaging applications so far. Fur-
thermore, the isolation of the nanocellulose of sugarcane
bagasse using the current method and its antibacterial
analysis are not attempted so far. The fruitful applica-
tions and the value addition of waste sugarcane bagasse
are the purposes of this work. These facts motivated to
extract the nanocellulose from sugarcane bagasse using
the chemo-mechanical method and characterize them for
packaging applications.

2 | MATERIALS AND METHODS

FIGURE 1 Nanocellulose of sugarcane bagasse
2.1 | Materials

Sugarcane bagasse was supplied by K.M Sugar Mills,

Faizabad, Uttar Pradesh, India. The raw bagasse fibers
consist of the following chemical composition and
mechanical properties: hemicellulose (20–25%), cellulose
(45–55%), pectin (0.6–0.8%), lignin (18–24%), and ash (1–
4%); and tensile modulus (17 GPa), tensile strength
(290 MPa), and density (1.25 g/cm3),1,44 respectively.
Sodium hydroxide (NaOH), distilled water, and concen-
trated hydrochloric acid (HCL) were purchased from Sci-
ence Corporation Johnstonganj, Prayagraj, UP, India.
Nutrient Agar (NM011, Sisco Research Laboratories,
India) with a pH of 7.4 ± 0.2 at 25
C was also procured
from Science Corporation Johnstonganj, Prayagraj, UP,
India. The standard culture of Bacillus and Escherichia
coli bacteria was obtained from the Department of Bio-
technology, Motilal Nehru National Institute of Technol-
ogy, Allahabad, Prayagraj—211,004, Uttar Pradesh,
India.
F I G U R E 2 Extraction process for the isolation of the
nanocellulose of sugarcane bagasse
2.2 | Extraction process for isolation of
nanocellulose of sugarcane bagasse
the alkali-treated fibers were soaked in 1 mol HCL solu-
Initially, sugarcane bagasse fibers (SBFs) were washed tion, prepared by adding 88.28 ml of HCl into 911.72 ml
with distilled water to remove any type of sugar present of distilled water, for 4 h at 80
C to break up the cell
on their surface; subsequently, it was air-dried in hot air walls and thus to separate microfibrils. Furthermore, the
oven for 24 to 36 h at 80
C to remove the absorbed mois- fibers were washed well with deionized water to get rid
ture. Dried SBFs were soaked in 15 wt% NaOH solution of acid HCL, and then, the treated SBFs were again
for 4 h at room temperature.28 After this treatment, SBFs soaked in 2 wt% NaOH solution for 4 h at 80
C to remove
were washed several times with distilled water to eradi- the remaining noncellulosic constituents. Finally, the
cate the rest of alkali from the fibers. This process is treated fibers were washed and dried in hot air oven.
effective to increase surface roughness and also to remove These fibers were ground using a high-speed grinder at a
the certain amount of lignin, wax, and natural oils that revolution of 1500 rpm for 20 min to obtain the
covered the external surface of the fibers. After this, SBFs nanocellulose of SBFs. Figure 1 shows the developed
were air-dried in hot air oven at 80
C for 24 h, and then, nanocellulose of SBFs, whereas Figure 2 shows the
4 GOND ET AL.

flowchart of the extraction process of the nanocellulose The little amount of nanocellulose of SBFs was well
of sugarcane bagasse. To isolate the nanocellulose from distributed in distilled water using a magnetic stirrer, and
natural fibers for packaging applications, some other pro- its average size was measured using DLS (model: DLS,
cesses such as steam explosion,45 microgrinding,46 Microtrac MRB's, Nanotrac Wave, USA).
microfluidization,47 and high-intensity ultrasonication48
were also used by the researchers instead of the chemo-
mechanical process. Notation used for fibers at various 3.3 | Morphology analysis
stages of treatments is provided as follows:
BF = untreated bagasse fiber, BF(T1) = 15% NaOH- Field emission scanning electron microscope (FE-SEM)
treated bagasse fiber, BF(T2) = 15% NaOH + HCl (model: Nova Nano SEM 450) was used for the morphologi-
(1 mol)-treated bagasse fiber, BF(T3) = 15% NaOH + cal analysis at a low range of accelerating voltage (i.e., 3–
HCl (1 mol) + 2% (NaOH)-treated bagasse fiber, and 5 kV) to obtain a good image resolution. These images were
NBF = bagasse nanocellulose. captured using secondary electrons emitted from the sur-
face of the fibers. The surface of the specimen was sputter-
coated with gold nanoparticles prior to observation.
3 | C H A R A C T E R I Z A T IO N

3.1 | Yield measurement 3.4 | X-ray diffraction (XRD)

The yield percentage of nanocellulose was measured X-ray diffractometer (model: SmartLab 3KW, Rigaku,
using the gravimetric analysis method. The yield in terms Tokyo, Japan) with Cu Kα radiation at 40 kV operating
of percentage of nanocellulose was computed using the voltage was used to obtain the XRD patterns of fibers and
following Equation (1)42,49: nanocellulose. The diffraction intensities were recorded
between 5 and 65
(2θ angle range) at a rate of 3
per
M2 minute.
Yield ð%Þ ¼  100 ð1Þ
M1 The crystallinity index was calculated using the Segal
method by the following Equations (3 and 4)50:
where M2 is the weight of the final dry nanocellulose and
M1 is the weight of the initial dry SBF. I200  Iam
Crystallinity index ð%Þ ¼  100 ð3Þ
I200

3.2 | Dynamic light scattering (DLS) Degree of crystallinity ¼

I200
ð4Þ
I200 þ Iam
DLS also called photon correlation spectroscopy is
one of the most suitable methods to measure the where I200 is the maximum intensity of the diffraction
nanosize of the particles and has been commonly used peak, and Iam is the intensity of the amorphous diffraction
to measure the nanosize of the cellulose of natural peak. The mean crystal size (in nm) was measured using
fibers. The particle size can be obtained by measuring Scherrer Equation (4) to analyze the effect of the chemical
the random changes in the intensity of light scattered treatments on the crystalline structure of the fibers.51
from a suspension/solution. Small particles in suspen-
sion undergo Brownian motion, which is modeled by kλ
Crystal size D ¼ ð5Þ
the Stokes–Einstein Equation (2). The Stokes– βcosθ
Einstein relates the diffusion coefficient measured
using DLS with the particle size, which is given as where k is the dimensionless shape factor, which is gen-
follows: erally taken as 0.9, λ (1.54Å) is the X-ray wavelength, β is
the full width at the half maximum peak, and θ is the dif-
kT fraction or Braggʼs angle at the peak (200).
D1 ¼ ð2Þ
3πηD2

where D1 is the particle size, D2 the translational diffu- 3.5 | FTIR spectroscopy
sion coefficient (can be determined by the DLS method),
k the Boltzmann's constant, T the temperature, and η the FT-IR spectra of the untreated, treated, and nanocellulose
dynamic viscosity. of SBFs were obtained using FT-IR spectroscope (model:
GOND ET AL. 5

Bruker, Tensor-27), in the range of 500–4000 cm1 with nanocellulose. The conversion of absorbance into transmit-
the resolution of 5 cm1. The peak heights were visible tance was carried out using the following equation:
from the absorbance spectra because of the presence of
different functional groups at the different wavenumbers. Absorbance
¼ 2  logð%TransmittanceÞ ð6Þ

3.8 | Antibacterial analysis

3.6 | Thermogravimetry analysis (TGA)
The antibacterial analysis of the nanocellulose of sugar-
TGA was carried out to study the degradation characteris- cane bagasse was performed against Bacillus and E. coli
tics with the temperature of untreated, treated, and bacteria by the disc diffusion method as shown in
nanocellulose of SBFs. The test was performed using TGA Figure 3. To prepare the agar medium, 2.8 g of agar nutri-
instrument (model: TGA, STA 409 Netzsch) at a tempera- ent was uniformly mixed into 100 mL of distilled water;
ture range of 30 to 500
C with a heating rate of 10
C/min subsequently, this mixture was placed in an autoclave at
in nitrogen atmosphere using 3–5 mg of dried samples. a pressure of 15 psi and a temperature of 121
C for 4 h
for sterilizing. Then, it was poured into the sterile Petri
dish of 90 mm diameter at ambient temperature for
3.7 | Optical properties 15 min to get solid. In a laminar box, fresh bacterial cul-
tures of E. coli and Bacillus were spread over the whole
The optical property was measured from 190 to 700 nm media; 18 mg of nanocellulose was poured into a punc-
using UV–Vis spectrometer (model: Shimadzu, UV-3600 tured disc of 5 mm developed using a micropipette tube.
Plus), in which, the wavelength of barium sulphate (BaSO4) Finally, the sealed Petri dish was incubated at 40
C for
set as a reference for optical absorbance of the fibers and 18 h. After incubation, the plate is allowed for the

F I G U R E 3 Antibacterial
analysis process for the
nanocellulose of sugarcane
bagasse
6 GOND ET AL.

found in the nanorange credited to homogeneous distri-

bution. This evidenced the effectiveness of the chemo-
mechanical method in isolating nanocellulose from SBFs.
The nanoparticles with a size of around 60 nm have the
maximum pass percentage (78%), whereas those with a
size of around 90 nm has the minimum pass percentage
(4%). This fact shows that nanoparticles with a larger size
close to 100 nm are less and ignorable. In addition, the
calculated average size for NBF is 53 nm. This method
has been already adopted by researchers to measure the
F I G U R E 4 Particle size distribution of the nanocellulose of nanosize of isora fibers,41 areca nut husk fibers,53 and
sugarcane bagasse pineapple leaf fibers.54

examination of inhibition zone growth. The microorgan- 4.3 | Morphology analysis

ism growth was observed by measuring the diameter of
the inhibition zone. The percentage of the inhibition Figure 5(A–E) shows the FE-SEM images of BF, BF (T1),
zone was calculated using the following equation49: BF (T2), BF (T3), and NBF, respectively. Figure 5(A)
shows the larger diameter of the untreated SBF, which
Diameter of the inhibition zone ðmmÞ might be due to the presence of many microfibrils. Mean-
Inhibition zone ð%Þ ¼  100
Diameter of the Petri disc ðmmÞ while, the smooth surface of the untreated fiber is evi-
ð7Þ dence of the presence of lignin, wax, and oil. Figure 5(B)
shows the partial removal of the surface impurities such
as hemicelluloses and lignin along with defibrillation
4 | R ES U L T S A N D D I S C U S S I O N after alkali treatment.55,56 Figure 5(C) shows the further
defibrillation of bagasse fibers by acid treatment. The
4.1 | Yield analysis reduced diameter and increased surface roughness by
the removal of noncellulosic elements can be seen from
To calculate the yield percentage, an initial weight of sug- Figure 5(D). The reduced diameter is due to the hemicel-
arcane the bagasse fiber was taken as 25 g. The final luloses soluble in water after hydrolysis.53 A high-magni-
weight of the nanocellulose obtained after several pro- fication image of NBF in a cylindrical shape with an
cesses was 16.25 g. Therefore, the calculated yield per- improved surface area to volume ratio can be seen from
centage of the presented nanocellulose was 65%. The Figure 5(E). Thus, the morphological analysis supports
alkali treatment was efficient to eliminate the hemicellu- the fact that nanoparticles could be isolated from sugar-
lose from sugarcane bagasse, whereas most of the hemi- cane bagasse with the method presented in this study.
cellulose and lignin were removed after acid treatment. The length, diameter, and aspect ratio of the isolated
Finally, complete removal of hemicellulose and lignin nanocellulose shown in Figure 5(E) were measured using
can be assumed after the alkali treatment and grinding ImageJ software. Ten particles were selected randomly
process. The maximum yield percentage of 48% for for the measurement of their length, diameter, and aspect
nanocellulose from the isora fiber was reported.41 In ratio, and their measured values are presented in Table 1.
another study, the maximum yield percentage of 24.5% The average values of diameter, length, and aspect ratio
for nanocellulose from the bamboo fiber was reported.52 were 57.07, 364.84, and 6.97 nm, respectively. The aspect
ratio reveals the reinforcing aptitude of the nanocellulose
to prepare its composites/films for various applications.53
4.2 | DLS analysis A good aspect ratio of the presented nanocellulose indi-
cates its better capacity for reinforcement.
The DLS technique has been popularly used to find the
statistical distribution of the nanosize particles. It gives
information about the variation in pass percentage 4.4 | Fourier-Transform (FTIR) infrared
(Q0) with respect to variations in diameters (d) of the spectroscopy analysis
particles. The particle size distribution of NBF was ana-
lyzed using the DLS analyzer, which is shown in This analysis was carried out to study the change in the
Figure 4. It can be observed that all the particles were chemical structure by recognizing the functional group
GOND ET AL. 7

F I G U R E 5 FE-SEM images of sugarcane bagasse: (A) untreated, (B) alkali-treated, (C) alkali- and acid-treated, (D) alkali-, acid-, and
alkali-treated, and (E) nanocellulose

T A B L E 1 Particle size and aspect ratio of nanocellulose of

sugarcane bagasse

Length (nm) Diameter (nm) Aspect ratio (L/D)

570.355 51.088 11.164
288.78 48.374 5.969
277.074 69.26 4.000
295.237 75.717 3.899
273.066 83.193 3.282
319.862 63.285 5.054
780.767 44.598 17.506
272.406 46.573 5.849
309.015 51.088 6.048
261.895 37.59 6.967
F I G U R E 6 FTIR spectra of untreated, treated, and
nanocellulose of sugarcane bagasse

present in the fibers after treatments and mechanical have main constituents (i.e., cellulose, hemicelluloses, and
grinding. The FTIR spectra of BF, BF (T1), BF (T2), BF lignin), which are composed of alkanes, ketones, esters,
(T3), and NBF are shown in Figure 6. The natural fibers alcohols, and aromatics with distinct oxygen-containing
8 GOND ET AL.

functional groups.57,58 There were mainly two absorption

regions: low-wavelength region (560–1731 cm1) and
high-wavelength region (2912–3342 cm1). A broad
absorption band in the region of 3333–3342 cm1
observed for all the fibers represents the O H stretching
vibration of the hydroxyl group in cellulose, hemicellu-
loses, and lignin.55,58,59 The absorption band in the range
of 2911–2916 cm1 seen in the spectra for all types of
fibers shows the C H group of cellulose, hemicellu-
lose, and lignin .59,60 The peaks at 1732 cm1 observed in
the spectrum of the untreated fiber is due to the C═O
stretching vibration of the acetyl and uronic ester group
of lignin and hemicelluloses.39 This peak was absent in
the treated fiber and nanocellulose, indicating the
removal of hemicelluloses and lignin. The peaks at 1238,
F I G U R E 7 XRD graph of untreated, treated, and nanocellulose
1370, and 1512 cm1 seen in the untreated fiber corre-
of sugarcane bagasse
spond to the aromatic skeletal vibration of lignin.53 The
peak at 1238 cm1 found to be disappeared in the treated
fiber and nanocellulose shows the complete removal of enhancement in cellulose after the removal of the amor-
lignin. The most significant absorption band at phous fraction of SBFs.
1035 cm1 found to be continuously increased from the The values of the degree of crystallinity, crystallinity
untreated fiber to nanocellulose revealed an improve- index (%), and crystallite size measured using X-ray dif-
ment in the cellulose and surface area of fraction curves are presented in Table 2. Stiffness and
nanocellulose.53,59,60 strength of the fibers can be predicted on the basis of
The peak at 1600–1640 cm1 in the untreated fiber their crystallinity index.63 The highest value (58.32%) and
shows the O H bending vibration of absorbed moisture lowest value (32.41%) of crystallinity index were found
due to the hydrophilic nature of SBFs.53,61 This peak found for NBF and BF, respectively, whereas intermediate
to be shifted in treated fibers and disappeared in values were shown using the treated fibers. There was an
nanocellulose shows the reduced tendency of water uptake improvement in the crystallinity index from 32.41% to
after treatments and grinding.53 The peak at around 53.02% after alkali and acid treatment. Further improve-
3342 cm1 could be due to the stretching vibration of ment in the crystallinity index up to 58.32% could be
O H and was found to be minor shifted towards a high credited to the mechanical grinding process resulting in
wavelength in treated fibers and nanocellulose also shows the breaking of the amorphous region and enrichment of
the decrease in moisture absorption.39,40,53 the crystalline region. The highest crystallinity for NBF
could be due to the formation of H-bonding, which
opposes the free movement of cellulose chains,
4.5 | X-ray diffraction analysis which results in the parallel arrangement of crystalline
structure .53,61 In the case of BF, the lowest crystallinity
Cellulose, hemicellulose, and lignin are the main constitu- was seen ascribed to the presence of noncellulosic ele-
ents of sugarcane bagasse, where cellulose has a crystalline ments.53 Furthermore, the alkali-treated fiber exhibited
structure due to its H-bonding, and hemicelluloses and lig- higher intensity, crystallinity index, and degree of crystal-
nin are amorphous in nature.31 Chemical treatments and linity than the untreated fiber because of the exclusion of
mechanical processes highly affected the crystallinity of amorphous noncellulosic elements. The improvement in
the fibers.40 The X-ray diffraction patterns in terms of crystallinity by alkali treatment was also reported in liter-
intensity versus 2θ of untreated, treated, and nanocellulose ature.61 The crystalline behavior of fiber BF (T2) was
of SBFs are shown in Figure 7. The highest values of inten- found to be higher than that of BF(T1) because of hydro-
sity, crystallinity index (%), and degree of crystallinity of lysis. An enhanced crystallinity index improves the ten-
BF, BF (T1), BF (T2), BF (T3), and NBF were found sile strength and modulus, and rigidity because of the
between 22 and 24
of the 2θ with different intensities. highly aligned and more compact structure among cellu-
Normally, a typical cellulose I exhibits a sharp diffraction lose molecules.63 Similar results for the crystalline behav-
peak at 22
of 2θ.62 This fact shows the presence of cellu- ior after acid treatment is already reported.47,48 The
lose I in all types of the fibers. An expected shoulder peak crystalline behavior of fiber BF (T3) was found to be
at 16
of 2θ was also observed for nanocellulose owing to higher than that of BF (T1) and BF(T2) because of the
GOND ET AL. 9

T A B L E 2 XRD results of untreated,

Sample Crystallinity index (%) Degree of crystallinity Crystal size (nm)
treated, and nanocellulose of sugarcane
bagasse BF 32.41 0.59 3.74
BF(T1) 41.32 0.63 2.47
BF(T2) 51.72 0.67 2.47
BF(T3) 53.02 0.69 2.47
NBF 58.32 0.72 3.48

further removal of noncellulosic elements and increase in

the aspect ratio of the fibers. Thus, it can be suggested
that the degree of crystallinity was found to improve after
treatments, and its maximum value was found for the
NBF as expected. The higher value of the crystal size was
seen in untreated fibers, whereas the lower but almost
same value was seen in treated fibers and nanocellulose.

4.6 | Thermogravimetric analysis (TGA)

The percentage change in weight with the temperature of

untreated, treated, and nanocellulose of SBFs examined
by TGA is illustrated in Figure 8, whereas the thermal
data were summarized in Table 3. The decomposition of
different components of natural fibers occurs at different F I G U R E 8 TGA graph of untreated, treated, and nanocellulose
temperatures: cellulose from 315 to 400
C, hemicellu- of sugarcane bagasse
loses from 220 to 315
C, and lignin from 200 to
700
C.58,64 In initial low temperature weight loss (up to
100
C), the maximum and minimum weight losses
against temperature were provided by BF (T1) and NBF, T A B L E 3 TGA results of untreated, treated, and nanocellulose
respectively, whereas intermediate weight losses were of sugarcane bagasse
shown by treated fibers BF (T1), BF (T2), and BF (T3). At Weight loss (%)
this stage, the highest thermal stability was shown by
Temperature BF Bf(T1) Bf(T2) Bf(T3) NBF
NBF. The initial weight loss was owing to the evapora-
tion of loosely bound moisture from the surface of the 50 3.98 5.83 3.87 4.07 2.74
SBFs.65–67 The highest weight loss for BF (T1) was attrib- 100 8.08 13.05 7.45 8.21 6.57
uted to the elimination of amorphous hemicelluloses and 200 8.67 15.77 7.65 9.90 7.42
lignin. The second stage of degradation occurs above 300 29.40 48.73 17.72 36.80 42.58
200
C owing to the degradation of SBF constituents such
400 77.45 61.72 73.48 70.96 72.16
as cellulose, hemicelluloses, and lignin.65–67 At 200
C,
500 93.60 66.49 86.30 82.44 85.98
minimum and maximum weight losses were shown by
NBF and BF (T1), respectively. At above 400
C tempera-
ture, minimum weight loss was shown by BF
(T1) followed by BF (T3) and NBF. From 500
C tempera- noncellulosic materials. However, the thermal stability of
ture onwards, the highest weight loss was shown by BF the isolated nanocellulose in this work was considerably
and BF (T2), followed by NBF, whereas better thermal great, which may broaden their suitability for its bio-
stability was shown by the treated fibers. According to lit- composites at high-temperature applications.
erature, lower thermal stability of nanocellulose than
that of the raw fiber had been reported due to the
removal of protective lignin and hemicellulose acting as a 4.7 | Optical properties
thermal barrier.68 The better thermal stability at evalu-
ated temperature of treated fibers could be due to the Optical properties of nanocellulose are one of the impor-
dense structure of cellulose formed by the exclusion of tant analyses for packaging applications, which is
10 GOND ET AL.

measured by UV–Visible spectroscope.69 It measures the to humans. All the fibers observed the higher amount of
intensity of light passing through a nanoparticle sample incident radiation at a wavelength of 240 nm, which is
and compares it with the intensity of incident light. UV very useful for packaging applications. On the other
light, having wavelength ranged from 10 to 400 nm, hand, a minimum amount of radiations was absorbed by
occurs between visible light and X-rays in the electro- the fibers at a wavelength of 600 nm, as shown in Table 4
magnetic spectrum. Due to its shorter wavelength, it can- and Figure 9(A). The absorbance peak of NBFs was lesser
not be seen by human eyes. The UV absorbance and than the other treated fibers, which indicates that NBF
transmittance spectra were measured from 200 to 700 nm was allowing to pass minimum amount of light. This
wavelength range, and the corresponding results are result might be due to the nanosize of cellulose with a
illustrated in Figure 9(A,B), respectively. At wavelengths higher surface area. On the other hand, the highest
of 240 nm (UV region) and 600 nm (visible region), the absorbance was shown by the untreated bagasse fiber,
absorbance and transmittance percentage for all due to the presence of cemented elements
the fibers and nanocellulose is also presented in Table 4. (i.e., hemicelluloses and lignin), followed by the treated
UV-C light ranging from 100 to 290 nm is very harmful fiber. Treated fibers exhibited the intermediate absor-
bance as a result of the removal of noncellulosic elements
by treatments. The absorbance spectra of all the fibers
and nanocellulose were found in the following order:
NBF < BF (T3) < BF (T2) < BF (T1) < BF. At the wave-
lengths of 240 and 600 nm, the percentage of transmit-
tance by fibers was found in the range of 5–14 and 43–65,
respectively. At every wavelength, minimum transmit-
tance was shown by the untreated fiber and higher was
shown by the treated fiber and nanocellulose as expected.
Another reason for the variation in transmittance might
be the variation in size of the particles. As per Rayleigh's
scattering theory, a scattering cross section is directly pro-
portional to the diameter of particles.

4.8 | Antibacterial analysis

Antibacterial analysis of the prepared nanocellulose with

E. coli and Bacillus bacteria was carried out using the disc
diffusion method. Figure 10 a shows the presence of
nanocellulose in the agar medium without any bacteria,
whereas Figure 10(B,C) shows the presence of E. coli and
Bacillus bacteria in the agar medium, respectively. The
diameter of the inhibition zone and the percentage of
F I G U R E 9 Optical properties of untreated, treated, and the inhibition zone were provided by these bacteria. . The
nanocellulose of sugarcane bagasse; (A) absorbance and sensitivity of antibiotic can be observed by measuring its
(B) transmittance diameter of the inhibition zone. No inhibition zone was

TABLE 4 Optical properties of untreated, treated, and nanocellulose of sugarcane bagasse

Absorbance (a.u) Transmittance (%)

Sample Wavelength (240 nm) Wavelength (600 nm) Wavelength (240 nm) Wavelength (600 nm)
BF 1.28 0.48 5.26 42.65
BF(T1) 1.19 0.42 8.87 47.86
BF(T2) 0.90 0.36 13.09 56.23
BF(T3) 0.88 0.27 14.52 62.08
NBF 0.55 0.17 13.89 65.31
GOND ET AL. 11

F I G U R E 1 0 Antibacterial
result of nanocellulose of sugarcane
bagasse: (A) without any bacteria,
(B) inhibition zone with E. coli, and
(C) inhibition zone with bacillus

found in the absence of bacteria as shown in Figure 10 roughness of fibers and a reduction in the diameter were
(A), whereas good inhibition zones could be seen in the confirmed by the FE-SEM analysis. FTIR revealed con-
presence of bacteria as depicted in Figure 10(B,C). siderable changes in chemical structures and exclusion of
The diameter of the inhibition zone increased to 14.6 mm noncellulosic constituents after the treatments. A higher
(for E. coli) from 12.2 mm (for Bacillus), whereas the per- degree of crystallinity and crystallinity index was seen for
centage of the inhibition zone increased to 16.22 (for the nanocellulose of bagasse, as confirmed by the XRD
E. coli) from 13.55 (for Bacillus). Thus, it can be con- analysis. Moreover, a better thermal stability and accept-
cluded that E. coli is more sensitive than Bacillus as it has able antibacterial performance were also shown by the
a larger inhibition zone. The variation in the susceptibil- nanocellulose. Based on higher crystallinity and thermal
ity of these bacteria could be due to the variation in their stability, the lowest UV absorption capacity, and excel-
composition and structure of the membrane cell wall. lent antibacterial performance, the isolated nanocellulose
The outer membrane acts as a protective layer, which can be considered appropriate reinforcement to develop
opposes the entrance of antibacterial agents. This result the polymeric composites and films for packaging
shows that the nanocellulose of sugarcane bagasse has applications.
superior antibacterial properties, desired in various pack-
aging application, due to the large specific surface area of ACKNOWLEDGMENT
nanocellulose having spherical/rod likes structure. As per The financial support for this work is provided by CST-
the standard (SNV 195920–1992) of the antibacterial test, UP, India.
materials having >1 mm inhibition zone could be consid-
ered as a good antibacterial agent.70Thus, on the basis of ORCID
the result of antibacterial analysis, it can be concluded M. K. Gupta https://orcid.org/0000-0001-6174-364X
that the presented nanocellulose may be considered as
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