Bioresource Technology 182 (2015) 364–367

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Bioresource Technology
journal homepage: www.elsevier.com/locate/biortech

Short Communication

Obtaining fermentable sugars by dilute acid hydrolysis of hemicellulose

and fast pyrolysis of cellulose
Liqun Jiang, Anqing Zheng, Zengli Zhao ⇑, Fang He, Haibin Li, Weiguo Liu
Key Laboratory of Renewable Energy, Guangzhou Institute of Energy Conversion, Chinese Academy of Sciences, Guangzhou 510640, China

h i g h l i g h t s

 Dilute acid hydrolysis was used to remove hemicellulose as sugars.

 Levoglucosan yields from crystalline cellulose could be as high as 61.47%.
 The crystalline structure has positive role in enhancing the yield of levoglucosan.
 The demineralization by dilute acid pretreatment can improve the yield of levoglucosan.

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

Article history: The objective of this study was to get fermentable sugars by dilute acid hydrolysis of hemicellulose and
Received 19 December 2014 fast pyrolysis of cellulose from sugarcane bagasse. Hemicellulose could be easily hydrolyzed by dilute
Received in revised form 8 January 2015 acid as sugars. The remained solid residue of acid hydrolysis was utilized to get levoglucosan by fast pyro-
Accepted 9 January 2015
lysis economically. Levoglucosan yield from crystalline cellulose could be as high as 61.47%. Dilute acid
Available online 7 February 2015
hydrolysis was also a promising pretreatment for levoglucosan production from lignocellulose. The dilute
acid pretreated sugarcane bagasse resulted in higher levoglucosan yield (40.50%) in fast pyrolysis by
Keywords:
micropyrolyzer, which was more effective than water washed (29.10%) and un-pretreated (12.84%). It
Dilute acid hydrolysis
Fast pyrolysis
was mainly ascribed to the effective removal of alkali and alkaline earth metals and the accumulation
Levoglucosan of crystalline cellulose. This strategy seems a promising route to achieve inexpensive fermentable sugars
Lignocellulose from lignocellulose for biorefinery.
Ó 2015 Elsevier Ltd. All rights reserved.

1. Introduction residue (mainly contained crystalline cellulose and lignin) is more

Nowadays, the production of chemicals and fuels from lignocel-
lulose has received a significant interest due to the forthcoming
scarcity of conventional resources (Luterbacher et al., 2014). Sugars
are one of the natural intermediates in the biological and chemical
conversion of biomass. The most common approach to recover sug-
ars from biomass is by enzymatic and acid hydrolysis. Nonetheless,
low rates of hydrolysis and the costs of both pretreatment and
enzyme are potential drawbacks to enzymatic hydrolysis. Mean-
while the hazards of handling concentrated acid and the complex-
ities of recycling it have limited the adoption of concentrated acid
hydrolysis (Binder and Raines, 2010). In dilute acid, hemicellulose
and amorphous cellulose are easily and nearly completely hydro-
lyzed to fermentable sugars accessible to microorganisms for bio-
fuels production (Jiang et al., 2013). But the remained solid
⇑ Corresponding author.
E-mail address: zhaozl@ms.giec.ac.cn (Z. Zhao).
difficult to be hydrolyzed (Jiang et al., 2012).
Although enzymatic and acid hydrolysis have received most of
the attention, a little-explored alternative method to obtain fer-
mentable sugars is fast pyrolysis (Rover et al., 2014). Pyrolysis
has a lower capital cost than that of biochemical pathways (Anex
et al., 2010). Fast pyrolysis can depolymerize cellulose to levoglu-
cosan (1,6-anhydro-b-D-glucopyranose) using high heating rates
and very short residence time at 450–550 °C without the use of
enzymes or catalysts (Kuzhiyil et al., 2012). Several microorgan-
isms have been identified that can use levoglucosan as substrate
for itaconic acid, citric acid and ethanol production (Nakagawa
et al., 1984; Zhuang and Zhang, 2002; Layton et al., 2011). Levoglu-
cosan can also be hydrolyzed to glucose easily, thereby providing a
potentially rapid and efficient route to the production of ethanol
(Bennett et al., 2009; Luque et al., 2014). Nevertheless, biomass fast
pyrolysis produces a very low amount of levoglucosan when com-
pared to the theoretical yield based on cellulose fraction and pre-
treatment prior to pyrolysis is essential (Chang et al., 2013;
Carpenter et al., 2014; Zheng et al., 2014).

http://dx.doi.org/10.1016/j.biortech.2015.01.032
0960-8524/Ó 2015 Elsevier Ltd. All rights reserved.
 L. Jiang et al. / Bioresource Technology 182 (2015) 364–367 365

Table 1
Components and elements of biomass used in the current study.

Un-pretreated sugarcane bagasse Water washed sugarcane bagasse Acid pretreated sugarcane bagasse
Components [wt%]
Cellulose 38.56 40.14 57.90
Hemicellulose 21.24 21.12 5.86
Elements [wt%]
C 46.58 46.89 47.13
H 6.37 6.42 6.23
N 0.26 0.18 0.16
AAEM [ppm]
Na 34.26 19.97 19.71
K 2632.36 54.56 20.24
Mg 241.25 116.65 9.15
Fe 291.14 307.36 126.41
Al 125.41 104.15 66.23
Ca 499.89 489.58 49.22
Total AAEM 3824.31 1092.27 290.96

In this study, the feasibility of combination dilute acid hydroly-
sis and fast pyrolysis for fermentable sugars was evaluated. Sugar-
cane bagasse was firstly hydrolyzed by dilute sulfuric acid to
remove hemicellulose as sugars. The remained cellulose in the
solid residue was further utilized to get levoglucosan by fast pyro-
lysis. Water washed sugarcane bagasse and cellulose (including
microcrystalline cellulose and ionic liquid (IL) pretreated cellulose)
were employed as control for comparison. The effects of compo-
nents, elements and crystalline structure on the products distribu-
tion of fast pyrolysis were investigated.
2. Methods
2.1. Materials
Sugarcane bagasse was obtained from Dehong in Yunnan,
China. It was air dried at 60 °C for 72 h, ground and sieved to the
particle size range 0.11–0.18 mm and then dried in an oven at
105 °C for 12 h. Microcrystalline cellulose (AvicelÒ PH-101, Cat.
No. 11363) was purchased from Sigma. 1-Butyl-3-methylimidazo-
lium chloride ([BMIM]Cl) (purity > 99%) was purchased from Lanz-
hou Institute of Chemical Physics, Chinese Academy of Sciences
(Lanzhou, China). Sulfuric acid was purchased from Chuandong
Chemical Co. Ltd., Chongqing.
2.2. Dilute acid and water washed pretreatment
The dilute acid and water washed pretreatment were carried
out in a 100 mL high pressure autoclave (HKY-3, Haian Petroleum
Research Co. Ltd., Jiangsu, China). For dilute acid pretreatment,
3.6 g sugarcane bagasse were loaded in the autoclave containing
60 mL 0.1 wt% dilute sulfuric acid solution, and reacted at 145 °C
for 0.5 h. Solid residue was collected by filtration and washed with
300 mL distilled water to remove residual sulfuric acid, then air
dried at 105 °C for 12 h. Sugarcane bagasse powders were also
washed with 60 mL fresh deionized water at 145 °C for 0.5 h, then
washed with 300 mL distilled water, air dried at 105 °C for 12 h
and recovered as water washed sugarcane bagasse.
2.3. IL pretreatment
Cellulose suspension in [BMIM]Cl was prepared by adding 4 g
(4 wt%) cellulose in a 500 mL flask containing 100 g IL. Then, with-
out removing air, the flask was sealed with a cork and placed in oil
bath at 110 °C with stirring at 200 rpm for 1 h. Deionized water at
90 °C (400 mL) was added to the mixture to precipitate biomass
with vigorous shaking for 10 s. The regenerated biomass was then
transferred into a 1000 mL beaker with deionized water at 75 °C,
washed with 75 °C water (700 mL) thoroughly for 15 times to
remove IL, freeze dried (EYELA 1200 freeze dryer, Tokyo Rikakikai
Co., Ltd.) and recovered as IL-pretreated cellulose (Jiang et al.,
2013).
2.4. Components and elements analysis
Characterization of biomass was carried out following the
National Renewable Energy Laboratory (NREL) procedure (Sluiter
et al., 2008). Carbon, hydrogen and nitrogen content were mea-
sured with an Organic Elemental Analyzer (Vario EL cube, Hanau,
Germany). The metal content was determined by using an induc-
tively coupled plasma optical emission spectrometry (ICP-OES)
(Optima 8000, PerkinElmer, USA).
2.5. Crystallinity measurement
Crystallinity index (CrI) of samples were determined by X-ray
diffraction (XRD) in a X’Pert PROMPD X-ray diffractmeter (PANa-
lytical V.B., Holland). X-ray diffractometer was set at 40 kV and
40 mA. Cu radiation (k = 1.54 Å) was scanned over diffraction angle
(2h°) of 5–45° with a step of 0.01°. CrI was determined by the equa-
tion (Segal et al., 1959):
CrI% ¼ ðI002  Iam Þ=I002  100%
where I002 was the highest peak intensity at an angle of diffraction
of 2h = 22.5°, whereas Iam was the peak for the amorphous cellulose
at 2h = 18°.
2.6. Fast pyrolysis of biomass
Analytical pyrolysis of biomass samples were performed by
using a CDS 5200 micropyrolyzer coupled to a gas chromatography
mass spectroscopy (GC/MS, 7890A, 7975C, Agilent, USA). Sample
weights in the range of 200–450 lg produced resolvable spectra
for compounds interest while minimizing mass transfer effects.
Pyrolysis was carried out at 500 °C with the interface line between
the pyrolyzer and GC maintained at 300 °C to prevent condensa-
tion of vapors. The volatile products were swept from micropyro-
lyzer directly to the 240 °C GC injector using helium as carrier
gas (1 mL/min). The column used for the chromatographic separa-
tion of chemical components was a HP-INNO wax capillary column
(Agilent 19091N-133, Polyethylene Glycol, 30 m length, 0.25 mm
ID, 0.25 lm film thickness). The GC oven temperature program
was 2 min at 50 °C, 10 °C/min to 90 °C, 4 °C/min to 129 °C, and then
8 °C/min to 230 °C with a dwell time of 29 min. The mass
366 L. Jiang et al. / Bioresource Technology 182 (2015) 364–367
Table 2
Crystallinity index and compounds yield of biomass.
Biomass CrI (%)
Un-pretreated cellulose 93.3
IL-pretreated cellulose 35.6
Un-pretreated sugarcane bagasse 49.7
Water washed sugarcane bagasse 52.4
Acid pretreated sugarcane bagasse 63.1
*
Data (%) were based on mass of cellulose.
spectrometer was operated in electron impact mode at 70 eV. Full
scan mass spectra were acquired from 12 to 500 m/z. The standard
compounds were used to confirm the peak identification based on
matching the retention time and mass spectrum.
3. Results and discussion
3.1. Hemicelluose removal by dilute acid hydrolysis
The components analysis indicated that the content of hemicel-
lulose decreased obviously after acid hydrolysis and that changed
slightly after water washed pretreatment (Table 1). In the case of
bulk chemicals and biofuels, the cost of the raw material mostly
affected the price of the final products and the efficient utilization
of hemicellulose and cellulose were essential. The dilute acid
hydrolysate of hemicellulose was evaluated the fermentability
and proved to be an attractive raw material for 2,3-butanediol fer-
mentation in previous study (Jiang et al., 2012, 2013).
3.2. Alkali and alkaline earth metals (AAEM) removal by dilute acid
The composition of C and H in sugarcane bagasse changed little
after dilute acid or water washed pretreatment (Table 1). The con-
tent of N decreased after water washed and dilute acid pretreat-
ment. This phenomenon may be due to the removal of protein
during pretreatments. AAEM consisting of Na, K, Mg, Al, Fe and
Ca in dilute acid pretreated sugarcane bagasse exhibited lower
content than that of original material. Water washed pretreatment
could remove K effectively. However Mg, Fe, Al and Ca were not
effectively removed by water washed pretreatment. The total con-
tent of AAEM decreased by 71% after water washed and that of
dilute acid pretreatment decreased by 92%. AAEM were powerful
catalysts to decrease the yield of anhydrosugars by catalyzing
pyranose and furanose ring fragmentation leading to increased
yields of less desirable light oxygenates and non-condensable
gases at the expense of levoglucosan and other anhydrosugars dur-
ing pyrolysis (Kuzhiyil et al., 2012; Dalluge et al., 2014). In this
study, AAEM could be removed effectively by dilute acid
pretreatment.
3.3. Crystalline structure of pretreated biomass
The crystallinity of biomass was evaluated as the most impor-
tant factor to interpret the structural evolution of biomass after
pretreatment. In Fig. S1(e), the intensity of crystalline peaks in sug-
arcane bagasse were increased after acid pretreatment (from 49.7%
to 63.1%). The reason was that in the case of dilute acid pretreat-
ment, the amorphous components broke down more but this pre-
treatment process was unable to break the inter- and intra-chain
hydrogen bonding in cellulose fibrils, and crystalline cellulose
broke down less. After water washed, the CrI increased from
49.7% to 52.4%. This phenomenon might be due to the slightly
accumulation of cellulose. In order to directly explain the effect
Yield of compounds (%)
Acetic acid Furfural 5-HMF Levoglucosan Levoglucosan*
0.47 0.43 0.77 61.47 61.47
1.55 0.92 2.52 40.76 40.76
5.66 0.70 0.23 4.95 12.84
4.73 0.68 0.35 11.68 29.10
1.41 0.50 0.68 23.45 40.50
of crystalline structure of cellulose on the products distribution
of fast pyrolysis, microcrystalline and IL-pretreated celluloses were
introduced. IL pretreatment could change the crystalline structure
and morphology of cellulose by disrupting hydrogen bonding. After
IL pretreatment, the CrI of cellulose decreased from 93.3% to 35.6%.
XRD profiles of biomass showed that microcrystalline cellulose
(curve a) has five distinct peaks with their diffraction angles (2h)
at around 13.9°, 15.0°, 18.8°, 20.5° and 30.1°, which decreased
remarkably or even disappeared with a flat and broad peak
remained at around 20.1° after IL pretreatment (curve b). The pos-
sible reason was due to the transformation of cellulose I to cellu-
lose II. The efficiencies of different pretreatments were examined
by the product distribution from fast pyrolysis.
3.4. Obtaining levoglucosan by fast pyrolysis of cellulose
Levoglucosan yields from pure crystalline cellulose could be as
high as 61.47%. However, the yield of levoglucosan was decreased
to 40.76% after IL pretreatment. Therefore, the increase of CrI had
positive role in enhancing the levoglucosan yield and a negative
influence on the formation of acetic acid, furfural and 5-hydroxym-
ethylfurfural (5-HMF) (Table 2). The yield of levoglucosan
increased by 126.64% for water washed sugarcane bagasse com-
pared to raw material, which was mainly due to the removal of
AAEM. The sugarcane bagasse exhibited significantly higher yield
of levoglucosan and increased by a remarkable 215.42% after dilute
acid pretreatment compared to raw material, which was mainly
due to the removal of AAEM and increase of crystalline cellulose.
The yield of acetic acid and furfural were gradually diminished
after water washed and dilute acid pretreatment, while the yield
of 5-HMF increased. Furfural and 5-HMF were known to be prod-
ucts of carbohydrate dehydration. The decrease of acetic acid and
furfural was likely due to the removal of hemicellulose. The
increase of 5-HMF was likely due to high content of cellulose in
dilute acid pretreated sugarcane bagasse. The experimental results
showed that dilute acid hydrolysis could serve as an effective pre-
treatment method to improve the yield of levoglucosan. In the next
work, the dilute acid hydrolysate and levoglucosan will be utilized
as substrate for fermentation. The strategy in this work seemed a
promising method to get inexpensive fermentable sugars from bio-
mass for biorefinery.
4. Conclusions
Fermentable sugars were obtained by dilute acid hydrolysis of
hemicellulose and fast pyrolysis of cellulose. Hemicellulose was
effectively hydrolyzed as sugars by dilute acid. The cellulose of
solid residue after acid hydrolysis was depolymerized to levogluco-
san economically by fast pyrolysis. Dilute acid hydrolysis could
effectively diminish the content of alkali and alkaline earth metals
and could accumulate crystalline cellulose to enhance the yield of
levoglucosan.
 L. Jiang et al. / Bioresource Technology 182 (2015) 364–367
Acknowledgement
The authors wish to acknowledge the financial support from the
National Natural and Science Foundation of China (No. 51376186
and No. 21406227).
Appendix A. Supplementary data
Supplementary data associated with this article can be found, in
the online version, at http://dx.doi.org/10.1016/j.biortech.2015.01.
032.
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