Carbohydrate Polymers 148 (2016) 69–77

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Carbohydrate Polymers
journal homepage: www.elsevier.com/locate/carbpol

Nanofibrillated cellulose from tobacco industry wastes

Glaiton Tuzzin, Marcelo Godinho, Aline Dettmer ∗ , Ademir José Zattera
Engineering of Processes and Technologies Post-Graduate Program, University of Caxias do Sul, Caxias do Sul, Rio Grande do Sul, Brazil

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

Article history: Tobacco stems waste underwent steam explosion pulping for nanofibrillated cellulose (NFC) production.
Received 4 February 2016 In order to obtain NFC hydrogels, the pulp obtained by steam explosion was bleached and refined in
Received in revised form 6 April 2016 a grinder employing specific energy of up to 5067 kWh/t. Eucalyptus kraft pulp was processed under
Accepted 9 April 2016
the same conditions to produce NFC hydrogels, later used in order to compare with NFC hydrogels from
Available online 12 April 2016
tobacco stems waste. According to statistical analysis, the optimum tobacco stems pulping condition was
obtained with a severity index of log 3.0 and active alkali of 16.25%. These conditions allowed obtaining
Keywords:
a bleached pulp with Schopper Riegler degree of 46. Electronic microscopy with field emission showed a
Tobacco
Waste higher presence of nanofibers in the tobacco stems pulp than in commercial eucalyptus kraft pulp, both
Steam explosion after refining. Thermal analysis indicated that tobacco stems pulp degrade at lower temperatures than
Nanocellulose eucalyptus kraft pulp. FTIR analysis did not indicate chemical bonding differences between the two pulps.
© 2016 Elsevier Ltd. All rights reserved.

1. Introduction content in weight, which makes it an interesting raw material for

The demographic and economic expansion has been demand-
ing higher amounts of natural resources worldwide. Hence, waste
generation grows and becomes a threat to ecological equilibrium
and to human survival.
Tobacco industry employs tobacco leaves for cigarette and cigar
production (Peševski et al., 2010). According to data of the Brazilian
Association of Tobacco Growers, Brazil produced 607 thousand tons
of tobacco in the 2015/16 harvest. About 4% of the tobacco leaves
processed become a waste, which is basically constituted of tobacco
stems (leaves midribs) and dust from processing.
Tobacco stems correspond to about 20% of the total leaves
weight (Browne, 1990) and present a higher cellulose content
(up to 23%) than the leaf lamina (Leffingwell, 2001; Wakeham &
Silberman, 1966). Since cellulose confers an unpleasant taste when
burnt, tobacco stems are not directly used in cigar and cigarette
production (Leffingwell, 2001). In order to allow tobacco stem use,
it is laminated and turned into reconstituted tobacco, to which
flavorings and humectants are added (Browne, 1990). However,
reconstituted tobacco is only used in low quality cigarette, thus
not consuming all the waste produced (Peševski et al., 2010).
Tobacco stalks, in their turn, are directly disposed on the land
used for tobacco cultivation. They can have up to 40% of cellulose
∗ Corresponding author at: Rua Francisco Getúlio Vargas, 1130, Caxias do Sul,
95070-560 Rio Grande do Sul, Brazil.
E-mail address: alinedettmer@gmail.com (A. Dettmer).
papermaking or for the production of other cellulose derivatives
(Shakhes et al., 2011; Agrupis, Maekawa, & Suzuki, 2000).
Among all the cellulose derivatives, one has been attract-
ing particular interest: nanocellulose. Some advantages of this
new material, which is obtained from renewable sources, is its
high area/volume ratio, high Young’s modulus, its capability of
being biodegraded, and its nontoxic characteristics (Flauzino Neto,
Silvério, Dantas, & Pasquinia, 2013). Nanocellulose can also be
obtained from agricultural wastes like pineapple leaves (Cherian,
Leão, Souza, Pothan, & Kottaisamy, 2010), banana tree stalk (Deepa
et al., 2011), wheat straw (Kaushik & Singh, 2011), rice husk (Kalita
et al., 2015), and pinecone (Rambabu, Panthapulakkal, Sain, & Dalai,
2016). Nanocellulose may be applied in nanocomposites, in elec-
tronic devices, in cosmetic products, and in membranes, among
others (Eichhorn et al., 2010).
Different processes may be used to produce nanocellulose. Acid
hydrolysis produces nanocrystalline cellulose—NCC (Klemm et al.,
2011; Johar, Ahmad, & Dufresne, 2012). Bacterial nanocellulose
(BNC) is produced from sugar consumption by bacteria of the genus
Gluconacetobacter (Dufresne, 2013; Klemm et al., 2011). Although
both processes present high yields, NCC production generates high
amounts of wastewater and BNC production is expensive (Klemm
et al., 2011).
In 1979, researchers at the ITT Rayonnier developed a mechan-
ical process that enables nanofibers production from sulfite pulp
employing high-pressure homogenizers. This material, known as
microfibrillated cellulose (MFC) or nanofibrillated cellulose (NFC),
displays a gel-like behavior. However, this process of obtaining

http://dx.doi.org/10.1016/j.carbpol.2016.04.045
0144-8617/© 2016 Elsevier Ltd. All rights reserved.
70 G. Tuzzin et al. / Carbohydrate Polymers 148 (2016) 69–77
nanocellulose has a high energy cost, which can reach 27 MWh for
each ton of MFC (Klemm et al., 2011).
In order to reduce this energy consumption, enzymatic hydrol-
ysis (Pääkko et al., 2007) and tetramethylpiperidine-N-oxyl
(TEMPO)-mediated oxidation (Kekäläinen, Liimatainen, Biale, &
Niinimäki, 2015) followed by refining process can be used to obtain
MFC hydrogels. Applying a steam explosion pretreatment in the
lignocellulosic material can also reduce nanocellulose production
costs (Cherian et al., 2010; Deepa et al., 2011; Kaushik &d Singh,
2011). This treatment consists in subjecting lignocellulosic materi-
als (with little or no chemical addition) to the action of water vapor
pressure for short periods, followed by rapid decompression. The
increase in specific volume of water vapor during decompression
breaks the lignocellulosic structure, partially hydrolyzing the hemi-
cellulose and allowing access to the cellulosic fibers (Kumar, Barret,
Delwiche, & Stroeve, 2009; Jedvert et al., 2012). According to Kokta
and Ahmed (1998), steam explosion can be considered an alterna-
tive process to thermomechanical and chemithermo-mechanical
processes since it separates and softens the fibers, thereby reduc-
ing energy consumption during refining. When combining the use
of chemical pulping additives and steam explosion, it is possible to
remove substantial quantities of lignin and obtain bleachable pulps
in shorter times than the ones required in a conventional chemical
pulping (Agrupis et al., 2000).
In this work, NFC was produced from tobacco stems waste. The
NFC was obtained by steam explosion of tobacco stems followed
by pulp bleaching and refining. The nanofibers were characterized
by TGA, FTIR and FE-SEM. For comparative data, NFC was produced
from a commercial sample of eucalyptus kraft pulp refined under
the same conditions that tobacco stems pulp were refined.
2. Materials and methods
2.1. Materials
Tobacco stems were obtained from a tobacco industry (Rio Pardo
Valley—RS, Brazil). The steam explosion reactor used in the experi-
ments was designed according to the concepts of Siropulper Process
(Mamers, Menz, & Yurita, 1981).
2.2. Methods
2.2.1. Chemical composition of tobacco stems
The chemical composition of tobacco stems was determined
according to the following TAPPI standards: extractives (T 204 cm-
97), ashes (T 211 om-02), insoluble lignin (T 222 om-02) and
␣-cellulose (T 203 cm-99). Hemicelluloses were determined by
mass difference.
2.2.2. Sample preparation
Raw tobacco stems (RTS) were subjected to a solid-liquid extrac-
tion to reduce their water soluble constituents. For this process,
5 l of water were added to 1.5 kg of RTS. The system was heated
at 90 ◦ C and kept in this temperature for 2 h. The solution was
drained and the extract was collected for other studies not pre-
sented in this work. The procedure was repeated three times. The
partially extracted stems (PES) were then dried under the sun to
avoid microorganisms proliferation. PES characterization was per-
formed according to the methodology presented in Section 2.2.1.
A factorial experimental design was used in order to identify
the best operating conditions for PES pulping by steam explosion.
Two factors were tested in two levels (2k ) and in a central point
(intermediate condition level). The experiments were performed
in duplicates, totalizing 10 tests. In order to evaluate the efficiency
of steam explosion process, the severity index was used. This index
was introduced by Overend and Chornet (1987) and can be calcu-
lated according to Eq. (1).
 T − 100 
R0 = e ×t (1)
14, 75
where R0 = Severity index; T = Water steam temperature (◦ C);
t = Time (min).
The temperature in the experiments was kept at 175 ◦ C to avoid
excessive cellulose degradation. The severity index (expressed
as log R0 ) varied between 3.0 and 3.8. The reaction times were
obtained from Eq. (1). Active alkali was ranged between 0 and
25% (g NaOH/g of PES, on a dry basis). Statistical analysis of the
data obtained from the experimental design was performed using
software Statistica 12.0, with a confidence level of 95%.
About 60 g of PES were placed in a beaker containing 500 ml of
deionized water and this system was kept at room temperature for
24 h. For the tests in which NaOH additions were required, 500 ml
of an impregnation solution were used instead. This solution con-
tained active alkali equivalent to 12.5 or 25.0% and 0.1% (by weight)
of anthraquinone. After 24 h of immersion, the suspensions were
drained.
2.2.3. Steam explosion pulping
The steam explosion reactor digester was charged with impreg-
nated PES (Section 2.2.2) and then fed with saturated steam at
175 ◦ C. After the experiment time, the material was suddenly
released to the blow tank through a nozzle. Due to this expan-
sion, two phases (liquid and solid) were formed inside the blow
tank. The solid phase (rich in fiber) was collected and placed onto a
200 mesh screen (Tyler mesh) where it was rinsed with water. The
liquid phase (black liquor) was discharged. The gross yield of the
process was calculated and the residual lignin content in the fibers
was determined by TAPPI standard T236 om-06 (kappa number).
The pulps obtained in each test were bleached by immersing
them, respectively, in NaClO (5% w/v), Na2 SO3 (5% w/v), NaClO (5%
w/v) and H2 O2 (10% w/v). The temperature used in the immersion
was 50 ◦ C and the pulps were kept for 10 min in each immersion
stage. The bleaching stage yield was calculated.
Through statistical analysis it was defined the optimum operat-
ing condition of the steam explosion reactor for bleachable pulp
production. In this condition, three tests were conducted using
200 g of PES each. The obtained pulps were bleached according to
the previously described methodology.
2.2.4. Refining process
For refining, the pulps obtained in the optimum condition (EBS)
were diluted to 5 l (which resulted in a concentration of 1.8% w/v
of fibers). The material was then loaded in a grinder (Supermass-
colloider Masuko Sangyo—MKCA6-2) operated under the following
conditions: 2500 rpm, current consumption of 1A, 0,1 mm between
discs and recycle flow of 0.6 l/min for 4 h. Eucalyptus kraft pulp
(EKP), supplied by Eldorado Brazil in the form of dry boards, was
refined under the same conditions for comparative data. Pulp
aliquots (100 ml) of both samples were collected every hour of
processing.
The specific refining energy applied to the fibers was calcu-
lated according to the equation proposed by Annergren and Hagen
(2009) as shown in Eq. (2).
(P − P0 ) × t
ENET = (2)
c×v
where ENET = Specific refining energy (kWh/t of fiber, dry
basis); P = Total refining power (kW); P0 = No load power (kW);
c = Concentration of fibers in the pulp (consistency) (t/m3 );
t = Refining time (h); v = Pulp volume (m3 ).
 G. Tuzzin et al. / Carbohydrate Polymers 148 (2016) 69–77 71

P and P0 were calculated based on the product of voltage Table 1

Tobacco stems characterization.
(380 V) and current consumption of the grinder loaded (1.0 A) and
unloaded (0.7 A). The degree of drainability of the pulp obtained in Constituents (% m/m) Tobacco stems
every hour of refining was measured by Schopper Riegler method
RTS EFS PES
(◦ SR) (ISO 5267-1: 1999).
O.S. extractives 3.39 ± 0.13a 0 1.24 ± 0.21a
Hot water extractives 46.53 ± 0.56a 0 21.91 ± 0.72a
2.2.5. Infrared fourier transform spectroscopy (FTIR) Ash 20.78 ± 0.20a 9.75 ± 0.17a 9.96 ± 0.27b
FTIR analyses were performed for RTS and for the samples col- Insoluble lignin 6.93 ± 0.13b 13.84 ± 0.26a 10.13 ± 0.24b
lected in different processing steps: partially extracted stems (PES), ␣-cellulose 20.85 ± 0.52b 41.75 ± 0.64a 35.42 ± 0.69b
Hemicelluloses 1.86 ± 0.81b 34.36 ± 0.42a 21.34 ± 0.34b
extractives free stems (EFS), steam exploded stems (SES), exploded
and bleached stems (EBS), and eucalyptus kraft pulp (EKP). The Note: O.S. = Organic solvents.
a
equipment used was a spectrometer (Thermo Scientific Nicolet, Values obtained according to Section 2.1.
b
Values previously obtained in extractives free basis (requirement of TAPPI stan-
model IS10, United States) with KBr technique (for RTS, PES, EFS,
dards) and corrected to as-received basis.
SES) and ATR technique (for EKP and EBS) in a wave number range
from 400 to 4000 cm−1 .
image analyzer software (ImageJ). The diameters frequency distri-
2.2.6. Thermal analysis (TGA/DTG) bution was then obtained.
Thermal analysis was performed to evaluate the thermal degra-
dation of RTS, PES, EFS, SES, EBS, and EKP. The thermal behavior 2.2.8. X-ray diffraction analysis (XRD)
of tobacco stems NFC and eucalyptus NFC (NFC-EBS and NFC-EKP, The crystallinity of EBS, EKP, NFC-EBS and NFC-EKP samples
respectively) after 4 h of refining was also evaluated. The tests were was evaluated in a X-Ray diffractometer (XRD-6000, Shimadzu,
performed under N2 atmosphere at a heating rate of 10 ◦ C/min from Japan) using the following conditions: Cu-k␣ 0,1542 nm, 40 kV,
room temperature to 600 ◦ C. The equipment used was a simul- 30 mA, 5◦ < 2␪ < 35◦ , 0.05◦ s−1 . The crystallinity index was calcu-
taneous thermal analyzer (STA, Model Jupiter 449, Netzsch, 119 lated according to the method proposed by Segal, Creely, Martin,
Germany). and Conrad (1959) (Eq. (3)) (Lengowski, Muniz, Nigovski, &
Magalhães, 2013):
2.2.7. Scanning electron microscopy with field emission (FE-SEM) I200 − Iam
Pulp samples from EBS and from EKP (obtained according to the I(%) = × 100 (3)
I200
methodology of Section 2.2.4 after 0, 2 and 4 h of refining) were
analyzed by FE-SEM in order to evidence the presence of cellu- were I200 = Peak intensity corresponding at (200) plane at diffrac-
losic fibers and nanofibers and to determine their morphology. The tion angle of 2␪ = 22.5◦ ; Iam = Intensity of the valley between (200)
equipment used was a Scanning Electron Microscope with Field and (100) planes at a diffraction angle of 2␪ = 18.0◦ .
Emission (Mira 3 model, TESCAN, Czech Republic). The samples
were dried by supercritical fluid extraction and then prepared by 3. Results and discussion
replacing the water present in the pulp for ethanol. The replace-
ment was accomplished by centrifuging 50 ml of pulp at 4500 rpm 3.1. Tobacco stems characterization
for 10 min, followed by collecting the supernatant and redissolv-
ing the fibers in ethanol. This procedure was repeated 6 times. Table 1 shows the characterization of the tobacco stems used
An aliquot of 25 ml of fiber suspension in ethanol was dried with for NFC production
carbon dioxide in a supercritical extractor (model SFT-150, Super- Tobacco stems have high amounts of extractives (both in organic
critical Fluids Technologies, United States) by dynamic extraction at solvents and in water), corresponding to approximately 50% of
1400 psi (9.52 MPa) and 45 ◦ C for 6 h. The process was followed by their total mass. The extractives contain mainly starch and pectin
static extraction at a rate of 8 psi/min (54.4 kPa/min) until room besides other constituents that are responsible for tobacco color,
pressure, at the same temperature. The dried fibers were then odor, and taste (Leffingwell, 2001). Tobacco stems have about 20%
placed in a desiccator. For FE-SEM analysis, a small fraction of (m/m) of ␣-cellulose. Wakeham and Silberman (1966) obtained cel-
these fibers was carefully deposited on a stub containing conduc- lulose concentrations between 13 and 23% in tobacco stems. The
tive adhesive tape for subsequent coating with gold. The evaluation concentration of insoluble lignin (13.84 ± 0.26% in an extractives
of micrographs allowed the determination of NFC-EBS and NFC-EKP free basis) is higher than the value obtained by Sewalt et al. (1997)
nanofibers diameters (for samples after 4 h of refining). For this, 200 (10.95 ± 0.2% (m/m)). These values may vary depending on the type
random measures of the fibers were taken in each image using an and on the position of the tobacco stems in relation to the plant

Table 2
Results for gross yield, kappa number, ␣-cellulose and removed lignin.

Test LogR0 A. A. Gross yield (%) Bleaching yield (%) Final yield (%) Kappa number Removed ligninb (%) ␣-cellulosea (%)

1 3.8 25.0 32.58 80.42 26.20 43.2 ± 2.2 81.92 ± 0.96 67.2 ± 0.9
2 3.8 0.0 55.08 58.27 32.10 102.1 ± 2.9 27.81 ± 2.07 64.6 ± 0.1
3 3.8 25.0 33.42 79.65 26.62 41.7 ± 1.1 82.09 ± 0.47 65.0 ± 0.5
4 3.0 0.0 69.10 49.19 33.99 107.7 ± 0.8 4.48 ± 0.75 60.0 ± 0.7
5 3.8 0.0 54.44 60.53 32.95 103.4 ± 2.1 27.78 ± 1.47 63.5 ± 0.3
6 3.4 12.5 44.41 66.74 29.64 74.4 ± 1.5 57.63 ± 0.87 62.7 ± 0.1
7 3.0 25.0 41.01 74.46 30.54 48.0 ± 0.9 74.72 ± 0.49 61.4 ± 0.3
8 3.0 25.0 41.67 75.50 31.46 46.4 ± 0.4 75.21 ± 0.24 61.0 ± 0.3
9 3.4 12.5 45.50 65.83 29.95 76.4 ± 0.8 55.37 ± 0.45 61.9 ± 0.2
10 3.0 0.0 67.76 49.85 33.77 106.2 ± 1.4 6.61 ± 0.64 59.1 ± 0.1

Notes: Times for severity index: log (3.0) = 7 min; log (3.4) = 16 min; log 3.8 = 39 min. A. A. = Active Alkali.
a
Results for bleached pulps. Yields expressed on a dry basis.
b
Values obtained through mass balance.
72 G. Tuzzin et al. / Carbohydrate Polymers 148 (2016) 69–77

Fig. 1. Pareto diagrams for experimental design. Kappa number (a) and gross yield (b).

Fig. 2. Charts obtained for evaluation of steam explosion process and desirability profiles for optimum condition determination.

height. The removal of extractives (even if partial) reduced the ash
content, indicating that most of the inorganic matter in the tobacco
stem is soluble in water. Low hemicellulose values in the raw stems
are explained by the calculation methodology (mass difference),
due to the percentage of ash present in the extractives.
3.2. Steam explosion
Table 2 presents the results of the tests performed according to
the experimental design of this study.
The results indicate a low gross yield for the tests performed
with high severity index and high active alkali (tests 1 and 3).
This behavior is justified by the combined action of water vapor
hydrolysis and of delignification promoted by pulping reagent. As
the lignin is removed, hemicellulose and cellulose are separated
from the lignocellulosic matrix, allowing the hemicellulose to sol-
ubilize and the cellulose fibrils to disperse in the liquor. The shorter
cellulose chains (low degree of polymerization) dispersed in the
liquor tend to be solubilized by the action of alkali and temperature
after some time. This phenomenon (alkaline solubilization) occurs
whenever any new region containing fibers becomes accessible or
when there is a significant reduction in fibers size (Sixta, 2006). The
low polymerization degree (between 600 and 1800) (Leffingwell,
2001) of tobacco stems fibers favors the alkaline solubilization phe-
nomenon.
In tests performed with 25% of active alkali and low severity
index (tests 7 and 8), the yield was found to be slightly higher
than the ones of the tests 1 and 3. Lower residence time in the
 G. Tuzzin et al. / Carbohydrate Polymers 148 (2016) 69–77
reactor resulted in smaller carbohydrate amounts solubilized. The
highest gross yields were obtained with a low severity index in the
absence of active alkali (tests 4 and 10). Under these operating con-
ditions, the solubilization of carbohydrates occurs only as a result
of water vapor hydrolysis. In the absence of active alkali and with
high severity index (tests 2 and 5), the gross yield reduced to about
55%. Intermediate conditions (center point, tests 6 and 9) resulted
in gross yield similar to tests 7 and 8.
Tests 1 and 3, conducted with high severity index and high active
alkali, resulted in the lowest kappa numbers. These tests resulted in
maximum tobacco stems delignification because of the high reac-
tion time and active alkali presence. Kappa numbers of tests 7 and 8
are similar to those obtained in the tests 1 and 3; therefore, the reac-
tion time did not contribute significantly to the removal of lignin.
This is confirmed by tests 6 and 9 (center point), in which the reduc-
tion of active alkali raised kappa number, even when severity index
was increased.
The tests without addition of alkali resulted in high kappa num-
bers (low delignification). For maximum severity index (tests 2 and
5), there was a considerable initial removal of lignin but other car-
bohydrates degradation (evidenced by gross yield of 55% shown
in Table 2). There was practically no lignin removal in tests 4 and
10, performed with low severity index and in the absence of active
alkali.
Lignin reactions in the lignocellulosic matrix, which happen due
to the steam explosion process, begin by the hydrolysis of acetyl
groups from hemicellulose, which acidify the medium. Acidic con-
ditions favor the formation of a carbocation that attacks benzene
ring ␤-O-4 bonds. This electrophilic attack breaks the ␤-aryl-ether
bonds, forming a ketone and a new phenolic group, starting the
lignin depolymerization. At the same time, in the presence of other
electron rich carbons, such as the C-2/C-6 of syringyl and guaia-
cyl groups, lignin condensation reactions may occur, competing
with acid hydrolysis. It results in penta-substituted aromatic rings,
which are less reactive (Li, Gellerstedt, & Toven, 2009). These com-
petitive reactions difficult ligning solubilization when it occurs only
by water vapor. According to Assumpção, Pinho, Cahen, and Phillip
(1988) and Pires, Kuan, Benazzi, and Bugajer (1988), in the presence
of alkali, the acetic acid formed is neutralized and the ␣-aryl-ether
bonds are cleaved by hydroxyl groups (the ␤-aryl ether bonds are
preferably cleaved in the presence of HS− ions in the kraft process).
This is why high concentrations of NaOH were more effective in
ligning removing. Fig. 1 shows the Pareto diagrams for gross yield
and kappa number. Variations in the active alkali values results in
more significant changes in kappa number and in gross yield than
variations in the severity index.
3.2.1. Steam explosion (optimum condition)
The optimum severity index and active alkali were determined
as the ones that led to the maximum gross yield and lowest kappa
number. As can be seen in Fig. 2, an active alkali concentration of
16.25% and a log3.0 severity factor are the optimum conditions for
the process. They result in a kappa number of 68.7 and in a gross
yield of 50.8%.
Three tests were conducted to check the optimum condition
obtained. They resulted in a gross yield of 52.2 ± 1.7% and in a
kappa number of 70.8 ± 2.3. It indicates a good experimental repro-
ducibility. The removal of lignin from tobacco stems (51.4 ± 1.3%)
was satisfactory considering the low reaction time (7 min). The
removal of lignin at a low reaction time indicated that the reac-
tion rate of delignification for tobacco stems is more significant
in the beginning of the process, fact confirmed by the pioneering
studies of Kleinert (1964). According to the author, lignin under-
goes structural changes over time and becomes less reactive than
the originally present lignin.
73
Fig. 3. NFC hydrogel for EBS after two hours of refining and its gel-like behavior.
Table 3
SR and specific refining energy for EBS and EKP.
◦
Time (h) Enet (kWh/t)a SR
EBS EKP
0 0 46 20
1 1267 90 54
2 2533 10 85
3 3800 10 10
4 5067 9 11
a
Value determined through Eq. (2).
3.3. Refining
EBS pulp absorbed high water quantities during the first pro-
cessing hour. After about 90 min, the pulp became gelatinous and
so remained until the end of the process. In the test conducted with
the EKP, the gelatinous material was obtained after the third pro-
cessing hour. Stelte and Sanadi (2009) also observed difficulties in
hardwood fibers delamination. The authors used the same grinder
model employed in this study and found that even after 75 passes,
the structure of the fibers remained not altered.
An EBS aliquot collected in the second hour of processing was
dissolved to a fiber concentration of 0.5% (w/v) and subjected to
ultrassonification in a sonicator (SONICS, model vibracell) for 1 min
to improve fibers dispersion and remove air bubbles. After the
dispersion, the material displayed a gel-like behavior and good
transparency, which are common features of the NFC hydrogels,
as can be seen in Fig. 3.
As it is shown in Table 3, EBS pulp has a higher initial ◦ SR than
EKP. This value changes as a function of the specific refining energy
applied. This may be related to the fibrillation effect caused by the
steam explosion process. According to Wagberg (2009), higher fibril
areas exposed leads to an increase in the water amount retained
by these fibrils and by their capillaries due to surface tension. EKP
fibers retained less water than EBS, probably because the first ones
were previously dried. Dry fibers adsorb less water than humid
fibers when processed into pulp since pores are irreversibly closed
during drying process. It restricts water penetration into the fibrils,
which do not swell easily.
When employed a specific refining energy of 1267 kWh/t, ◦ SR
of EBS increased to 90. The method used for ◦ SR determination is
valid for samples that present ◦ SR between 10 and 90. Therefore, the
value found is in the validation and reliability limit of the method.
For the same specific energy employed, ◦ SR of EKP increased to 54.
With specific refining energy of 2533 kWh/t, the EBS pulp was
not retained in the machine screen, causing a low ◦ SR and invali-
dating the measurement. In this condition, the eucalyptus cellulose
pulp had its ◦ SR increased to 85. For specific refining energies of
3800 and 5067 kWh/t, due to the cutting and fibrillation effect,
fibers of both pulps were dragged through the equipment screen,
resulting in low ◦ SR that could not be validated. Nevertheless, it
confirmed that the tobacco stem fibers were more susceptible to
74 G. Tuzzin et al. / Carbohydrate Polymers 148 (2016) 69–77

Fig. 4. FE-SEM micrographs for EBS (a–c) and EKP (d–f) pulps obtained after 0 h (a and d), 2 h (e and b) and 4 h (c and f) of refinnig. Diameter distribution of NFC-EBS (g) and
NFC-EKP (h) samples after 4 h of refining.

fibrillation action than the eucalyptus fibers when the same spe-
cific energy was employed. This was only observed up to specific
refining energies of 2533 kWh/t.
3.4. Scanning electron microscopy with field emission (FE-SEM)
The micrographs shown in Fig. 4 indicate that EBS (4-a) have
partial fibrillation on the external layer, an effect that may be
attributed to the steam explosion process. EKP (4-d) are more cohe-
sive and do not present fibrillation on the external layer (primary
wall). Partial fibrillation on the external walls of EBS fibers is one
 G. Tuzzin et al. / Carbohydrate Polymers 148 (2016) 69–77 75

Fig. 5. (a) Thermal analysis (TGA/DTG) of RTS, PES and EFS. (b) Thermal analysis (TGA/DTG) of SES, EBS, EKP, NFC-EBS and NFC-EKP samples.

Fig. 6. (a) FTIR spectra for RTS, PES, EFS and SES samples. (b) FTIR spectra of EBS and EKP samples.

of the factors responsible for them to have higher ◦ SR than EKP
fibers. This delamination increases the area of exposed fibrils and
the amount of water retained by them and by their capillaries
due to the surface tension. The disruption of the outer layers also
facilitates the entry of water into the inner layers of the fibers, pro-
moting their swelling because of the new hydrogen bonds created
between water and cellulose/hemicellulose amorphous fractions.
This is called internal fibrillation, commonly associated with the
refining effects (Annergren & Hagen, 2009).
When applied 2533 kWh/t of specific refining energy to EBS (4-
b), the effects of fibrillation also reached the inside walls, exposing
some nanofibrils. When the same condition was applied to EKP
(4-e), the fibrillation effect was less pronounced and restricted to
the external layers of the fibers. The EBS was almost completely
fibrillated and nanofibers were produced when a specific refining
energy of 5067 kWh/t (4-c) was employed. In this condition, the
presence of some partially delaminated fibers in EKP (4-f) indicates
that fibrillation was not homogeneous.
Fig. 4g and f shows the diameter distribution for, respectively,
the NFC-EBS and NFC-EKP fibers. Most of the fiber diameters were
found to be between 20 and 70 nm for both samples. For the NFC-
EBS sample, 77% of the fibers have diameters between 20 and
50 nm, while 65% of NFC-EKP fibers have diameters between 20
and 50 nm.
3.5. Thermogravimetric analysis
The thermogravimetric curves and their derivative (TGA/DTG)
for the RTS, PES and EFS samples are shown in Fig. 5a.
RTS showed four different thermal degradation peaks in the
DTG. The first peak (80 ◦ C) refers to the maximum rate of mois-
ture loss (Sung & Seo, 2009). The second peak (170 ◦ C) is associated
with the release of volatile and non-polymeric compounds (Sung
& Seo, 2009; Gómez-Siurana et al., 2013). The third peak (250 ◦ C)
indicates the maximum rate of hemicellulose and pectin degrada-
tion (Muramatsu, 1979; Sung & Seo, 2009). The last peak observed
(around 300 ◦ C) represents the maximum rate of cellulose degra-
dation (Gómez-Siurana et al., 2013; Sung & Seo, 2009).
For the PES and EFS samples, the maximum rate of moisture loss
is similar to the one of RTS. However, for these samples, there was
no volatile compounds release peak. In the region of pectins and
hemicelluloses thermal degradation, it is possible to observe two
small shoulders at 249 ◦ C and 265 ◦ C. The absence of the volatile
compounds thermal decomposition peak, as well as the intensity
reduction of the pectins thermal degradation peak, indicate that
these compounds were removed during the solid-liquid extraction,
to which the samples were submitted before steam explosion.
The EBS and EKP showed a maximum of cellulose degradation
rate at around 360 ◦ C (Fig. 5b). The maximum SES degradation
occurred at a lower temperature (340 ◦ C), indicating the presence of
76 G. Tuzzin et al. / Carbohydrate Polymers 148 (2016) 69–77
non-cellulosic material in the sample (hemicellulose and residual
lignin). In the bleaching process these constituents are removed,
giving a higher structural order degree to the remaining pulp and,
consequently, higher thermal stability (Kaushik & Singh, 2011).
The NFC-EBS showed a thermal degradation peak at approxi-
mately 350 ◦ C, revealing the influence of the refining process in
decreasing thermal stability. The maximum thermal degradation
of the NFC-EKP (approximately 360 ◦ C) was similar to the one
observed for EKP, indicating that the refining process did not sig-
nificantly change thermal stability of this sample, due its partial
fibrillation.
3.6. Infrared fourier transform spectroscopy (FTIR)
The FTIR spectra of the samples RTS, PES, EFS and SES are
presented in Fig. 6a. Similar spectra for the four samples were
observed. Between 3700 and 3100 cm−1 , vibrations of OH bonds
occur (Kaushik and Singh, 2011; Prasad et al., 2011) in the sam-
ples. The band around 2900 cm−1 is related to asymmetric and
symmetric stretching of C H bonds (Gómez-Siurana et al., 2013).
Between 1600 and 1680 cm−1 , elongation of S = saturated C bonds
occurs (Gómez-Siurana et al., 2013.), while the band at 1630 cm−1 is
associated with vibrations of the carboxylic groups (McCann, Shi,
Roberts, & Carpita, 1994; Prasad et al., 2011). With regard to the
possible effects of extractions on the transmittance bands, it was
observed smoothed peaks in the region below 1500 cm−1 , where
it is not possible to attributed signatures for particular vibrations,
since, in this region, complex overlap effects may occur (McCann
et al., 1994).
With respect to the SES, it was noted the softening of a small
peak at 1738 cm−1 , which can be attributed both to hemicellulloses
acetyl and uronic ester groups or to carboxyl groups ester bonds
of the of lignin p-coumaric acid (Kaushik & Singh, 2011). These
constituents were partially removed during the steam explosion
process.
The FTIR spectra for EBS and EKP samples are shown in Fig. 6b.
The bands for both samples are similar, which shows that the
bleached pulp obtained after steam explosion did not change in its
chemical nature if compared to EKP. The spectra of both samples is
typical of cellulose, presenting bands between 3700 and 3100 cm−1
and 2900 cm−1 (Kaushik & Singh, 2011). The band at 1450 cm−1 is
associated with the bending vibration of the CH bonds. Between
1200 and 1050 cm−1 it is shown the elongation of carbonyl groups,
while the band at 894 cm−1 is related to the elongation of cellu-
lose typical pulp ␤-glycoside bonds (Ciolacu, Ciolacu, & Popa, 2011;
Kaushik & Singh, 2011).
3.7. X-ray diffraction (XRD)
EBS, NFC-EBS, EKP, and NFC-EKP samples presented degrees
of crystallinity of, respectively, 66.4, 63.8, 82.5, and 72.8%. EKP’s
degree of crystallinity was similar to the ones obtained by Lavoratti,
Scienza & Zattera (2016) and Lengowski et al. (2013) for Eucalyptus
Kraft Pulps (83.5 and 79.2%, respectively). The EBS sample pre-
sented lower degree of crystallinity than the EKP sample. This may
have happened due to the different enzymatic processes that are
responsible for the cellulose bio polymerization on the two species
(Sixta, 2006).
For the NFC-EKP and NFC-EBS samples, it was observed a reduc-
tion in the degree of crystallinity, which was also observed by
Iwamoto, Nakagaito, and Yano (2007), Lengowski et al. (2013),
and Lavoratti et al. (2016). These authors used the same grinder
employed in this work. According to them, the mechanical pro-
cessing fractures the crystalline domains due to high shear rates,
favoring the formation of amorphous structures.
4. Conclusions
It was concluded that it was possible to obtain NFC through
tobacco stems pulping by steam explosion followed by bleaching
and refining, which was the objective of this work. FE-SEM analyses
indicated higher amounts of nanofibers in the cellulose obtained
from these wastes than for the one obtained from eucalyptus kraft
pulp for the same specific refining energy. FTIR analysis showed no
differences in chemical nature between the two cellulose sources.
Therefore, it can be concluded that nanofibrillated cellulose pro-
duction from tobacco stems can be a promising alternative for the
reuse of these organic residues.
Acknowledgements
The authors thank to the Engineering of Processes and Technol-
ogy Post-Graduate Program of the University of Caxias do Sul, to the
Brazilian Research and Project Financial Agency (FINEP—agreement
01.12.0359.00), to the Central Microscopy Laboratory Prof. Israel
Baumvol for the FE-SEM analysis and to the Department of Science
and Technology of the Rio Grande do Sul State.
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