Bioresource Technology 132 (2013) 361364

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

Short Communication

Effect of ionic liquid pretreatment on the composition, structure and

biogas production of water hyacinth (Eichhornia crassipes)
Jing Gao, Li Chen, Zongcheng Yan , Lin Wang
School of Chemistry and Chemical Engineering, South China University of Technology, Guangzhou 510640, Guangdong, PR China

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

Article history: The effect of the pretreatment of water hyacinth with ionic liquid and co-solvent on the lignocellulosic
Received 17 August 2012 composition, structural change and biogas production was evaluated in this study. The results from
Received in revised form 26 October 2012 regenerated water hyacinth indicate that, the content of the lignocellulosic composition was changed,
Accepted 26 October 2012
the crystallinity of the structure was decreased, and the surface became more porous. After the pretreat-
Available online 5 November 2012
ment with 1-N-butyl-3-methyimidazolium chloride ([Bmim]Cl)/dimethyl sulfoxide (DMSO) under 120 !C
for 120 min, the cellulose content of regenerated water hyacinth was increased by 27.9%, 49.2% of the lig-
Keywords:
nin was removed, and the biogas yield was increased by 97.6% as compared with unpretreated water hya-
Water hyacinth
Biogas production
cinth. The ionic liquids and co-solvents were successfully recovered by forming aqueous biphasic systems
Pretreatment with K3PO4.
Ionic liquid " 2012 Elsevier Ltd. All rights reserved.
Aqueous biphasic system

1. Introduction
Water hyacinth (Eichhornia crassipes) is lignocellulosic biomass
consisting of a complex mixture of lignin, hemicelluloses and cel-
lulose. Both the cellulose and hemicelluloses are polymers of sug-
ars, and are thereby a potential source of sugars (Xie et al., 2012).
As a result, the conversion of water hyacinth to fuels has received
significant interest in the last few decades (Kumar et al., 2009;
Mishima et al., 2008). However, the cellulose content of the water
hyacinth was much lower when compared with wood and straw
(Snchez, 2009). The lignin, an amorphous heteropolymer consist-
ing of three different phenylpropane units, physically protects cel-
lulose and hemicelluloses from degradation (Hendriks and
Zeeman, 2009). Moreover, the crystalline structure and available
surface area make hydrolysis processes difficult and expensive
(Teghammar et al., 2010). Consequently, pretreatment to remove
the lignin and enhance the hydrolysis of cellulose is essential. Xu
et al. (2011) reported that pretreatment of Bermuda grass with
3% NaOH solution could improve methane yield by 20%. Dilute acid
pretreatment could also improve the reducing sugar yield of sugar-
cane tops (Sindhu et al., 2011). However, the pretreatment meth-
ods have significant drawbacks including high energy demands,
toxic byproducts and unrecyclable reagents (Rafique et al., 2010).
Ionic liquids (ILs) have been used as pretreated solvents to re-
duce the crystallinity of cellulose and further enhance its hydroly-
sis (Li et al., 2010). ILs have the advantages of having low volatility,
being non-flammable and easily recyclable (Gremos et al., 2011),
and are less energy demanding (Qiu et al., 2012). Biomass digest-
ibility was 65% for [Emim]Ac pretreated cotton stalk after 72 h of
enzymatic hydrolysis, which was 9-fold higher compared to un-
treated cotton stalk (Haykir et al., 2013). Bahcegul et al. (2011)
and Guragain et al. (2011) reported that ionic liquid pretreatments
gave higher glucose yields than dilute acid pretreatment. However,
nearly all ionic liquids have higher viscosity than organic solvents,
which is a major barrier to the commercial application of ILs
(Pinkert et al., 2009).
Here, polar aprotic solvents were added as co-solvent to reduce
the viscosity of ionic liquids, and the effect of pretreatment with
ionic liquid/co-solvent on the changes of lignocellulosic composi-
tion and structure of water hyacinth and its biogas production
was evaluated. Additionally, the recovery of the solvents by form-
ing aqueous biphasic systems was also studied.
2. Methods
2.1. Materials

Fresh water hyacinth stems were harvested from the Liuxi

River in Guangzhou. The collected stems were washed manually
Corresponding author. Tel./fax: +86 20 87111109. to remove adhering dirt and chopped in small pieces, dried at
E-mail address: zcyan@scut.edu.cn (Z. Yan). 45 !C for 48 h, and powdered. ILs 1-ethyl-3-methlyimidazolium

0960-8524/$ - see front matter " 2012 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.biortech.2012.10.136
362 J. Gao et al. / Bioresource Technology 132 (2013) 361364

acetate ([Emim]Ac) and 1-N-butyl-3-methyimidazolium chloride
([Bmim]Cl) were provided by Centre of Green Chemistry and Catal-
ysis, Lanzhou Institute of Chemical Physics, China. Dimethyl ace-
tylamide (DMAC) and dimethyl sulfoxide (DMSO) were used as
co-solvents of the ILs and purchased from SigmaAldrich, Inc. All
other chemicals were from commercial source and of reagent
grade.
2.2. Pretreatment and regeneration of water hyacinth
A 5% (w/w) water hyacinth solution was prepared by combining
3.0 g of water hyacinth with 51.0 g IL and 11.0 g co-solvent in a
250 mL round-bottom flask. The solution was heated and stirred
in an oil bath pan at varied temperatures from 20 to 140 !C and
incubated for 60, 120, 150 or 240 min. Equal volume of deionized
water was added to the water hyacinth/solvents solution for
regenerating water hyacinth. A precipitate immediately formed.
The sample was briefly centrifuged. The supernatant containing
IL and co-solvent was removed and used in the solvents recovery,
while the precipitate was washed for three times with additions of
deionized water. Finally, the regenerated water hyacinth was fil-
tered and oven-dried at 45 !C for 48 h in a vacuum for the use of
biogas production.
2.3. Anaerobic batch digestions
water hyacinth, the supernatant mainly containing the solvent was
removed and placed to 25 !C. 30.0 g K3PO4 was added into the
solution, vibrated by a vortex mixer (XW-80A, Jingke, Inc., China),
and placed over night.
2.6. Analytical methods
Cellulose, hemicelluloses and lignin contents of the pretreated
or untreated water hyacinth species were determined according
to NREL procedures (Sluiter et al., 2008). Biogas production was
measured by water displacement. Methane and carbon dioxide
were analyzed using a gas chromatograph (GC 9560, Huaai Inc.,
Shanghai, China) equipped with a thermal conductivity detector.
The gas samples were analyzed twice in a week. The concentration
of ILs and K3PO4 in the upper and lower phases were analyzed by
an ion chromatography (Basic IC 792, Methohm, Switzerland)
equipped with electric conductivity detector. Karl-Fisher titration
was used to measure the water content in all samples. The weight
fractions of co-solvents were calculated by material balance
method.
3. Results and discussion
3.1. Lignocellulosic composition and structural property of regenerated
samples

The anaerobic batch digestions were carried out at 35 !C in
250 mL glass bottles, closed with rubber seals. The inoculum was
obtained from sludge digester operating at 35 !C. For each flask
containing 100 mL inoculum, 8 g untreated water hyacinth were
added together with 2 g pretreated water hyacinth samples.
2.4. Structure characterization of the regenerated water hyacinth
samples
XRD analysis of the regenerated water hyacinth samples were
conducted with a X-ray diffractometer (D8 Advance, Bruker Inc.,
Germany) operated at 40 kV and 50 mA with Ni-filtered Cu-Ka
radiation. The surface properties of were observed by a scanning
electron microscopy (S-3700N, Hitachi Ltd., Japan) equipped with
an alpha ray spectrometer (Quantax, Bruker Inc., Germany).
2.5. Recovery of pretreatment solvent
Pretreatment solvent was recovered by separated from an aque-
ous biphasic system (ABS). After pretreatment and regeneration of
The structure swelling of the water hyacinth was observed dur-
ing the pretreatment, and the high number of particles evidenced
disruption and exhibited loose structure. Amount of smaller mole-
cules such as fructose, oligosaccharide and lignin were produced
and dissolved in ionic liquid solutions during the pretreatment
process. In general, 27.160.4% of the initial lignin was removed
and the mass loss of water hyacinth was 4.925.1% as compared
with the unpretreated sample (Table 1). Although the addition of
DMSO and DMAC could reduce the viscosity of ionic liquid and in-
crease the flowability of the system, DMAC weakened the deligni-
fication capability of both [Bmim]Cl and [Emim]Cl. The result
indicates that the dissolving and delignification capability of the
solvents studied for pretreatment of water hyacinth follows the
order: [Emim]Ac > [Emim]Ac/DMSO ! [Bmim]Cl/DMSO > [Bmim]-
Cl ! DMSO > [Emim]Ac/DMAC ! [Bmim]Cl/DMAC " DMAC.
The pretreatments make the surface of water hyacinth more
porous, and [Emim]Ac showed better ability of crystallinity
reduction than [Bmim]Cl. Moreover, the formation of aggregates
could be observed in pretreated water hyacinth, especially in
the samples pretreated at 140 !C for 120 min or at 120 !C for

Table 1
Lignocellulosic composition and degradation of unpretreated and pretreated water hyacinth samples based on the dry weight.

Pretreated conditions Cellulose (%) Hemicellulose (%) Lignin (%) Degradation (%)
Solvents Temperature (!C) Time (min)
a. None None None 18.3 (0.5) 23.3 (0.3) 17.7 (0.1)
b. DMSO 120 150 24.3 (1.0) 22.5 (0.8) 8.6 (0.5) 13.5
c. DMAC 120 150 19.5 (0.1) 24.2 (0.5) 14.3 (0.8) 4.9
d. [Bmim]Cl 120 150 23.3 (1.0) 22.9 (0.3) 9.1 (0.5) 17.3
e. [Emim]Ac 120 150 26.8 (0.2) 25.1 (0.7) 7.0 (0.1) 25.1
f. [Bmim]Cl/DMSO 120 150 24.0 (0.3) 24.6 (0.1) 8.0 (0.5) 15.8
g. [Emim]Ac/DMSO 120 150 24.8 (0.6) 24.1 (0.8) 7.5 (0.5) 19.8
h. [Bmim]Cl/DMAC 120 150 22.9 (0.6) 21.8 (0.1) 11.4 (0.1) 23.8
i. [Emim]Ac/DMAC 120 150 21.6 (0.3) 24.5 (0.5) 12.9 (0.1) 23.0
j. [Bmim]Cl/DMSO 100 120 21.7 (0.1) 23.1 (0.5) 11.7 (0.8) 13.6
k. [Bmim]Cl/DMSO 120 120 23.4 (0.5) 24.1 (0.6) 9.0 (0.5) 13.9
l. [Bmim]Cl/DMSO 140 120 25.0 (0.5) 20.6 (0.2) 8.3 (0.2) 18.6
m. [Bmim]Cl/DMSO 120 60 21.7 (0.3) 21.9 (0.1) 10.0 (0.4) 5.0
n. [Bmim]Cl/DMSO 120 240 31.7 (0.2) 23.0 (1.0) 7.5 (0.8) 24.3
 J. Gao et al. / Bioresource Technology 132 (2013) 361364
180
160
Cumulative biogas production
140
120
(L kg-1 VS)
100
80
60
40
20
0
0 5 10 15 20 25
Time (d)
Fig. 1. Biogas production of the water hyacinth unpretreated and pretreated under
various conditions: !, none; j, 100 !C for 120 min; N, 120 !C for 120 min; h, 140 !C
for 120 min; s, 120 !C for 60 min; 4, 120 !C for 240 min.
Fig. 2. Methane concentration of the biogas from water hyacinth unpretreated and
pretreated under various conditions.
240 min. The reason for such a fact can be explained by the
association of positively charged molecules released during pre-
treatment and negative sites of microorganisms or polysaccha-
rides (Fernndez et al., 2012).
3.2. Biogas production
The effect of pretreated temperature and time on cumulative
biogas production of water hyacinth is shown in Fig. 1. Untreated
biomass started to produce biogas until the 8th day, but a couple
of days were enough for the pretreated water hyacinth to convert.
This indicates that the pretreatment of ionic liquid can accelerate
biogas production and reduce lag phase. The water hyacinth pre-
treated at 100 !C for 120 min, 120 !C for 60 min, and 120 !C for
120 min showed higher biogas yield (100, 104, 170 L kg#1 VS) than
untreated water hyacinth (86 L kg#1 VS). However, the acids such
as formic acid and levulinic acid, which are toxic for anaerobic
microorganisms (Chen et al., 2008), were produced from the pre-
treatment and acetogen at the initial digestion and then the pH va-
lue of the systems was quickly dropped within the first 3 days. On
the other hand, the formation of aggregates may make the diges-
tion difficult. Therefore, the samples pretreated at 140 !C for
120 min and at 120 !C for 240 min showed less production of bio-
gas (32 and 30 L kg#1 VS), and only lasted for about 8 days,
although higher pretreated temperature or longer pretreated time
could cause more loss of lignin.
363
Methane concentration of the biogas from each sample is pre-
sented in Fig. 2. The initial methane concentration from pretreated
water hyacinth samples was 1535% until the fourth day, but there
was no methane produced from untreated sample. On the 16th
day, treatments of 100 !C for 120 min, 120 !C for 120 min and
120 !C for 60 min resulted in higher methane concentration of
65%, 68% and 60%, respectively, compared to the untreated water
hyacinth with 53%. Treatments at 140 !C for 120 min and at
120 !C for 240 min resulted in the highest methane concentration
of 35% and 32%, respectively. However, after 7 days the methane
concentration dropped to 15% and 10%, respectively, and no further
methane was produced. The result indicates that the pretreat-
ments of 140 !C for 120 min and at 120 !C for 240 min against
30 35 the anaerobic digestion.
3.3. Solvents recovery
In the present work, separation processes were observed to
compare separation speed which follows the order DMA ! DM-
SO > [Bmim]Cl/DMA ! [Bmim]Cl/DMSO > [Emim]Ac/DMA ! [Emim]-
Ac/DMSO > [Emim]Ac " [Bmim]Cl. The recovery rates for DMAC,
DMSO, [Emim]Ac and [Bmim]Cl were 96%, 95%, 92%, and 90%,
respectively. This result was in accordance with the order of the
separation speed, which indicated that separation speed of ABS
,
could reflect its separation ability.
,
4. Conclusions
,
, After the pretreatment with ionic liquid and co-solvent, 27.1
, 60.4% of the lignin was removed from the water hyacinth, the crys-
tallinity of the structure was decreased, and the biogas yield and
methane concentration were increased by 16.397.6% and 13.2
28.3%, respectively. When the water hyacinth was pretreated at
120 !C for 120 min by [Bmim]Cl/DMSO, the biogas production
was the highest and the lag phase was one day. Higher pretreated
temperature and longer pretreated time against the anaerobic
digestion of the water hyacinth. The recovery rates of ionic liquid
and co-solvent were all over 90%.
Acknowledgements
The authors would gratefully acknowledge the support from the
Guangdong Provincial Laboratory of Green Chemical Technology.
They would also like to acknowledge the support from the Key So-
cial Development Program. Foundation of Guangdong Province
(No.: 2011A030600011).
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