Cellulose

DOI 10.1007/s10570-009-9288-y

Cellulose liquefaction in acidified ethylene glycol

Edita Jasiukaityt_e Matjaz Kunaver
Matija Strlic

Received: 14 March 2008 / Accepted: 15 February 2009

Springer Science+Business Media B.V. 2009

Abstract Wood pulp cellulose was used in a study

of its catalyzed liquefaction in the presence of
ethylene glycol, p-toluene sulfonic acid monohydrate
or sulphuric acid being the catalysts. For this study,
microcrystalline cellulose, Whatman filter paper no. 1
and cotton linters with molar masses of 76,000,
699,000 and 1,910,000 g mol-1, respectively were
used. This liquefaction was studied by gravimetric
determinations, by X-ray diffraction analysis of the
residual cellulose and by monitoring of the molar
mass decrease over different time intervals, using
size-exclusion chromatography. The disordered
regions, even of cellulose with the highest molar
mass degraded in the initial minute of liquefaction.
However, the highly ordered cellulose regions
remained relatively stable for a longer time. None
the less, partial degradation of the highly ordered
regions of the cellulose was achieved.
Keywords Cellulose
Catalyzed liquefaction

Size-exclusion chromatography
Ethylene glycol

Crystallinity
X-ray
E. Jasiukaityt_e (&)
M. Kunaver
National Institute of Chemistry, Hajdrihova 19,
1000 Ljubljana, Slovenia
e-mail: edita.jasiukaityte@ki.si
M. Strlic
Faculty of Chemistry and Chemical Technology,
University of Ljubljana, Askerceva 5, 1000 Ljubljana,
Slovenia

Introduction
Renewable plant biomass sources, such as lignocellulosics and other polysaccharides, are gaining
importance as a suitable replacement for fossil-fuel
resources.
Wood is one of the more important natural
products. With respect to both its structural and
chemical properties, wood is a complex, non-uniform
material. However, the depolymerization of macromolecular wood components in a liquefaction
process, followed by reaction with specific organic
reagents, enables one to convert wood into a potential
feedstock for the synthesis of new, environmentally
friendly polymers (Wei et al. 2004; Kishi et al. 2006).
The depolymerization of wood components can be
achieved with phenol or polyhydric alcohols, under
acid-catalyzed conditions (Lin et al. 1994, 1995;
Alma et al. 1998; Kobayashi et al. 2004).
The liquefaction of wood, as a unit, is not
completely understood. Despite the fact, extensive
studies to elucidate the mechanisms were initiated
more than 10 years ago (Emsley and Stevens 1994).
Cellulose powder, steamed lignin and the mixtures of
these two components have been used by Kobayashi
et al. (2004) as a model compounds. They studied the
reaction process during liquefaction using polyhydric
alcohol. The liquefaction of lignin with phenol has
been studied on the basis of the behaviour of model
substances, such as guaiacylglycerolb-guaiacyl
ether (Lin et al. 2001a, b). Thus, these authors

123

Cellulose

attempted to clarify the mechanism of the reaction of

cellulose with phenol under the acid-catalyzed conditions, using cellobiose as a model compound.
A mechanism for the degradation of cellulose during
ethylene glycol supported liquefaction was proposed
by Yamada and Ono. These authors demonstrated the
formation of a large quantity of hydroxyethyl glucosides, in the early stage of reaction. These glucosides
subsequently decomposed into the 2-hydroxyethyl
levulinate (Yamada and Ono 2001).
The high crystallinity and/or high degree of
cellulose polymerization (DP) tended to constrain
cellulose depolymerization during the liquefaction
reaction. The disordered, amorphous zones of cellulose fibers were more accessible to the penetration of
certain solvents, while the crystalline regions
remained unaffected. In addition, only a prolonged
treatment with an appropriate diluted acid can reduce
the DP to a certain value, the leveling-off degree of
polymerization (LODP; Fengel and Wegener 1989).
It has been reported that the rate of hydrolysis of the
amorphous cellulose regions is much higher than
the rate of hydrolysis of the crystalline regions. The
products of such hydrolysis can be even removed
from a cellulose fiber, leaving the crystalline regions
almost untouched (Zhao et al. 2006). Thus, the tight
packing of cellulose in the crystalline regions causes
the reaction kinetics to be dependent on the diffusion
rate of the liquefying agents into the highly packed
macromolecular system.
The aim of the current study was to clarify the
influence of the cellulose DP on its liquefaction in
ethylene glycol under acid catalysis. Due to better
affinity of organic sulfonic acids to cellulose (Mun
et al. 2006), p-toluene sulfonic acid monohydrate and
sulphuric acid were chosen as the catalysts in our
study of the liquefaction of cellulose in ethylene
glycol.
In this study, the liquefaction of microcrystalline
cellulose, of Whatman filter paper no. 1 and of cotton
linters, with molar masses of 76,000, 699,000 and
1,910,000 g mol-1, respectively was carried out. The
objective was to determine the influence of cellulose
DP, structural features of the microfibrils and surface
morphology on the cellulose liquefaction in the
presence of ethylene glycol as a liquefying reagent,
with sulphuric acid or p-toluene sulfonic acid monohydrate as catalysts.

123

Experimental
Materials
Microcrystalline cellulose (MC, M W 76,000 g mol-1,
Acros Organics), purified cellulose sheets (Whatman
filter paper no. 1, WH, M W 699,000 g mol-1),
cotton linters (CT, M W 1,910,000 g mol-1, Radece,
Slovenia) were used without any pretreatment. Ethylene glycol (E.G, Merck), 97% sulphuric acid (H2SO4,
Merck), p-toluene sulfonic acid monohydrate (PTSA,
Acros Organics), N,N-dimethylacetamide for HPLC
(DMAc, Fluka) lithium chloride (LiCl, Acros Organics, dried before use at 180 C in vacuo for 24 h and
kept in desiccator) and sodium hydroxide (NaOH,
Merck) were of reagent grade and were used without
further purification.
Cellulose liquefaction
Thirty gram of cellulose (dried at 105 C for 24 h),
150 g of E.G and 4.5 g (3% w/w based on the E.G) of
H2SO4 or PTSA as a catalyst were placed in a
500 cm3 glass reactor (three necks), equipped with a
mechanical stirrer. The liquefaction was carried out at
150 C over 4 h. Samples from reaction mixture were
taken at different time intervals and immediately
cooled in an ice-bath. The acidity component was
neutralized with dilute NaOH solution to prevent
further cellulose degradation prior to characterization
of the products.
Measurement of residual cellulose content
The yield of liquefaction was evaluated by determining the residual cellulose content. Each sample was
diluted with an excess of distilled water and vacuumfiltered through filter paper. Insoluble residues were
rinsed several times with DMAc and acetone. The
samples were dried in vacuum at 50 C for 24 h. The
residue content was determined as the weight of
the obtained solids relative to the starting amount of
cellulose (Eq. 1).
Residual cellulose %

Wt
 100
W0

W0 is the weight of starting cellulose and Wt is the

weight of residual cellulose.

Cellulose

The residual cellulose samples were analyzed using a

PerkinElmer Spectrum-1 FTIR spectrophotometer.
The transmittance measurements were conducted
using the KBr pellet method in frequency range from
4,000 to 500 cm-1.

Intensity, a.u.

FT-IR spectroscopic analysis

I200

NMR spectrometry
13

C NMR spectra of derivatives were recorded using

a Unity Inova 300 Varian NMR spectrometer operating at 75 MHz. The measurements were conducted
in DMSO-d6 at 25 C and tetramethylsilane (TMS)
was used as an internal standard.
X-ray diffraction
XRD measurements were performed on a Siemens
D5000 PANalytical XPert PRO system in order to
estimate the crystallineamorphous ratio of the initial
cellulose samples and of the residues. The diffracted
) was measured
intensity of CuKa radiation (1.5406 A
in 2h intervals between 10 and 30. The X-ray
diffractograms of all samples were analyzed using the
empirical procedure of Segal et al. (Segal et al. 1959).
The calculation of the crystallinity index (Cr
I.)
followed the Eq. 2:
Cr:I:%

I200  Iam
 100
I200

Here, I200 is the maximum intensity of the diffraction

from the (200) plane at 2h = 22.8 and Iam is the
intensity of the amorphous background scatter measured at 2h = 18.
Figure 1 shows the approach taken in the determination of the crystalline cellulose content in the
microcrystalline cellulose by the Segal method.
The crystallite widths of both the original and the
residual cellulose were estimated and evaluated using
Scherrer equation (Eq. 3; Klug and Alexander 1974)
from the line profile of the (200) reflection at
2h = 22.8 that refers to the width of crystallite
(Andersson et al. 2003).
L Kk=b cos h

Here, L is the crystallite width, h is the Bragg angle,

k is the wavelength of the radiation, K is a constant
and b is the corrected width of the line given by the

Iam
5

10

15

20

2 ,

25

30

Fig. 1 Diffractogram of microcrystalline cellulose

specimen. A value of K of 0.9 for half-width of the

line profiles was used.
The measured line widths include the effects of
crystallite size and instrumental broadening. The
effect of instrumental broadening on the line widths
was assessed by measuring the width of line of a
large undistorted crystal under identical operating
geometries. The instrumental broadening amounted
to 0.059 for the half-width where the silicon crystal
was used as a sample (Warren and Biscoe 1938). The
line profiles were assumed to follow a Gaussian
distribution.
Size-exclusion chromatography
Samples for size-exclusion chromatography were
prepared as follows: the aliquot of neutralized
reaction mixture was mixed with distilled water and
any suspension of residual cellulose was filtered
through a 0.45 lm polyamide membrane filter
(Milipore). The obtained cellulose residue was rinsed
several times with distilled water and dissolved in a
LiCl/DMAc solvent system.
All the solutions of residual cellulose in LiCl/
DMAc were filtered through the PTFE filters
(0.45 lm) prior to injection. The LiCl concentration
in the sample solutions was 1% (0.1 g of LiCl in
10 cm3 of DMAc).
The HP-AGILENT system consisted of an isocratic pump HP 1100, refractive index detector
AGILENT 1100 (detection cell temperature: 40 C)
and column thermostat (temperature: 80 C). The
sample injection volume was 100 lL, and the sample

123

Cellulose

concentration 0.1%. The column used was PLgel

5 lm MIXED C 7.5 9 300 mm. The eluent (1%
LiCl/DMAc), filtered through the 0.45 lm polyamide
membrane filter (Supelco), was pumped into the
system at a flow rate of 0.5 cm3 min-1. The
chromatographic data were processed with PSS
(Polymer Standards Service) WinGPC Unity
software.
Pullulan standards (Polymer Laboratories) with
peak molecular masses of 1,660,000, 788,000,
404,000, 212,000, 22,800, 5,900 and 667 g mol-1
were used for calibration. The standards with molecular masses of 1,660,000, 788,000 and 404,000
g mol-1 were prepared in individual 0.1% solutions,
while pullulan standards with molecular masses of
212,000, 22,800, 5,900 and 667 g mol-1, were prepared as mixed standards in a solution containing
0.025% of each standard. The standards were weighed,
transferred into 2 and 5-cm3 volumetric flasks and
dissolved in DMAc. Finally, appropriate volumes of
8% LiCl in DMAc were added in order to achieve 1%
solution of LiCl (Strlic and Kolar 2003; Strlic et al.
2002).

Results and discussion

FT-IR analysis
FT-IR analyses of the residues that were obtained
after 4 h of MC, WH and CT liquefaction, were

performed to ensure that residual cellulose was being

dealt with and not insoluble cellulose derivatives. All
of the residues that were analyzed exhibited the
characteristic bands of cellulose: 3,4193,342 cm-1
(OH stretching), 2,901 cm-1 (CH stretching),
1,430 cm-1 (CH2 bending), 1,3711,373 cm-1 (CH
bending), 1,0601,031 cm-1 (CO stretching). The
obtained results suggested the presence of residual,
not derivatized, cellulose. Since all of the FT-IR
spectra were very similar, that of the residue from
the WH cellulose is given as an example (Fig. 2).
Cellulose liquefaction reaction in acid-catalyzed
E.G
Cellulose liquefaction in an acidic, non-aqueous E.G
medium seemed to be analogous to hydrolysis. The
effects of H2O cannot be completely excluded from
the system as H2O may be preferentially adsorbed by
the cellulose (Valley 1955). Accordingly, it was taken
into account that approximately 5% (w/w based on
cellulose) of H2O is adsorbed by cellulose and
additional 3% (w/w based on H2SO4) or 9.4% (w/w
based on PTSA) of H2O is provided by catalysts used
for the liquefaction. Therefore, it can be assumed that
the primary reaction is hydrolysis, followed by
glycosidation of the new reducing groups. Cellulose
degradation in acidified E.G begins with glycosidic
oxygen protonation, followed by carbonium ion
formation and scission of the glycosidic bond.
After the glycosidic bond is broken, cellulose

b
1,0

0,8

%T (normalized)

%T (normalized)

1,0

1
0,6

0,4

0,2

0,0
4000

0,8

1
0,6

0,4

0,2

3500

3000

2500

2000
-1

cm

1500

1000

500

0,0
4000

3500

3000

2500

2000

1500

1000

500

-1

cm

Fig. 2 a FT-IR spectrum of the WH (1), FT-IR spectrum of the residual WH after 240 min of degradation under catalysis of PTSA
(2). b FT-IR spectrum of the residual WH after 240 min of degradation under catalysis of H2SO4 (3)

123

Cellulose

Cellulose weight loss

depolymerization could occur at both the reducing

(O1H) and the non-reducing (O4H) chain ends.
Reducing chain ends are especially reactive due to
the stabilized proton in the carbonium ion (I), this
carbonium ion, initially being attacked by E.G to give
the 2-hydroxyethyl-D-glucopyranoside and the reformation of the 2-hydroxyethyl oxonium ion (II)
(Scheme 1).
Due to a higher carbonium ion (I) than the
carbonium ion (Ia) reactivity with E.G, the reaction
between the non-reducing ends of the chain and
E.G is negligible. The structure of 2-hydroxyethyl-Dglucopyranoside was confirmed by 13C NMR (Fig. 3;
Table 1). Thus, it can be concluded that cellulose
degradation by acid catalyzed glycolysis occurs at the
reducing end of the chains.

H OH

H OH
H O

HO

H O
O

+
+ H3O

HO

Figure 4 shows the percentage of residual celluloses

as a function of the reaction time. With the cellulose
samples, catalyzed by H2SO4, *50% of the initial
celluloses was liquefied and converted into the
soluble products during the initial 30 min of
treatment.
Cellulose liquefaction that was catalyzed by PTSA
proceeded significantly slower. Only 2240% of the
initial cellulose was liquefied in the first 30 min. The
celluloses with molar masses of 76,000, 699,000 and
1,910,000 g mol-1 under H2SO4 catalysis, were
converted to soluble products in extents up to 98.9,
95 and 85%, respectively. In the case of PTSA
catalysis, the cellulose series representatives were

OH

HO
H

OH

OH
n

H OH

H OH
H

HO

+
O

OH

O
H

HO

HO
H

OH

OH
n

OH

H OH

H OH

H O

HO
+

HO
HO

HO

OH

OH

OH
H

+
O
H

H
H

O
H

HO
H

OH

+
H

O
H

HO
H

OH

OH

OH
HO

H OH

OH

OH

Ia

n-2

HO

OH
+
H2O

H O
+

HO
HO
H
H

H O
H

HO

OH2

HO
OH

OH

OH

OH

OH

OH

HO

HO

H OH

H
H O

HO
H
H

+ OH2

HO

OH
OH

H
O

HO

OH

H
+

OH

II

HO
H
H

+ OH2
OH

OH
OH

II

Scheme 1 Cellulose liquefaction reaction scheme in acid-catalyzed E.G

123

Cellulose

OH

H2C
HO

H2
C

HO
H

C
H2

OH

Table 2 Amounts of H2SO4 and PTSA and equivalent

hydronium ion concentrations used in liquefaction of MC, WH
and CT

2'

1'

Catalyst

Amounta (mol)

[H?] (mol/L)

H2SO4

0.045

0.09

PTSA

0.023

0.023

OH

Fig. 3 Structure of 2-hydroxyethyl-D-glucopyranoside

4.5 g of each catalyst used for the liquefaction, that made 3%

w/w based on E.G

Table 1 13C NMR shifts of 2-hydroxyethyl-D-glucopyranoside in dimethylsulfoxided6

d (ppm)
C10

C20

C1

C2

C3

C4

C5

C6

69.09

60.11

98.73

72.18

72.54

70.24

73.45

60.89

Carbon atom positions as indicated in Fig. 3

liquefied in extents up to 98.7, 85 and 72%. A greater

rate of cellulose weight loss is observed under
catalysis of strong mineral acid such as in the
E.G-H2SO4 system. Figure 4 shows that the weight
loss of MC under H2SO4 catalysis follows a pseudo
first-order kinetics, while both WH and CT under
H2SO4 catalysis appears to follow a bi-exponential
model, which consists of two parallel reactions: the
fast one corresponding to the depolymerization of
disordered inter-crystalline regions and the slow one
corresponding to the degradation of crystalline
regions. In contrast to the WH and CT weight losses
under H2SO4 catalysis, for MC, WH and CT the
weight loss under PTSA catalysis follows pseudofirst-order kinetics. The bi-exponential model in the

case of WH and CT depolymerization under H2SO4

catalysis is a result of the greater hydronium ion
concentration. The dissociation of 1 mol of PTSA
provides 1 mol of hydronium ion that is involved in
glycosidic oxygen protonation followed by carbonium
ion formation (Scheme 1), while after dissociation of
1 mol of H2SO4, consequently 2 mol of hydronium ion
are provided for the further reaction. Accordingly, the
dissociation of 0.045 mol of H2SO4 present in the
liquefaction mixture depolymerized WH and CT
amorphous regions with the higher rate than the
dissociation of 0.023 mol of PTSA (Table 2).
Consequently, the amorphous cellulose regions
present in WH and CT samples, were depolymerized
with the twice higher rate under H2SO4 catalysis than
in the case of PTSA. When the hydronium ions
started to interfere with the accessible reducing ends
in the crystalline WH and CT cellulose regions, the
rate of the weight loss became dependent only on
the amount of the accessible reducing ends in the
crystalline cellulose regions and not on the hydronium ion concentration. The weight loss of the MC

100

Residual cellulose (%)

Residual cellulose (%)

100

10

MC + H2SO4
WH + H2SO4
CT + H2SO4

50

10

MC + PTSA
WH + PTSA
CT + PTSA

100

150

Time, min

200

250

50

100

Fig. 4 Percentage of weight loss (Eq. 1) during the acid-catalyzed glycolysis of cellulose, with time

123

150

Time, min

200

250

Cellulose

699,000 g mol-1) and of residual celluloses from this

sample, taken at 1 min and 240 min of liquefaction in
the E.G-H2SO4 system and in the E.G-PTSA system,
are shown in the Fig. 5.
Compared with the chromatogram of the initial
WH sample, the chromatogram of the WH, taken
after 1 min of liquefaction, shows that the main peak
with narrow weight distribution is shifted to a lower
molecular weight. The increased intensity of the main
peak and the appearance of a low-molecular weight
shoulder can be interpreted in the terms of the
simultaneous hydrolysis and crystallization of the
amorphous fraction in the microfibrils (Wood et al.
1989). After 1 min of the liquefaction of WH, the
newly formed cellulose crystallites were composed
from approximately 12 glucose units. About 240 min
of the treatment resulted in a loss of the low molar
mass shoulder. This loss was due to the fast
accumulation and slow degradation of the crystallites,
as indicated by the increase of the main peak
intensity, observed in the chromatogram of the WH
sample, taken after 240 min.
During the first minute of cellulose liquefaction in
E.G, the initial polymerization degree of WH and CT,
decreased to 261 and 308 under catalysis by H2SO4
and to 336 and 342 under catalysis by PTSA,
respectively (Table 3).
Due to the sharp initial drop of DPw , the assumption could be made that cellulose degradation occurs
predominantly at chain centers. However, due to

sample differed from the WH and CT samples.

Presumably, this was the reason why the rate of the
weight loss of the highly crystalline MC sample in
both cases, under H2SO4 or PTSA catalysis, did not
depend on the hydronium ion concentration and
followed the first-order kinetics. Thus, the greater
hydronium ion concentration is a determinant factor
for WH and CT weight loss to follow bi-exponential
model under H2SO4 catalysis.
Molecular weight distribution and crystallinity
change
The consequences of the cellulose liquefaction, in
E.G under catalysis by H2SO4 or PTSA were
monitored using size-exclusion chromatography
(SEC). Molar mass averages (M W ) of the residual
cellulose samples, taken at different time intervals,
were determined by SEC, relative to pullulan
standards.
Molecular weight distributions obtained by sizeexclusion chromatography enable one to monitor the
cellulose degradation process that occurs by acidcatalyzed glycolysis. The SEC chromatograms of the
residual cellulose samples, demonstrated the presence
of an analogous degradation pathway for WH (M W
699,000 g mol-1) and for CT (M W 1,910,000
g mol-1), while MC (M W 76,000 g mol-1) behaves
differently. As an example, chromatograms (normalized to the sample weight) of WH (M W

25

25

240 min

20

15

RI detector responce, a.u.

RI detector responce, a.u.

240 min

1 min
0 min

10

1 min

20

15

0 min

10

log M

log M

Fig. 5 SEC chromatograms of residual cellulose (M W 699,000 g mol-1) from the samples, taken at 0, 1 and 240 min of liquefaction
catalyzed by H2SO4 (a), PTSA (b)

123

Cellulose
Table 3 Average polymerization degree DPw of residual
cellulose samples with time of reaction with H2SO4 and PTSA
as catalysts
Time
(min)

H2SO4

PTSA

MC

WH

CT

MC

WH

CT

470

4,320

264

261

11,790

470

4,320

11,790

308

330

336

256

342

267

305

275

279

306

5
15

230

264

291

254

278

299

186

253

285

211

278

283

60

174

250

274

167

271

270

120

162

242

265

146

276

262

240

149

239

246

143

272

264

increased crystallinity of WH and CT after 5 min of

liquefaction (Table 3), the initial drop in the cellulose
molar mass has to be considered in terms of the
primary breakdown of covalent bonds and glycosidic
linkages in the amorphous cellulose regions together
with breakdown of inter-chain hydrogen bonds and
intra-chain hydrogen bonds. The glycosidic linkages
in the less ordered cellulose regions, at 150 C in the
acid-catalyzed E.G system, undergo rapid scission,
forming new reducing end-groups that immediately
give rise to the end-wise degradation at the ends of
the remaining inaccessible (crystalline) regions
(Sharples 1957; Table 4).
During the first minute of MC liquefaction, the
initial DP of 470, decreased to 264 under catalysis
with H2SO4 and to 330 under catalysis with PTSA
(Table 2). The degradation of the MC in acidified
E.G proceeds differently. This can be deduced from
the SEC chromatograms of the MC initial sample

Table 4 Crystallinity (%) of residual MC, WH and CT cellulose with time

Time
(min)

H2SO4
MC

WH

CT

MC

WH

CT

83.26

78.19

79.77

83.26

78.19

79.77

87.76

90.23

92.74

87.52

92.64

91.91

60

86.54

92.28

91.88

86.58

92.28

91.96

120

83.93

89.30

91.00

86.53

90.87

91.96

240

79.54

74.23

90.16

85.05

71.30

90.50

123

PTSA

where it can be seen that the shoulder at the low

molecular-weight is already present, meaning that
WH and CT behave similar to MC once the less
ordered regions have been hydrolyzed (Fig. 6). After
1 min of the MC liquefaction, the cellulose crystallites that gave rise to the low molecular-weight
shoulder, tended rapidly to accumulate and subsequently to increase the intensity of the main peak in
the chromatogram of MC sample that had been
subjected to 240 min of treatment.
During the next 4 h, cellulose liquefaction in the
acid-catalyzed E.G systems proceeds at a significantly slower rate as the ordered (crystalline) regions
began to degrade. This period represents the random
glycolysis of the glycosidic bonds in the accessible
regions of the polysaccharide (Nelson and Tripp
1953). Only small amounts of the residual celluloses,
with relatively high DP values from 272 to 143 were
obtained. Similar behaviour of crystallite degradation
during strong acid hydrolysis has been demonstrated
by several authors (Nelson and Tripp 1953; Bouchard
et al. 1992), where some chains in the crystalline
regions degrade completely and become soluble
causing great weight loss, while others remain quite
untouched, maintaining the high DP values of the
insoluble residue, where the LODP does not significantly change over longer times. In addition, after
the LODP is reached, due to the slow E.G penetration
into the crystalline cellulose regions, small amounts
of soluble fragments could be produced, causing a
continuous weight loss in residual cellulose.
Cellulose degradation during acid-catalyzed
liquefaction
A plot of the number of scissions per cellulose chain
(DP0/DP - 1) against time (Fig. 7) was made in
order (Calvini et al. 2008) to minimize errors due to
the polydispersity ratios. It indicates that MC cellulose suffered approximately two scissions per chain,
while WH and CT experienced 16 and 45 scissions,
respectively. MC degraded more slowly, as expected.
Microcrystalline cellulose (MC) powder is produced by the acid reflux hydrolysis of wood, where
amorphous regions of the cellulose fibers are hydrolyzed, leaving a very highly crystalline residue with
its leveling-off degree of polymerization (Battista
1971). Thus, the depolymerization of MC in acidified
E.G should represent the breakdown pathway of

RI detector responce, a.u.

25

20

240 min

1 min

15

0 min
10

25

RI detector responce, a.u.

Cellulose

20

240 min

1 min
0 min

15

10

0
3

log M

log M

Fig. 6 SEC chromatograms of the residual cellulose (M W 76,000 g mol-1) from the samples, taken at 0, 1 and 240 min of
liquefaction, catalyzed by H2SO4 (a) and PTSA (b)

1,5

1,0
0,5

12
8
4

W H + H 2SO 4

MC + H 2SO 4
MC + PTSA

0,0
0

50

100

150

200

250

40
30

DP /DP - 1

50

16

2,0

DP /DP - 1

2,5

DP /DP - 1

W H + PTSA

0
0

50

Time, min

100

150

200

250

Time, min

20
10

CT + H2SO4
CT + PTSA

0
0

50

100

150

200

250

Time, min

Fig. 7 Scissions per chain (DP0/DP - 1), with time during depolymerization in acidified E.G; MC (a), WH (b) and CT (c)

crystalline regions only. In the current study, after

initial 1 min treatment a slightly increased crystallinity, crystallite width and significant decrease in DP
were observed. Taking into account that in MC some
of the surface cellulose material may be disordered,
such MC could be used as a reference to study
degradation of crystalline cellulose regions.
Zou et al. (1994) have suggested that the ratio
DPz/DPw is a good guide to the homogeneity of the
degradation process of Whatman filter paper and
cotton linters. The DPz/DPw ratios and DPw associated with the current study are presented in Fig. 8.
The ratios, after the initial 1 min treatment, remain
fairly constant, indicating that the degradation proceeded gradually by attack by the E.G at the
accessible cellulose reducing ends, presumably on
the crystallite surface.

Rate of cellulose depolymerization

The rate of cellulose depolymerization was calculated
using the Ekenstam equation (Ekenstam 1936):
1=DPt  1=DP0 kt

Here, k reaction rate constant, DPt and DP0 are the

polymerization degrees at times t and 0, respectively.
Very high rates of the depolymerization and strong
deviations from linearity in plots were observed in
the current study. The rates of glucoside bond
breakage during the first 5 min of MC depolymerization were determined to be 7.90 9 10-4 and
9.67 9 10-4 min-1 in E.G-H2SO4 and E.G-PTSA
systems, respectively. The beginning of the WH and
CT depolymerization was especially fast. Therefore,
due to the limitation of the Ekenstam equation, the

123

Cellulose

b 350

4,0

4,0
3,5

3,5

2,5

DPw

250

2,0
200

3,0

3,0

2,5

250

DPw

MC + H2SO4

MC + PTSA

300

DPz/DPw

2,0

200

1,5

1,5
1,0

1,0

150

150

0,5

0,5
100

100

0,0

10

0,0

100

340

4,0

340

4,0

3,5
3,0
2,5

DPw

300

WH + PTSA

320

2,0
280

2,5

300

2,0

280

1,5

1,5

1,0

260

1,0

260

0,5

0,5

240

240

0,0

10

0,0

100

Time, min

10

100

Time, min

4,0
340

4,0

340

3,5

3,5

CT + H2SO4

CT + PTSA
3,0

2,0
1,5

280

2,5

DPw

2,5
300

3,0

320

DPz/DPw

320

DPw

3,5
3,0

DPw

WH + H2SO 4

DPz/DPw

320

100

Time, min

Time, min

10

DPz/DPw

300

DPz/DPw

350

300
2,0
1,5

280

1,0
260

DPz/DPw

1,0

260
0,5

240

0,0

10

1 00

Time, min

0,5

240

0,0

10

1 00

Time, min

Fig. 8 The change of the DPw (filled squares) and DPz/DPw

(squares) of MC, WH and CT residual cellulose samples with
time of reaction with H2SO4 and PTSA as catalysts;

MC ? H2SO4 (a), MC ? PTSA (b), WH ? H2SO4 (c),

WH ? PTSA (d), CT ? H2SO4 (e) and CT ? PTSA (f)

rates of glucoside bond breakage were determined

only for the first minute of depolymerization. The
WH depolymerization during the first minute proceeded with the rates of 11.46 9 10-3 and
9.23 9 10-3 min-1 in the E.G-H2SO4 and E.G-

PTSA systems, respectively. The initial rates of

glucosidic bond breakage in CT were determined to
be 10.41 9 10-3 and 9.68 9 10-3 min-1 and in
E.G-H2SO4 and E.G-PTSA systems, respectively.
From these results, it is not possible to determine

123

Cellulose

weather or not the initial cellulose depolymerization

is random or organised. After the amorphous cellulose regions have been hydrolyzed, one can analyse
the slower depolymerization of cellulose crystallites.
Cellulose crystallite degradation during
acid-catalyzed liquefaction
In order better to understand the degradation that
occurs in the MC, WH and CT crystalline regions and
to determine the influence of simultaneous hydrolysis
and crystallization of amorphous fraction in the
microfibrils (Wood et al. 1989) the average widths
of crystallites were evaluated using the Scherrer
equation (Klug and Alexander 1974).
MC crystallites during the initial 5 min of treatment tended to accumulate (crystallized) by slightly
increasing crystallite widths from 5.1 to 5.2 nm and
from 5.1 to 5.3 nm in E.G-H2SO4 and E.G-PTSA
system, respectively (Table 5). The accumulation of
MC crystallites can be also interpreted from the SEC
chromatograms in Fig. 6a. Here, the low-molecular
weight shoulder, which is observed for the beginning
of the reaction, disappears with time.
The crystallization resulted either from the formation of new crystallites or by crystallization on the
surface of existing crystallites. Since in MC there is
only a small amount of the disordered surface
material, it is rapidly hydrolyzed and consequently,
due to crystallization a slight increase in crystallites
width is observed. After the crystallization of amorphous regions is over, the crystallites are exposed to
the degradation, thereafter at the end of the treatment the width of MC crystallites was reduced to
4.65.0 nm in the presence of H2SO4 and PTSA as
catalysts, respectively. Considering the results
Table 5 Width changes of the crystallites (nm) of residual
MC, WH and CT cellulose with treatment time, evaluated
using Scherrer equation
Time
(min)

H2SO4

PTSA

MC

WH

CT

MC

WH

CT

5.1

6.8

6.4

5.1

6.8

6.4

5.2

6.6

6.9

5.3

7.1

6.7

60

5.1

6.9

6.7

5.1

6.9

6.7

120

4.8

6.7

6.7

5.2

6.9

6.6

240

4.6

5.9

6.6

5.0

6.1

6.6

obtained, one may claim that due to the greater

hydronium ion concentration, H2SO4 degrades MC
crystallites more extensively than PTSA. The more
extensive degradation of MC crystalline regions was
confirmed by obtained reduced degrees of crystallinity from 83.26 to 79.54% and from 83.26 to 85.05%
in the presence of H2SO4 and PTSA as catalysts,
respectively.
The WH crystallite degradation during the initial
minute of the liquefaction in both of the E.G-H2SO4
and E.G-PTSA systems, suffered simultaneous
hydrolysis and crystallization of the amorphous
fraction in the microfibrils in the same manner as
the MC. In the first 60 min, the width of the residual
WH crystallites increased from 6.8 to 6.9 nm,
simultaneously increasing the crystallinity of WH
residual sample to 92.28%. Therefore, one may claim
that the accumulation of the crystallites during the
first hour proceeded faster, than the crystallite
degradation. On completion of the accumulation of
new WH crystallites, the degradation of remaining
crystallites was induced. During the final 2 h of WH
liquefaction, the crystallinity decreased by about
1520% analogously in both E.G-H2SO4 and in
E.G-PTSA systems. In the mean time, due to induced
crystallite degradation, the width of the crystallites
decreased from 6.9 to 5.9 nm and from 6.9 to 6.1 nm,
respectively. According to the obtained results, the
more extensive degradation of WH crystallites using
H2SO4 as a catalyst due to the greater hydronium ion
concentration was observed.
The degradation of CT in acidified E.G proceeded
relatively in the same manner as that of the WH
degradation. The amorphous regions in CT were
hydrolyzed during the first 5 min of CT liquefaction
and, consequently, the greatest degrees of crystallinity were achieved. The increased widths of
crystallites, confirmed the initial crystallization and
accumulation of the amorphous fractions as well as
during the degradation of both the MC and the WH.
After the accumulation of crystallized amorphous
fraction was over, the degradation of remaining
crystallites was induced, as confirmed by the reduction of crystallite width from 6.9 to 6.7 nm and from
6.9 to 6.6 nm, in the E.G-H2SO4 system and in the
E.G-PTSA system, respectively.
By comparing MC, WH and CT crystallites width
at the end of the liquefaction, the MC crystallites
were observed to be the smallest in size with the

123

Cellulose

largest reduction of width (4.65.0 nm) after degradation in E.G-H2SO4 and in E.G-PTSA systems,
respectively. The more extensive MC crystallites
degradation is a result of more reducing ends
available for the reaction than in the case of WH
and CT. Thereof, the assumption could be made that
the crystallite degradation occurs on the surface of
the crystallites by attacking accessible MC, WH and
CT reducing ends by E.G.
The cellulose degradation during acid-catalyzed
liquefaction was evaluated by plotting the number of
scissions per cellulose chain (DP0/DP - 1) against
time (Fig. 7). Figure 7 shows that MC cellulose
suffered approximately 2 scissions per chain, while
WH and CT suffered 16 and 45 scissions per chain,
respectively. Considering that the CT suffered much
more scissions per chain than WH and MC, it would
be logical to expect the lowest residual amounts. The
residual MC, WH and CT amounts as well as the
width of crystallites differed also due to the number
of reducing ends available for the acid-catalyzed
glycolysis. Since the number of reducing ends is
proportional to reciprocal DP, the residual WH and
CT consequently contains less reducing ends than
MC, that can also explain their slower weight loss
and formation of wider crystallites. However, due to
the smallest number of reducing ends, the greatest
amounts of residual cellulose and the largest crystallites were obtained after CT liquefaction in both the
E.G-H2SO4 and in the E.G-PTSA systems. In addition, the amorphous cellulose in CT was hydrolyzed
during the initial period, whereas some was immediately converted into the soluble fragments, some
crystallized on the surface of the crystallites, consequently increasing their average width. In comparison
to the MC and WH crystallites, due to the most
increased average width of the CT crystallites, the
smallest total surface area was exposed to the E.G
attack.

Conclusions
Microcrystalline cellulose, Whatman filter paper no.
1 and cotton linters with molar masses of 76,000,
699,000 and 1,910,000 g mol-1, respectively were
liquefied at 150 C in the presence of ethylene glycol
and under catalysis of sulphuric acid or p-toluene
sulfonic acid monohydrate. After four hours the

123

achieved yield of cellulose liquefaction was

7298.9%. The lower molar mass cellulose chains,
with higher number of reducing ends, were liquefied
and consequently lesser amounts of residual cellulose
were obtained.
From the gravimetric determination of cellulose
residue, we have concluded that the molar mass of the
initial cellulose and the physical structure are important parameters in cellulose liquefaction, under acidcatalyzed glycolysis. It was determined that the rate
of cellulose weight loss depends on hydronium ion
concentration and on the number of reducing ends.
The cellulose degradation was monitored using
size-exclusion chromatography. The observed
changes in the molecular weight distribution of the
cellulose samples (Microcrystalline cellulose,
Whatman filter paper no. 1, cotton linters) followed
the same degradation trend, independent of the
starting molar mass and physical structure. The
combination of the high temperature, glycol concentration and the amount of catalysts caused
particularly rapid depolymerization of the less
ordered cellulose regions. In addition, partial degradation of crystalline cellulose regions was achieved.
The molecular weight distributions and the change of
crystallites width with time showed that the degradation of cellulose crystallites proceeded at the
accessible cellulose reducing ends on the surface of
the cellulose crystallites.
Acknowledgments The authors gratefully acknowledge
financial support of the Ministry of Higher Education,
Science and Technology of the Republic of Slovenia and
Slovenian Research Agency (programme P2-0145).

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