Biomass Conversion and Biorefinery

https://doi.org/10.1007/s13399-022-03171-z

ORIGINAL ARTICLE

Colocasia esculenta stems for the isolation of cellulose nanofibers:

a chlorine‑free method for the biomass conversion
C. D. Midhun Dominic1 · S. Maheswary1,2 · K. V. Neenu3 · S. Mohammad Sajadi4,5 · Derval dos Santos Rosa6 ·
P. M. Sabura Begum3 · Mariya Mathew1 · T. G. Ajithkumar7 · Jyotishkumar Parameswaranpillai8 ·
Tresa Sunitha George9 · V. C. Resmi10 · R. A. Ilyas11,12 · Michael Badawi13

Received: 6 May 2022 / Revised: 18 July 2022 / Accepted: 31 July 2022

© The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature 2022

Abstract
The reuse of waste products is the green key to sustainability. The extraction of cellulose nanofibers from Colocasia esculenta
stems is presented in the paper. The study proved that the waste biomass could be effectively re-engineered into highly valued
cellulose nanofibers (CNFs). Cellulose nanofibers were extracted via a chemo-mechanical route. The pre-treatments included
mild alkali hydrolysis (2% NaOH) and chlorine-free bleaching (peroxide bleaching in an alkaline medium). Cellulose ­Iβ
structure was confirmed using 13C solid-state nuclear magnetic resonance spectroscopy and X-ray diffraction analysis. The
elemental analysis of CNFs detected the elements, carbon and oxygen. The CNFs had a crystallinity and transmittance of
71.72% and 60%, respectively. Microscopic studies verified the elimination of non-cellulosic components and the fibrous
nature of CNFs. Moreover, the fiber diameter of CNFs was 20–40 nm. Thermal analysis revealed good thermal stability
of 335.8 °C ­(T50) for nanofibers. Long-term aids are numerous in eco-friendly technology. Developing an eco-design will
support zero waste ideals, lowers carbon dioxide emissions, and encourages a circular economy. Owing to the merits of
natural fibers, they can be adopted in various sectors including packaging, automobile, aerospace, electronics, biomedical,
construction, and furniture.

Keywords Acid hydrolysis · Cellulose nanofibers · Colocasia esculenta stems · High crystallinity

8
* C. D. Midhun Dominic Department of Science, Faculty of Science & Technology,
[email protected] Alliance University, Chandapura‑Anekal Main Road,
Bengaluru, Karnataka, India Pin‑562106
1
Department of Chemistry, Sacred Heart College 9
Department of Chemistry, St. Paul’s College,
(Autonomous), Kochi, Kerala, India Pin‑682013
Pin‑683503 Kalamassery, Ernakulam, Kerala, India
2
Department of Chemistry, PSGR Krishnammal College 10
Department of Chemistry, SNM College, Maliankara, Kerala,
for Women, Tamil Nadu, Coimbatore, Peelamedu,
India Pin‑683516
India Pin‑641004
11
3 School of Chemical and Energy Engineering, Faculty
Department of Applied Chemistry, Cochin University
of Engineering, Universiti Teknologi Malaysia, UTM,
of Science and Technology (CUSAT), Kochi, Kerala,
81310 Johor Bahru, Malaysia
India Pin‑682022
12
4 Centre for Advanced Composite Materials (CACM),
Department of Nutrition, Cihan University-Erbil, Kurdistan
Universiti Teknologi Malaysia, UTM, 81310 Johor Bahru,
Region, Erbil, Iraq
Malaysia
5
Department of Phytochemistry, SRC, Soran University, KRG, 13
Laboratoire de Physique Et Chimie Théoriques, UMR CNRS
Iraq
7019, Université de Lorraine, Nancy, France
6
Center for Engineering, Modeling and Applied Social
Sciences (CECS), Federal University of ABC (UFABC),
Santo André, São Paulo, Brazil
7
Central NMR Facility and Physical/Materials Chemistry
Division, CSIR-National Chemical Laboratory, Pune,
India Pin‑411008

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 Biomass Conversion and Biorefinery

1 Introduction Natural fibers have gained importance owing to the fact,

they are successful substitutes for synthetic fibers in all
The future wave is on sustainability. In many domains regards including abundance, safety, durability, affordabil-
of modern industry, the quest for eco-efficient products ity, and reinforcing capability [10, 11]. Thyavihalli et al.,
is eroding the allure of synthetic materials. From recent reviewed several natural fibers as an ecofriendly equiva-
research, undeniably bio-compactable products are a step lent to synthetic fibers for polymer composites [12]. It was
ahead in grabbing markets. The environmental issues of concluded that the degradation property of natural fibers
synthetic-based materials necessitate revolutionary bio- compared to conventional fillers diminishes the deleterious
material breakthroughs. Indeed, a fossil-free future is impact on the environment. As natural fibers are from renew-
required for positive environmental change. As a ubiq- able sources, sustainable development can be fully realized
uitous organic compound, the use of cellulose for value- by their practical application. Cellulose, the primary con-
added products increases the wide prospects of a green era. stituent of cell walls, has unique properties that make it an
The occurrence of cellulose in living terrestrial biomass inevitable modern material. For potential use, the effective
and bio-residues can be considered as a portfolio of per- molecular interactions and glycosidic linkages of cellulose
manency. The isolation of nanocellulose from a variety of moieties have been intensively investigated. Cellulose is usu-
bio-resources is one of the current study fields, highlight- ally bound along with hemicellulose and lignin, and the rea-
ing the diversity of natural sources in this biosphere. Being son behind the stiffness and strength of these natural fibers
a perfect precursor, entire plant parts can be considered for is due to the presence of cellulose chains [13]. Nano-dimen-
the isolation of nanocellulose. sional extraction of cellulose gives nanocrystals, nanofibers,
Herein, the study investigates the utilization of Colo- and nanowhiskers with enhanced features compared to the
casia esculenta stems in the isolation of nanocellulose. native fibers. Nanocellulose outperforms other biopolymers
Colocasia esculenta is a tropical plant species belonging owing to their renewability, biodegradability, non-toxicity,
to the Araceae family and is widely available all over the crystallinity, cost efficiency, environmental friendliness,
world. It is the world’s fourth most consumed root crop by lightweight nature, high aspect ratio, physico-mechanical,
weight behind cassava, potato, and sweet potato, with an and thermal properties [14]. As enormous features subsist
annual production of around 229,088 tones [1]. It is usually in a single material, they have a superior stand in various
cultivated for its nutritious leaves and to obtain 70–80% of fields such as polymer composites, drug delivery, cosmetics,
starch from its corms [2]. The cultivation of staple crops automotive parts, packaging films, electronics, solar cells,
like Colocasia esculenta can generate waste, especially its smart films, and bio-printing [15].
stem since its leaves and tubers are used for consumption. As the reinforcing ability of these natural fibers greatly
To a limited extent, agricultural residues can be considered depends on their morphology and the employed method-
as a good source to increase the fertility of the soil. On the ology, caution should be there while choosing a method
other hand, these wastes may pose a substantial hazard to of synthesis. Strong acid hydrolysis is usually adopted to
human health due to environmental contamination. Unfor- isolate nanocellulose from native fiber because of its easy
tunately, poor waste management can cause major financial processing, simplicity, and remarkable efficiency. The grow-
loss to small-scale farmers. India is one of the major coun- ing apprehension of environmental security demands the
tries having Colocasia esculenta cultivation and the waste replacement of strong acids with green methods. Mechanical
that arises after this farming should be properly utilized. methods, enzyme hydrolysis, ionic liquids, mild acid treat-
Accordingly, this research offers a superior option for the ments, and other similar methods are more environmentally
complete utilization of this biomass and its conversion to friendly. By utilizing low-energy technology (grinding and
a value-added product with rich applicability. It is worth homogenization), Zhuo et al. produced nanocellulose from
noting that the idea of utilizing these non-wood stems as a Amorpha fruticosa with a dimension of 10 nm and thermal
feedstock for nanocellulose is a great drive that reduces the stability of 337 °C [16]. The use of endoglucanase enzyme
agro-waste produced after the harvesting of the colocasia as a green method for the isolation of nanocellulose is pre-
tubers. Even though different biomass is widely explored sented by Filson et al. [17]. The milder and greener acid
for nanocellulose isolation, the study on the isolation of hydrolysis alone is not effective in producing nanocellulose
nanofiber from secondary/waste fibers is seldom reported. from a native source. As a result, for improved isolation,
The other reported isolation of nanocellulose from the stem mechanical aid should be provided. Ravindran et al. used
in recent literatures are cassava stem [3], Cardiospermum lime juice as a green alternative to conventional acids for
halicacabum stem [4], Shwetark stem [5], Cola lepidota the isolation of nanocellulose assisted with ball milling.
stem [6], pineapple stem [7], banana pseudo-stem [8], The study produces nanofibers with a thermal stability of
water hyacinth stem [9], and so on. 344.5 °C and a crystallinity of 77% [18]. Chirayil et al. con-
veyed the efficacy of steam explosion along with oxalic acid

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Biomass Conversion and Biorefinery

hydrolysis in the isolation of nanofibers from the Helicteres were purchased from Nice Chemicals Private Limited,
isora plant. The study reported nanofibers with a crystallin- Kerala, India.
ity of 90%, diameter of 10–20 nm, and good thermal stability
using this approach [13]. Steam explosion is an effective
method for the isolation of nanofibers with a high aspect 2.2 Methods
ratio and good yield. This approach aids as a perfect solu-
tion for reduced thermal stability and crystallinity by using 2.2.1 Pre‑treatments
strong acid hydrolysis and high mechanical stress. Conse-
quently, cellulose nanofibers from lignocellulosic biomass The raw sample was prepared from the stems of Colocasia
were successfully isolated via acid-coupled steam explosion. esculenta. The stems were separated from the collected
The treatment of raw material in high pressurized steam and plant and cut into small fragments of length 2 cm each.
the explosion (decompression or pressure release) causes The separation was then followed by water washing for
mulching (fiberization) as a result of chemi-mechanical four times to get rid of the impurities stuck on the stems.
combination [19]. The sample was dried with an air oven (Rotek Plus, India),
Creating a green circle with waste natural materials at 100 °C for 4 days. Finally, the cut dried stems were
can significantly minimize pollution. As environmentally made into granulated powder using a grinder (Sujata Pow-
friendly products are green building blocks for eco-secu- ermatic Plus 900 W 900 Mixer Grinder) and stored. From
rity, endorsing these products will be a safeguard for our the prepared sample, further steps were carried out.
sole earth. As a renewable raw material, Colocasia escu- About 100 g of the raw Colocasia esculenta (CER) was
lenta stems are new insights into the cellulose world eco- subjected to alkaline treatment with 2% (w/w) NaOH solu-
logically and economically. In essence, nanocellulose was tion in 1:10 g/mL (fiber to liquid ratio) in an autoclave
isolated from Colocasia esculenta stems via acid-coupled at 121 °C and 15 psi for 15 min [20]. Two more subse-
steam explosion. It is noteworthy that the study suggests an quent cycles were performed under the same conditions.
eco-friendly policy for nanocellulose isolation as the conven- The alkali-treated fibers of Colocasia esculenta (CEA)
tional strong acid was substituted with mild acid along with were collected and washed well with distilled water to
mechanical assistance. Even though progressive researches remove excess alkali. Afterward, CEA fibers were sub-
on nanocellulose were ongoing, studies that describe the jected to a bleaching process with 15% (w/w) ­H2O2 and
extraction of nanofibers using chlorine-free purification 10% (w/w) NaOH solution in a ratio of 1:5 g/mL for 3 h
technique are fewer. Consequently, the study utilizes the using a magnetic stirrer at room temperature. The process
pre-treatments such as low concentration mercerization was repeated for two more cycles (until the fibers became
(2% NaOH), chlorite-free bleaching ­(H2O2/NaOH), and mild white) and washed thoroughly using distilled water until
acid hydrolysis (5% ­C2H2O4), for the isolation of nanofibers. it was free from the reagent mixture to get the bleached
Also, the cellulose type I­ α allotrope was a most discoursed Colocasia esculenta fibers (CEB).
topic among investigates in nanocellulose. Herein, for the
first time, we present cellulose type ­Iβ allotrope from Colo-
casia esculenta stems for future applications. The promising 2.2.2 Isolation of cellulose nanofibers (CNFs)
products (bio-derived) with high-performance applications
are desirable for future materials. Moreover, the ease of pro- The CEB fibers were acid treated with 5% (w/w) oxalic
cessing and reduced production cost unlocks the sustain- acid in a ratio of 1:10 g/mL. The treated fibers were then
able goals. The proper design for this agro-waste (Colocasia subjected to undergo steam explosion in an autoclave at a
esculenta stems) management formulates a zero-waste lifes- temperature of 121 °C and a pressure of 15 psi [21]. The
pan and can help to balance the ecosystem. Thus, the study process was repeated two more times. The acid-treated
underlines the transformation of “trash to treasure” which fibers were then washed thoroughly with distilled water
makes the circular economy a reality. to remove excess acid (to neutralize the pH) and homog-
enized (D lab-made homogenizer) at 10,000 rpm speed
for 30 min with distilled water to obtain a fine suspension
2 Materials and methods of Colocasia esculenta cellulose nanofibers (CECNFs).
The nanofibers were stored in the refrigerator as suspen-
2.1 Materials sion. At each stage of the chemical treatment, a small
portion of the sample was taken from the bulk and dried
The raw material Colocasia esculenta stems were obtained for the evaluation of chemical composition and a portion
from Thrissur, Kerala, India. The reagents such as NaOH, of the homogenized sample was freeze-dried for further
­H 2O 2, and ­H 2C 2O 4 for the chemical treatment of fibers characterizations.

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 Biomass Conversion and Biorefinery

2.3 Characterization experimental section was according to our previous study

[21].
The chemical composition analyses of CER, CEA, and CEB
were conducted according to the ASTM standards, cellulose
(ASTM D 1103-55 T), hemicellulose (ASTM D 1104–56), 3 Result and discussion
and lignin (ASTM D 1106–56).
The yield of CECNFs was calculated using the following 3.1 Yield of CECNFs
Eq. (1) explained by [22]:
The yield of the nanofibers is critical when choosing a proper
W2 isolation process and in practical applications. In addition, the
YieldoftheextractedCECNFs(%) = × 100 (1)
W1 yield of the product is an essential element as per green prin-
where W2 is the weight of extracted CECNFs and W1 is the ciples. The calculated yield of CECNFs was 59 ± 4%. Debiagi
initial weight of CER. et al. presented a study with a yield of 60% for nanocellulose
The chemical structures of all the samples (CER, CEA, which was consistent with the current study [23]. The effect
CEB, and CECNFs) were studied using Thermo Nicolet of acid hydrolysis on the yield of nanocellulose from sugar
Avatar 370 model FTIR spectrophotometer (Waltham, MA, palm was investigated by Ilyas et al. in which the study dis-
USA), in a wavenumber range of 4000 to 400 ­cm−1 at a played a trend of decreasing yield with increasing hydrolysis
resolution of 4 ­cm−1. time [24]. It was also found that the yield of nanocellulose
Bruker D8 Advance X-ray powder diffractometer (Bruker, was poor while using stronger acid hydrolysis. In developing
MA, USA) was used for the X-ray diffraction analysis of all nanocellulose from corncob residue, Liu et al. compared the
the fibers. The crystallinity indexes of these samples were yields of nanocellulose using different isolation approaches
calculated using the Segal Eq. (2) [9]: like sulfuric acid hydrolysis, formic acid hydrolysis, TEMPO
( ) oxidation, and pulp refining. From the study, it was inferred
Iam that sulfuric acid hydrolysis yields less nanocellulose (34.5%)
IC (%) = 1 − × 100% (2)
I200 compared to others [25]. Moreover, Trilokesh et al. in devel-
oping nanocellulose from jackfruit peels reported a yield of
where Iam is the minimum intensity of the peak contributing 20.08% using strong acid hydrolysis [26]. The reduction in the
to the amorphous phase (~ 2θ = 18.6°) and I200 is the maxi- yield of the product was due to the over-hydrolysis of crystal-
mum intensity of the peak contributing to the crystalline line regions along with the amorphous region in a strongly
phase (~ 2θ = 22.8°). acidic medium [24]. A similar observation was presented by
Thermo Scientific, Evolution 201 UV–Visible Spectro- Fitriani et al. in isolating nanocellulose from pineapple crown.
photometer with dual silicon photodiode detector and 1 nm With the increase in hydrolysis time, the yield of the nanocel-
spectral bandwidth measured the absorbance of the CNFs lulose decreased due to the excessive break-down of β glyco-
suspensions. In a quartz cuvette, dilute CNFs suspension of sidic linkage [27]. Thus, it could be inferred that rather than
0.1 wt% was taken, and absorbance is measured between 200 using acid hydrolysis alone, a chemi-mechanical method will
and 800 nm. The blank used was distilled water. be preferable in the isolation process to yield more nanofibers.
The scanning electron microscope, JEOL 6390LA/ Utilizing softwood kraft pulp, Wang et al. extracted nanocel-
OXFORD XMX N, was used to examine the morphological lulose with a high yield (65–78%) through chemi-mechanical
changes of fibers before and after chemical treatment. The synergy [28]. Hu et al. adopted a chemi-mechanical process to
purity of CECNFs was determined using energy dispersive isolate nanofibers from bamboo and obtained a similar yield
X-ray analysis (EDX), using a JEOL 6390LA/OXFORD (65–70%). According to Chen et al. the combination of differ-
XMX N instrument. A high-resolution transmission electron ent processes will impart a synergic effect which reduces the
microscope, JEOL/JEM 2100 (JEOL USA, Inc., Peabody, drawback of a single method alone and the severity of strong
MA, USA), was used to find the diameter of the prepared acid hydrolysis [29]. Consequently, the adopted method in the
CECNFs. current study is favorable as it produces nanocellulose with
Thermogravimetric analyzer, Perkin Elmer STA600 a better yield.
(Waltham, USA), at a temperature range of 0 °C to 500 °C
­ 2 atmosphere, at a heating rate of 10 °C/min in N
in the N ­ 2 3.2 Chemical composition analysis
flow 20 m­ Lmin−1 was used to investigate the thermal char- and the mechanism of pre‑treatments and acid
acteristics of the fibers. hydrolysis
Bruker AV NEO500 MHz wide-bore spectrom-
eter equipped with a 4-mm solid-state MAS probe was Cellulose is found along with biopolymers such as lignin
used to study the solid-state 13C NMR of CECNFs. The and hemicellulose in the cell walls of plants. Thus, while

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Biomass Conversion and Biorefinery

extracting cellulose from lignocellulosic biomass, proper Initially, a higher amount of lignin can be seen in CER
pre-treatments should be employed for the removal of these (23.7%). Upon chemical treatments, there was a remarkable
residues. The conventional methodologies for non-cellulosic decrease in the lignin content in CEB (3.5%). The useful-
components removal and extraction of purified cellulosic fib- ness of the alkali treatment and bleaching in the successful
ers are alkali treatment and bleaching. The chemical compo- removal of lignin was revealed here. Lignin is the amor-
sition analyses of raw, alkali-treated, and bleached samples phous macromolecule containing syringyl units, guaiacyl
(CER, CEA, and CEB) were determined according to the units, and p-hydroxy phenyl units which act as a cementing
ASTM standards and are tabulated in Table 1. Initially, the material within the cell wall [37]. Wu et al. found that ­H2O2
raw sample consisted of 40.6% cellulose. Upon alkali treat- treatment in presence of alkali caused the oxidative degrada-
ment, there is a noticeable increase of cellulose to 72.8%, tion of the carbonyl and quinoid structure of lignin resulted
and a further increase to 86.2% on bleaching was observed. in their removal [38]. Also, the peroxide ion (nucleophile
On the other hand, the concentrations of hemicellulose and intermediate) attacked the chromophore structure of the
lignin varied considerably after each stage of pre-treatments lignin causing the destruction of its corresponding structure
as 24.4, 12.5, 7.4% as well as 23.7, 9, 3.5%, respectively. leading to delignification. The destruction of the chromo-
Also, it can be noted that there will be some other non- phore structure led to the increased brightness of the sample.
cellulosic components such as pectin and wax in the sample It was found that the formation of hydroxyl, carboxylic, and
in minor amounts (which will be discussed in the upcoming carbonyl groups resulted in the oxidation of lignin, making
XRD section). It is worth mentioning that initially, the raw them soluble in an alkaline medium [20]. Herein, the strive
fiber contained a higher amount of cellulose (40.6%), and toward the green methodology (chlorite-free bleaching) is
the value was comparable with Cuscuta reflexa (41%) [30], also fulfilled.
Humulus japonicus stems (43.11%) [31] and higher than that After bleaching, the dimension of fibers is reduced, but
of banana pseudo-stem (23.8%) [32], lotus leaf stalk (34.6%) still there exists adhesion between cell walls and the fibers.
[33], and apple stems (35.8%) [34]. The sufficient cellulose There was 7.4% hemicellulose and 3.5% lignin after bleach-
content in the sample can impart better tensile, thermal, and ing. This was further removed by acid hydrolysis leading to
crystalline properties (which will be explained in forthcom- the formation of purified cellulose. During acid hydrolysis,
ing sections) which serve as the basic criterion for reinforc- the amorphous matrix was eluted by the attack of hydronium
ing ability. Moreover, the enhanced cellulose content (46% ions into the glycosidic linkage. Thus, the protonation reac-
higher than the raw fibers) after pre-treatments (even as a tion to the oxygen components in the anhydroglucose units
non-wood source) made them good bio-alternatives for non- led to the bond cleavage [39].
conventional cellulose sources.
Upon mercerization, the hemicellulose content was 3.3 Chemical functionality analysis
almost reduced to half (24.4 to 12.5%), and it was fur-
ther reduced on bleaching (7.4%). It was found that alkali As the isolation of nanofibers from the raw samples
hydrolysis led to ester bond cleavage within the hemicellu- includes pre-treatments and acid hydrolysis, it is cru-
lose molecules along with the breakage of alkali susceptible cial to identify the modification in the chemical struc-
lignin bonds. This increases the dissolution of hemicellulose ture of the sample at each stage of extraction to confirm
and lignin during alkali treatment and is the reason for the the leaching out of non-cellulosic components. Making
increase in cellulose content to 72.8% after NaOH treatment use of Fourier transform infrared spectroscopy (FTIR),
[35, 36]. As expected, some remains of hemicellulose were the change in chemical functionalities can be effectively
left after alkali treatment indicating strong hydrogen bonds resolved. The FTIR spectra of native Colocasia escu-
with the cellulose fibrils and hemicellulose. It was further lenta stems, intermediate samples, and CECNFs are given
removed by bleaching, usually adopted for the degradation in Fig. 1A. The predominant broad band at 3392 ­c m −1
of lignin. in all the samples implied the hydrophilic nature arising
from the –OH groups [40]. The characteristic s­ p 3 -CH
stretching vibrations of alkyl groups in cellulose, hemi-
cellulose, and lignin are given by the band at 2927 ­cm−1
Table 1  Chemical composition of fibers [29]. Sango et al. present the peak at 2858 ­c m −1 as the
Colocasia esculenta fibers Cellulose (%) Hemicel- Lignin (%) wax coating which are usually esters presents in the
lulose (%) plants [41]. Similar wax coatings in native plant fibers
and their successful elimination on chemical treatment
Untreated fibers (CER) 40.6 24.4 23.7
were reported by Astruc et al. and Dominic et al. [30,
Alkali-treated fibers (CEA) 72.8 12.5 9.0
42]. The band at 1735 ­c m −1 corresponded to the C = O
Bleached fibers (CEB) 86.2 7.4 3.5
stretching frequency of acetyl and uronic ester groups

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 Biomass Conversion and Biorefinery

Fig. 1  FTIR spectra of CER,

CEA, CEB, and CECNFs (A),
13
C Solid-state NMR spectra of
CECNFs (B), EDX Spectra of
CECNFs (C)

in hemicellulose and can also be due to the vibrations 3.4 Structural analysis

of p-coumaric units of the lignin [43]. A similar shoul-
der peak corresponds to hemicellulose and lignin was For the isolation of nanocellulose, the chemical treatment
reported elsewhere in the extraction of nanocellulose should be effective in producing the same without chang-
from plant stems [6, 44]. The FTIR spectra of the treated ing the structural features. 13C solid-state NMR is used as
Colocasia esculenta fibers (CEA, CEB, and CECNFs) a strong tool for studying the atomic structure of nanocel-
clearly showed the disappearance of this specific peak. lulose, and it is given in Fig. 1B. As the cellulose structure
As the applied pre-treatment is very much effective, it consists of 6 carbon atoms corresponding to glucopyra-
facilitated the effectual removal of the non-cellulosic nose units, their corresponding resonance frequencies were
components within the sample. Moreover, the lessening obtained at 105.31 ppm for ­C1; 88.96 ppm and 84.41 ppm for
of this peak intensity can be considered as validation for ­C4; 75.33 ppm and 72.77 ppm for ­C2, ­C3, and ­C5 (unresolved
chemical composition analysis which demonstrated the peaks); and 65.03 ppm and 62.84 ppm for ­C6 [48].
elution of non-cellulosic components at each stage of From the studies of Ma et al. it can be inferred that the
chemical treatment. The chemically adsorbed water on chemical shift of aromatic carbonyl groups, esters, ketones,
the fiber and its bending angular deformation was repre- and proteins (which consist of hemicellulose and lignin)
sented by the peak at 1623 ­cm−1 [25]. According to Zheng arise at a higher chemical shift value usually at 155.2,
et al. the peak at 1298 ­cm−1 represents the aromatic skel- 144.8, and 131.4 ppm [49]. The absence of these chemical
etal vibration of lignin [45]. The absence of this lignin shift values for the CECNFs implied the chemical treatment
characteristic peak is also consistent with the chemical applied for the dissolution of hemicellulose and lignin was
composition analysis. As per the study of Zaki et al. the effective. This further validated the FTIR, XRD, and EDX
peak at 1030 ­cm−1 reflected the C–OH stretching vibra- results which confirmed the existence of nanofibers with
tions of primary and secondary alcohol groups in cellu- high purity. Moreover, the narrower peaks in the 13C solid-
lose [44]. Besides, Benini et al. reported a similar peak at state NMR spectrum indicated the ordered arrangement of
1030 ­cm−1 and represented this as the characteristic peak the fibers which can be the reason for the increased crystal-
C-O stretching and C-H rocking vibrations of the pyra- linity and thermal stability of the CECNFs obtained from the
nose ring of cellulose [46]. The peak at 892 ­cm−1 shows XRD and thermal analysis which will be discussed in detail
the intensive hydrogen bonding between the –OH and O in the forthcoming sections. According to Foster et al. the
groups (on third and fifth positions) near to β glycosidic type of polymorph present in cellulose is determined by the
linkage in the cellulose structure [47]. distinctive peak of anomeric carbon ­C1. The characteristic

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Biomass Conversion and Biorefinery

resonance peak at 105 ppm for ­C1 implied the presence of makes them ideal ones to be used in advanced materials.
cellulose type ­Iβ structure. Furthermore, the proximity of the The peaks of carbon and oxygen in the EDX spectrum were
peak, ­C4 at approximately 88.7 ppm suggested native ­Iβ crys- consistent with the study reported by Hussin et al. in which
talline cellulose [48]. The 13C solid-state NMR spectrum of the nanofibers from raw oil palm leaves pose a carbon and
CECNFs also posed the above-mentioned resonance wings oxygen of 56.7 and 43.3%, respectively [52]. Making use of
of ­Iβ structure. This supported the findings of XRD study EDX spectra, the presence of other elements such as sodium,
which revealed the presence of cellulose type I­ β. Alike XRD potassium, silicon, sulfur, nitrogen, and more was reported
analysis, the crystalline behavior of the nanocellulose can elsewhere [53–55]. As the EDX spectrum of CECNFs did
also be investigated using 13C solid-state NMR. C ­ 4 and C
­6 not show any traces of other elements, the result validated
were the unique peaks (usually exist as a doublet) that pro- the FTIR and 13C solid-state NMR results.
vide information about the crystalline and amorphous zones.
From the doublets of C ­ 4, the peak at 88.96 ppm represented 3.6 Crystallinity studies
the crystalline region and 84.41 ppm gives the amorphous
region. Similarly, the doublet wings of C ­ 6 at 65.03 ppm and The diffractogram was used to study the crystalline behavior
62.84 ppm represent the crystalline and amorphous regions, of all fibers and is given in Fig. 2A. The samples showed two
respectively [50]. 13C solid-state NMR can also be used to diffraction peaks, the first peak in-between 14 and 16° and
evaluate the conformations present in the sample. The ­C6 the second sharp peak at 22° which corresponded to the lat-
carbon of the C ­ H2OH group in the cellulose molecules has tice planes (1–10)/(110) overlapping and (200), respectively.
a relaxation time lower than that of the rest of the ring car- Additionally, the CECNFs showed a reflection plane at (004)
bon atoms. Consequently, C ­ 6 was assessed for the confor- corresponding to the diffraction angle of 34°. All these peaks
mational studies. Herein, the chemical shift of C ­ 6 at 65 ppm showed the cellulose type I­ β in the fibers [43]. Moreover, the
validated the existence of a trans-gauge system within the study of Nisar et al. concluded that the peak at ~ 25° corre-
molecule [51]. sponds to that of the compound, pectin [56]. The Colocasia
esculenta showed a shoulder peak at this lattice, and this can
3.5 Elemental analysis be attributed to the presence of pectin. The study of cellulose
nanofibers by Jongaroontaprangsee et al. on lime residues
As the commercial application of nanocellulose is depend- and Jayaprada et al. on orange peels also reported a similar
ent on chemical purity, elemental analysis was performed XRD pattern [57, 58].
to ensure its pureness. The energy dispersive X-ray spec- As summarized in Table 2, nanocellulose is an important
troscopy was conducted to study the elemental composi- material with numerous applications. Crystalline behavior
tion of the nanocellulose (Fig. 1C), and the spectra con- is a crucial factor to be discussed while considering nano-
firmed 54.55% carbon and 45.45% oxygen in the CECNFs, cellulose in advanced applications. Cellulose usually has
which were the basic constituents of the cellulose. Mostly, a well-defined crystalline structure which generally arises
traces of some minor elements can occur along with these from the strong hydrogen bonding and van der Waals inter-
constituent elements which can affect the integrity of the action between the cellulose molecules within themselves
nanofibers. Herein, the EDX spectra of CECNFs lacked the compared to the non-cellulosic components [29]. The
presence of other elements which implied the adequacy of crystallinity index of CER, CEA, CEB, and CECNFs were
the applied chemical treatment in isolating nanocellulose 47.33, 57.39, 65.13, and 71.72%, respectively. Notably, the
and significantly reducing the non-cellulosic part. This CECNFs posed a superior crystallinity than the rest of the

Fig. 2  XRD diffractogram of

CER, CEA, CEB, and CECNFs
(A), UV Absorbance and trans-
mittance spectra of CECNFs
(B)

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 Biomass Conversion and Biorefinery

Table 2  A comparative study of crystallinity index (using Segal’s method), dimension, and applications of cellulose nanofibers from various
researches
Source Method of synthesis Crystallinity Diameter (nm) Application Reference
index (%)

Carrot Homogenization 70 18 Cellulose films [68]

Cuscuta reflexa Oxalic acid hydrolysis 67 10–30 Natural rubber reinforcement [30]
Jute fibers Oxalic acid hydrolysis 82.22 50 Natural rubber reinforcement [69]
Oil palm empty fruit bunch Sulfuric acid hydrolysis 73 4–15 PVA/starch film reinforcement [70]
Rice hulls Sulfuric acid hydrolysis 75 < 100 Biodegradable starch films [71]
Kenaf bast fibers Mechanical shearing 81.5 6.03 TPS biofilm reinforcement [72]
Sugarcane bagasse Disc grinding 72.9 39 ± 13 Food packaging [73]
Saw dust Blending and grinding - 2–27 Food packaging [74]
Grape fruit peel TEMPO oxidation 55.5 40–80 Fat Substitute in ice creams [75]
Pineapple leaves Oxalic acid hydrolysis - 5–15 Medical implants [76]
Palm Sulfuric acid hydrolysis 60 20–50 Drug delivery [77]
Cotton Sulfuric acid hydrolysis 86.2 5–15 Drug delivery [78]

fibers, remarkably 24.39% higher than the native fiber. Also, higher crystallinity caused the better transmittance of the
the intensity of the crystalline peak increased considerably nanofibers [59]. The CECNFs had a better transmittance,
with progressive reaction conditions. The penetration of pro- which was supported by the lower particle dimension and
tons into the glycosidic linkage resulted in the weakening of higher crystallinity. Also, the removal of the non-cellulosic
C–O–C bonding and thus bond cleavage made the dissolu- components which acted as adhesives within the cellulose
tion of the amorphous region. Consequently, the enhance- molecules imparted better transmittance.
ment of crystallites increased the crystallinity of the chemi-
cally treated fibers. The increase in the crystallinity further 3.8 Morphological analysis
increased the strength and stiffness of the nanofibers due to
the refined molecular orientation [43]. A comparative study The morphological studies of untreated and treated fib-
of crystallinity index of nanocellulose from various plant ers were studied using SEM analysis. The change in
sources is discussed in Table 2. Furthermore, the crystallin- morphology of the fiber structure on each processing
ity index of nanocellulose obtained from various plant stems stage is depicted in Fig. 3. Figure 3 (A, B, C, and D) rep-
were banana pseudo-stem (80.13%) [59], banana pseudo- resents the SEM micrograms of CER, CEA, CEB, and
stem (75%) [32], Shwetark stems (72.06%) [5], apple stems CECNFs, respectively. Usually, cellulose fibers exist as
(69.1%) [34], ginger stems (67.05%) [44], water hyacinth aggregates of long fibrils with compact structures [63].
stems (67%) [9], and rose stems (22.2%) [22]. Interestingly, The irregularity in the surface of the fiber in the case of
the CECNFs exhibited comparable crystallinity index values raw fibers (CER) is clearly visible in Fig. 3A. According
with the afore-said nanocellulose. to Astruc et al., the presence of pectin, lignin, hemicel-
lulose, and other plant residues accounts for this [42].
3.7 Optical analysis The strong adhesion of hemicellulose and lignin with the
cellulose fibers (through non-covalent bond) formed a
The optical properties of the nanofibers were studied, and tight wrap around the fibers, and their removal attrib-
the corresponding absorbance and transmittance spectra uted to the formation of individualized fibers with more
of the nanofibers was given in Fig. 2 Yan et al. presented roughened surface. The fiber bundles tended to loosen
a similar UV absorbance spectra [60]. The transmittance on alkali treatment, and Fig. 3B represents the initia-
of the CECNFs was nearly 60%. The transmittance of the tion of individual fiber formation from the native fiber.
nanofibers was largely dependent on the dimension and crys- In fact, after alkali treatment, the compact structure of
tallinity. Sun et al. discovered that the smaller fibers result in the fibers did not have any appreciable change due to
lesser backscattering of light and better transmittance will the presence of lignin, and the result was in agreement
be obtained, in accordance with Rayleigh scattering theory with the chemical composition analysis that only a small
[61]. Furthermore, as crystallinity increases, light scattering fraction of lignin was susceptible to mercerization. After
reduces, and the transmittance increases due to the ordered bleaching (Fig. 3C), reduced firmness of the fiber was
arrangement of crystallites [62]. The lower dimension and observed and the fiber become more defibrillated. The

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Biomass Conversion and Biorefinery

Fig. 3  SEM microgram of CER

(A), CEA (B), CEB (C), and
CECNFs (D)

delignification process and ejection of remaining hemi- 3.9 Dimensional analysis

cellulose caused the formation of discrete fibers [64].
The morphological difference between the raw and pre- The TEM micrograph of CECNFs is shown in Fig. 4, and
treated fibers pointed out the successful removal of the the fibers possessed a width of 20–40 nm. A comparable
cementing materials (mainly lignin and hemicellulose) nano diameter was obtained for the CECNFs with the tab-
embedded on the fiber surface. The ambient temperature ulated nanofibers in Table 2. The nanofibers were clearly
and pressure applied also contributed to the cleavage of defined, and the result could be the indication of solubiliza-
hydrogen bonds of microfibrils and the individualiza- tion of unwanted plant residues on chemical treatment. A
tion of fibers from bundles [31]. Figure 3D represents well-arranged network of fibers with some aggregates was
the fine filament-shaped nanofibers after acid hydrolysis observed in the figure. The presence of hydrogen bonds in-
and homogenization. It was found that homogenization between the fibers were revealed here [65]. Gadzama et al.
was effective in the formation of nanofibers [13]. Thus, explained the hydrophobic behavior of untreated fibers
in relation to the study of Chirayil et al., the thin fila- which can affect their reinforcing ability [66]. The enhanced
ments in the case of CECNFs exhibited nanodimension hydrophilicity of nanofibers on chemical treatments makes
that was further confirmed using TEM analysis. them ideal ones among various fillers. The destruction of the
structure of cellulose fibers on various methods (especially

Fig. 4  TEM images of CECNFs

at different magnifications:
2 μm (A), 200 nm (B)

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 Biomass Conversion and Biorefinery

From the TGA curves, the samples demonstrated mainly

three weight loss regions, a minor weight loss at ~ 100 °C,
major weight loss at a temperature range of 280 to 380 °C,
and the degradation around 400 to 450 °C. The crude sam-
ple (CER) showed less weight loss contributed to the pres-
ence of different plant residues in it. A small mass decrease at
100 °C in all samples indicated the moisture loss from each
sample and the hydrophilic behavior of cellulose samples
[79]. According to Wang et al. the degradation in between the
region 110 °C to 200 °C corresponds to the pectin degradation
[81]. As Colocasia esculenta exhibited a peak in this region
(Fig. 5B), the presence of pectin is ascribed here. This result
is in agreement with the XRD result of Colocasia esculenta
fibers. As per studies of Chirayil et al., hemicellulose degra-
dation took place at ~ 260 °C, and the pyrolysis of cellulose
was occurred at ~ 375 °C. Thus, the principal degradation
was (280 to 380 °C) attributed to the thermal depolymeriza-
Fig. 5  TGA curves of CER, CEA, CEB, and CECNFs tion of hemicellulose and cleavage of glycosidic linkages in
cellulose. The final degradation peak in the thermograms (400
to 450 °C) showed the lignin degradation [13].
Table 3  Thermal analysis data Ton ­(T10) values of untreated, treated (alkali, bleached), and
Samples T10 (°C) T50 (°C) Residue at
nanofibers were 146.3, 199.1, 156.1, and 196.3 °C respectively.
400 °C (%) Comparing the T ­ 10 values of CER and CECNFs, a delayed
degradation for CECNFs (about 50 °C increase in T ­ onset value)
CER 146.3 310.0 39.2
was found. The increase in cellulose content and removal of
CEA 199.1 351.9 36.9
amorphous non-cellulosic components were responsible for
CEB 156.1 329.8 30.2
this behavior. A similar trend was observed for ­T50, and the
CECNFs 196.3 335.8 18.4
corresponding values are 310.0, 351.9, 329.8, and 335.8 °C,
respectively, for CER, CEA, CEB, and CECNFs. Residue or
ash content is an important factor that determines thermal sta-
while adopting high mechanical stress) was reported by Xie bility. From the study conducted by Meng et al., the research-
et al. [67]. From the TEM image, it was clear that the fibrous ers concluded that high residue had a negative effect on ther-
structure of the nanocellulose was undisturbed and bundles mal stability [32]. From the thermal data (Table 3), the residue
of several nanofibers were obtained. Thus, mild acid hydrol- at 400 °C of the CECNFs was very much lower (18.4%) than
ysis assisted with steam explosion and homogenization was CEB (30.2%), CEA (36.9%), and CER (39.2%) samples. Thus,
effective in developing nanocellulose while maintaining its the lower residue content of CECNFs is a contributing factor
fibrous nature intact. to its increased thermal stability primarily due to the effective
solubilization of hemicellulose and lignin on pre-treatments
3.10 Thermal analysis which was corroborated by compositional analysis (Table 1).
According to Nascimento et al. the increased crystallinity con-
The thermal stability studies of cellulose derivatives are very tributes to the elevated thermal stability of the CNFs [71].
important in developing bio-composites with high thermal Increased crystallinity of CECNFs from XRD analysis can be
stability. The thermograms of respective samples (CER, related here. High crystallinity (ordered arrangement) of the
CEA, CEB, and CECNFs) are given in Fig. 5, and the cor- cellulose molecules by the removal of the amorphous region
responding data are provided in Table 3. The TGA curve by acid hydrolysis resisted the heat transfer through the glyco-
is plotted as the change in mass (weight loss) as a function sidic chains. It thus boosted the thermal stability [82].
of temperature. All the chemically treated samples exhib-
ited similar TGA curves (3 degradation peaks), while the
untreated fiber exhibited several degradations. Nurul et al., 4 Conclusion
Legrand et al., and Naduparambath et al. presented similar
degradation curves on developing nanofibers from kenaf Considering the environmental consequences, an appeal-
plant, sago seed shells, and Cola lepidota stems, respec- ing method is crucial to turn bio-wastes to useful product.
tively [6, 79, 80]. This work discusses the potential for producing cellulose

13
Biomass Conversion and Biorefinery

nanofibers from Colocasia esculenta stems, an agro-waste. Declarations

For nanofiber isolation, chlorine-free purification and acid
hydrolysis (assisted with steam explosion and homogeni- Conflict of interest The authors declare no competing interests.
zation) were applied in accordance with the environmental
standpoint. The effectual pre-treatments productively reduce
the lignin and hemicellulose within the native Colocasia
References
esculenta stems and reduced the need for strong acids while
isolating the nanofibers. The collective impact of eco- 1. Legesse T, Bekele T (2021) Evaluation of improved taro (Colo-
acceptable pre-treatments and mild acid hydrolysis prompted casia esculenta(L.) Schott) genotypes on growth and yield per-
a worthier strategic approach for nanocellulose isolation. formance in North-Bench woreda of Bench-Sheko zone, South-
Western Ethiopia. Heliyon 7:e08630. https://​doi.​org/​10.​1016/j.​
The successful isolation of nanofibers with high yield
heliy​on.​2021.​e08630
(59 ± 4%) and high crystallinity (71.72%) can be obtained 2. Rashmi DR, Raghu N, Gopenath TS et al (2018) Taro (Colocasia
using the recommended process. Interestingly, the nanofiber esculenta): An overview. J Med Plants Stud 6:156–161
possessed cellulose type ­Iβ structure, and it was confirmed 3. Obele CM, Ejimofor MI, Atuanya CU, Ibenta M (2021) Cassava
stem cellulose (CSC)Nanocrystal for optimal methylene BlueBio
using XRD and 13C solid–solid state NMR studies. Also, the
sorption with response surface design. Curr Res Green Sustain
high elemental purity of the nanofibers indicates the efficacy Chem 4:100067. https://​doi.​org/​10.​1016/j.​crgsc.​2021.​100067
of mild chemical treatment. The fibrous morphology with 4. Vinod A, Vijay R, Singaravelu DL et al (2021) Extraction and
a diameter of 20–40 nm was obtained for the nanocellulose characterization of natural fiber from stem of Cardiospermum
halicacabum. J Nat Fibers 18:898–908. https://​doi.​org/​10.​1080/​
from TEM analysis. The acid hydrolysis was also capable in
15440​478.​2019.​16695​14
increasing the thermal stability of the nanofibers as the T ­ 50 5. Raja K, Prabu B, Ganeshan P et al (2020) Characterization studies
of nanofibers was 335.8 °C, which was 25.8 °C greater than of natural cellulosic fibers extracted from Shwetark stem. J Nat
the native fiber. In general, the study provides an overview of Fibers 00:1–12. https://​doi.​org/​10.​1080/​15440​478.​2019.​17106​50
6. Legrand NBR, Lucien M, Pierre O et al (2020) Physico-chemical
chlorine-free purification method for the pre-treatment and
and thermal characterization of a lignocellulosic fiber, extracted
mild chemical treatment of cellulose sources for nanocellu- from the Bast of <i>Cola lepidota</i> Stem. J Miner Mater Char-
lose extraction which can amend the conventional methods. act Eng 08:377–392. https://​doi.​org/​10.​4236/​jmmce.​2020.​85024
Accordingly, mechanically aided acid hydrolysis was found 7. Rigg-Aguilar P, Moya R, Oporto-Velasquez GS, et al (2020)
Micro- and nanofibrillated cellulose (MNFC) from pineap-
to be effective in forming nanofibers. Evidently as presented
ple (Ananas comosus) stems and their application on polyvinyl
in the research, Colocasia esculenta stems derived cellu- acetate (PVAc) and urea-formaldehyde (UF) wood adhesives. J
lose nanofibers are a viable option for uplifting the natural Nanomater 2020. https://​doi.​org/​10.​1155/​2020/​13931​60
resource production since they are safe and acceptable in 8. Mumthas ACSI, Wickramasinghe GLD, Gunasekera USW (2019)
Effect of physical, chemical and biological extraction methods
practical applications. In fact, using the adopted method,
on the physical behaviour of banana pseudo-stem fibres: based
nanocellulose with superior properties can be extracted for on fibres extracted from five common Sri Lankan cultivars. J Eng
beneficial use in a wide range of industries. Fiber Fabr 14:1–15. https://​doi.​org/​10.​1177/​15589​25019​865697
9. Juárez-Luna GN, Favela-Torres E, Quevedo IR, Batina N (2019)
Acknowledgements The authors acknowledge SAIF STIC, CUSAT Enzymatically assisted isolation of high-quality cellulose nano-
for the chemical analysis. particles from water hyacinth stems. Carbohydr Polym 220:110–
117. https://​doi.​org/​10.​1016/j.​carbp​ol.​2019.​05.​058
10. Gapsari F, Purnowidodo A, Setyarini PH et al (2022) Properties
Author contribution Midhun Dominic C. D.: Conceptualization,
of organic and inorganic filler hybridization on timoho fiber-rein-
methodology, visualization, supervision, project administration, writ-
forced polyester polymer composites. Polym Compos 43:1147–
ing—original draft. Maheswary. S.: Investigation. Neenu K. V.: Inves-
1156. https://​doi.​org/​10.​1002/​pc.​26443
tigation. S. Mohammad Sajadi: Writing, reviewing and editing. Der-
11. Gapsari F, Mudjib Sulaiman A, Putri TM, et al (2022) Influence of
val dos Santos Rosa: Writing, reviewing and editing. P. M. Sabura
calcium carbonate fillers on pine fiber reinforced polyester com-
Begum: Supervision. Mariya Mathew: Data curation. T. G. Ajithku-
posites. Polym Compos 1–12. https://​doi.​org/​10.​1002/​pc.​26692
mar: Resources. Jyotishkumar Parameswaranpillai: Conceptualization,
12. Thyavihalli Girijappa YG, Mavinkere Rangappa S, Parameswaran-
methodology, reviewing and editing. Tresa Sunitha George: Resources.
pillai J, Siengchin S (2019) Natural fibers as sustainable and
Resmi V. C.: Resources. Ilyas R. A.: Writing, reviewing and editing.
renewable resource for development of eco-friendly composites:
Michael Badawi: Writing, reviewing, editing, supervision, funding
a comprehensive review. Front Mater 6:1–14. https://​doi.​org/​10.​
acquisition, and resources.
3389/​fmats.​2019.​00226
13. Chirayil CJ, Joy J, Mathew L et al (2014) Isolation and charac-
Funding This work was financially supported by Sacred Heart Col- terization of cellulose nanofibrils from Helicteres isora plant. Ind
lege (Autonomous), Thevara under the basic research funding scheme Crops Prod 59:27–34. https://​doi.​org/​10.​1016/j.​indcr​op.​2014.​04.​
(No.1/RES/2020). The authors greatly acknowledge the financial sup- 020
port from Université de Lorraine, France. 14. Trache D, Tarchoun AF, Derradji M, et al (2020) Nanocellulose:
from fundamentals to advanced applications
Data availability All of the data sets that were included and/or exam- 15. Yadav C, Saini A, Zhang W et al (2021) Plant-based nanocel-
ined in the present study can be obtained from the corresponding author lulose: a review of routine and recent preparation methods with
upon request. current progress in its applications as rheology modifier and 3D

13
 Biomass Conversion and Biorefinery

bioprinting. Int J Biol Macromol 166:1586–1616. https://​doi.​org/​ high temperature pretreatment. Carbohydr Polym 164:284–293.
10.​1016/j.​ijbio​mac.​2020.​11.​038 https://​doi.​org/​10.​1016/j.​carbp​ol.​2017.​02.​021
16. Zhuo X, Liu C, Pan R et al (2017) Nanocellulose mechanically 32. Meng F, Wang G, Du X, et al (2018) Preparation and charac-
isolated from Amorpha fruticosa Linn. ACS Sustain Chem Eng terization of cellulose nanofibers and nanocrystals from liquefied
5:4414–4420. https://​doi.​org/​10.​1021/​acssu​schem​eng.​7b004​78 banana pseudo-stem residue. Compos Part B https://​doi.​org/​10.​
17. Filson PB, Dawson-Andoh BE, Schwegler-Berry D (2009) Enzy- 1016/j.​compo​sitesb.​2018.​08.​048
matic-mediated production of cellulose nanocrystals from recy- 33. Chen Y, Wu Q, Huang B et al (2015) Isolation and characteristics
cled pulp. Green Chem 11:1808–1814. https://​doi.​org/​10.​1039/​ of cellulose and nanocellulose from lotus leaf stalk agro-wastes.
b9157​46h BioResources 10:684–696. https://​doi.​org/​10.​15376/​biores.​10.1.​
18. Ravindran L, Sreekala MS, Thomas S (2019) Novel processing 684-​696
parameters for the extraction of cellulose nanofibres (CNF) from 34. Hanthong PP, Yufei MA, Uan GG, Budula AA (2015) Extrac-
environmentally benign pineapple leaf fibres (PALF): Structure- tion of nanocellulose from raw apple stem. J Japan Inst Energy
property relationships. Int J Biol Macromol 131:858–870. https://​ 94(8):787–793
doi.​org/​10.​1016/j.​ijbio​mac.​2019.​03.​134 35. Oriez V, Peydecastaing J, Pontalier P-Y (2020) Lignocellulosic
19. Abdul Khalil HPS, Bhat AH, Ireana Yusra AF (2012) Green com- biomass mild alkaline fractionation and resulting extract purifica-
posites from sustainable cellulose nanofibrils: a review. Carbohydr tion processes: Conditions, Yields, and Purities. Clean Technol
Polym 87:963–979. https://d​ oi.o​ rg/1​ 0.1​ 016/j.c​ arbpo​ l.2​ 011.0​ 8.0​ 78 2:91–115. https://​doi.​org/​10.​3390/​clean​techn​ol201​0007
20. Cherian BM, Leão AL, de Souza SF et al (2010) Isolation of nano- 36. Nie S, Zhang C, Zhang Q et al (2018) Enzymatic and cold alka-
cellulose from pineapple leaf fibres by steam explosion. Carbo- line pretreatments of sugarcane bagasse pulp to produce cellulose
hydr Polym 81:720–725. https://​doi.​org/​10.​1016/j.​carbp​ol.​2010.​ nanofibrils using a mechanical method. Ind Crops Prod 124:435–
03.​046 441. https://​doi.​org/​10.​1016/j.​indcr​op.​2018.​08.​033
21. Dominic CDM, Raj V, Neenu KV et al (2022) International Jour- 37. Ramesh S, Doddipatla P, Pamidipati S (2021) Optimization of
nal of Biological Macromolecules Chlorine-free extraction and parameters for biological pre-treatment route for the production
structural characterization of cellulose nanofibers from waste husk of nanocellulose from sugarcane bagasse. Biomass Convers Biore-
of millet (Pennisetum glaucum ). Int J Biol Macromol 206:92– finery https://​doi.​org/​10.​1007/​s13399-​021-​01306-2
104. https://​doi.​org/​10.​1016/j.​ijbio​mac.​2022.​02.​078 38. Wu Y, Wu J, Yang F et al (2019) Effect of H2O2 bleaching treat-
22. Ventura-cruz S, Tecante A (2019) Extraction and characterization ment on the properties of finished transparent wood. Polymers
of cellulose nanofibers from rose stems (Rosa spp.). Carbohydr (Basel) 11:1–13. https://​doi.​org/​10.​3390/​polym​11050​776
Polym 220:53–59. https://​doi.​org/​10.​1016/j.​carbp​ol.​2019.​05.​053 39. Ng HM, Sin LT, Tee TT et al (2015) Extraction of cellulose
23. Debiagi F, Faria-Tischer PCS, Mali S (2021) A green approach nanocrystals from plant sources for application as reinforcing
based on reactive extrusion to produce nanofibrillated cellulose agent in polymers. Compos Part B Eng 75:176–200. https://​doi.​
from oat hull. Waste and Biomass Valorization 12:1051–1060. org/​10.​1016/j.​compo​sitesb.​2015.​01.​008
https://​doi.​org/​10.​1007/​s12649-​020-​01025-1 40. Ventura-Cruz S, Flores-Alamo N, Tecante A (2020) Preparation
24. Ilyas RA, Sapuan SM, Atikah MSN et al (2021) Effect of hydroly- of microcrystalline cellulose from residual rose stems (Rosa spp.)
sis time on the morphological, physical, chemical, and thermal by successive delignification with alkaline hydrogen peroxide. Int
behavior of sugar palm nanocrystalline cellulose (Arenga pinnata J Biol Macromol 155:324–329. https://d​ oi.o​ rg/1​ 0.1​ 016/j.i​ jbiom ​ ac.​
(Wurmb.) Merr). Text Res J 91:152–167. https://​doi.​org/​10.​1177/​ 2020.​03.​222
00405​17520​932393 41. Sango T, Cheumani Yona AM, Duchatel L et al (2018) Step–
25. Liu C, Li B, Du H et al (2016) Properties of nanocellulose isolated wise multi–scale deconstruction of banana pseudo–stem (Musa
from corncob residue using sulfuric acid, formic acid, oxidative acuminata) biomass and morpho–mechanical characterization of
and mechanical methods. Carbohydr Polym 151:716–724. https://​ extracted long fibres for sustainable applications. Ind Crops Prod
doi.​org/​10.​1016/j.​carbp​ol.​2016.​06.​025 122:657–668. https://​doi.​org/​10.​1016/j.​indcr​op.​2018.​06.​050
26. Trilokesh C, Uppuluri KB (2019) Isolation and characterization of 42. Astruc J, Nagalakshmaiah M, Laroche G et al (2017) Isolation of
cellulose nanocrystals from jackfruit peel. Sci Rep 9:1–8. https://​ cellulose-II nanospheres from flax stems and their physical and
doi.​org/​10.​1038/​s41598-​019-​53412-x morphological properties. Carbohydr Polym 178:352–359. https://​
27. Fitriani, Aprilia NAS, Arahman N (2020) Properties of nanocrys- doi.​org/​10.​1016/j.​carbp​ol.​2017.​08.​138
talline cellulose from pineapple crown leaf waste. IOP Conf Ser 43. Dai H, Ou S, Huang Y, Huang H (2018) Utilization of pineapple
Mater Sci Eng 796. https://​doi.​org/​10.​1088/​1757-​899X/​796/1/​ peel for production of nanocellulose and film application. Cel-
012007 lulose 25:1743–1756. https://d​ oi.o​ rg/1​ 0.1​ 007/s​ 10570-0​ 18-1​ 671-0
28. Wang J, Xu J, Zhu S et al (2021) Preparation of nanocellulose in 44. Zaki FA, Abdullah I, Ahmad I (2014) The physical and chemi-
high yield via chemi-mechanical synergy Preparation of nano- cal natures of cellulose extracted from torch ginger stems. Int J
cellulose in high yield via chemi-mechanical synergy. Carbo- Mater Eng Innov 5:48–60. https://d​ oi.o​ rg/1​ 0.1​ 504/I​ JMATE ​ I.2​ 014.​
hydr Polym 251:117094. https://​doi.​org/​10.​1016/j.​carbp​ol.​2020.​ 059484
117094 45. Zheng D, Zhang Y, Guo Y, Yue J (2019) Isolation and characteri-
29. Chen YW, Lee HV (2018) Revalorization of selected municipal zation of nanocellulose with a novel shape from walnut (Juglans
solid wastes as new precursors of “green” nanocellulose via a Regia L.) shell agricultural waste. Polymers (Basel) 11(7):1130.
novel one-pot isolation system: a source perspective. Int J Biol https://​doi.​org/​10.​3390/​polym​11071​130
Macromol 107:78–92. https://d​ oi.o​ rg/1​ 0.1​ 016/j.i​ jbiom​ ac.2​ 017.0​ 8.​ 46. de C. Benini KCC, Voorwald HJC, Cioffi MOH et al (2018) Prep-
143 aration of nanocellulose from Imperata brasiliensis grass using
30. Dominic MCD, Joseph R, Begum PMS et al (2020) Cellulose Taguchi method. Carbohydr Polym 192:337–346. https://​doi.​org/​
nanofibers isolated from the Cuscuta reflexa plant as a green rein- 10.​1016/j.​carbp​ol.​2018.​03.​055
forcement of natural rubber. Polymers (Basel) 12:814. https://​doi.​ 47. Souza AG, Santos DF, Ferreira RR et al (2020) Innovative process
org/​10.​3390/​POLYM​12040​814 for obtaining modified nanocellulose from soybean straw. Int J
31. Jiang Y, Zhou J, Zhang Q et al (2017) Preparation of cellulose Biol Macromol 165:1803–1812. https://​doi.​org/​10.​1016/j.​ijbio​
nanocrystals from Humulus japonicus stem and the influence of mac.​2020.​10.​036

13
Biomass Conversion and Biorefinery

48. Foster EJ, Moon RJ, Heux L et al (2018) Current characteri- using high-pressure homogenization. Cellulose 25:1757–1768.
zation methods for cellulose nanomaterials. Chem Soc Rev https://​doi.​org/​10.​1007/​s10570-​018-​1686-6
47:2609–2679. https://​doi.​org/​10.​1039/​c6cs0​0895j 66. Gadzama SW, Sunmonu OK, Isiaku US, Danladi A (2020) Isola-
49. Ma H, Hsiao BS (2018) Nanocellulose extracted from defolia- tion and characterization of nanocellulose from pineapple leaf
tion of ginkgo leaves. MRS Adv 3:2077–2088. https://​doi.​org/​ fibres via chemo-mechanical. Sci World J 15:100–105
10.​1557/​adv.​2018.​148 67. Xie H, Du H, Yang X, Si C (2018) Recent strategies in preparation
50. Mariño M, Lopes L, Durán N, Tasic L (2015) Enhanced materi- of cellulose nanocrystals and cellulose nanofibrils derived from
als from nature: nanocellulose from citrus waste. 20:5908–5923. raw cellulose materials. Int J Polym Sci 2018. https://​doi.​org/​10.​
https://​doi.​org/​10.​3390/​molec​ules2​00459​08 1155/​2018/​79230​68
51. Horii F, Hirai A, Kitamaru R (1983) Solid-state 13C-NMR 68. Varanasi S, Henzel L, Sharman S et al (2018) Producing nanofi-
study of conformations of oligosaccharides and cellulose. bres from carrots with a chemical-free process. Carbohydr Polym
Polym Bull 10:357–361. https://​doi.​org/​10.​1007/​bf002​81948 184:307–314. https://​doi.​org/​10.​1016/j.​carbp​ol.​2017.​12.​056
52. Hussin FNNM, Attan N, Wahab RA (2020) Extraction and char- 69. Thomas MG, Abraham E, Jyotishkumar P et al (2015) Nanocel-
acterization of nanocellulose from raw oil palm leaves (Elaeis luloses from jute fibers and their nanocomposites with natural
guineensis). Arab J Sci Eng 45:175–186. https://​d oi.​o rg/​1 0.​ rubber: preparation and characterization. Int J Biol Macromol
1007/​s13369-​019-​04131-y 81:768–777. https://​doi.​org/​10.​1016/j.​ijbio​mac.​2015.​08.​053
53. Kumar A, Singh Negi Y, Choudhary V, Kant Bhardwaj N (2020) 70. Lani NS, Ngadi N, Johari A, Jusoh M (2014) Isolation, characteri-
Characterization of cellulose nanocrystals produced by acid- zation, and application of nanocellulose from oil palm empty fruit
hydrolysis from sugarcane bagasse as agro-waste. J Mater Phys bunch fiber as nanocomposites. J Nanomater 2014:1–7. https://​
Chem 2:1–8. https://​doi.​org/​10.​12691/​jmpc-2-​1-1 doi.​org/​10.​1155/​2014/​702538
54. Xiao Y, Rong L, Wang B et al (2018) A light-weight and high- 71. Nascimento P, Marim R, Carvalho G, Mali S (2016) Nanocel-
efficacy antibacterial nanocellulose-based sponge via covalent lulose produced from rice hulls and its effect on the properties of
immobilization of gentamicin. Carbohydr Polym 200:595–601. biodegradable starch films. Mater Res 19:167–174. https://​doi.​
https://​doi.​org/​10.​1016/j.​carbp​ol.​2018.​07.​091 org/​10.​1590/​1980-​5373-​MR-​2015-​0423
55. Ali JB, Danladi A, Bukhari MM et al (2020) Extraction and 72. Md Karimi S, Tahir P, Dufresne A et al (2014) A compara-
characterization of cellulose nanofibres and cellulose nanocrys- tive study on characteristics of nanocellulose reinforced ther-
tals from Sammaz-14 maize cobs. J Nat Fibers 00:1–16. https://​ moplastic starch biofilms prepared with different techniques.
doi.​org/​10.​1080/​15440​478.​2020.​18562​79 Nord Pulp Pap Res J 29:41–45. https:// ​ d oi. ​ o rg/ ​ 1 0. ​ 3 183/​
56. Nisar T, Wang ZC, Alim A et al (2019) Citrus pectin films npprj-​2014-​29-​01-​p041-​045
enriched with thinned young apple polyphenols for potential 73. Ghaderi M, Mousavi M, Yousefi H, Labbafi M (2014) All-cellu-
use as bio-based active packaging. CYTA - J Food 17:695–705. lose nanocomposite film made from bagasse cellulose nanofibers
https://​doi.​org/​10.​1080/​19476​337.​2019.​16407​98 for food packaging application. Carbohydr Polym 104:59–65.
57. Jayaprada et al (2020) Preparation and properties of a micro- https://​doi.​org/​10.​1016/j.​carbp​ol.​2014.​01.​013
fibrillated cellulose reinforced pectin/fenugreek gum biocom- 74. John MJ, Dyanti N, Mokhena T et al (2021) Design and devel-
posite. New J Chem 44:18792–18802. https://​doi.​org/​10.​1039/​ opment of cellulosic bionanocomposites from forestry waste
d0nj0​3101a residues for 3D printing applications. Materials (Basel) 14:3462.
58. Jongaroontaprangsee S, Chiewchan N, Devahastin S (2018) Pro- https://​doi.​org/​10.​3390/​ma141​33462
duction of nanocellulose from lime residues using chemical-free 75. Yu B, Zeng X, Wang L, Regenstein JM (2020) Preparation of
technology. Mater Today Proc 5:11095–11100. https://​doi.​org/​ nanofibrillated cellulose from grapefruit peel and its application
10.​1016/j.​matpr.​2018.​01.​027 as fat substitute in ice cream. Carbohydr Polym 117415. https://​
59. Kumar R, Kumari S, Surah SS, et al (2019) A simple approach doi.​org/​10.​1016/j.​carbp​ol.​2020.​117415
for the isolation of cellulose nanofibers from banana fibers. 76. Mathew B, Lopes A, Ferreira S, et al (2011) Cellulose nanocom-
Mater Res Express 6. https://​d oi.​o rg/​1 0.​1 088/​2 053-​1 591/​ posites with nanofibres isolated from pineapple leaf fibers for
ab3511 medical applications. 86:1790–1798 https://​doi.​org/​10.​1016/j.​
60. Yan N, Chai XS, Runge T (2021) A simple multi-wavelength carbp​ol.​2011.​07.​009
spectroscopic method for the determination of carboxyl group 77. Evelyna A, Astifanni TK, Ruth I, et al (2019) Preparation of nano-
content in nanocellulose. Cellulose 28:2805–2811. https://​doi.​ cellulose-alginate nanocomposites for chlorhexidine digluconate
org/​10.​1007/​s10570-​021-​03731-y drug carrier. IOP Conf Ser Mater Sci Eng 547. https://​doi.​org/​10.​
61. Sun X, Wu Q, Ren S, Lei T (2015) Comparison of highly trans- 1088/​1757-​899X/​547/1/​012046
parent all-cellulose nanopaper prepared using sulfuric acid and 78. Galkina OL, Ivanov VK, Agafonov AV et al (2015) Cellulose
TEMPO-mediated oxidation methods. Cellulose 22:1123–1133. nanofiber-titania nanocomposites as potential drug delivery sys-
https://​doi.​org/​10.​1007/​s10570-​015-​0574-6 tems for dermal applications. J Mater Chem B 3:1688–1698.
62. Zhang Z, Chang H, Xue B et al (2017) New transparent flexible https://​doi.​org/​10.​1039/​c4tb0​1823k
nanopaper as ultraviolet filter based on red emissive Eu(III) 79. Nurul Atiqah MS, Gopakumar DA, Owolabi FAT et al (2019)
nanofibrillated cellulose. Opt Mater (Amst) 73:747–753. https://​ Extraction of cellulose nanofibers via eco-friendly supercritical
doi.​org/​10.​1016/j.​optmat.​2017.​09.​039 carbon dioxide treatment followed by mild acid hydrolysis and the
63. Chen YW, Lee HV, Hamid SBA (2016) Preparation of nano- fabrication of cellulose nanopapers. Polymers (Basel) 11:1813.
structured cellulose via Cr(III)- and Mn(II)-transition metal salt https://​doi.​org/​10.​3390/​polym​11111​813
catalyzed acid hydrolysis approach. BioResources 11:7224– 80. Naduparambath S, T.V. J, Shaniba V et al (2018) Isolation and
7241. https://​doi.​org/​10.​15376/​biores.​11.3.​7224-​7241 characterisation of cellulose nanocrystals from sago seed shells.
64. Debiagi F, Faria-Tischer PCS, Mali S (2020) Nanofibrillated Carbohydr Polym 180:13–20. https://​doi.​org/​10.​1016/j.​carbp​ol.​
cellulose obtained from soybean hull using simple and eco- 2017.​09.​088
friendly processes based on reactive extrusion. Cellulose 81. Wang Z, Yao Z, Zhou J et al (2019) Isolation and characterization
27:1975–1988. https://​doi.​org/​10.​1007/​s10570-​019-​02893-0 of cellulose nanocrystals from pueraria root residue. Int J Biol
65. Panyasiri P, Yingkamhaeng N, Lam NT, Sukyai P (2018) Extrac- Macromol 129:1081–1089. https://​doi.​org/​10.​1016/j.​ijbio​mac.​
tion of cellulose nanofibrils from amylase-treated cassava bagasse 2018.​07.​055

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 Biomass Conversion and Biorefinery

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