ASIA-PACIFIC JOURNAL OF CHEMICAL ENGINEERING

Asia-Pac. J. Chem. Eng. (2014)

Published online in Wiley Online Library
(wileyonlinelibrary.com) DOI: 10.1002/apj.1812

Research article
Effects of acid treatments on bamboo cellulose nanocrystals
Peng Peng Zhang,1 Dong Shen Tong,1 Chun Xiang Lin,2 Hui Min Yang,3 Zhe Ke Zhong,3 Wei Hua Yu,1 Hao Wang4
and Chun Hui Zhou1,4*
1
Research Group for Advanced Materials & Sustainable Catalysis (AMSC), College of Chemical Engineering, Zhejiang University of Technology, Hangzhou
310014, China
2
Australian Institute of Bioengineering and Nanotechnology, The University of Queensland, Brisbane, Queensland 4067, Australia
3
China National Bamboo Research Center, Hangzhou 310012, China
4
The Institute for Agriculture and the Environment, University of Southern Queensland, Toowoomba, Queensland 4350, Australia

Received 10 December 2013; Revised 31 January 2014; Accepted 2 March 2014

ABSTRACT: The production of nanocellulose with controlled structure from plant sources with easy availability and good
sustainability is very important in science and engineering. In this work, the isolation of cellulose from bamboo and the
effects of the hydrolysis of different acids on the morphology, structure, and properties of the resultant cellulose and cellulose
nanocrystals were investigated. Bamboo cellulose (PHC) was first isolated from raw bamboo powder by alkali and
consecutive bleaching treatments. Cellulose crystallites were then prepared by the controlled hydrolysis of PHC in acid
solutions. The samples were characterized by using thermogravimetric analysis, Fourier transform infrared spectroscopy,
powder X-ray diffraction, laser particle size analyzer, Scanning electron microscopy, and Transmission electron microscopy.
Cellulose nanocrystals prepared from the hydrolysis of the isolated bamboo cellulose in the sulfate acid, hydrochloric acid,
phosphoric acid, or acetic acid solution had the length of 3–200 nm, 20–85 nm, 20–40 nm, and 6.5–20 nm, respectively.
Nanocrystals made by the uses of sulfuric acid and phosphoric acid possessed higher crystallinity and lower thermal stability
than those by the uses of hydrochloric acid and a mixture of acetic acid and nitric acid. Results suggested the type of acids
significantly influenced the structure, morphology, and thermal stability of cellulose crystallites. The anions in the acid
solution also contributed to those differences because they affected the swelling of cellulose chains and breakage of hydrogen
bonds in cellulose. The findings indicate that a judicious choice of pretreatment of the cellulose and its moieties can be used
for the production of different cellulose nanocrystals. © 2014 Curtin University of Technology and John Wiley & Sons, Ltd.

Keywords: bamboo; cellulose; nanocrystals; hydrolysis; acid treatment; extraction

INTRODUCTION is the first important step for further processing to make

Cellulose is the main component of plant cell walls and is
regarded as the most abundant natural bioresource.[1]
Cellulose has remarkable practical and potential appli-
cations in the development of new high-performance
materials with low environmental impact. In particular,
cellulose nanocrystals have large surface areas, relative
high strength and stiffness, and low density (about 1.6 g
cm
3).[2] They also have many reactive surfaces with –
OH groups that allow functional chemical species to be
grafted on them. As a result, cellulose nanocrystals can
be used to make green nanocomposite polymer,[3]
catalysts,[4] drug carriers,[5] membranes,[6] and so forth.
Extraction of cellulose from lignocellulosic biore-
sources to yield pure cellulose fibers and nanocrystals
*Correspondence to: Chun Hui Zhou, Research Group for Advanced
Materials & Sustainable Catalysis (AMSC), College of Chemical
Engineering, Zhejiang University of Technology, Hangzhou 310014,
China. E-mail: clay@zjut.edu.cn; Chun.Zhou@usq.edu.au
advanced functional materials. Until now, several
methods for isolating cellulose fibers, such as alkali ex-
traction with a bleaching process,[7] ionic liquid,[8]
diglyme-HCl methods,[9] alcoholic extraction[10] were
developed. Among them, alkali extraction with a
bleaching process is still considered as the most efficient
way to remove hemicellulose and lignin from
lignocellulosic bioresources.
For the extracted cellulose, under a well-controlled
chemical hydrolysis or biological enzyme hydrolysis to
break the amorphous region of it, cellulose nanocrystals
can be obtained.[11] Up till now, many methods for
isolating cellulose nanocrystals from various plant fibers,
such as acid hydrolysis,[12] enzymatic procedures,[13]
oxidation,[14] high pressure homogenization,[15] and
steam explosion processes,[16] had been explored. Acid
treatment is a conventional process for preparation of
cellulose nanocrystals. Especially sulfuric acid hydrolysis
is often used.[17] However, because the structure and
distribution of crystalline and amorphous region of

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Curtin University is a trademark of Curtin University of Technology
P. P. ZHANG
cellulose change dramatically between species, habitats,
and even within different parts of the same plant, there is
no ubiquitous conditions for the extraction of cellulose
and nanocrystals.[18] Up to now, a variety of cellulose
and cellulose nanocrystals have been obtained from
jute,[19] mulberry,[20] sisal,[21] pandanus tectorius,[22] rice
husk,[23] and milkweed stems.[24] However, the effects of
different acid treatment on the morphology and structure
of the resultant cellulose nanocrystals has not been fully
explored yet.
Theoretically, nanocellulose can be prepared from any
cellulose source material. Yet, for practical uses, the
cellulose source material must be well considered with
availability and sustainability. One of the most attractive
sources is bamboo, which is widely cultured in tropical
and subtropical regions and has regarded as ‘the world’s
second largest forest’. Remarkably, in its growth period,
bamboo can grow around 8 cm per day. In comparison to
most woods, the harvest period of bamboo is much
shorter, about 3 years, and it has higher yield and lower
cost. In this context, it is of significance to have a better
understanding of the extraction and of how the structure
and properties of the extracted bamboo cellulose
crystallites are affected by the acid treatment. Hence, in
the present work, four different acid solutions were used
for the hydrolysis of extracted bamboo cellulose to
produce its nanocrystals, and the characteristics of the
products were compared. Alkali and bleaching treatment
was first carried out to extract bamboo cellulose. Cel-
lulose crystallites were then prepared by the controlled
hydrolysis of bamboo cellulose in acid solutions. Four
kinds of acid solution systems: sulfuric acid, hydro-
chloric acid, phosphoric acid, and a mixture of acetic
and nitric acid were used, respectively. The chemical
groups, morphology, and thermal stability of bamboo
cellulose were characterized. The influences of different
acid treatments on cellulose nanocrystals structure and
morphology were investigated, and the possible reasons
behind it were discussed.
Figure 1. Structure of PH and the process of chemical treatment. This figure is
available in colour online at www.apjChemEng.com.
Copyright © 2014 Curtin University of Technology and John Wiley & Sons,
Ltd.
Asia-Pacific Journal of Chemical Engineering
EXPERIMENTAL
Raw materials
Sodium hydroxide, sulfuric acid, hydrochloric acid,
phosphoric acid, acetic acid, nitric acid, and commercial
cellulose for control tests were purchased from Sino-
pharm Chemical Reagent Co., Ltd, Shanghai, China; so-
dium chlorite was purchased from Aladdin Chemical
Regent Inc., Shanghai, China. All these reagents were
of analytical grade and used without further purification.
Phyllostachys heterocycla chips were made from P.
heterocycla species gathered in Linan (Hangzhou,
China) and in chips provided by China National Bamboo
Research Center, Hangzhou.
Phyllostachys heterocycla cellulose extraction
Phyllostachys heterocycla chips were first washed with
distilled water and dried in an oven at 70 °C for 24 h then
smashed into 60 mesh powders with a micronizer (DF-20,
Wenling, China). The color of the raw powder was
brown. The raw powder was labeled as PH. A total of
8 g of bamboo powder was added into a cellulose
extraction apparatus (VELP, Italy). For the delignification
and the removal of hemicellulose, the powder was soaked
in a 2 wt% NaOH solution and reflux for 4 h at 90 °C. The
ratio of powder to liquid used for the alkali treatment was
1:25 (g/mL). Such an operation was repeated four times.
Bleaching treatment was carried out to remove lignin
residues by using a solution made up of aqueous chlorite
(1.7 wt% NaClO2 in water) and an acetate buffer
(27 g NaOH and 75 mL glacial acetic acid, diluted to
1 L using distilled water).[25] The ratio of aqueous chlorite
to acetate buffer was 1:1 (v/v). The conditions used in
bleaching steps were set as follows: the reaction was
conducted at 80 °C for 6 h, and the ratio powder to liquid
was 1:20 (g/mL). Such an operation for the removal of
Asia-Pac. J. Chem. Eng. (2014)
DOI: 10.1002/apj
Asia-Pacific Journal of Chemical Engineering EFFECTS OF ACID TREATMENTS ON BAMBOO CELLULOSE NANOCRYSTALS
lignin was repeated two times. The bleached powder was
washed with distilled water three times and dried. Finally,
the extracted cellulose was dried in an air-circulating oven
at 50 °C until constant mass. In this way, bamboo
cellulose was extracted (Fig. 1B), and the sample was
labeled as PHC (P. heterocycla bamboo cellulose).
Cellulose nanocrystals preparation
Bamboo cellulose (PHC) sample was milled. Then 1 g
milled PHC was added into 25 mL of sulfuric acid
solution, hydrochloric acid solution, phosphoric acid
solution, and a mixture of acetic and nitric acid solution
(the ratio of 99% (v/v) acetic acid to 68% (v/v) nitric acid
is 10:1 (v/v)), respectively. All acid solutions con-
centration was set as 6.5 M. Acid hydrolysis was carried
out under strong agitation at a temperature of 60 °C for
2 h (Fig. 1 (b)). After that, the reaction was stopped by
diluting with 10-fold cold water. The diluted hydrolyzed
cellulose suspension was then centrifuged at 8000 rpm
for 15 min at 10 °C with a centrifuge (3–30 K, Sigma,
Germany) to remove the excess of acids, followed by a
prolonged dialysis (cellulose membrane, cut-off 8000–
14 000 Da) against distilled water until a constant pH in
the range of 5–6 was reached. The resultant nanocrystals
were further dispersed in distilled water by an ultrasonic
treatment for 30 min (Ultrasonic equipment, KQ-500DE,
Kunshan, China) and then stored in a refrigerator with
the addition of chloroform for further characterization
and analysis. The cellulose nanocrystals prepared by
using sulfuric, hydrochloric, phosphoric, and the mixture
of acetic and nitric acid solutions were labeled as CNS,
CNH, CNP and CNA, respectively.
Characterization of bamboo powder, bamboo
cellulose and cellulose nanocrystals
Fourier transform infrared spectroscopy
The Fourier transform infrared(FTIR) spectra of all
samples were recorded on an attenuated total reflection
FTIR spectroscopy (Nicolet Is 10, Thermo Scientific,
USA) equipped with smart omni-transmission in the
range of 400–4000 cm
1 with a resolution of 4 cm
1.
PH, PHC, CNS, CNH, CNP, and CNA were dried in an
air-circulating oven at 50 °C over night. All samples were
finely ground and mixed with potassium bromide (the
mass ratio of sample to potassium bromide is 1:200)
followed by pressing the mixture into ultra-thin pellets.
A laser particle size analyzer
The length of PH and PHC were recorded at room tem-
perature on a laser particle size analyzer (WinnerR2308B,
Winner, China). In each experiment, a small quantity of
the sample was dispersed in distilled water under soni-
cation until the concentration of 0.03 M was reached.
Copyright © 2014 Curtin University of Technology and John Wiley & Sons,
Ltd.
Scanning electron microscope of cellulose
nanocrystals
The morphology of PH and PHC was examined on a
scanning electron microscope(S-4700, Hitachi, Japan).
In each experiment, a small quantity of the sample
was put on a pin stub, and then it was coated with gold
prior to recording the micrographs. The samples were
observed and imaged at a 6 mm working distance and
15 kV accelerating voltage.
Transmission electron microscopy of cellulose
nanocrystals
Transmission electron microscope (TEM) (Tecnai G2
F30 S-Twin, Philips-FEI, Netherlands) was used to
image the cellulose nanocrystals obtained from PHC. A
10 μL drop of 0.1 wt% cellulose nanocrystals dispersion
was mounted on a glow-discharged carbon-coated Cu
grid. Excess solution on it was absorbed by filter paper,
and the grid was then kept under infrared heat lamp for
drying. The diameter of bamboo cellulose nanocrystals
were measured on about 100 representative cellulose
nanocrystals by using Adobe Photoshop CS5 V12.0.
Thermogravimetric analysis
The thermodynamic properties of the different samples
were determined on a Mettler Toledo thermogravimetric
analyzer (TGA/DSC 1A 85-F, Mettler Toledo,
Switzerland). The amount of samples used for each
measurement was about 10 mg. All measurements were
performed under a nitrogen atmosphere at a gas flow rate
of 60 mL min
1. The samples were heated from 30 to
700 °C at a heating rate of 10 °C min
1.
Powder X-ray diffraction
Powder X-ray diffraction was used to determine the
crystallinity of the samples obtained from different
treatments. Each sample in the form of milled powder
was placed on the sample holder and leveled to obtain
total and uniform X-ray exposure. The samples were
analyzed on an X-ray diffractometer (X’Pert Pro,
PANalytical, Netherlands) at room temperature with a
monochromatic CuKα radiation source (λ = 0.154 nm)
in the step-scan mode with a 2θ angle ranging from 5°
to 40° with a step of 0.04 and scanning time of 5.0 min.
To characterize the crystallinity of the different samples,
the crystallinity index was determined on the basis of the
reflected intensity data and calculated by using the
following equation[26]:
I 002
I am
CrI ð%Þ ¼  100 (1)
I 002
Where I002 is the maximum intensity of the (002)
lattice diffraction peak, and Iam is the intensity scattered
by the amorphous part of the sample. The diffraction
peak for plane (002) is located at a diffraction angle
Asia-Pac. J. Chem. Eng. (2014)
DOI: 10.1002/apj
P. P. ZHANG Asia-Pacific Journal of Chemical Engineering

around 2θ = 22◦, and the intensity scattered by the
amorphous part is measured as the lowest intensity at
a diffraction angle around 2θ = 18◦.
RESULTS AND DISCUSSION
Fourier transform infrared spectroscopy analyses
As many other natural plant materials, cellulose in the
cell walls of bamboo also exists in the form of
microfibrils enclosed by hemicellulose and lignin
(Fig. 1A). Alkali and bleaching treatments are usually
implemented to remove hemicellulose and lignin.
Then, during acid hydrolysis, amorphous domains of
cellulose are preferentially hydrolyzed, because
crystalline regions have a higher resistance to acid
attacking.[27] Table 1 summarized the typical
functional groups of cellulose, hemicellulose, and
lignin detected on FTIR spectra.[14] Figure 2A shows
the FTIR spectra of (a) PH, (b) PHC, and (c) CNS.
Dotted lines highlight the changes before and after
chemical treatments. First, some absorbance peaks
can be observed on the FTIR spectra of PH, PHC,
and CNS. Three samples all exhibited absorbance in
the regions as follows: the broad band located from
3000 to 3750 cm
1, the peak centered at 2885 cm
1
and the small narrow peak at 900 cm
1 (Fig. 2A (a),
(b), and (c)), which can be ascribed to the stretching-
vibration of hydroxyl groups (O–H) (Rosa et al.
2012) (Table 1, entries 1, 7, 11), the strain of C–H
(Table 1, entries 2, 8, 12),[28] and stretching-vibration
of C–O (entries 6, 10, 19),[13,15] respectively.
Nevertheless, alkali and bleaching treatments
obviously brought about changes of the structure of
the solid samples as observed from FTIR spectra
(Fig. 2A). A shoulder peak around 1721 cm
1 in the
spectrum of PH (Fig. 2A (a)) was ascribed to the

Table 1. Typical Fourier transform infrared absorbance bands of functional groups of cellulose, hemicellulose, and lignin.

Entry Fiber component Wavenumber (cm
1) Involved groups References

[28]
1 Cellulose 3428 Strain O–H in hydroxy
[28]
2 2885 Strain C–H
[28]
3 2358 Acetals
[16]
4 1640 O–H bond of water absorption
[14]
5 1170, 1035

[28]
6 900 C–H vibration, C–O stretching
[28]
7 Hemicellulose 3428 Strain O–H in hydroxy
[28]
8 2885 Strain C–H
[16]
9 1721 C═O stretching
[28]
10 900 C–H vibration or C–O stretching
[28]
11 Lignin 3428 Strain O–H in hydroxy
[28]
12 2885 Strain C–H
[16]
13 1721 C═O stretching
[14]
14 1600 Strain of

15 1640, 1516, 1463 Stretching of [16]

in lignin

[14]
16 1430, 1423 Stretching of O–CH3 in

[15]
17 1240 Corresponds to C–O–C (aryl-alkyl ether)

in

[14]
18 832 Strain O–H in

[28]
19 900 C–H vibration or C–O stretching

Copyright © 2014 Curtin University of Technology and John Wiley & Sons, Asia-Pac. J. Chem. Eng. (2014)
Ltd. DOI: 10.1002/apj
Asia-Pacific Journal of Chemical Engineering EFFECTS OF ACID TREATMENTS ON BAMBOO CELLULOSE NANOCRYSTALS
Figure 2. Fourier transform infrared spectra of A: (a) PH,
(b) PHC, and (c) CNS; B: (a) commercial cellulose, (b)
CNS, (c) CNH, (d) CNP, and (e) CNA. Dotted lines point
out the changes before and after chemical treatments.
This figure is available in colour online at www.
apjChemEng.com.
stretching-vibration of C═O (Table 1, entries 9, 13), which
derived from and in hemicellulose or
the in the ferulic and p-coumaric components of
lignin.[13] This peak disappeared in the spectrum of PHC
(Fig. 2A (b)) owing to the removal of hemicellulose and
lignin from PH under the aforementioned chemical
treatments. In addition, the absorbance peaks at 1640,
1600, 1516, and 1463 cm
1, which are caused by the
C═C stretching of aromatic rings in lignin (Fig. 2A (a))
(Table 1, entries 14, 15), also disappeared after alkali and
bleaching treatments. The results indicated the successful
removal of hemicellulose and lignin after four-step alkali
treatments and two-step bleaching treatments.
The band at 1640 cm
1 could be assigned to the C═C
stretching of aromatic rings of lignin, but it was
also present in PHC and CNS. This band was caused by
the bending vibration of adsorbed water (Table 1,
entry 4).[14] Because of the strong interaction between
adsorbed water and the hydrophilic surface group –OH
of cellulose, it was found to be difficult to dry cellulose
completely. Moreover, after the chemical extraction, a
weak broad peak at 2358 cm
1 appeared. It was caused
Copyright © 2014 Curtin University of Technology and John Wiley & Sons,
Ltd.
by the hemiacetals in cellulose units. This weak broad peak
proved the enrichment of cellulose fibers. The peaks at
1170 and 1035 cm
1 in each spectrum (Fig.2A (a), (b),
and (c)) were ascribed to the vibration of pyranose ring
skeletal (Table 1, entry 5) in cellulose units.[29] All these
results indicated that cellulose was extracted from PH.
Finally, it can be also observed from Fig. 2A that
there was no significant difference in the spectra of
PHC and CNS. Moreover, the FTIR spectrum of the
extracted bamboo cellulose obtained in this work was
identical with that of commercial cellulose (Fig. 2B).
Morphology of bamboo powder, cellulose and
cellulose nanocrystals
The scanning electron microscope micrograph of the PH
sample (Fig. 3) indicated that PH possessed a hierarchical
structure on the microscopic level, as reported by Moon
et al.[30] The average length of the raw fibers was in the
range of about 102.1 μm while that of PHC was in the
range of 91.4 μm. It was also observed that the fibers of
PHC became thinner and shorter after being treated with
a sodium hydroxide solution and a chlorite solution
(Fig. 3). For the original fibers in the PH, fibers were
bonded together by hemicellulose and lignin (Fig. 1A).
The bonds were then diminished after the alkali and
bleaching treatments (Fig. 1 (a)), and individual cellulose
fibers were released accordingly (Fig. 1 (1-1′, 2-2′).
After the hydrolysis of the extracted bamboo cellulose
in four kinds of acid solutions, respectively, the fibers of
PHC were defibrillated to thinner and shorter fibers, as
shown by TEM images in Fig. 4 ((a) CNS, (b) CNP,
(c) CNH, and (d) CNA). CNS cellulose sample exhibited
needlelike, wormlike, and small spherical crystals and
these nanometer-scaled crystals were 4–9 nm in width
and 6–200 nm in length (Fig. 4 (a)). However, as for
CNP made from the hydrolysis of the extracted bamboo
cellulose in H3PO4 solution, the obtained nanocrystals
exhibited in the length of 20–85 nm (Fig. 4 (b)). Clearly,
the length distribution of CNP was narrower than that of
CNS. The differences could be explained by the fact that
the hydrogen ions attacked amorphous regions of bamboo
cellulose with a high hydrolysis rate until the cellulose
was hydrolyzed to release soluble sugars, such as
cellobiose and polysaccharides. The hydrolysis of the
extracted bamboo cellulose in different acids led to
cellulose nanocrystals with different morphologies and
sizes as observed in TEM images. H2SO4 solution had
more hydrogen ions than H3PO4 solution when both were
set and used at the same concentration. Therefore, in the
case of H3PO4 solution, less hydrogen ion attacked
amorphous regions, resulting in a lower hydrolysis rate
and larger particles.
Under the hydrolysis of the extracted bamboo
cellulose in hydrochloric acid, the resulting CNH
cellulose crystallites appeared to be wormlike and were
20–40 nm in length (Fig. 4 (c)), whereas the hydrolysis
Asia-Pac. J. Chem. Eng. (2014)
DOI: 10.1002/apj
P. P. ZHANG Asia-Pacific Journal of Chemical Engineering

Figure 3. Scanning electron microscope micrographs (scale bar: 500 microns; inset, scale
bar: 50 microns) and size distribution of (a) PH and (b) PHC.

Figure 4. Transmission electron microscope micrographs of (a) CNS, (b) CNP, (c) CNH, and
(d) CAN (scale bar: 200 nm). The resulting colloidal suspensions (2 wt%) of (e) CNS, (f)
CNP, (g) CNH, and (h) CNA, which were allowed to stand for 12 h after treated with
ultrasonic dispersion. This figure is available in colour online at www.apjChemEng.com.

of the extracted bamboo cellulose in the mixture of (c)). This was due to the aggregating of nanocrystals.
acetic and nitric acids resulted in CNA cellulose Araki et al.[31] reported that there was no surface charge
crystallites in the form of spherical particles and in on the surface of CNH nanocrystals. Therefore, here, the
the length of 6.5–20 nm (Fig. 4 (d)). Moreover, for aggregating of CNH nanocrystals was thought to be
CNH samples, some bundles could be observed (Fig. 4 caused possibly by the hydrogen bonds on the surface
Copyright © 2014 Curtin University of Technology and John Wiley & Sons, Asia-Pac. J. Chem. Eng. (2014)
Ltd. DOI: 10.1002/apj
Asia-Pacific Journal of Chemical Engineering EFFECTS OF ACID TREATMENTS ON BAMBOO CELLULOSE NANOCRYSTALS
of individual cellulose crystallites. In addition to CNH,
many aggregations of spherical cellulose nanocrystals
could also be clearly observed in Fig. 4 (d). Besides, it
was found for the first time that bamboo cellulose
nanocrystals, as evidenced by CNA samples, could also
be prepared by the hydrolysis of the extracted bamboo
cellulose in the mixed acetic and nitric acid systems
under strong agitation at temperature of 60 °C.
The width of all cellulose nanocrystals was in the
range of 2 to 12 nm (Table 2). The measurement of the
length and the width of the four samples demonstrated
that, by the hydrolysis of the extracted bamboo cellulose
in different acids, cellulose nanocrystals in remarkably
different length can be obtained, but there were no
significant influences on the width of the crystal.[11]
Colloidal suspensions of bamboo cellulose
nanocrystals
The characteristics of colloidal suspensions of (e) CNS,
(f) CNP, (g) CNH, and (h) CNA were also analyzed
(Fig. 4). The content of cellulose nanocrystals in the four
colloidal suspensions was about 2 wt% and ultrasonic
dispersion was carried out. Stability of colloidal
suspensions was observed after 12 h. Colloidal
suspensions of CNS exhibited excellent stability (Fig. 4
(e)). On the one hand, the charged surface sulfate esters
that can promote dispersion of the long cellulose
nanocrystals in water, thereby leading to good stability.[24]
On the other hand, charged small spherical cellulose
nanocrystals could act as a matrix that could hold up the
long nanocrystals in water. By contrast, the stability of
CNP colloidal suspensions was lower (Fig. 4 (f)),
although specific chemical adsorption may occur between
the surface of CNP nanocrystals and the charged
phosphorylated group and thus prevent the aggregation
of the cellulose nanocrystals.[32] But no small spherical
nanoparticles could be seen in CNP samples (Fig. 4 (b).
In addition, from the phenomena of CNS and CNP
colloidal suspensions together, it was found that the
Table 2. The diameter and aspect ratio of cellulose
nanocrystals.
Preparation Length Width Aspect
Samples conditions (nm) (nm) ratio†a
CNS H2SO4 6.5 M, 3–200 3–12 1–28
60 °C, 2 h
CNP H3PO4 6.5 M, 20–85 4–10 3–11
60 °C, 2 h
CNH HCl 6.5 M, 20–40 5–9 3–6
60 °C, 2 h
CNA HAc 6.5 M, 6.5–20 5–7 1–3
60 °C, 2 h
a
: Aspect ratio, defined as the length-to-width (L/W), is a predominant
factor in the morphology characterization of cellulose nanocrystals.
Usually, sphere-tended nanoparticles exhibit a low aspect ratio value
(≥1), which is close to 1.
Copyright © 2014 Curtin University of Technology and John Wiley & Sons,
Ltd.
charged small needlelike cellulose nanocrystals played a
major important role in stabilizing colloidal suspensions.
As for the colloidal suspensions of CNH and CNA,
both colloidal systems were much less stable than the
colloidal suspension of CNS, which meant easier
aggregation of wormlike CNH and spherical CNA.
The small particles CNH and CNA possessed high
specific surface area. As a result, the high exposure of
surface hydroxyl groups of CNH and CNA made it
easy to form hydrogen bonds among individual
cellulose crystallites.
X-ray diffraction analysis
Generally, bamboo cellulose elementary fibrils contains
alternate crystalline and amorphous regions.[19] More-
over, there exists both intra- and intermolecular hy-
drogen bonding in amorphous regions and crystalline
regions. Additionally, hemicellulose and lignin are
amorphous polymers[17] and act as adhesives among
cellulose elementary fibrils (Fig. 1). Powder XRD
patterns of PH and PHC showed reflections at
2θ = 14.7°, 16.4°, and 22.6° (Fig.5A), which are
characteristic of cellulose. And the crystallinity index
of samples can be calculated according to the Eqn (1).
The crystallinity index of PH was 48.2% while that of
PHC was 57.8% (Table 3). Compared with PH, the
increased crystallinity of PHC was ascribed to the
removal of amorphous polymers hemicellulose and
lignin by alkali and bleaching treatments (Fig. 1 (a)).
The crystallinity of cellulose nanocrystals was
obviously related to acid treatment (Fig. 5B and
Table 3). The crystallinity index of CNH, CNA, CNS,
and CNP was found to be 61.0%, 61.8%, 73.6%, and
67.2%, respectively. The crystallinity index of these
cellulose nanocrystals was higher than that of PHC.
The results were in agreement with the report in the
previous literature that acid treatment on natural fibers
could significantly affect the crystallinity of cellulose.
Particularly, proper acid treatment can break the intra-
and intermolecular hydrogen bonds in the amorphous
region of cellulose.[7] In other words, acid treatment
resulted in hydrolysis of cellulose partly and even
completely.[33] The hydrogen ions could penetrate into
the amorphous regions of cellulose, promoting the
hydrolytic cleavage of glycosidic bonds and finally
releasing the individual cellulose crystallites.[33,34] In
addition, slight realignment of crystals may also occur,
thus leading to the further increase of crystallinity of
the resultant nanocrystals. Moreover, the reflections of
CNS and CNP around 22.6° are stronger than that of
CNH and CNA (Fig. 5B). These results clearly
demonstrated that hydrolyzing cellulose by sulfuric
and phosphoric acids could result in cellulose
nanocrystals with high crystallinity whereas CNH and
CNA possessed low crystallinity. Besides the effects
of hydrogen cations on the resultant crystallites, the
Asia-Pac. J. Chem. Eng. (2014)
DOI: 10.1002/apj
P. P. ZHANG Asia-Pacific Journal of Chemical Engineering

Thermogravimetric analysis

The thermal behaviors of samples during different che-

mical treatment were measured by thermalgravimetric
(TG) and derivative thermogravimetric (DTG) technique
(Fig. 6). All the TG curves showed an initial slight weight
loss at around 100 °C, which corresponded to the removal
of absorbed water on the surfaces of these materials and
the intermolecular H-bonded water. [22,36] As for TG
and DTG curves of PH and PHC, it was noticed that the
decomposition of PH started at about 250 °C and that
of PHC started at about 280 °C (Fig. 6B). This
indicated that the alkali and bleaching treatments led
to an increase of thermal stability in the extracted
bamboo cellulose PHC. The temperatures of
hemicellulose and lignin decomposition are lower than
that of cellulose.[16] Thus, their removal in the
PHC sample exhibited better thermal stability. In
addition, the rate of weight loss of PH in the range of
250–380 °C was lower than PHC (Fig. 6B). It was
because that decomposition of lignin happened in a
wider temperature range than cellulose.[29]
The different thermal behaviors of cellulose nano-
crystals made from different acid systems can be clearly
observed from the TG and DTG curves. The decom-
position of commercial cellulose whiskers started at

Figure 5. Powder X-ray diffraction patterns of A: (a)

PHC and (b) PH; B: (a) CNA, (b) CNP, (c) CNS, (d)
CNH, and (e) commercial cellulose. This figure is
available in colour online at www.apjChemEng.com.

Table 3. Crystallinity indexes of all samples and com-

mercial cellulose.

2θ (am) 2θ (002) Crystallinity

Samples (°) (°) index (%)
PH 18.1 21.8 48.2
PHC 18.9 22.3 57.8
CNS 18.7 22.4 73.6
CNP 18.6 22.3 67.2
CNH 18.4 22.3 61.0
CNA 18.3 22.3 61.8
Commercial 18.2 22.7 87.4
cellulose

anions might also have a remarkable effect. For

example, compared with SO4 2
and PO4 3
, Cl
and
CH3COO
were thought to have better capability to
swell cellulose, thereby facilitating the breakage of
intra- and intermolecular hydrogen bonds in crystalline
regions of cellulose attacked by hydrogen cations.[35]
Hence, cellulose nanocrystals CNH and CNA appeared
to have low crystallinity (Table 3).
Copyright © 2014 Curtin University of Technology and John Wiley & Sons,
Ltd.
Figure 6. Thermalgravimetric and derivative thermo-
gravimetric curves of (a) PH, (b) CNP, (c) CNH, (d) CNA,
(e) CNS, (f)PHC, and (g) commercial cellulose. This figure
is available in colour online at www.apjChemEng.com.
Asia-Pac. J. Chem. Eng. (2014)
DOI: 10.1002/apj
Asia-Pacific Journal of Chemical Engineering EFFECTS OF ACID TREATMENTS ON BAMBOO CELLULOSE NANOCRYSTALS
about 320 °C and the main decomposition peak was at
about 342 °C. By contrast, the decomposition of CNS
and CNP started at about 270 °C and reached a
maximum at about 310 °C, whereas that of CNH started
at about 310 °C and reached a maximum at 342 °C
(Fig. 6B). This suggested that the cellulose nanocrystals
obtained by sulfuric and phosphoric acids led to a slight
decrease in the thermal stability. Although the
crystallinity index of CNS and CNP is higher than that
of others, actually, the introduction of sulfated groups
and phosphorylated groups into the crystals during the
acid hydrolysis process could reduce the thermal stability
of nanocrystals as reported earlier in the literature.[36] The
SO4 2
and PO4 3
can weaken the hydrogen bonds
between neighboring cellulose chains and intramolecular
hydrogen bonds. As a result, such a role play by SO4 2

and PO3
4 led to decreased thermal stability. In addition,
SO4 2
and PO4 3
can act as thermal conductor to
decompose cellulose.[37] Furthermore, the residual mass
of PHC was less than that of raw PH can be observed in
Fig. 6A. This resulted from the removal of hemicellulose
and lignin.[38,39] In short, the thermal stability of extracted
bamboo cellulose and nanocrystals was affected by
factors such as crystallinity, size, and components, which
in turn depended on acid treatment conditions.
CONCLUSION
Bamboo cellulose was able to be extracted from raw P.
heterocycla bamboo fibers by consecutive treatments,
which involved alkali extraction four times and
bleaching treatment two times. After such alkali
extraction and bleaching treatment, cellulose nano-
crystals were successfully prepared by the hydrolysis of
the extracted bamboo cellulose in acid solution. The type
of acid used had a significant influence on the structure,
morphology, and thermal stability of cellulose nano-
crystals. When sulfuric acid was used to hydrolyze
bamboo cellulose, the resulting cellulose nanocrystals
were mixed needlelike, wormlike, and small spherical
crystallites, whereas the cellulose nanocrystals made
from the hydrolysis of the extracted bamboo cellulose
in phosphoric acid, hydrochloric acid, or the mixed
acetic and nitric acid solution were needlelike, wormlike,
and spherical particles, respectively. In addition, the
anions from the acid used affected the hydrolysis of
cellulose and the structure and properties of the resulting
crystallites. Compared with SO4 2
and PO4 3
, Cl
and
CH3COO
showed higher inclination to promote the
breakage of the hydrogen bonds in crystalline regions
of cellulose through facilitating the swelling of cellulose.
As a result, the resultant cellulose nanocrystals
demonstrated low crystallinity index. Besides, they can
also help heat transfer during thermal degradation. Thus
the resulting extracted cellulose crystallites exhibited a
low thermal stability. These findings offer a new
Copyright © 2014 Curtin University of Technology and John Wiley & Sons,
Ltd.
understanding on acid treatment for tuning the structure
and properties of extracted cellulose crystallites.
Abbreviations
PH: Phyllostachys heterocycla fibers.
PHC: Cellulose isolated from Phyllostachys
heterocycla fibers.
CNS: Cellulose nanocrystals prepared from
bamboo cellulose by the hydrolysis of
bamboo cellulose in the sulfuric acid
solution.
CNP: Cellulose nanocrystals prepared from
bamboo cellulose by the hydrolysis of
bamboo cellulose in the phosphoric acid
solution.
CNH: Cellulose nanocrystals prepared from
bamboo cellulose by the hydrolysis of
bamboo cellulose in the hydrochloric
acid solution.
CNA: Cellulose nanocrystals prepared from
bamboo cellulose by the hydrolysis of
bamboo cellulose in the mixed solution
of acetic acid and nitric acid.
Acknowledgements
The authors wish to acknowledge the financial support
from the National Natural Scientific Foundation of China
(21373185), the Distinguished Young Scholar Grants
from the Natural Scientific Foundation of Zhejiang
Province (ZJNSF, R4100436), ZJNSF (LQ12B03004),
Zhejiang ‘151 Talents Project’, and the projects
(2010C14013 and 2009R50020-12) from Science and
Technology Department of Zhejiang Provincial
Government and the financial support by the open fund
from breeding base of state key laboratory of green
chemistry and synthesis technology. CHZ conceived the
project and PPZ mainly conducted the experiments and
collected the data. PPZ wrote the manuscript with support
and revisions from CHZ. All coauthors provided
assistance during the work.
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