Industrial Crops and Products 44 (2013) 542–548

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Industrial Crops and Products

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Fractionation and characterization of physicochemical and structural features of

corn stalk hemicelluloses from yellow liquor of active oxygen cooking
Jian-Bin Shi a,∗ , Qiu-Lin Yang a , Lu Lin b,∗ , Lin-Cai Peng a
a
State Key Laboratory of Pulp and Paper Engineering, South China University of Technology, Guangzhou 510640, Guangdong Province, China
b
School of Energy Research, Xiamen University, Xiamen 361005, Fujian Province, China

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

Article history: The hemicellulosic fractions were obtained by graded ethanol precipitation from the yellow liquor of
Received 20 July 2012 active oxygen cooking. The chemical composition, structural features, and thermal stability were inves-
Received in revised form tigated by a combination of HPAEC, GPC, FT-IR spectra, NMR spectra and TG. Sugar analysis suggested
20 September 2012
that the xylose was primary sugar unit in the hemicellulosic fractions and the final residue contained
Accepted 22 September 2012
high amounts of uronic acids. The GPC results showed that only 5.31% hemicelluloses in the liquor
showed high molecular weight (24,824 g/mol), the remainder of the hemicellulosic fractions were signifi-
Keywords:
cantly degraded during the cooking process and presented low molecular weight (2020–4574 g/mol). The
Hemicelluloses
Corn stalk
corn stalk hemicellulosic fractions were composed of (1 → 4)-ˇ-d-xylopyranosyl backbones, but the side
Yellow liquor chains attached to the xylopyranosyl units were diverse, depending on the precipitation process. In addi-
Active oxygen tion, hemicelluloses with infrequent side chains were more thermally stable and could be precipitated
Degradation with a low concentration of ethanol.
© 2012 Elsevier B.V. All rights reserved.

1. Introduction Pulping is one of conventional methods of pretreatment pro-

The depletion of fossil fuel and the concern of environmental
protection have prompted the research for alternative and sustain-
able energy resources from the lingo-cellulosic biomass. The source
of biomass can be divided into two domains (Jeguirim and Trouve,
2009). The first domain is the growing of energy crops to gener-
ate electricity and produce biofuel. The second consists on the use
of organic fraction of municipal wastes as well as residual mate-
rial from forestry to agriculture, such as wood, straw. Since 1970s,
biomass crops have attracted increasing interest in energy supplied
because they can satisfy a relevant part of the energy demand and at
same time reduce CO2 emission (Peng et al., 2012). Corn stalk is an
agriculture residue and has a huge potential to biomass conversion
due to the high production. It has been partly used for producing
useful chemicals in China, but the scale is small, and the techniques
can cause serious environmental pollution (Pang et al., 2012).
Abbreviations: X, (1 → 4)-ˇ-d-Xylp; XU, (1 → 4)-ˇ-d-Xylp-2-O-(4-O-Me-␣-d-
GlcpA); A, ␣-l-Araf; U, 4-O-Me-␣-d-GlcpA; G, (1 → 4)-ˇ-d-glucp; ax, axial; eq,
equatorial; Xyl/Ara, the molar ratio of xylose to arabinose; Xyl/Ura, the molar ratio
of xylose to uronic acids; Nd, not detectable; o.d., oven dry weight.
∗ Corresponding authors at: State Key Laboratory of Pulp and Paper Engineering,
South China University of Technology, Guangzhou 510640, Guangdong Province,
China. Tel.: +86 020 22236719; fax: +86 020 22236719.
E-mail addresses: shijianbin1022@126.com (J.-B. Shi), lulin@xmu.edu.cn (L. Lin).
cess of biomass conversion. But pulping and paper production has
long been recognized as a significant point source of pollution.
Since the development of modern pulp mills, the reduction of efflu-
ents has been one of the main target in improving this technology
(López et al., 2001). The environmental pressures have confronted
the pulping and paper industry. This industry has responded by
developing pulp bleaching process that use non-chlorinated chem-
icals such as oxygen, hydrogen peroxide, and ozone (López et al.,
2001; Gamelas et al., 2005; Zhang et al., 2005). The conventional
cooking process, such as, sod and kraft cooking, has been modi-
fied (Huang et al., 2008; González-García et al., 2010; Garcia et al.,
2011; Loacutepez et al., 2011). But such approaches still cannot
fundamentally solve the pollution caused by the cooking chemicals.
In an effort to reduce the pollution, an active oxygen cooking
process involving solid alkali from corn stalks was developed by
our group (Shi et al., 2012; Yang et al., 2012), in which clean active
oxygen (O2 and H2 O2 ) and solid alkali (MgO) were added as cooking
chemicals. The O2 and H2 O2 have been used in the total chloride
free (TCF) bleaching system due to high activity to the residual
lignin in pulp (Salmela et al., 2008; Egüés et al., 2012). Many types
of solid alkali also have been applied in trans-esterification reac-
tions, biodiesel, and other industries (Cantrell et al., 2005; Ebiura
et al., 2005; Xie et al., 2006; Villaverde et al., 2009; Sun et al.,
2010). But there is no published paper about the cooking pro-
cess involving active oxygen and solid alkali as a pretreatment of
biomass conversion. In the active oxygen cooking process, active

0926-6690/$ – see front matter © 2012 Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.indcrop.2012.09.026
 J.-B. Shi et al. / Industrial Crops and Products 44 (2013) 542–548
oxygen was used to remove the lignin, and solid alkali was applied
for protecting the carbohydrate from carbonizing and hydrolyzing.
Comparison with conventional cooking processes, the pollution
caused by the cooking chemicals can be completely eliminated in
the active oxygen cooking process. In practice, no frowzy gas and
effluent containing high consistency alkali were discarded to the
environment after cooking. Moreover, the solid alkali, hemicellu-
loses, and lignin could be recycled from weak alkaline (pH ca. 8.0)
yellow liquor. After cooking, it was found that 85.5% lignin and
73.4% hemicelluloses were removed from the raw material, and the
yield of the pulp was 50.2% (Shi et al., 2012; Yang et al., 2012). The
above facts showed that the active oxygen cooking was an efficient
and clean pretreatment process of biomass conversion.
Hemicelluloses are the most abundant hetero-polysaccharide
in the corn stalk cell wall and composed of various sugar units and
uronic acids (Jin et al., 2009). And they were degraded and dissolved
in the coking liquor (called yellow liquor after cooking) during
the cooking process. To clarify the physicochemical characteriza-
tions of hemicelluloses in yellow liquor and further understand
cooking mechanism, the hemicelluloses were precipitated with dif-
ferent concentration ethanol and characterized by a combination
of destructive and nondestructive methods.
2. Materials and methods
2.1. Materials
Corn stalk was supplied by China BBCA Group Corp., China.
It was first dried in the sunlight and cut into small pieces (less
than 10 cm). The main components of corn stalk were determined
according to the literature (Qi et al., 2006) and Tappi standards T
204 cm-97, T211-02 (Tappi, 2002). It was found that the component
(w/w) of corn stalk was cellulose 39.12%, hemicelluloses 23.96%,
lignin 21.73%, ash 7.95%, and benzene/ethanol extraction 4.22% on
oven dry weight (o.d.). All standard chemicals were purchased from
Sigma Chemical Co. (Beijing). Other chemicals used were analytical
grade.
2.2. Methods
2.2.1. The cooking operations
The cooking operations were described in the our previous study
(Pang et al., 2012; Shi et al., 2012; Yang et al., 2012), and the detailed
operations were given as follow: 50 g corn stalk (o.d.), 7.5 g MgO and
3% wt dosage of H2 O2 (based on the o.d. of corn stalk) were placed
in a 2 L stainless, rotating autoclave at a solid-to-liquor ratio of 1:6
(g/mL). After being sealed, the autoclave was filled with1.0 MPa O2 .
Cooking was performed at 165 ◦ C for 2 h. After cooking, the yellow
liquor was extruded from the raw stock and stored in a refrigerator,
and the resulting pulp was washed with the deionized water and
then dried in air.
2.2.2. Fractionation of hemicelluloses by the graded ethanol
The scheme for fractionation of hemicelluloses by graded
ethanol precipitation from the yellow liquor is illustrated in Fig. 1.
The pH of yellow liquor (185 mL) was adjusted to 1.5–2.0 with
HCl in a beaker to precipitate the lignin. The resulting liquor was
centrifuged at 5000 rpm for 10 min. The supernatant was neutral-
ized to pH 5.5 with NaOH, then poured it into one volume of 95%
ethanol. The precipitation were centrifuged at 5000 rpm for 10 min
and freeze-dried. And the supernatant was concentrated to 100, 50,
70, 30 mL at vacuum, and poured into two, three, five, seven vol-
umes of 95% ethanol, respectively. The rest of process occurred as
described above without adjusting the pH. The final supernatant
was concentrated to 20 mL and directly freeze-dried. The hemi-
cellulosic fractions precipitated with one, two, three, five, seven
543
Yellow liquor 185 ml
Adjustion pH to 1.5-2.0 with HCl
Supernatant Lignin
Neutralized to pH 5.5 with NaOH, and
precipitation in one volumes 95 %
Freeze-dried
Supernatant Pellet C1
Concentration at reduced pressure to 100 ml,
and precipitation in two volumes 95 % ethanol
Freeze-dried
Supernatant Pellet C2
Concentration at reduced pressure to 70 ml, and
precipitation in three volumes 95 % ethanol
Freeze-dried
Supernatant Pellet C3
Concentration at reduced pressure to 50 ml, and
precipitation in five volumes 95 % ethanol
Freeze-dried
Supernatant Pellet C5
Concentration at reduced pressure to 30 ml, and
precipitation in seven volumes 95 % ethanol
Freeze-dried
Supernatant Pellet C7
Concentration at reduced pressure to 20 ml,
freeze-dried
RC
Fig. 1. The scheme for fractionation of hemicelluloses by graded ethanol precipita-
tion from the yellow liquor.
volumes ethanol were labeled as C1 , C2 , C3 , C5 , C7 , respectively; the
final residue was named RC .
2.2.3. Physiochemical characterization of the hemicelluloses
2.2.3.1. Sugar composition analysis. The monosaccharides and the
uronic acids in the hemicelluloses were liberated by hydrolyz-
ing approximate 10 mg sample using 5.5 mL of 6.5% H2 SO4 for
2.5 h at 105 ◦ C. The hydrolyzates were diluted to 50-fold (after
adjusted the pH value to 7.0), filtered, and injected into the high-
performance anion-exchange chromatography (HPAEC) (Dionex
ICS-3000, USA) with pulsed amperometric detection, a CarboPacTM
PA20 column (3 mm × 150 mm) and a CarboPacTM Guard column
(3 mm × 30 mm). The operations were run at 30 ◦ C for 30 min, and
the flow rate during the analysis process was 0.5 mL/min. When
eluting the monosaccharide, the eluent was 2 mM NaOH. However,
due to the weak eluting power of the NaOH, the eluent was 100 mM
NaAc and 2 mM NaOH when eluting the uronic acids.
2.2.3.2. Molecular weight analysis. The molecular weights of the
hemicelluloses were determined by gel permeation chromatogra-
phy (GPC). The GPC system comprises a Waters 1525 binary HPLC
pump, a Waters 717 plus Auto-sampler, a Waters 2414 refractive
index detector, and a Breeze (V3.3) GPC work station (Waters, USA).
The samples were dissolved in the eluent and injected into the
TSk-GELG-5000PW xL column (7.8 mm × 300 mm) and TSk-GELG-
3000 PW xL column (7.8 mm × 300 mm) (TOSOH, Japan) in a series.
The eluent flow (0.02 M pH 6.0 KH2 PO4 ) was 0.6 mL/min. The GPC
544 J.-B. Shi et al. / Industrial Crops and Products 44 (2013) 542–548

Table 1 Table 2
Yield of hemicelluloses fractions (%, relative to the total amount of hemicelluloses The content (%) of neutral sugars and uronic acids in the hemeicelluosic fractions
obtained from the yellow liquor) from the yellow liquor. and final residue.

Hemicelluloses C1 C2 C3 C5 C7 RC Sugar Hemicellulosic fractions (%)

Yield/% 5.31 15.63 33.66 9.45 6.34 29.61 C1 C2 C3 C5 C7 RC

Arabinose 14.82 12.01 11.35 12.83 14.63 7.72

Galactose 8.60 5.36 6.14 5.37 4.14 1.97
columns and injection system were maintained at 35 ◦ C
during the Glucose 8.26 9.82 13.99 13.37 15.04 26.26
analysis. Glucan was applied as a reference substance. Xylose 57.92 68.44 64.54 50.76 39.48 19.35
Mannose 3.05 Ndc Nd Nd Nd Nd
Uronic acids 7.35 4.37 3.98 17.68 26.70 44.70
2.2.3.3. Fourier-transform infrared spectroscopy (FT-IR) analysis. FT- Xyl/Araa 3.91 5.70 5.69 3.96 2.70 2.51
IR spectra of the hemicellulosic fractions were obtained on a FT- Xyl/Urab 10.19 20.24 20.97 3.71 1.91 0.56
IR spectrophotometer (Bruker Tensor 27, Germany). The samples a
Xyl/Ara, the molar ratio of xylose to arabinose.
b
were combined with KBr in slices containing 1% hemicelluloses. Xyl/Ura, the molar ratio of xylose to uronic acids.
The scan range was 4000–400 cm−1 in the transmission mode.
c
Nd, not detectable.

2.2.3.4. Nuclear magnetic resonance (NMR) analysis. The 1 H–13 C 2D
hetero-nuclear single quantum coherence (HSQC) spectra of the
hemicelluloses (80 mg in 1 mL D2 O) were obtained on Bruker AV
600 instrument in the HSQC experiment mode at 25 ◦ C. The spectral
widths were 6000 and 25,000 Hz for the 1 H- and 13 C-dimensions,
respectively. The acquired time per scan was 0.0984 s, the relax-
ation delay time was 1.5 s, and the pulse width was 12.1 Hz.
2.2.3.5. Thermo-gravimetric (TG) analysis. The thermo-gravimetric
analysis of the hemicellulosic fractions was performed on the TGA
Q500 (TA, USA) thermal analysis instrument. Approximately 10 mg
hemicelluloses was evenly distributed in the platinum crucible and
heated form the room temperature to 600 ◦ C at a heating rate of
10 ◦ C/min under high purity nitrogen gas flow at 60 mL/min.
3. Results and discussions
3.1. Yield of hemicellulosic fractions
The hemicelluloses in the corn stalk were degraded during the
active oxygen cooking process, and the polymers in the yellow
liquor were a mixture of a number of different degree of polymer-
ization (DP) polysaccharides. The yields of hemicellulosic fractions
obtained from yellow liquor with different concentration ethanol
are listed in Table 1. The precipitates collected with one, two, three,
five, and seven volumes ethanol, yielded 5.31%, 15.63%, 33.66%,
9.45% and 6.34% hemicelluloses, respectively. Obviously, the results
showed that the yields of hemicellulosic fractions increased firstly
(5.31–33.66%) and then decreased (33.66–6.34%) with ethanol con-
centration increasing. The H3 precipitated with three volumes
ethanol presented the highest yield (33.66%). The hemicelluloses
in the yellow liquor were water-soluble. Increase of the ethanol
concentration can precipitate more hemicelluloses, but the hemi-
celluloses in the yellow liquor were less and less, and the yield
decreased even in the high concentration ethanol. However, it
should note that 29.61% solid content was still contained in the
final supernatant, which might contain highly degraded hemicel-
luloses, lignin and cellulose, degradation products of corn stalk, and
magnesium salt.
content of uronic acids (17.68–26.70%), mainly 4-O-methyl-a-d-
glucuronic acid and galacturonic acid, were enriched in the H5
and H7 . Mannose was only detected in H1 (3.05%), suggested
that the hemicelluloses in yellow liquor contained minor of man-
nose. The amounts of glucose came from cellulose increased with
increasing of ethanol concentration, showed the hemicellulosic
fractions obtained in the higher concentration ethanol contained
more degraded cellulose. The sugar analysis demonstrated that
all hemicellulosic fractions might composed of xylopyranosyl
(Xylp) backbones, which was substituted with residues or short
chains contained arabinose, and glucuronic acid or 4-O-methyl-a-
d-glucuronic acid. But the side chains attached to the Xylp were
diverse, depending on the precipitation process. Interestingly, the
molar ratios of xylose to arabinose (Xyl/Ara) (5.70–2.70) and xylose
to uronic acids (Xyl/Ura) (20.24–1.91) were decreased with increas-
ing ethanol concentration from two volumes to seven volumes,
which indicated that the hemicellulosic fractions precipitated with
high concentration of ethanol were more highly branched than
that precipitated with low concentration ethanol. In addition, the
sugar residues in RC mainly contained uronic acids (44.70%), glu-
cose (26.26%) and xylose (19.35%), which indicated that the content
of hemicelluloses was low in the RC .
3.3. The molecular weight of hemicellulosic fractions
In order to estimate the degradation extent of hemicelluloses
during the cooking process, five hemicellulosic fractions and final
residue were further analyzed by GPC. The weight-average (MW )
and number-average (MN ) molecular weight (g/mol), as well as
polydispersity (MW /MN ) are listed in Table 3. Obviously, the MW of
all hemicellulosic fractions and residue decreased with the increase
of ethanol concentration, but the MW of H3 , H5 and H7 was barely
changed. The hemicelluloses in the yellow liquor were water-
soluble, the hemicellulosic fractions of high molecular weight can
be precipitated by low concentration ethanol solution. The GPC
results and yields of hemicellulosic fractions showed that only
5.31% hemicelluloses in the liquor owned high molecular weight
(24,824 g/mol), the remainder of hemicellulosic fractions and final
residue presented low molecular weight (2020–4574 g/mol), which

3.2. Sugar composition and uronic acids content Table 3

The weight-average (MW ) and number-average (MN ) molecular weights (g/mol) and
The neutral sugar composition and content of uronic acids of the polydispersity (MW /MN ) of all polysaccharide fractions and the residue.
five hemicellulosic fractions and final residue are given in Table 2. Hemicellulosic fractions
As seen from Table 2, xylose was the predominant sugar unit
in the all of hemicellulosic fractions, comprising 39.48–68.44% of C1 C2 C3 C5 C7 RC

the total sugar and uronic acids; while arabinose (12.01–14.82%), MN 19,778 4437 2517 2514 2569 1953
galactose (4.14–8.60%) and glucose (8.26–15.04%) presented in MW 24,824 4574 2540 2537 2588 2020
MW /MN 1.255 1.031 1.009 1.009 1.007 1.034
small amounts. In comparison with the H1 , H2 , and H3 , high
 J.-B. Shi et al. / Industrial Crops and Products 44 (2013) 542–548 545

to the skeletal vibration of lignin (Sun et al., 2012). The sharp

Fig. 2. FT-IR spectra of hemicellulosic fractions and final residue.
indicated that most of hemicelluloses in the yellow liquor were sig-
nificantly degraded under the cooking conditions comparing with
the MW of hemicelluloses in the raw material (Shi et al., 2012). Fur-
thermore, all hemicellulosic fractions showed a relatively low index
of polydispersity (MW /MN ) (1.007–1.255), indicating a chemical and
structural homogeneity within each fraction.
bands at 1322 cm−1 were assigned to the C O stretching vibration
of syringyl units (Yuan et al., 2009). The intensities of this band
increased when the concentration of ethanol was increased, which
indicated that lignin was not completely removed in the acidifi-
cation process. The sharp bands at 1384 cm−1 and weak peak at
1244 cm−1 represented C H stretching, OH or C O (in ester
group) bending vibration (Jin et al., 2009; Bian et al., 2012). Bands
between 1130 and 1000 cm−1 were typical absorbance of hemi-
celluloses (Xiao et al., 2001). The prominent band at 1044 cm−1
was attributed to the C O, C C, stretching or C OH bending typi-
cal xylan (Xu et al., 2006). The sharp band at 896 cm−1 indicated
that hemicellulosic fractions C1 and C2 were dominant for the
ˇ-glycosidic linkages between the sugar units. The presence of ara-
binosyl units in hemicellulosic fractions C1 and C2 , which attach
to xylopyranosyl constituents, were identified by a low intensity
shoulder at 1157 and 988 cm−1 (Sun et al., 2005). The intensity
changes above two bands could reflect the arabinosyl substituent
and identify the arabinoxylan structure (Peng et al., 2009). The
disappearance of signals at 988 and 896 cm−1 indicated that the
structures of the hemicellulosic fractions C3 , C5 , C7 and RC were
significantly damaged during the cooking process, and the same
conclusion was also got from GPC results.
3.5. 1 H–13 C HSQC NMR analysis

To further study the structure of the hemicelluloses in yellow

3.4. FT-IR spectra
liquor, the hemicellulosic fractions C1 , C3 , C7 and RC were investi-
gated using 1 H–13 C HSQC NMR spectroscopy. The NMR spectra of
The FT-IR spectra of all hemicellulosic fractions and final residue
three hemicellulosic fractions and final residue are given in Fig. 3,
obtained from the yellow liquor are shown in Fig. 2. The typical
and the assignment of chemical shift to 1 H and 13 C of C1 are listed
functional groups and the FT-IR signals with the possible com-
in Table 5 based on reported NMR data (Yuan et al., 2011; Bian
pounds are listed in Table 4. The absorption at 3405 cm−1 (not
et al., 2012; Prozil et al., 2012; Xue et al., 2012). The hemicellu-
shown in the spectra) was attributed to the stretching of OH
loses obtained from the yellow liquor contained some lignin, so the
groups in the sugar residues. The C H stretching vibration gave
HSQC spectra of hemicelluloses could be divided into three regions
signals at 2978 and 2931 cm−1 (not shown in the spectra). The
of 1 H–13 C correlations: corresponding to aliphatic region (ıC /ıH
signals of C O stretching vibration (at c.a. 1732 cm−1 ) in acetyl,
0–50/0–2.8), lignin side chains and polysaccharides region (ıC /ıH
uronic and ferulic ester groups were not observed (Jin et al., 2009),
50–110/2.8–5.5), and aromatic region (ıC /ıH 100–160/5.5–9.0).
which suggested that ester bonds might be completely destroyed
Considering the signals usually originate from the certain impu-
during the cooking process. The small signals at 1690 cm−1 in the
rities, the aromatic region and aliphatic region was not detailed
all hemicellulosic fractions were attributed to stretching vibra-
discussed in this study.
tion of conjugate carbonyl with the benzene rings of lignin (Wang
In the polysaccharides region of C1 , the cross peak at
et al., 2012), which indicated that the lignin in the yellow liquor
101.56/4.46 ppm corresponded to the C-1/H-1 anomeric proton
was oxidized. The presence of uronic acids in the hemicellulosic
and carbon of the (1 → 4)-linked-ˇ-d-Xylp, while the major signals
fractions were also manifested mainly by the appearance of anti-
at 73.63/3.27, 73.56/3.54, 76.18/3.77, 62.79/4.09, 62.77/3.60 ppm
symmetric and symmetric stretching from the COO− groups at
were attributed to C-2/H-2, C-3/H-3, C-4/H-4, C-5/H-5ax, C-5/H-
1616 and 1422 cm−1 , respectively, which was consistent with the
5eq of D-Xylp, respectively. Concerning the arabinofuranose (Araf)
sugar analysis. In additionally, the absorbance at 1507 cm−1 related
units, which were decorated at C-2 and/or C-3 position of Xylp
(Bian et al., 2012), the signals of C-1/H-1, C-2/H-2, C-3/H-3, C-
Table 4 4/H-4, C-5/H-5ax, C-5/H-5eq appeared at 107.56/5.37, 76.99/3.90,
The main functional groups of corn stalk hemicelluloses in the yellow liquor.
80.64/4.14, 84.38/4.26, 60.93/3.78, 60.95/3.71 ppm, respectively.
Wavenumbers (cm−1 ) Functional groups Compounds Moreover the weak signals at 97.46/5.28, 72.33/3.72, 82.49/3.20
3800–3000 O H stretch Hydroxyl group
and 59.72/3.45 ppm were characteristic respectively of C-1/H-1,
2980–2840 C H stretch Alkyl C-3/H-3, C-4/H-4 and OCH3 of 4-O-methyl-␣-d-glucuronic acid
1690 C O stretch Lignin residues, which attached to the (1 → 4)-ˇ-d-Xylp at position of
1616, 1421 COO- anti- and Uronic acids C-2 (Peng et al., 2010). Additionally, the (1 → 4)-ˇ-d-Xylp related
symmetric stretch
with 4-O-methyl-␣-d-glucuronic acid were determined by signals
1507 Aromatic skeleton Lignin
vibration at 102.32/4.55 (C-1/H-1), 75.71/3.41 (C-2/H-2), 72.21/3.63 (C-
1322 C O stretch Lignin 3/H-3), 73.19/3.79 (C-4/H-4), 64.83/3.96 (C-5/H-5ax), 65.06/3.29
1384, 1244 C H stretch, OH or Hemicelluloses (C-5/H-5eq) ppm, respectively. A strong contour at 60.71/3.94 ppm
C O bending stretch corresponded to the methyl of methoxyl in lignin. Besides the
1700–980 Arabinoxylan
1157, 998 Arabinose
aforementioned signals of sugar residue, weak signals corre-
1044 C O stretch Hydroxyl group sponding to glucose were also observed. The above results can
896 ˇ-d-Xylose demonstrate that the C1 precipitated with one volume ethanol
788–400 C C stretch The ring of the sugar and from the yellow liquor was composed of (1 → 4)-ˇ-d-Xylp back-
lignin
bones substituted with ␣-L-Araf and 4-O-methyl-␣-d-glucuronic
546 J.-B. Shi et al. / Industrial Crops and Products 44 (2013) 542–548

Fig. 3. The HSQC of C1 , C3 , C7 and RC . The designations are as follows: X (1 → 4)-ˇ-d-Xylp; XU, (1→4)-ˇ-d-Xylp-2-O-(4-O-Me-␣-d-GlcpA); U, 4-O-Me-␣-d-GlcpA; A, ␣-l-Araf;
G, (1 → 4)-ˇ-d-glucp; ax, axial; eq, equatorial.

acid. Some unlabeled signals in the lignin side chain informed
side chain inter-unit linkages of lignin (Martínez et al., 2008). In
the aliphatic region, the signals at 1.19/20.08 and 1.03/16.72 ppm
were attributed to the methylene and methyl groups in the propyl
side chains, respectively. The peak at 1.78/23.11 ppm represented
methyl from acetyl groups (Sun et al., 2011a,b). The signals in
the aromatic region were originated from the aromatic carbons
and protons in syringyl (S), guaiacyl (G), and p-hydroxyphenyl (H)
units, p-coumarate and ferulate acids (Sun et al., 2012).
The spectra of C1 , C3 , C7 and RC could illustrate the differences in
structure and component. The decrease of the sugar residues spec-
tral complexity in the polysaccharides region and the increase of
spectral complexity in the lignin chain region indicated that the
concentration of ethanol solution could affect the sugar residues
constituent and the content of lignin in the hemicellulosic frac-
tions. The spectral contour and relative intensities of C1 and C3 were
similar in the polysaccharides region, indicating similar structure
of hemicellulosic fractions obtained in different concentrations of
ethanol. The signals of sugar residues in spectra C7 and RC disap-
peared and intensities were much lower than that in C1 and C3
showed that the contents of hemicelluloses in the C7 and RC were
lower than in C1 and C3 . The signals in the aliphatic region and
lignin side chain region of C1 , C3 and C7 showed more lignin was
precipitated when the high concentration of ethanol solution was
used. The spectrum RC indicated that the most of component of RC
was lignin.
3.6. Thermo-gravimetric analysis
The thermo-gravimetric analysis can rapidly investigate the
mass loss and compare the thermal behavior of hemicellulosic
fractions. The pyrolysis process of biomass could be divided into

Table 5
1
H and 13 C chemical shift (ppm) assignments for C1 .

Sugars residue Chemical shift (ppm) H/C

1 2 3 4 5ax 5eq 6 OCH3

101.56 73.63 73.56 76.18 62.79 62.77 – –

X
4.46 3.27 3.54 3.77 4.09 3.60 – –
107.56 76.99 80.64 84.38 60.93 60.95 – –
A
5.37 3.90 4.14 4.26 3.78 3.71 – –
102.32 75.71 72.21 73.19 64.83 65.06 – –
XU
4.55 3.41 3.63 3.79 3.96 3.29 – –
97.46 72.33 82.49 – 59.72
U
5.28 3.72 3.20 – 3.45
101.03 76.58 72.96 69.19 – 62.02 –
G
4.61 3.54 3.42 3.61 – 3.64 –

The designations are as follows: X (1 → 4)-ˇ-d-Xylp; XU, (1 → 4)-ˇ-d-Xylp-2-O-(4-O-Me-␣-d-GlcpA); U, 4-O-Me-␣-d-GlcpA; A, ␣-l-Araf; G, (1 → 4)-ˇ-d-glucp; ax, axial; eq,
equatorial.
 J.-B. Shi et al. / Industrial Crops and Products 44 (2013) 542–548
Fig. 4. The TGA/DTG curves of C1 , C3 and RC .
four stages according the DTG curve: at the first stage (room
temperature to 200 ◦ C), the sample just lost water; at the sec-
ond stage (200–315 ◦ C), hemicelluloses pyrolyze rapidly; the third
stage (315–400 ◦ C), cellulose decomposition; and the last stage
(400–700 ◦ C), degradation of volatile components and further char-
ring processes of the residues with lignin decomposition occurring
over a wide temperature range, forming char (Yang et al., 2007).
The hemicelluloses obtained from the yellow liquor may con-
tain some other organics, such as, lignin, cellulose and organic
acids. The hemicellulosic fractions C1 , C3 and, RC were investigated
with the thermo-gravimetric analysis (TGA) and derivative thermo-
gravimetry (DTG) between room temperature and 600 ◦ C. The TGA
and DTG curves of C1 , C3 and RC are given in Fig. 4.
The DTG curves were divided in to three regions: Region A, B
and C. In the Region A (30–200 ◦ C), the weight loss was mainly due
to the removal of moisture and some lower molecular weight com-
ponents (Zhang et al., 2011; Bian et al., 2012). Two weight loss rate
peaks at 101 and 128 ◦ C in the curve C3 , and five peaks at 135, 142,
159, 164, and 181 ◦ C were observed in the curve RC , which may
be mainly caused by the degradation products of hemicelluloses,
lignin and cellulose, such as, uronic acids, low molecular organic
acids. But the sharp peaks at 188 and 201 ◦ C in the curve C3 could
not be identified. The region B (200–360 ◦ C) was the mainly weight
loss happened range of hemicelluloses. The gas products included
CO2 , CH4 , CO and some organism (acids, aldehydes, alkanes, etc.)
with some water (Sun et al., 2011a,b). The C1 , C3 and RC got the max-
imum mass loss rate at 249, 262, and 273 ◦ C, respectively, while RC
has another loss peak at 300 ◦ C, which may come from the degraded
cellulose. The decomposition of lignin, volatile components and
further charring processes of the residues took place in the Region
C. And C1 , C3 and RC got the maximum mass loss rate at 426, 437
and 361 ◦ C, respectively. The RC contained more degraded lignin
and low molecular weight degraded products of corn stalk. There-
fore, the temperature of maximum mass loss rate of RC was lowered
and there were two shoulder peaks at 400 and 486 ◦ C in main peak.
Evidently, the thermal stability of C3 appeared to be more stable
than C1 ; and C1 was more stability RC in the main decomposition
region from TG curves. The sugar analysis showed the C3 contained
less degree of side chain substitutions than C1 and RC . RC was the
final residue, which could not be precipitated by seven volumes
ethanol. In other words, the RC was the strongly degraded products
of corn stalk. This revealed that the hemicelluloses with infrequent
side chains were more thermally stable and could be precipitated
with a low concentration of ethanol, while molecules containing
a high degree of side chains substitutions were less thermally sta-
ble and were difficultly precipitated under low concentration of
ethanol.
547
4. Conclusions
It was concluded that xylose was primary sugar residue in
the corn stalk hemicellulosic fractions obtained from active oxy-
gen cooking process. However, the final residue contained high
amounts of uronic acids. Only 5.31% hemicelluloses in the yellow
liquor owned high molecular weight (24,824 g/mol), the remainder
of the hemicellulosic fractions were significantly degraded during
the cooking process. The corn stalk hemicellulosic fractions were
composed of (1 → 4)-linked-ˇ-d-Xylp backbones, but the side chain
attached to the Xylp units were diverse, depending on the precipi-
tation process. In addition, hemicellulosic fractions with infrequent
side chains were more thermally stable and could be precipitated
with a low concentration of ethanol.
Acknowledgment
The authors are grateful for the financial support from the
National Key Basic Research Program (2010CB732201) from the
Ministry of Science.
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