Philippine e-Journal

Philippine e-Journal for Applied Researchfor

Website: pejard.slu.edu.ph
Applied Research and Development 12(2022), 1-11
and Development
ISSN 2449-3694 (Online)
ISSN 2449-3694 (Online)
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Comparative Characterization of Nanocellulose Produced from

Kawayan Kiling (Bambusa vulgaris Schrad ex. Wendl) Pulp Using
Different Pretreatment Methods

Herbert Dunhill R. Aumentado1,*, Ramon A. Razal1, Ruel C. Nacario2, and Marlon N. Manalo2
1
Department of Forest Products and Paper Science, College of Forestry and Natural Resources, University of the
Philippines Los Baños, College, Laguna
2
Institute of Chemistry, College of Arts and Sciences, University of the Philippines Los Baños, College, Laguna
* Corresponding author ([email protected])

Received, 05 June 2021; Accepted, 15 December 2021; Published, 31 January 2022

Copyright @ 2022 H.D.R. Aumentado, R.A. Razal, R.C. Nacario, and M.N. Manalo. This is an open access article
distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and
reproduction in any medium, provided the original work is properly cited.

Abstract

Kraft pulp from kawayan kiling (Bambusa vulgaris Schrad ex. Wendl) were subjected to two
separate pre-treatment procedures (namely, chemical and mechanical) prior to sulfuric acid
hydrolysis for nanocellulose production. Experimental nanocellulose yields were 26.94% for
the bleached and alkali pretreated pulp and 1.20% for the mechanically refined pulp. FTIR
spectroscopy showed that the chemically pre-treated bamboo pulp yielded nanocellulose with
no residual lignin while the ground, acid-hydrolyzed pulp retained some lignin. FIB-FESEM
imaging showed that nanofibrils with average widths of 39.13 + 34.11 nm were formed as
hydrolysis products from the friction grinder treated material while nanometer-sized crystals
with average widths of 134.2 + 34.33 nm were produced from the chemically treated bamboo
pulp. EDS analysis showed no impurities in the products from the chemical treatment, whereas
the presence of silicon was detected in the mechanically refined pulp. Based on XRD analysis,
the degree of crystallinity of the nanofibrils and crystals were 49.47% and 56.92%, respectively.
In future works, B. vulgaris pulp that will be mechanically treated may require bleaching to
further remove residual lignin and thus improve acid hydrolysis.

Keywords: methods of nanocellulose production, acid hydrolyzed production of nanocellulose,

bamboo pulp bleaching and alkali digestion, friction grinder supermass colloider, nanofibrils

Introduction 2010), corn stover (Costa et al., 2015), cacao pod

husk (Hutomo et al., 2015), sugarcane bagasse
Nanocelluloses have applications in (Wulandari et al., 2016), wood pulp (Thanh
nanocoats, bioimaging, biomedicine, gas barrier et al., 2019), and bamboo (Razal, 2016; Wang
films and optically-transparent functional et al., 2015; Chitbanyong et al., 2018). In the
materials (Jiang & Hsieh, 2013). Nanocellulose Philippines, bamboos are a promising source of
can be obtained from plants especially from nanocellulose, as they are ubiquitous, easy to
lignocellulosic-rich fibers found primarily in grow in almost all climate types, and have the
the stems and branches of trees and the culms ability to regenerate quickly when cut.
of bamboos. Several plants have been explored The common local varieties of bamboo used
for nanocellulose production like cotton stalks for pulping and nanocellulose production are
(Soni et al., 2015), grain straws (Oun & Rhim, the kawayan kiling (Bambusa vulgaris Schrad
2016), pineapple leaf fibers (Cherian et al., ex Wendl) and the kawayan tinik (Bambusa
2 H.D.R. Aumentado, R.A. Razal, R.C. Nacario, and M.N. Manalo

blumeana J.A. & J.H. Schultes). Kawayan kiling study will explore on two methods of pretreating
culms contain a host of natural products with the Kraft bamboo pulp prior to acid hydrolysis.
potential for commercial application (Fei et al., The first will be an all-chemical sequence
2016), and several studies were conducted to involving bleaching and alkali digestion while the
determine the suitability of this for pulp and second will dispense of the chemical treatments
paper (Ameh et al., 2017; Sekyere, 1994; Escolano in favor of mechanical disintegration by multiple
and Semana, 1970). Altenatively, Bayani et passes of the pulp in a friction grinder supermass
al. (2016) tested the use of acid-hydrolyzed colloider. The nanocellulose resulting from the
cellulose nanowhiskers from kawayan tinik as two different pretreatment methods will then be
reinforcement for xylan films. Villegas (2015) also analyzed and compared.
attempted to incorporate cellulose nanowhiskers
from kawayan tinik as a reinforcement in
thermoplastic starch. Meanwhile, Jara (2019) Materials & Methods
used newly emerging bamboo shoots of B. vulgaris
as feedstock for nanocellulose production without Materials
prior pulping, capitalizing on the low lignin
content of young bamboo shoots. The kawayan kiling culms were obtained
Several preparatory steps such as pulping from the Makiling Forest Reserve, University of
and removal of residual lignin are employed the Philippines Los Baños (UPLB). All reagent-
to separate cellulose from the lignocellulosic grade chemicals were purchased from commercial
matrix. However, these methods commonly use sources and were used without further purification
chlorine containing compounds that could cause unless noted. The apparatus and equipment
instability and irregularities in the properties used in nanocellulose isolation are located at
of the nanocrystals produced (Dufresne, 2018). the Wood Chemistry Laboratory, Department
Moreover, the many steps required could affect of Forest Products and Paper Science, College of
production cost and energy requirement, and Forestry and Natural Resources, University of
likewise have environmental impact. To address the Philippines Los Baños and the Institute of
these problems, it would be helpful to search for Chemistry, College of Arts and Sciences, UPLB.
methods of producing nanocellulose that can
overcome the disadvantages and limitations of Sample Preparation
production processes that are highly dependent
on the use of chemicals. Preparation of bamboo culms for pulping was
Acid hydrolysis is by far the most common done in accordance with the procedure reported
method of producing nanocellulose from by Razal (2016). Bamboo culms were examined
lignocellulosic materials, with either sulfuric for blemishes or damage and those found to be
acid (Mascheroni et al., 2016; Zhang et al., 2020) defective were discarded. The selected culm was
or hydrochloric acid (Hastuti et al., 2018), or both cut into 1 to 1.5-inch rings, leaving out portions
(Yu et al., 2013; Hutomo et al., 2015) serving that contain the nodes. The bamboo skin was
as the inorganic catalyst to break down the removed and then the bamboo rings were further
isolated cellulose. Other reagents and methods subdivided into chunks using a utility cutter.
tested are ammonium persulfate reaction The bamboo chunks were air-dried for three
(Mascheroni et al., 2016; Zhang et al., 2020), days and were made to pass through a Pallmann
TEMPO (2,2,6,6-tetramethylpiperidine-1-oxyl PHM3 Model 10180015 (Germany) hammermill
radical) oxidation (Isogai et al., 2011), and FeCl3- for further breakdown.
catalyzed hydrolysis assisted by ultrasonication
(Lu et al., 2014). Kraft Pulping
This study aims to find a method of
preparing nanocellulose from kawayan kiling Weighed amounts of NaOH and Na2S were
that minimizes the issues attendant to the use sequentially dissolved in a predetermined volume
of chemicals in pretreatment procedures prior of water and added to hammermilled bamboo
to acid hydrolysis of cellulose. Towards this, the chips in a large glass container. The quantity of
 3

92 subdivided into chunks using a utility cutter. The bamboo chunks were air-dried for three days and
93 were made to pass through a Pallmann PHM3 Model 10180015 (Germany) hammermill for further
94 breakdown.Philippine e-Journal for Applied Research and Development 12(2022), 1-11
Philippine e-Journal for Applied Research and Development
3
Website: pejard.slu.edu.ph ISSN 2449-3694 (Online)
95
96 Kraft Pulping
97
pulping chemicals used, and the water volume the flask with subsequent addition (3 mL each) of
98 Weighed amounts of NaOH and Na2S were sequentially dissolved in a predetermined volume of
99 were
water based
and added on calculations
to hammermilled bamboothat provided
chips in 25%
a large glass container.the
TheNaClO
quantity2ofsolution
pulping after 0.5, 1.0, 1.75, and 2.5
100 sulfidity, 13% effective alkali, and a liquor to hr., with swirling
chemicals used, and the water volume were based on calculations that provided 25% sulfidity, after every addition. After 3.25
101 13%bamboo chip ratio
effective alkali, (ovendry
and a liquor basis)
to bamboo chipofratio
10:1,
(ovendry basis) ofhr., the mixture was suction-filtered and washed
10:1,
102 with portions of chilled distilled water, four 75-
𝑁𝑁𝑁𝑁2𝑆𝑆
103 where: %𝑆𝑆𝑆𝑆𝑆𝑆𝑆𝑆𝑆𝑆𝑆𝑆𝑆𝑆𝑆𝑆𝑆𝑆 =
where: (𝑁𝑁𝑁𝑁𝑁𝑁𝑁𝑁+𝑁𝑁𝑁𝑁2𝑆𝑆)
EquationEquation
× 100 1 mL 1 quantities of 1% CH COOH and two 30-mL
3
104 quantities of methanol.
and and 𝐸𝐸𝐸𝐸𝐸𝐸𝐸𝐸𝐸𝐸𝐸𝐸𝐸𝐸𝐸𝐸𝐸𝐸 𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴 = 1/2𝑁𝑁𝑁𝑁2𝑆𝑆 EquationEquation
2
𝑁𝑁𝑁𝑁𝑁𝑁𝑁𝑁
105 2
106 2. Alkali-insoluble Cellulose Preparation
107 and NaOH and NaOH andand NaNa S are the weights (g)
2 2S are the weights (g) of sodium sulfide and sodium
108 of sodium sulfide
hydroxide, and sodium
respectively,
expressed as Na2O (Bajpai, 2018). Cellulose was prepared from the holocellulose
109 hydroxide, respectively, expressed as fraction following the modified TAPPI procedure
110 The improvised
Na2O pulping
(Bajpai, digester
2018). that was assembled for the pulping of thefor bamboo chips consisted
alpha-cellulose determination (TAPPI
111 of a large glass vessel, a pressure cooker, and burner. The glass vessel containing the chips and
112 Test Method
“liquor” was covered with cheesecloth and placed inside the pressure cooker partially filled with 203 cm-09, 2009). To the bamboo
113 The improvised pulping digester that was o holocellulose
distilled water, then sealed. It was heated up to a temperature of 121 C and 15 psi pressure, which (3 g oven-dry weight) preparation
114 wasassembled
then maintainedforforthe pulping2 hours.
an additional of theThebamboo chips
pressure was released,inand
a the
flask,
cooked 60bamboo
mL of 17.5% NaOH solution was
115 pulpconsisted
was placedofover
a large glass vessel,
cheesecloth a pressure
and thoroughly washed cooker, added
with tap water withand heated
squeezing. Theat 20oC in a water bath for 30
116 resulting pulp was then
and burner. Thefurther
glassdisintegrated using an Oster
vessel containing (U.S.A.) commercial
the chips min. The blender Model
alkali-treated holocellulose was then
117 4172 for at least 15 seconds.
and “liquor” was covered with cheesecloth and suction-filtered and sequentially washed with
118
119 placed
Chemical inside the pressure cooker partially filled
Treatment 750 mL distilled water, 10% CH3COOH (with
120 with distilled water, then sealed. It was heated prior soaking for 5 min), and 250 mL distilled
121 up to a temperature
1. Bleaching and Holocellulose ofPreparation
121oC and 15 psi pressure, water until the pH of the filtrate was neutral.
122 which was then maintained for an additional
123 Two-stage
2 hours. bleaching
Thefollowing
pressure the procedure of (Villegas,
was released, and2015) was employed
the Mechanical using 0.5% (w/v)
Treatment
124 hypochlorite solution (3% consistency [dry mass to total solution], 70 oC, 30 min) followed by
125
cooked
soaking in 5% bamboo
(v/v) H2pulp
O2 solutionwas placed over cheesecloth
(3% consistency, 70 oC, 120 min). The bleached pulp was
126 and thoroughly washed with
washed several times with water under vacuum filtration tap waterto remove
with excess
bleaching
The Kraft pulp (without prior bleaching) was
chemicals.
127 squeezing. The resulting pulp was then further subjected
Holocellulose was prepared from bleached pulp by acid chlorite delignification (Erickson, 1962). to mechanical refining using a friction
128 disintegrated
Solution A (60 mL CH using
3COOH anand Oster
1.3 g (U.S.A.)
NaOH in 1commercial
L H2O) was added grinder supermass
to the bleached pulp in a colloider Model MKCA6-2
129 flaskblender
at a ratioModel 4172
of 2 g pulp perfor at least
30 mL 15A.seconds.
solution The flasks were placed(Masuko Sangyo
in a water-filled Co. Ltd, Japan) for 15 passes at
metal
130 pan mounted on a hot plate and heated to a temperature of 75oC monitored with a thermometer.
1500 rpm. The grinder clearance was gradually
131 Then, 3 mL of 20% sodium chlorite (NaClO2) solution was placed in the flask with subsequent
132 Chemical
addition Treatment
(3 mL each) of the NaClO2 solution after 0.5, 1.0, 1.75, and 2.5 decreased fromafter
hr., with swirling -1 during the first pass, until
every addition. After 3.25 hr., the mixture was suction-filtered and washed with portions of chilled -10 in the 15 pass. After
the setting reached th
133
134 1. Bleaching
distilled and Holocellulose
water, four 75-mL CH3COOH and two 30-mL every
quantities of 1% Preparation quantitiespass, grinding was discontinued, and the
of methanol.
135 grinder housing was opened. This was done to
136 2. Alkali-insoluble
Two-stage Cellulose
bleachingPreparationfollowing the procedure recover the residues stuck around the periphery
of (Villegas, 2015) was employed using 0.5% (w/v) of the grinding stones by washing with water.
hypochlorite solution (3% consistency [dry mass to However, pulp retained on the outer surfaces
total solution], 70 oC, 30 min) followed by soaking of the grinding stones was discarded. This step
in 5% (v/v) H2O2 solution (3% consistency, 70 oC, was done to ensure that for the experiment,
120 min). The bleached pulp was washed several there was homogeneity in the ground pulp
times with water under vacuum filtration to recovered from the colloider. By removing the
remove excess bleaching chemicals. Holocellulose pulp that were stuck outside the grinding discs
was prepared from bleached pulp by acid chlorite and collecting only the pulp that was uniformly
delignification (Erickson, 1962). Solution A (60 pressed between the discs, the experiment had
mL CH3COOH and 1.3 g NaOH in 1 L H2O) was minimized the variability in the pulp recovered
added to the bleached pulp in a flask at a ratio of for subsequent tests. However, this would result
2 g pulp per 30 mL solution A. The flasks were in low yield recovery from this experimental
placed in a water-filled metal pan mounted on procedure. The opening of the grinder housing to
a hot plate and heated to a temperature of 75oC recover undrained pulp was repeated for every
monitored with a thermometer. Then, 3 mL of 20% pass until there were no observable differences
sodium chlorite (NaClO2) solution was placed in between the pulp in and outside the grinding
4 H.D.R. Aumentado, R.A. Razal, R.C. Nacario, and M.N. Manalo

stones. After 15 passes, 500 mL of the ground characterization tests.

pulp suspension was obtained.
Characterization Tests
Acid Hydrolysis
1. Focused Ion Beam - Field Emission Scanning
In a two-neck round bottom flask, 1 g of Electron Microscopy (FIB-FESEM)
air-dried alkali-insoluble cellulose was added,
followed by 50 mL of 46% v/v sulfuric acid (Jara, A drop of suspension containing the bamboo
2019; Villegas, 2015). The flask was placed in nanocellulose product was casted and spread on
a water bath on a hot plate and was constantly the surface of aluminum foil. The casted samples
stirred with a magnetic stirrer. The reaction were dried in a covered glass petri dish with
was carried out for 30 min while maintaining desiccant under dark condition before analysis
the water bath temperature at 45oC. After 30 using a Dual Beam Helios Nanolab 600i (Field
min, 500 mL of cold distilled water was added Electron and Ion Company, OR, USA). The
to the mixture and subsequently neutralized analysis was performed using an accelerated
using 10% NaOH. The mixture was transferred voltage of 2.0 kV (SE/TLD) and a beam current
to a separatory funnel and allowed to settle of 86 pA (SE/TLD). Data processing of the
until a distinct boundary between two layers images was done through an image processing
was observed, where one layer contained the software using the Dual Beam Helios Nanolab
hydrolysis products. The acid-hydrolyzed 660i xT Microscope Server. The measurement of
cellulose produced from the chemically treated the widths of the nanocellulose was done using
kawayan kiling pulp settled at the bottom layer ImageJ software.
and was subsequently separated by draining
from the funnel. 2. Energy Dispersive Spectroscopy (EDS)
For the colloider-treated pulps in water
suspension, the water content was reduced in a This analysis was performed alongside FIB-
Stuart RE301P rotary evaporator (Cole Parmer FESEM imaging to determine the elemental
Ltd., USA). A portion of the thick suspension composition of the sample. Both methods used
containing approximately 1 g of oven dried the same equipment, with the EDS analysis being
ground pulp was taken, and then concentrated conducted using an accelerated voltage of 15.0 kV
H2SO4 was added to bring to a final concentration and a beam current of 0.69 nA. Data processing
of 46% (v/v). The same conditions employed and elemental composition determination were
for the acid hydrolysis of the alkali insoluble done using the imaging software Oxford EDS
cellulose and the recovery of the acid-hydrolyzed AZtecEnergy.
cellulose were followed for the colloider-treated
Kraft pulp. 3. Fourier Transform Infrared (FTIR)
The solution containing the hydrolysis Spectroscopy
products was transferred in falcon tubes and
was washed repeatedly with distilled water and The FTIR spectra of freeze-dried samples of
centrifuged at 5oC and 9500 rpm for 10 min using kawayan kiling nanocellulose products Pin KBr
Allegra X-22R centrifuge (Beckman Coulter Inc., pellets (1:20 w/w) were obtained using a Nicolet
USA). The pellets were combined and dialyzed, 6700 FT-IR spectrometer (Thermo Scientific,
with stirring, using a dialysis tubing cellulose Waltham, MA, USA). The spectra were collected
membrane (16 mm diameter, 14,000 molecular at ambient conditions in transmittance mode,
weight cut off) for 24 hr. Following dialysis, the from an accumulation of 16 scans at a 4 cm-1
samples were homogenized using Cole Palmer® resolution over the region of 4000-400 cm-1.
ultrasonic cleaner Model 08895-20 at 50-60 Hz
for 30 min. The yield of the resulting products 4. Dynamic Light Scattering (DLS)
(for both the alkali-insoluble cellulose and
ground Kraft pulp) was determined and a portion The Z-average (d.nm) and polydispersity
of the product was freeze-dried for various indices (PdI) of the samples in pure water
 Philippine e-Journal for Applied Research and Development 12(2022), 1-11
Philippine e-Journal for Applied Research and Development
Website: pejard.slu.edu.ph ISSN 2449-3694 (Online) 5

were measured with Malvern® ZS90 Zetasizer to produce the nanocellulose. In our study, the
(Malvern Panalytical Ltd, Worcestershire, UK). yields of nanocellulose from acid hydrolysis were
Suspensions of 0.01 wt. % concentration were 61.76% and 11.24% from the solely chemically
previously homogenized using CP EW-08895-20 treated material and the mechanically treated
ultrasonicator (Cole-Parmer, Ltd. USA) for 5 min pulp preparation, respectively.
before analysis.
The lower yield from the mechanically
5. X-ray Diffraction (XRD) Analysis treated pulp can be attributed to the non-
accessibility of the ground cellulose to the acid.
The X-ray diffraction spectra of the Kawayan Even after 15 passes through the colloider, the
kiling nanocellulose products were measured cellulose may not have been fully liberated from
in a Shimadzu Lab-X XRD-6000 diffractometer the hemicelluloses and lignin to allow the acid
(Shimadzu, Columbia, MD, US) using a Ni- to penetrate the non-cellulosic matrices to reach
filtered Cu Kα radiation (λ = 1.54050 A˚) at the cellulose and break6 it down. This is evident
an anode voltage of 40 kV and a current of 30 in the observed color of the ground cellulose
mA. Freeze-dried samples were compressed in and the peaks obtained in the FTIR spectra
iation (λ = 1.54050 A˚) atholder.
a sample an anode voltage of 40
Diffractograms kV recorded
were and a current of 30 mA. Freeze-
corresponding to functional groups found in
◦
from 10
les were compressed in◦ a to 80◦ at
sample a scan
holder. rate of 2.00◦were
Diffractograms /min. recorded
lignin,from
which to 80◦ the presence of residual
10suggests
Crystallinity
ate of 2.00◦/min. Crystallinityindex (CrI)
index was
(CrI) wascalculated using
calculated lignin
using Eq. (1)in(Segal
the preparation.
et al.,
Equation 3 (Segal et al., 1959).
Nanocellulose particle size
𝐼𝐼002 −𝐼𝐼𝑎𝑎𝑎𝑎
𝐶𝐶𝐶𝐶𝐶𝐶 = 𝑥𝑥100 Equation 3
Using dynamic (1) light scattering, the
𝐼𝐼002
Z-averages and polydispersity indices (PdI) of
where Iam is the intensity of diffraction of the the nanocellulose products were determined to
s the intensity ofamorphous
diffraction region
of the amorphous
at 2θ angle region
and at I0022θisangle
the andestimate
I002 is thethe
maximum
size of the particles. It can be seen
f the 002-latticemaximum
diffractionintensity
peak at of 2θ.the 002-lattice diffraction in Table 1 that on average, the dimensions of the
peak at 2θ. samples appear to exceed 100 nm. Most of the
d Discussion mechanically prepared nanocellulose appeared
to be fibrillar which could be explained by the
mination Results and Discussion incomplete acid hydrolysis reaction. Refining
could not fully release the cellulose from the
Yield determination associated lignin and hemicelluloses. Based
l yield obtained from the chemically treated cellulose (i.e., bleached on followed
the PdI by alkalithe samples were broadly
values,
was 26.94% while for
Thethe pulpyield
overall that obtained
was submitted
from the tochemically
grinding in a refiner, the yield was
polydisperse.
%. The low yieldtreated
from the latter was
cellulose (i.e.,largely
bleached due to the removal
followed by alkaliof unground material for
treatment)
during the mechanical was 26.94%
refining (16.1%while
yield).for This
the pulp that
was designed to ensure that only
was submitted to grinding in a refiner, the yield
was uniformly ground was collected for further treatment and analysis. This ensured
ty of the ground wasmaterial
only 1.20%. The low to
subjected yield
thefrom the latter
property testswasand this need not be the
largely due to the removal of unground material
in actual production. In comparison, Jara et al.
for every pass during the mechanical refining
(2020) found the acid hydrolyzed yield
e nanocrystals from kawayan kiling
(16.1% yield). This was designed to ensure that studies reported yields
shoots to be 9.16%; other
ydrolysis or oxidation
only pulpreactions
that wastouniformly
produce ground
the nanocellulose.
was collectedIn our study, the yields of
ose from acid hydrolysis
for furtherwere 61.76%
treatment andand 11.24%
analysis. from
This the solely chemically treated
ensured
homogeneity of the ground
nd the mechanically treated pulp preparation, respectively.material subjected
to the property tests and this need not be the
yield from the procedure
mechanically in actual production. In comparison,
treated pulp can be attributed to the non-accessibility of
Jara et al. (2020) found the acid hydrolyzed
cellulose to theyield
acid. ofEven
cellulose 15
after passes through
nanocrystals fromthekawayan
colloider, the cellulose may not
fully liberated from
kilingtheshoots
hemicelluloses
to be 9.16%; and lignin
other to allow
studies the acid to penetrate the non-
reported
matrices to reach the cellulose
yields from the and break it
hydrolysis or down.
oxidation Thisreactions
is evident in the observed color
und cellulose and the peaks obtained in the FTIR spectra corresponding to functional
nd in lignin, which suggests the presence of residual lignin in the preparation.

ose particle size

 279 Figure 1 shows the images of the nanocellulose products from the acid-hydrolysis of differently
280 treated Kawayan kiling pulp. It was observed that the products from the combined bleaching and
281 alkali digestion gave crystal-like structures which were aggregated, while those that underwent
282 grinding were fibrillar in nature. The aggregation could be due to hydrogen bonding of the
283 hydroxyl groups of cellulose as water was removed during drying prior to microscopy. This
284 partially accounts for the broader widths (134.20 ± 34.33 nm) of the nanocrystals from the
285 cellulose that was subjected to combined bleaching and alkali digestion as compared with that of
286 the mechanically refined (39.13 ± 34.11 nm) product. In the latter, mechanical fibrillation greatly
6 287 H.D.R.the
contributed to shearing Aumentado, R.A.
microfibrils butRazal, R.C.
did not Nacario,
remove and M.N. Manalo
the lignin.
288
289
Table 1. Z-average and polydispersity index of
acid-hydrolyzed nanocellulose products from the
chemically and mechanically pretreated bamboo
acid hydrolysis feedstock.
Pretreatment
of the Bamboo Z-average Polydisper-
Acid Hydroly- (d.nm)* sity Index*
sis Feedstock
Combined 176.8 ± 7.5 0.501 ± 0.067
bleaching and
alkali digestion
(a) 8
Refining with 389.5 ± 74.7 0.561 ± 0.169
friction grinder 290
supermass col-
loider
* Values are mean ± s.d. of 5 measurements.

Nanocellulose morphologies and elemental

composition

Figure 1 shows the images of the nanocellulose

products from the acid-hydrolysis of differently
treated Kawayan kiling pulp. It was observed
that the products from the combined bleaching
and alkali digestion gave crystal-like structures (b)
which were aggregated, while those that
291
underwent grinding were fibrillar
292 in nature. The
aggregation could be due to hydrogen
293 Fig 1.bonding
FESEM images showing Figurethe 1. FESEMmorphology
crystal-like images showing the vulgaris nanocellulose
of Bambusa
of the hydroxyl groups of cellulose
294 as waterfrom
products crystal-like morphology of Bambusa vulgaris
was(a) pulp subjected to bleaching and alkali digestion and (b) pulp refined in the
removed during drying prior to295microscopy. nanocellulose
This supermass
friction grinder colloider. products from (a) pulp subjected
partially accounts for the broader
296 widths (134.20 to bleaching and alkali digestion and (b) pulp
± 34.33 nm) of the nanocrystals from the cellulose
297 refined in the friction grinder supermass
298 Table
that was subjected to combined bleaching and 2. Widths (nm) obtained by FESEM colloider.
imaging of acid hydrolysis products from variously
299 pretreated kawayan kiling (Bambusa vulgaris Schrad.) pulp.
alkali digestion as compared 300with that of the
mechanically refined (39.13 ± 34.11 nm) Pretreatment
product. Table 2. Widths (nm) obtained by FESEM
of the bamboo
In the latter, mechanical fibrillation greatly imaging of acid Mean hydrolysis
± SD products
Min from Max
acid hydrolysis feedstock
contributed to shearing the microfibrils but variously
did bleaching and
Combined pretreated kawayan kiling (Bambusa
not remove the lignin. vulgaris Schrad.)134.2
alkali digestion pulp.± 34.33 84.00 214.0
Pretreatment
Refining with friction grinder
39.13 ± 34.11 12.00 171.0
supermass colloider of the
301 Mean ±
bamboo acid Min Max
302 Table 3. Elemental analysis by EDS analysis of SD acid hydrolysis products from variously pretreated
hydrolysis
303 kawayan kiling (Bambusa vulgaris Schrad.) pulp.
304
feedstock
Pretreatment of theCombined
bamboo
acid hydrolysis bleaching
feedstock and
134.2
%C ± %O % Si
84.00 214.0
34.33
alkaliand
Combined bleaching digestion 23.93 77.07 -
alkali digestion
Refining
Refining with friction grinderwith
supermass colloider grinder
friction 31.73
39.13 ± 62.87 5.40
12.00 171.0
305 supermass 34.11
306 Elemental analysis (Tablecolloider
3) showed that both nanocellulose products contained carbon and
307 oxygen; however, the nanofibrils contained a significant amount of silicon. This could be attributed
308 to the mechanical grinding process. It can be inferred that in the refining process, small fragments
309 of the grinder stone, which is mainly composed of silicon carbide (SiC), could have been chipped
310 away and mixed with the sample.
 Philippine e-Journal for Applied Research and Development 12(2022), 1-11
Philippine e-Journal for Applied Research and Development
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Table 3. Elemental analysis by EDS analysis Chemical and crystalline structures of

of acid hydrolysis products from variously nanocellulose
pretreated kawayan kiling (Bambusa vulgaris
Schrad.) pulp. Freeze-dried nanocellulose suspensions from
Pretreat- the chemically treated kawayan kiling pulp
ment of the produced white solids while the mechanically
bamboo acid %C %O % Si treated kawayan kiling pulp gave brownish flaky
hydrolysis solids. As seen in Figure 2, both samples showed
feedstock the same characteristic peaks for cellulose, such
as the O-H vibration at 3413.29 cm-1 and 3439.93
Combined
cm-1 (Costa et al., 2015; Liu et al., 2016), C-O
bleaching and 23.93 77.07 -
stretching vibration at 1063.67 cm-1 and 1064.92
alkali digestion
cm-1, aliphatic C-H bonds at around 2920.12 9 cm-1
Refining with and 2898.87 cm , C-O-C stretching vibration of
-1

friction grinder the pyranose ring at around 1064.36 cm-1 and

31.73 62.87 5.40
supermass
311 1063.67 cm-1 (Liu et al., 2016). Furthermore,
312 Chemical
colloider and crystalline structures of nanocellulose
the presence of the cellulose backbone was also
313
confirmed by the peaks at 1645.54 cm-1 and
314 Freeze-dried nanocellulose
Elemental analysis (Table 3) showed that suspensions from the chemically treated kawayan kiling pulp produced
1633.55 cm-1 due to the O-H bending of the
315 white solids while the mechanically
both nanocellulose products contained carbon treated kawayan kiling pulp gave brownish flaky solids. As
absorbed water and the peaks at 898.86 cm-1 and
316 seen in Figure 2, both samples showed the same characteristic peaks for cellulose, such as the O-
and oxygen; however, the nanofibrils contained 896.92
et al.,cm inLiu
theet al.,
anomeric region which are
-1
317 H vibration at 3413.29 cm-1 and 3439.93 cm-1 (Costa 2015; 2016), C-O stretching
a significant amount of silicon.-1 This could be-1 attributed to the linkages between sugar -1 units in
318 vibration at 1063.67 cm and 1064.92 cm , aliphatic C-H bonds at around 2920.12 cm and
attributed to the mechanical
-1 grinding process. It cellulose (Jiang & Hsieh, 2013; -1 Liu et al., 2016)
319 2898.87 cm , C-O-C stretching vibration of the pyranose ring at around 1064.36 cm and 1063.67
can be
320
inferred thatetinal.,
cm-1 (Liu the2016).
refining process, the
Furthermore, smallpresence ofOne notablebackbone
the cellulose FTIR peak for confirmed
was also the hydrolysis
fragments
321 of the grinder stone, which
-1 is mainly -1 product from the refined pulp is at 1727.47 cm-1
by the peaks at 1645.54 cm and 1633.55 cm due to the O-H bending of the absorbed water and
composed
322 of peaks
the siliconat carbide
898.86 cm (SiC),
-1
and could
896.92 have
cm-1 in thewhich is associated
anomeric withare
region which acetyl ester to
attributed groups
the and
been323chipped away and mixed with the sample. uronic groups of
linkages between sugar units in cellulose (Jiang & Hsieh, 2013; Liu et al., 2016). hemicelluloses or ester bond of
324
1064.92

898.86
1645.01
2920.12

1064.36
3,413.29
% Transmittance

1727.47

1063.67
3439.93

896.92
1633.55
2898.87

3,500 3,000 2,500 2,000 1,500 1,000 500

Wavenumber (cm-1)
325
326
Figure 2. FTIR spectra of acid hydrolysis products from Bambusa vulgaris Schrad. pulp subjected
327 Fig 2. FTIR spectra of acid hydrolysis products from Bambusa vulgaris Schrad. pulp
to bleaching and alkali digestion ( in red) and of pulp refined with the friction grinder supermass
328 subjected to bleaching and alkali digestion ( in red) and of pulp refined with the friction
329 grinder supermass colloider (in colloider
blue). (in blue).
330
331
332 One notable FTIR peak for the hydrolysis product from the refined pulp is at 1727.47 cm-1 which
333 is associated with acetyl ester groups and uronic groups of hemicelluloses or ester bond of carboxyl
334 group of ferulic and p-coumaric acid of lignin and/or hemicellulose. This was not seen in the
 10

338 8pretreatedpulp suggests that the binding substance lignin, was

H.D.R. Aumentado, R.A. completely removed
Razal, R.C. Nacario, from
and M.N. the
Manalo
339 chemically pretreated bamboo pulp.
340
carboxyl group of ferulic and p-coumaric acid of removed from the chemically pretreated bamboo
341 The XRD
lignin and/orspectra are shownThis
hemicellulose. in Figure 3. Based
was not seen on pulp.
the peak height calculation using Segal’s (1959)
342 inmethod, the crystallinity
the hydrolysis productsindices
of the (CrI)
bambooof the
pulpchemically
TheandXRDmechanically shownkawayan
spectra aretreated in Figurekiling
3.
343 pulpswas
that are 56.92%
bleachedand and49.47%,
alkali respectively.
digested. This The lower
Basedcrystallinity
on the peakof the nanocellulose
height calculationfrom the
using
344 indicates that treated
mechanically residual pulp
hemicelluloses
could be dueand/orto theSegal’s (1959)
presence ofmethod, the crystallinity
amorphous indices
residual lignin. In
345 lignin remained in the hydrolysis products from (CrI) of the chemically and mechanically
comparison, cellulose nanocrystals prepared by sulfuric acid hydrolysis of commercial bamboo treated
346 the friction
pulp had agrinder
71.98% supermass colloider
crystallinity (Yutreated kawayan
et al. 2012) whilekiling pulpsstudy
a recent are 56.92% anda 49.47%,
reported 68.53%
pulp; the absence of peaks associated with lignin respectively. The lower crystallinity of the
347 crystallinity for nanocellulose extracted from isolated
in the chemically pretreated pulp suggests that
bamboo parenchyma using deep eutectic
nanocellulose from the mechanically treated
348 solvent (Gu et al. 2021).
the binding substance lignin, was completely pulp could be due to the presence of amorphous
349
350
18 o 21 o 23 o

I002
Intensity (a.u.)

Iam

11

18 o 21 o 23 o a

351
352 2 theta (2θ, o)
Intensity (a.u.)

I002

Iam

353 o
Figure
354 3. X-ray diffraction spectra of Bambusa 2 theta (2θ, nanocellulose
vulgaris ) products from (a) combined
bleaching
355 and3.alkali
Fig X-ray digestion
diffraction and (b)ofpulp
spectra subjected
Bambusa to nanocellulose
vulgaris grinding in aproducts
frictionfrom
grinder
(a) supermass
356 combined bleaching and alkali digestioncolloider.
and (b) pulp subjected to grinding in a friction
357 grinder supermass colloider.
358
359
360 Conclusion
361
362 The study compared the nanocellulose from the kraft pulp derived from kawayan kiling culms
 Philippine e-Journal for Applied Research and Development 12(2022), 1-11
Philippine e-Journal for Applied Research and Development
Website: pejard.slu.edu.ph ISSN 2449-3694 (Online) 9

residual lignin. In comparison, cellulose Forest Products and Paper Science, University
nanocrystals prepared by sulfuric acid hydrolysis of the Philippines Los Baños College of Forestry
of commercial bamboo pulp had a 71.98% and Natural Resources, College, Laguna for
crystallinity (Yu et al. 2012) while a recent study the assistance in sample preparation. Much
reported a 68.53% crystallinity for nanocellulose appreciation for DOST-PCIEERD for funding
extracted from isolated bamboo parenchyma the testing of FIB-FESEM and XRD analysis
using deep eutectic solvent (Gu et al. 2021). through the ADMATEL-EPDC Grant, and the
staff of ADMATEL and MSD-ITDI for assistance
in the characterization tests. The authors
Conclusion also acknowledge the UPLB Nanotechnology
Laboratory for the use of the particle size
The study compared the nanocellulose from analyzer for the DLS analysis and the De La
the kraft pulp derived from kawayan kiling Salle University Chemistry Department, Taft
culms subjected to two different pretreatment Ave., Manila for the FTIR analysis.
methods. FIB-FESEM analysis showed
crystal-like structures of chemically treated
acid-hydrolyzed nanocellulose while for the References
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