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Flow Properties of Microcrystalline

Cellulose Suspension Prepared by Acid
Treatment of Native Cellulose
Article in Colloids and Surfaces A Physicochemical and Engineering Aspects November 1998
DOI: 10.1016/S0927-7757(98)00404-X

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Colloids and Surfaces

A: Physicochemical and Engineering Aspects 142 (1998) 7582

Flow properties of microcrystalline cellulose suspension

prepared by acid treatment of native cellulose
Jun Araki *, Masahisa Wada, Shigenori Kuga, Takeshi Okano
Department of Biomaterial Sciences, Graduate School of Agricultural and Life Sciences, The University of Tokyo,
Yayoi 1-1-1, Bunkyo-ku, Tokyo 113, Japan
Received 5 December 1997; accepted 24 March 1998

Abstract
The properties of cellulose microcrystals and their colloidal suspensions obtained by acid treatments of native
cellulose have been investigated. Bleached softwood kraft pulp was treated with 65% (w/w) sulfuric acid or 4 N
hydrochloric acid and the resulting particle morphology and flow properties of the aqueous suspensions were studied.
Colloidal dispersion of the HCl-treated sample was obtained by more thorough removal of acid from the hydrolysate
than in the case of H SO treatment. The H SO -treated and HCl-treated samples had similar particle sizes and
2 4
2 4
shapes when observed by electron microscopy. Conductometric titration of suspensions revealed that the
H SO -treated sample had a surface charge of 84 m-equiv kg1 dry material due to the introduction of sulfate groups,
2 4
while that of the HCl-treated sample was undetectable. The H SO -treated suspension showed no time dependence in
2 4
viscosity, while the HCl-treated suspension was thixotropic at concentrations >0.5% (w/v) and anti-thixotropic
<0.3%. The inclination of the relative viscosityconcentration plot (Einstein coefficient) for the H SO - and HCl2 4
treated suspensions were 135 and 1380, respectively. The former value, when used in Simhas equation for rod-like
particle suspension, roughly corresponds to an axis ratio of 50 estimated from the micrographs. 1998 Elsevier
Science B.V. All rights reserved.
Keywords: Acid treatment; Cellulose microcrystals; Rod-like particles; Surface charge; Thixotropy/anti-thixotropy

1. Introduction
Treatment of native cellulose with 65 wt% sulfuric acid gives suspension of microcrystalline cellulose [17]. This is a colloidal suspension of rodlike particles (whisker) nanometers in size showing
chiral
nematic
phase
separation
[35].
Marchessault et al. studied the viscosity of
H SO -treated suspensions and found its
2 4
Newtonian behavior and dependence on particle
size [6 ].
During the preparation of this system a certain
* Corresponding author.

amount of sulfuric ester groups is introduced to

the surface of microcrystals. The amount of sulfuric esters on H SO -treated microcrystals reaches
2 4
0.52% in sulfur content [3,4]. A value of
0.155 e nm2 was reported for the surface charge
of cotton cellulose microcrystals due to sulfate
ester and carboxylic groups [5]. This surface
charge contributes to the stability of the colloidal
dispersion [27].
While these cellulose particle systems seem to
be suitable for the study of enzymatic degradation
of cellulose, the extrinsic surface groups can hinder
the substrate recognition by enzymes. In fact, a
remarkable difference was observed between the

0927-7757/98/$ see front matter 1998 Elsevier Science B.V. All rights reserved.
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J. Araki et al. / Colloids Surfaces A: Physicochem. Eng. Aspects 142 (1998) 7582

degree of degradation of two kinds of microcrystalline cellulose samples prepared by different acids
[8]. Sulfate groups may also affect the enzyme
activity through changes in the pH of the system.
To evaluate the influences of surface charge on
the rheological behavior of the suspension, and on
the enzymatic action on cellulose particles, it is
desirable to prepare microcrystalline cellulose with
a minimal and/or a known amount of surface
charge. Use of hydrochloric acid might meet this
requirement, but no report of this kind was found.
This is simply due to the difficulty in obtaining
good dispersion of particles treated with HCl.
During the removal of acid from the H SO
2 4
hydrolysate by centrifugation [3], the fine particle
components become non-sedimenting when the
pH reaches 13. The coarse component (pellet)
can be removed at this stage conveniently. In the
case of HCl treatment, however, such a condition
is not readily attained.
In the present study the initial aim was to
establish a method for preparing colloidal suspensions of cellulose microcrystals by HCl treatment
in order to investigate difference in rheological
behavior between the samples with different levels
of surface charge.

The HCl-treated suspension was prepared by

treating 5 g of the kraft pulp with 175 ml of 4 N
HCl at 80C for 225 min. Repeated centrifugation
up to more than pH 4 was necessary to obtain a
turbid supernatant under 1600g. The supernatant
was collected, purified by dialysis and sonicated as
described already.
The suspension, either H SO - or HCl-treated,
2 4
was concentrated by a high-speed centrifuge at
18 800g at 4C for 60 min. The suspension was
dialyzed against deionized water until the surrounding water became neutral (the pH of the
suspension itself, however, leveled off at 23 for
the H SO -treated sample due to sulfate attached
2 4
to cellulose; that of HCl-treated sample was ~6).
The suspension was then sonicated similarly and
kept in a refrigerator until use.

2. Materials and methods

2.3. X-ray analysis

2.1. Preparation of suspension

Approximately 150 mg of the freeze-dried microcrystal sample was pressed at 200 kg cm2 into a
pellet 20 mm wide and 1 mm thick. The X-ray
diffraction pattern was obtained with Ni-filtered
Cu Ka radiation using an X-ray generator (Rigaku
RINT2000) at 30 kV, 40 mA.

Unless specified, all reagents used in this study

were purchased from Wako Pure Chemical Ind.,
Ltd. Bleached softwood kraft pulp was used as the
cellulose material. The method by Revol et al. [3]
was used to prepare the H SO -treated suspension.
2 4
Ten grams of dried pulp was treated with 100 ml
of 65% (w/w) H SO at 70C for 10 min with
2 4
continuous stirring. The sample was then repeatedly washed with centrifugation (1600g, at 10C
for 5 min). When the pH of the sample reached
13, the fine particle component did not sediment
and the supernatant became turbid. The turbid
supernatant was collected and thoroughly dialyzed
against distilled water. The suspension was finally
sonicated with a rod-type sonicator (Nissei
Ultrasonic homogenizer) for 1 min.

2.2. Transmission electron microscopy (TEM)

Dilute suspension was dropped onto a thin
carbon film spread on a copper grid. Dried microcrystals were examined with a JEOL 2000EX transmission electron microscope at 200 kV in brightfield mode by defocus contrast. Images of microcrystals were recorded on Mitsubishi MEM films.

2.4. Infrared spectroscopy

The colloidal cellulose suspension was freezedried and ca 3 mg of dried sample was mixed with
300 mg of KBr ( UvasolA, MERCK ) and pressed
into a tablet at 200 kg cm2. The infrared (IR)
spectra were recorded by a Shimadzu
FTIR-8100M spectrometer. Spectra in the range
of 4004000 cm1 were obtained with a resolution
of 2 cm1 and the signal was accumulated from
100 scans.

J. Araki et al. / Colloids Surfaces A: Physicochem. Eng. Aspects 142 (1998) 7582

77

2.5. Conductometric titration

The surface charge on cellulose particles was
determined using the method described by Katz
et al. [9]. Forty-five milliliters of the suspension
was mixed with 5 ml of 0.01 M NaCl before measurement. NaOH (0.01 M ) was added to the suspension at a rate of 0.5 ml every 5 min with
continuous stirring. The change in conductivity
was recorded by a conductometer ( TOA electronic
CM-2A).
2.6. Viscosity measurement
A double-cylinder type viscometer with a small
volume sample adapter (Brookfield LVDV-I+)
was used to measure the viscosity of the suspensions. An ultra-low viscosity sample adapter was
used when necessary (<3 mPa s). The sample was
thermostated at 200.1C.

3. Results and discussion

3.1. Morphology and surface charge of particles
Apparently stable and turbid colloidal suspensions were obtained by treating the cellulose
sample with either H SO or HCl. Both suspen2 4
sions showed remarkable flow birefringence
through crossed nicols ( Fig. 1) as in the cases of
ramie, bacterial cellulose [6 ] and wheat straw [10].
TEM observations gave almost the same appearance for the two samples ( Fig. 2). The individual
microcrystals were ca 3.5 nm wide and 18075 nm
long for both H SO - and HCl-treated samples.
2 4
Electron microscopy showed that the size and
shape of the smallest rod-like units (microcrystal )
were the same for the two samples (arrowheads in
Fig. 2). The state of aggregation is apparently
different for Figs. 2(a) and (b), but it is not clear
at present whether this aggregation was present in
the suspension or caused by drying on the specimen grid.
Fig. 3 shows the X-ray diffraction patterns for
the HCl- and H SO -treated cellulose samples;
2 4
both showing the cellulose I crystalline form and
are nearly identical. Also no difference was found

Fig. 1. Suspensions prepared by (a) H SO and (b) HCl treat2 4

ments viewed through a pair of crossed nicols. Both suspensions
shows remarkable flow birefringence by stirring. Solid content:
(a) 0.75%; and (b) 0.17%.

between the IR spectra of the two microcrystal

samples. The amount of sulfate groups was so
small that the absorbance due to sulfate groups at
14001600 and 1200 cm1 was not detected.
Fig. 4 shows the conductometric titration curves.
The curve for the H SO -treated suspension
2 4
[Fig. 4(a)] first showed a decrease in conductivity
corresponding to the neutralization of strong acid
groups on microcrystals. The horizontal portion
after the first inflection point corresponds to the
neutralization of weak acid groups (probably carboxylic acid). After complete neutralization of the
weak acids, the conductivity increased due to
excess NaOH. By contrast, the titration curve for
the HCl-treated suspension [Fig. 4(b)] rose from
the beginning and did not show the first decrease
in conductivity. To ensure that no acid group was
present, 5 ml of 0.01 M HCl was added to HCltreated suspension prior to titration and the curve
shown in Fig. 4(c) was obtained. The neutralization of strong acid was completed by 5 ml of
0.01 M NaOH, confirming that no strong acid
groups except the 5 ml of 0.01 M HCl was present
in the original HCl-treated suspension. There
appears to be a small horizontal portion corresponding to weak acid in both Figs. 4(b) and (c).
The amount of weak acid could not be precisely
determined due to experimental limitation, but it
was estimated to be no more than 18 mmol kg1.
The weak acid groups in the original pulp were

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J. Araki et al. / Colloids Surfaces A: Physicochem. Eng. Aspects 142 (1998) 7582

Fig. 3. X-ray diffraction pattern (reflection mode) of microcrystals obtained by H SO and HCl treatments.
2 4

Fig. 2. Transmission electron micrograph of cellulose microcrystals prepared by (a) H SO and (b) HCl treatments. Typical
2 4
individual microcrystals are shown by arrowheads. Scale bars
show 500 nm.

determined to be 57 mmol kg1 by conductometric

titration, possibly originating from uronic acids
and oxidation products in the original kraft pulp.
Table 1 summarizes the amounts of strong and
weak acid groups in the two microcrystalline cellulose samples. The strong acid groups of the
H SO -treated sample are apparently introduced
2 4
by esterification of hydroxyl groups by sulfate
ions, which are naturally absent in the HCl-treated
sample. Earlier studies have shown that the
H SO -treated cellulose suspension forms a stable
2 4
colloid system owing to the negative surface charge
[27]. The amount of strong acid determined here,

Fig. 4. Conductometric titration curves of (a) 698 mg of

H SO -treated microcrystal, (b) 298 mg of HCl treated micro2 4
crystal and (c) 265 mg of HCl treated microcrystal with initial
addition of 5 ml of 0.01 M HCl.

J. Araki et al. / Colloids Surfaces A: Physicochem. Eng. Aspects 142 (1998) 7582

79

Table 1
The amounts of strong and weak acidic groups on microcrystalline cellulose
Sample

NBKP/H SO
2 4
NBKP/HCl

Amounts of acidic groups (mmol kg1)

Strong acid groups

Weak acid groups

84
0

26
<18

84 mmol kg1, roughly corresponds to the previously reported value of surface charge density
of 0.155 e nm2 [5] for a H SO -treated cotton
2 4
sample (see Appendix A).
Though the HCl-treated suspension contained
only a small amount of weak acid groups, it could
form a stable colloid at the very low ionic strength
attained by dialysis. Its colloidal stability was
much lower than that of the H SO -treated sample;
2 4
while the latter was precipitated by electrolyte
addition only at >20 mM, the former was precipitated at ~0.4 mM. The formation of stable cellulose colloid without introduction of strong acid
groups is a novel phenomenon and leads to the
anomalous viscosity behavior described in
Section 3.2.
3.2. Viscosity behaviors
Fig. 5 shows the time dependence of viscosity of
the cellulose suspensions. While the viscosity of
H SO -treated suspension showed no time depen2 4
dence [Fig. 5(a)], that of HCl-treated suspension
showed typical thixotropy at concentrations higher
than ca 5 g l1 [Fig. 5(b)]; on the other hand, the
dilute suspensions ( less than ca 3 g l1) showed
an increase in viscosity with time (anti-thixotropy).
The viscosity of the HCl-treated suspension equilibrated after 30 min of rotation in either case.
When the rotation was stopped for a short time
and re-started, the viscosity resumed the previous
value. The viscosity, however, returned to the
initial value with elongated interruption, in ca
70 min. In this respect the HCl-treated suspension
is a typical thixotropic system at higher
concentrations.
Thixotropy of the HCl-treated suspension indicate the effect of significant inter-particle aggrega-

Fig. 5. Time dependence of viscosity of (a) H SO -treated and

2 4
(b) HCl-treated suspensions. Note the difference in viscosity
scale.

tion [11,12], which forms in static condition and

is destroyed by shear flow. The behavior at lower
concentrations suggests that the increased interparticle distances reduce the contribution of such
aggregation to thixotropy, and that conversely the
interaction between particles may be enhanced by
their mutual alignment due to shear flow and
results in anti-thixotropy. This interpretation is
corroborated by that the behavior of a dilute HCltreated suspension (1.71 g l1) turned from anti-

80

J. Araki et al. / Colloids Surfaces A: Physicochem. Eng. Aspects 142 (1998) 7582

thixotropy to thixotropy by the addition of 3.5 mM

of NaCl; the suspension after the addition of salt
contained visually apparent flocculation. A somewhat similar phenomenon of concentration-dependent thixotropy-anti-thixotropy has been reported
for a claywater system [13]. The HCl-treated
cellulose suspension described here seems to be the
first instance to show this kind of behavior as a
rod-like particle system.
Both cellulose suspensions showed shear-thinning behavior ( Fig. 6; the viscosity at 30 min after
the start of measurement are plotted). Similar
behavior was observed with Avicel, a commercial
microcrystalline cellulose [14]. The steep decline
in viscosity of the HCl-treated suspension apparently corresponds to the breakdown of flocculates
which causes thixotropic behaviors described
already. The H SO -treated suspension, on the
2 4
other hand, showed only weak shear-rate dependence in viscosity, which is possibly due to flowinduced orientation of the rod-like particles.
Fig. 7 shows the relative viscosity of suspensions
plotted against the volume fraction, w, of cellulose
(value at 30 min after start as above). The viscosity
of suspension of rod-like particles, with an axis
ratio of f, can be estimated by Simhas formula
[15]:
n=

f2
15( log 2f3)
2

f2
5( log 2f1)
2

14
15

(1)

where n is the coefficient in Einsteins viscosity

equation [16 ]:
g =1+nw
rel

(2)

where g is the relative viscosity of suspension

rel
with volume fraction w; n equals 2.5 for spherical
particles. From the inclination of the curve at w=
0 in Fig. 7(a) for the H SO -treated suspension,
2 4
the coefficient n is determined as 135, which
translates to f=47 according to Eq. (1). This value
is in good agreement with the aspect ratio of
5020 determined from the electron micrographs
[Fig. 2(a)]. The same procedure with the HCltreated suspension gives n=1380 and f=172. These
values seem to reflect an anomaly in viscosity by
flocculation, because the size and shape of indivi-

Fig. 6. Shear rate dependence of viscosity of (a)

H SO -treated and (b) HCl-treated suspensions. Note the
2 4
difference in viscosity scale.

dual particles were similar in the two samples

in Fig. 2.
The conductometric titration and viscosity
measurements suggest that HCl-treated microcrystals form loose aggregates in water due to van der
Waals attraction because of the lack of surface
charge. Because the rod-like particles can contact
and interact extensively with each other, this interaction can lead to the formation of loose and
bulky aggregates. Such a system will show compli-

J. Araki et al. / Colloids Surfaces A: Physicochem. Eng. Aspects 142 (1998) 7582

81

exhibiting interesting properties that may find new

applications. Also its lower degree of modification
of surface molecules makes it more suitable for
substrate of cellulolytic enzymes, providing a
useful material for enzyme studies.

4. Conclusions

Fig. 7. Relative viscosity as a function of volume fraction w of

suspended solid of (a) H SO -treated and (b) HCl-treated sus2 4
pensions. The dotted lines show the tangent at w=0.

cated viscosity behavior depending on solid

content, shear velocity, electrolyte concentration
and history of shear application. The steady-state
viscosity is several folds greater than that of the
same concentration of H SO -treated suspension,
2 4
in which microcrystals are stabilized by surface
charge introduced during acid treatment.
Thus our preparation of microcrystalline cellulose suspension with lower levels of surface charge
seems to give a new type of rod-like particle system

Cellulose microcrystal suspensions were prepared by treating bleached softwood kraft pulp
with 65% (w/w) sulfuric acid or 4 N hydrochloric
acid. To obtain a good dispersion of HCl-treated
microcrystals, a more thorough removal of acid
from the hydrolysate was necessary than in the
case of H SO . Conductometric titration showed
2 4
that the H SO -treated sample had a surface
2 4
charge of 84 mmol kg1 of strong acid and
26 mmol kg1 of weak acid, ascribed to sulfate
ester and carboxyl groups, respectively. No strong
acid groups and a small amount of weak acid
groups (<18 mmol kg1) were present in the HCltreated sample.
Though particle morphology, X-ray diffraction
pattern and IR spectrum were the same for these
microcrystal samples, the viscosity behavior of the
two suspensions showed remarkable differences.
The H SO -treated suspension showed no time
2 4
dependence in viscosity, while the HCl-treated
suspension was thixotropic at concentrations
>0.5% (w/v) and rheopectic <0.3%. Both suspensions were shear-thinning, and its degree for HCltreated suspension was much greater than that
of the H SO -treated suspension. The Einstein
2 4
coefficient for the H SO -treated suspension at
2 4
135, corresponded to an aspect ratio of 50 calculated from Simhas equation. The same coefficient
for the HCl-treated suspension at 1380, reflects
aggregation of particles resulting in thixotropy or
anti-thixotropy, according to the concentration.

Acknowledgement
This work was supported in part by the Grantin-Aid for a Scientific Research from Ministry of

82

J. Araki et al. / Colloids Surfaces A: Physicochem. Eng. Aspects 142 (1998) 7582

Education, Science, Sports and Culture, Japan to

T.O. (No. 07406018) and to S.K. (No. 9460073).

Appendix A. Evaluation of the amounts of surface

sulfate groups from the charge density
Dong et al. [5] calculated the surface charge
density as 0.155 e nm2 for the H SO -treated
2 4
cellulose microcrystal. This calculation was based
on their evaluation of microcrystal size of
77115 nm, which in turn gives the volume and
the surface area of single particle as 5635 nm3 and
3318 nm2, respectively. This leads to the surface
charge of single particle of 0.1553318=514.3 e.
With the density of cellulose crystal as 1.6, the
numbers of microcrystal particles in 1 kg of microcrystalline cellulose is:

A B
1000
1.6

106

56351027

=1.111020

So the surface sulfate groups in 1 kg of cellulose

microcrystals are:
514.31.111020=5.711022(e kg1)
=

5.711022
6.031023

(mol kg1)=94.7(mmol kg1)

This value agrees well with the authors result of

84 mmol kg1.

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