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Article history: Lignocellulosic material (LCM) is promising alternative resource for sustainable energy production such
Received 10 June 2015 as ethanol and butanol and biohydrogen. Cellulose is an abundant renewable polymer of LCM found in
Received in revised form plant cell walls (30e50%). The high crystallinity of cellulose makes it recalcitrant to hydrolysis into its
31 October 2015
individual sugar subunits for biofuels production. Moreover, ionic liquids are considered as green sol-
Accepted 5 December 2015
Available online 22 December 2015
vents and have been used for biomass solubilization. The present work describes three properties:
KamleteTaft (KeT) parameters, viscosity and surface tension of ve imidazolium-based ionic liquids
(ILs); namely [C2mim][OAc], [C4mim][OAc], [C2mim][Cl], [C4mim][Cl] and [C4mims][BF4], and their ef-
Keywords:
Ionic liquids
ciency in the cellulose structural transformation for improved enzymatic glucose recovery. Crystalline
KeT parameter cellulose was treated with ILs at two different temperatures, i.e. 100 and 130 C for 5 and 2 h, respec-
Kinematic viscosity tively, with 10% solid loading followed by enzymatic saccharication using 10 and 20 FPU/g substrate of
Surface tension commercial cellulases. ILs treatment of crystalline cellulose signicantly reduces the crystallinity, which
Cellulose transformation resulted in a very sharp increase of sugar yields after enzymatic saccharication. Cellulose treated for
Saccharication 130 C/2 h resulted in better glucose yields as compared to 100 C/5 h. ILs comprising acetate anion
resulted in highest glucose yields and chloride based ILs performed moderately, whereas BF 4 based IL
was ineffective in transforming the cellulose structure. In order to decipher the possible reasons of
varying efciency of these ILs, the KeT parameters; hydrogen bond acidity (a), hydrogen bond basicity
(b), solvent polarizability (p*), kinematic viscosity (h) and surface tension (s) were calculated for 100 and
130 C. These results show that, among all the properties of ILs, hydrogen bond basicity (b) is relatively
more important than kinematic viscosity and surface tension for impacting the structural transformation
and subsequent enzymatic hydrolysis. [C2mim][OAc] with high b value (1.32) and lower viscosity (4.4 cSt/
s) and surface tension (30.3 mN/m) was found to be most efcient in cellulose transformation resulting
in higher glucose yields (89.8%) upon saccharication. Effect of size of cation and anion of ILs and
properties of regenerated cellulose is also examined by PXRD and FT-IR to further support the ndings.
2015 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.jclepro.2015.12.037
0959-6526/ 2015 Elsevier Ltd. All rights reserved.
1006 T. Raj et al. / Journal of Cleaner Production 113 (2016) 1005e1014
of ethanol from LCM follows three step process: pretreatment, cellulose microbrils. The imidazolium cation has been proposed to
enzymatic saccharication and fermentation (Hassan et al., 2013; have hydrophobic interactions with the hydrophobic face of the
Sharma et al., 2015). In lignocellulosic biomass, cellulose is tightly cellulose. The C-2 proton in the imidazolium has been simulated to
bound to lignin and hemicellulose through strong inter/intra- interact as a weak hydrogen bond donor with the cellulose hy-
molecular hydrogen and covalent bonding, which makes it recal- droxyl groups during dissolution and cation acidity has recently
citrant for chemical or biochemical degradation (Brandt et al., 2011; been suggested to be an important parameter for predicting cel-
Sissine, 2007; Virunanon et al., 2013). lulose solubility in certain cases (Brandt et al., 2013).
Pretreatment is the key step to overcome the lignocellulosic The ability of ILs to dissolve and transform cellulose efciently
stringent seal around cellulose and make them susceptible for hy- depends on their physicochemical and solvent properties such as,
drolysis (Haghighi Mood et al., 2013; Hou et al., 2015). Further steps size of cation and anion, KamleteTaft parameters: hydrogen bond
involve isolation and enzymatic hydrolysis of cellulose and hemi- acidity (a), hydrogen bond basicity (b) and polarizability (p*) and
cellulose to generate monomeric sugars followed by microbial viscosity (h) along with surface tension (s). There are few reports
fermentation and distillation to produce fuel ethanol (Bian et al., on the properties of ILs and their impact on cellulose structural
2014; Peleteiro et al., 2014). Over the years, many pretreatment transformation and enzymatic saccharication and are limited to
methods have been intensively studied and are discussed in liter- individual property such as KeT parameters (Parviainen et al.,
ature including chemical (e.g. acid and organosolv process), phys- 2013; Sun et al., 2014; Xia et al., 2014).
icochemical (e.g. steam or carbon dioxide explosion, AFEX), In this study, ve ILs namely; 1-ethyl-3-methylimidazolium
biological (e.g. fungi and actinomycete), and ozonolysis or hot acetate [C2mim][OAc], 1-butyl-3-methylimidazolium acetate
water treatment (Amiri et al., 2014; Jiang et al., 2013; Kapoor et al., [C4mim][OAc], 1-ethyl-3-methylimidazolium chloride [C2mim][Cl],
2015; Kim et al., 2011). Very often these methods require high 1-butyl-3-methylimidazolium chloride [C4mim][Cl] and 1-butyl-3-
temperature and pressure or alternatively strong acids and bases methylimidazolium tetrauoroborate [C4mim][BF4] were analyzed
(Goshadrou et al., 2013). Additionally, these pretreatment processes for their KeT parameters, viscosity and surface tension. We have
result in the formation of microbial inhibiting byproducts such as focused on Avicel PH 101 cellulose as a model substrate which re-
furfural (2-furaldehyde), 5-hydroxymethylfurfural (5-HMF), acetic ects the properties of biomass. After treatment and regeneration
acid along with lignin derived phenolic compounds (4- of cellulose (Avicel PH 101) using all these ILs, the regenerated
hydroxybenzaldehyde, syringaldehyde, etc.) (Bensah and Mensah, cellulose was subjected to enzymatic hydrolysis and the efforts
2013; Haghighi Mood et al., 2013; Kim et al., 2011; Larsson et al., were made to nd the co-relation of the properties of ILs with
1999). Moreover, all these treatment processes result in the envi- enzymatic hydrolysis. Cellulose, thus obtained was characterized
ronmental issues (Goshadrou et al., 2013; Ximenes et al., 2010, by PXRD and FT-IR to nd out structural modications. This is one
2011). To circumvent these drawbacks, ILs are being explored for of the few such studies which will help to choose best IL for
pretreatment of biomass (Dadi et al., 2007). biomass pretreatment, wherein a comprehensive analysis of ILs
ILs are molten salts below 100 C comprising organic cations properties, i.e. KeT parameters, viscosity and surface tension were
and organic or inorganic anions (Hou et al., 2012). ILs have proven conducted under treatment conditions and their impact on the
to be potential, more sustainable and environmentally responsible structure of cellulose and enzymatic hydrolysis was evaluated.
alternatives to organic solvents for many industrial applications
such as: in chemicals synthesis, catalysis, bio catalysis, electro- 2. Material and methods
chemical devices and as engineering uids (Brandt et al., 2013; da
Costa Lopes et al., 2013; Plechkova and Seddon, 2008). These can 2.1. Material
be recycled for consecutive cycles to reduce their high cost and
make the process more practicable (Weerachanchai and Lee, 2014; 1-Ethyl-3-methylimidazolium acetate [C2mim][OAc], (98%), 1-
Xia et al., 2014). Certain imidazolium-based ILs have demonstrated ethyl-3-methylimidazolium chloride [C2mim][Cl], (97%), and 1-
a promising ability for efcient dissolution of biomass and cellulose butyl-3-methylimidazolium acetate [C4mim][OAc], (98%), 1-
regeneration upon addition of an anti-solvent such as water/ butyl-3-methylimidazolium tetrauoroborate [C4mim][BF4],
acetone/ethanol (Brandt et al., 2013; Goshadrou et al., 2013; Xia (98%), 4-Nitroaniline (4NA) (99%) and Reichardt's dye (RD) (dye
et al., 2014). IL pretreatment does not produce degradation prod- content 90%), imidazole (97%), butyl chloride and Avicel (PH
ucts that inhibit enzymes or fermenting microorganisms (Dadi 101) (>98%) were purchased from SigmaeAldrich (India) and used
et al., 2007). The initial hydrolysis rate for regenerated cellulose without any further purication. N,N-Diethyl-4-nitroaniline
obtained after pretreatment with [C4mim][Cl ] was approximately (DENA) (97%), was purchased from Oakwood Chemical Products
50-fold higher as compared to untreated cellulose (Dadi et al., (West Columbia, USA). 1-Butyl-3-methylimidazolium chloride
2007). Li et al. (2010) have reported that, [C2mim][OAc] treatment [C4mim][Cl] was synthesized as per as the synthetic protocol
of switchgrass gave more than 90% of hydrolysis yield after 12 h (Dupont et al., 2003). All experiments were conducted using a
saccharication time as compared to dilute acid treatment, which single lot of Avicel (PH 101). Cellulase enzyme was obtained from
gave 80% of yield after 72 h of saccharication time. Similarly, Singh M/S Advanced Enzymes Technologies Ltd. (Mumbai, India).
et al. (2009) have examined the structural disturbance of switch-
grass after pretreatment with [C2mim][OAc], which enhance the 2.2. Ionic liquid treatment
enzymatic saccharication from 16.5% to 72.5%. In one previous
work, energy cane treated with [C2mim][OAc], resulted in 87.0% of A 10% (w/w) Avicel (PH 101) was prepared by combining 1 g of
enzymatic hydrolysis (Qiu et al., 2012). Table SI 1 shows the Avicel with 9 g of IL in a 50 ml round bottom ask under stirring.
comparative sugar yield from biomass pretreated with different ILs The mixture was then placed in a preheated silicone oil bath on a
(Sant'Ana da Silva et al., 2011; Xiao et al., 2012). hot plate (IKA C-MAG H7 basic, Germany) equipped with a stirrer at
ILs are able to interact with cellulose through a variety of 200e300 rpm for given temperature (100 and 130 C) for desired
different interactions, including dispersive, pep, nep, hydrogen time (5 and 2 h) under sealed conditions, so as to avoid moisture
bonding, dipolar, and ionic/chargeecharge interactions. These in- uptake. All experiments were conducted in triplicate. After treat-
teractions between the basic anion and the cellulose hydroxyl ment, the regeneration of the dissolved cellulose was carried out by
groups helps to break up the hydrogen bond network between the the precipitation of cellulose/IL slurry with 50 ml of hot deionized
T. Raj et al. / Journal of Cleaner Production 113 (2016) 1005e1014 1007
Table 1
Solid recovery and glucose yield % of regenerated cellulose obtained.
T/t Solid recovery (%)a Glucose yield (%)b Overall glucose recoveryc (g/g avicel) Crystallinity index (CrI (%)
Untreated avicel N/A N/A 33.86 0.5 46.19 0.9 0.46 0.01 89.0
[C2mim][OAc] 100/5 93.30 0.5A 83.28 0.9A 88.80 1.0A 0.83 0.02A 67.3
130/2 89.69 0.5A* 89.83 0.8A* 94.48 1.0A* 0.85 0.02A* 57.4
[C4mim][OAc] 100/5 92.23 0.2B 64.92 0.2B 77.77 0.9B 0.72 0.01B 75.3
130/2 86.32 0.9B* 73.87 1.2B* 80.10 0.7B* 0.69 0.03B* 70.6
[C2mim][Cl] 100/5 96.00 0.4C 61.67 0.7B 63.44 1.2C 0.61 0.02C 73.4
130/2 94.74 0.4C* 63.27 1.2C* 71.24 1.3C* 0.67 0.02C* 72.5
[C4mim][Cl] 100/5 97.41 0.1D 45.96 1.5C 50.54 1.0D 0.49 0.01D 75.0
130/2 94.50 0.5C* 50.82 1.0D* 63.44 1.6D* 0.60 0.02D* 73.0
[C4mim][BF4] 100/5 97.83 0.3E 25.53 2.0D 32.72 0.8E 0.32 0.02E 88.5
130/2 95.45 0.3C* 27.33 1.9E* 33.20 1.9E* 0.32 0.03E* 87.8
100 100
130 C/ 2 h
60 60
40 40
20 20
0
0
0 10 20 30 40 50 60 70
0 10 20 30 40 50 60 70
Time (h)
Time (h)
a b
Fig. 1. Time courses for enzymatic hydrolysis and glucose yield (%) of untreated and ILs treated cellulose. Conditions: 100 mg untreated/regenerated cellulose, 0.8 ml of sodium
citrate buffer 0.05 M, 10 FPU/g substrate, 50 C, 150 rpm, time, 72 h.
treatment efciency. Hence, in order to nd out the role of cation were in the order of: [C2mim][OAc] > [C4mim][OAc] > [C2mim]
and anion, we have measured KeT parameters, viscosity and sur- [Cl] > [C4mim][Cl] > [C4mim][BF4]. In a plot of KeT parameters vs.
face tension to nd the correlation of these properties with glucose 1000/temperature (Fig. 2), the slop of [OAc] containing ILs is
yields. In order to differentiate better and for economic aspects, steeper, showing that b values of ILs increases with increase in
lower enzyme loading, i.e. 10 FPU/g substrate was considered for temperature as compared to p* and a values (Fig. 2a, b and SI 2).
further studies and in detailed discussion. Cellulose treatment in ILs with b > 0.72 resulted in more than
55% glucose yield after 24 h of hydrolysis and ILs with b 0.72,
neither decrease cellulose crystallinity (discussed in later part) nor
3.2. Impact of KamleteTaft (KeT) parameters on enzymatic improve glucose yields. For instance, higher hydrogen bond basicity
saccharication of [C2mim][OAc] (b 1.27) gave highest glucose yield (83.3%) as
compared to [C4mim][OAc] (b 1.19), which resulted in 64.9%
KeT solvent parameters have been in past used to measure the glucose yields. Similarly, [C4mim][Cl] with b 1.16 gave 45.9% of
ability of a solvent to donate a hydrogen bond (a), accept a glucose yield whereas, [C4mim][BF4] having lower b value (0.67)
hydrogen bond (b) (Kamlet and Taft, 1976). It has earlier been gave only 25.5% glucose yield at 100 C/5 h. Similar trends were also
shown (Brandt et al., 2010) that basicity (b) correlates well with an observed for 130 C/2 h (Tables 1 and 2). Fig. 3 shows the correla-
IL's ability to dissolve lignocellulose. A recent study on the range of tion between glucose yield and b values concluding that, cellulose
cations in combination with same anion demonstrated that cation treated with ILs with higher b value leads to higher glucose yields.
acidity is also important for cellulose solubility (Parviainen et al., Higher hydrogen bond basicity (b) of [C2mim][OAc] could stabilize
2013). The KeT parameters describes the hydrogen bond acidity the solvation ion causing further higher solvation of cellulose and
(a), hydrogen bond basicity (b) and polarizability (p*) of ILs (Lee lower in case of [C4mim][BF4] due to low b value. In literature,
et al., 2008). KeT parameters of all ILs were measured at 100 and (Crowhurst et al., 2003; Welton, 1999); it is mentioned that
130 C and presented in Table 2. Since, [C2mim][Cl] has a high hydrogen bond basicity (b) of anion attracts hydroxyl protons of
melting point (ca. 77 C), its KeT parameters could not be deter- cellulose, disrupting the crystal lattice and hydrogen bonding
mined. The polarizability (p*) of ILs slightly decreased with an in- interaction of crystalline cellulose.
crease in temperature except for [C4mim][BF4] and was found to be
in the range of 0.88e0.99. With an increasing alkyl chain length
attached to cation, decrease in p* values was observed. [BF4] 3.3. Impact of viscosity (h) on enzymatic saccharication
containing IL are considered as poor solvents due to low acidity and
basicity (Ma ki-Arvela et al., 2010). Hydrogen bond acidity (a) and Kinematic viscosity (h) of ILs could also be an important factor
hydrogen bond basicity (b) were found to increase with increasing which can inuence the glucose yields. Viscosity of all ve ILs was
temperature (Table 2). The b values of ILs at both temperatures analyzed at 100 and 130 C and the data is presented in Table 3. The
Table 2
Solvatochromic KeT parameter of the ILs.
ILs a b p*
100 C 130 C 100 C 130 C 100 C 130 C
A A* A A* A
[C2mim][OAc] 0.72 0.03 0.79 0.02 1.27 0.0 1.32 0.02 0.99 0.01 0.98 0.01A*
[C4mim][OAc] 0.60 0.04B 0.63 0.01B* 1.19 0.0B 1.23 0.01B* 0.95 0.01B 0.88 0.02B*
[C4mim][Cl] 0.72 0.01A 0.77 0.01A* 1.16 0.0B 1.22 0.01B* 0.91 0.01C 0.90 0.02B*
[C2mim][Cl] N/Aa N/Aa N/Aa N/Aa N/Aa N/Aa
[C4mim][BF4] 0.59 0.01B 0.63 0.2B* 0.67 0.0C 0.72 0.01C* 0.99 0.02A 0.99 0.01A*
KeT parameters using the following set of dye: Reichardt's dye, N,N-diethyl-4-nitroaniline and 4-nitroaniline: (a), hydrogen bond acidity, (b), hydrogen bond basicity, (p*),
polarizability. All experiments were done in triplicate and the mean is reported with S.D. Values in the same column for same temperature with different superscript letters
indicate the signicance difference at P 0.05. Letter with star (*) superscript shows statistical signicance at 130 C.
a
N/A means not applicable, KeT parameter for [C2mim][Cl] could not be determined since the melting point of 1-ethyl-3-methyl imidazolium chloride is 77e75 C.
1010 T. Raj et al. / Journal of Cleaner Production 113 (2016) 1005e1014
1.1
[C2mim][OAc] [C2mim][OAc]
[C4mim][OAc] [C4mim][OAc]
[C4mim][Cl]
[C4mim][Cl]
[C4mim][BF4]
[C4mim][BF4]
0.7
2.4 2.6 2.8 3.0 3.2 3.4
1000/T(K) 1000/T(K)
a b
Fig. 2. KamleteTaft parameters of various ILs using the following set of dye: Reichardt's dye, N,N-diethyl-4-nitroaniline and 4-nitroaniline. (a) Hydrogen bond acidity (a), (b)
polarizability (p*).
data shows that, as expected an increase in temperature, a signif- hence almost no solubilization. It can be argued, based on the effect
icant decrease in viscosity of ILs was obtained, i.e. viscosity of of viscosity and b value on glucose yields, b value is preferred than
[C2mim][OAc] at 100 C is 8.4 cSt/s, whereas, 4.4 cSt/s at 130 C. the viscosity when comparing the ILs treatment efcacy. From this
Viscosity of ILs at 100 C and also at 130 C was following the same study it is also observed that, ILs having b > 0.72, with low viscosity
order as: [C2mim][OAc] [C4mim][BF4] < [C4mim][OAc] < [C2mim] results in higher glucose yields.
[Cl] < [C4mim][Cl]. An increase in viscosity of ILs with various
anion/cation combinations may be attributed to the increased of 3.4. Impact of surface tension on enzymatic saccharication
van der Waals forces over hydrogen bonding. Viscosity of ILs was
also increased with increasing alkyl chain length of cation of ILs Surface tension of all the ILs was measured at 25, 50 and 75 C
with same anion. Similar results are reported by Cao et al. (2014) and extrapolated by the linear t to get values for 100 and 130 C
and Huddleston et al. (2001). (Fig. SI 3). As shown in Table 3, the surface tension decreases with
In order to nd any correlation between viscosity of ILs and an increase in temperature. ILs with same anion and cation of
glucose yields, Fig. 4 was drawn, which shows that, the glucose different alkyl chain length, show an increase in surface tension
yield was in reverse order of viscosity. Highest glucose yield (83.2 with increasing alkyl chain length. Coming to the effect of anion,
and 89.8%) was achieved with [C2mim][OAc], having a lower vis- comparing the surface tension of ILs with butyl substituted on
cosity (8.3 and 4.4 cSt/s) at 100 and 130 C respectively. Similarly, cation the order is [OAc] > [Cl] > [BF4].
[C4mim][Cl] and [C2mim][Cl] showed highest viscosity, i.e. 25.71 Glucose yields after enzymatic saccharication of ILs treated and
and 21.71 cSt/s at 100 C leading to 45.9 and 61.7% glucose yields regenerated cellulose was drawn against the surface tension in
respectively. This is in consistent with the previous results (Ma ki- Fig. 5 and it showed negative correlation, i.e. ILs with lower surface
Arvela et al., 2010) that, ILs with lower viscosity can dissolve tension gave higher glucose yields except that of [C4mim][BF4].
more amount of cellulose. However, [C4mim][BF4] having a very [C2mim][OAc] with low surface tension, i.e. 32.5 mN/m gave 83.3%,
low viscosity 8.2 and 4.5 cSt/s at 100 and 130 C does not improve whereas, [C4mim][OAc] (37.3 mN/m) gave 64.9% of glucose yield at
the enzymatic hydrolysis. This may be attributed to the low 100 C/5 h. Similarly, [C2mim][Cl] with a surface tension of
hydrogen bond basicity (b < 0.72), [BF4] being non coordinating 42.4 mN/m gave 61.7% and [C4mim][Cl] (45.1 mN/m) gave 45.9%
anion, which can not interrupt the hydrogen bonding in cellulose glucose yield at 100 C/5 h. Similar trends of surface tension vis-
a-vis glucose yields were observed at 130 C/2 h. However, surface
tension of [C4mim][BF4] is 41.7 mN/m, lower than [C4mim][Cl]
(45.17 mN/m), which gave poor glucose yield (25.5%). It may be
1.8
90
1.6
80 Table 3
Viscosity and surface tension of different ionic liquids.
Glucose yield (%) after 72 h with
70 1.4 ILs Kinematic viscosity (h, cSt/s)a Surface tension (s, mN/m)b
10 FPU/g substrate
Beta value ()
100 100 order of glucose yields, thus anion with small size gives higher
Glucose yield (%) (130 C/ 2h) yields in enzymatic hydrolysis. [C2mim][OAc] and [C4mim][OAc]
90 Glucose yield (%) (100 C/ 5h)
Viscosity (cSt/ sec) (130 C/ 2h)
with same anion, i.e. [OAc] with different cation, i.e. [C2mim] and
80 Viscosity (cSt/ sec) (130 C/ 5h) 80 [C4mim] results in 89.8 and 73.9% glucose yields after 72 h of
enzymatic hydrolysis respectively. Similarly, [C4mim][OAc],
Glucose yield (%) after 72 h with
60 60
50
27.3% of glucose yields respectively at 130 C/2 h. Thus, [C2mim]
[OAc] with the best combination of smaller size anion and smaller
40 40 chain length on cation is quite powerful in cellulose structural
30 modication leading to highest glucose yields in enzymatic hy-
drolysis. The size of cation or anion is connected to its degree of
20 20 interaction with cellulose chain and disrupting inter and intra-
10 molecular hydrogen bonding in cellulose causing higher glucose
yields.
0 0
[C2mim][OAc] [C4mim][OAc] [C2mim][Cl] C4mim][Cl] [C4mim][BF4]
Fig. 4. Effect of viscosity of ILs and correlation with glucose yield (%) of the treated 3.6. Structural changes in treated cellulose
cellulose at 100 C/5 h and 130 C/2 h.
PXRD and FT-IR analysis of native and treated cellulose was
carried out to determine the structural transformations with
attributed to the low hydrogen bond basicity (b 0.72) of IL. respect to treatment temperature, time and ILs. Native cellulose
Therefore, it is clear from the above discussion that, surface tension showed a characteristic peak of cellulose I polymorph at 2q 15.2
and viscosity, both are showing negative correlation with glucose (101) plane and 22.6 (002) plane. Peaks at 2q 15.2 (101), 16.4
yields and a positive correlation is obtained between natural log- 101 and 22.6 (002) are characteristic of cellulose I. Peaks at 12.1
arithms of surface tension and viscosity as given in Fig. SI 4 (Singh (110), 20.0 (021) plane represents amorphous cellulose, herein
and Singh, 2011). From the above data it can be noticed that, the called as cellulose II. After ILs treatment at 100 C/5 h, [C2mim]
viscosity and surface tension has a signicant impact on glucose [OAc] and [C4mim][OAc] displayed the reduction in CrI, which may
yield if the variation is induced either in anion or cation. And a be due to conversion of cellulose I to cellulose II crystal structure as
possible explanation for the efcacy of ILs with lower surface ten- shown in Fig. 6. Cellulose treated with [C2mim][OAc] at 130 C/2 h
sion is that, liquids with lower surface tension has low surface has the lowest CrI (57.4%), compared with [C4mim][OAc] (70.6),
forces and could easily penetrate into the material resulting in more [C2mim][Cl] (72.5), [C4mim][Cl] (73.0), [C4mim][BF4] (87.8) and
interaction with cellulose hydroxyls. native cellulose (89.0%) as well. This decrease in CrI clearly shows
the transformation of cellulose I to cellulose II. Similar trends were
observed at 100 C/5 h, although the crystallinity index is higher at
3.5. Effect of size of cation and anion 100 C/5 h. Cellulose treated with [C4mim][Cl] and [C4mim][BF4]
retains the crystalline cellulose I structure exhibiting high CrI value,
The impact of different anion [OAc], [Cl], [BF4] and cations i.e. 73.0 and 87.8%, even at 130 C/2 h (Table 1), showing no
having two different alkyl chain lengths, [C2mim] and [C4mim] structural changes and hence, resulted to lower enzymatic hydro-
were analyzed to nd out the impact on glucose yields. Results of lysis (Fig. 7). Decrease in the X-ray peak intensity with the shift of
enzymatic saccharication of regenerated cellulose (Table 1) indi- the peak position to the left is observed with an increase in the
cate that, with increasing alkyl chain length of cation the glucose treatment temperature. For cellulose treated at 130 C/2 h, the in-
yields decreases. The size of anion of ve ILs follows the order of tensity at 22.6 decreases signicantly and disappeared at 15.2 and
[OAc] < [Cl] < [BF4] (Hou et al., 2015); which is reverse to the 16.4 and appearing a peak at 20.0 (110) plane for cellulose II.
Cellulose treated with [C2mim][OAc] at 130 C/2 h shows highly
amorphous nature with a broad peak around 21.4 assign to the
50 (I002) plane of cellulose II lattice with peaks at ~15.2 , 21.5 . It may
90 be argued that, ILs treatment improves the enzymatic hydrolysis by
transformation of cellulose I to cellulose II which is amorphous in
80 nature. Similar observation for cellulose I and II is also reported in
literature (Cheng et al., 2011).
Glucose yield (%) after 72 h
with 10 FPU/g of substrate
(101) (101)
(021) (021)
Intensity (a.u.)
Intensity (a.u.)
[C mim][BF ]
[C mim][BF ]
[C mim][Cl] [C mim][Cl]
[C mim][Cl] [C mim][Cl]
[C mim][OAc] [C mim][OAc]
[C mim][OAc]
[C mim][OAc]
5 10 15 20 25 30 35 40
5 10 15 20 25 30 35 40
Diffraction angle (2 )
Diffraction angle (2)
Fig. 6. PXRD diffraction patterns for untreated and ILs treated cellulose treated by ve ILs at 100 C/5 h and 130 C/2 h.
FT-IR spectrum is given in Fig. 8 for untreated and IL treated modication, hence, only 27.3% of glucose yield was achieved.
cellulose. A strong absorption band at 3400 cm1 is assigned to However, the absorption band at 898 cm1 assigned to CeOeC
different OeH stretching modes due to inter and intramolecular stretching at the b-(1 / 4) glycosidic linkage is weak and broad in
hydrogen bonding. This band is very intense in untreated cellulose cellulose I but strong and sharp in cellulose II. Therefore, infrared
and the intensity of these bands decreases with ILs treatment and spectra in the range of 850e1500 cm1 were characteristic of cel-
regeneration. Two bands at 2920 and 2850 cm1 are related to lulose samples regenerated from different treatments.
asymmetric and symmetric methyl and methylene stretching. The It was observed that the untreated cellulose possesses cellulose I
overlapping features from 1480 to 1290 cm1 region consist of a crystal type and transformed into cellulose II crystal structure after
number of bands such as 1419, 1328 and 1312 cm1 mainly due to the treatment and regeneration of cellulose. The absorption bands
various deformation modes of CeH bonds in the methylene groups. at 1367 and 2900 cm1 are for CeH and CH2 stretching, respectively
The absorption band at 1419 cm1 assigned to the CH2 scissoring and the band at 1631 cm1 relates to the bending mode of the
motion was strong in untreated cellulose and is the characteristic water. The peak at 1382 cm1 is attributed to the OeH bending and
peak of crystalline cellulose I. This band is reduced in [C2mim][OAc] absorption band at 1122 and 1161 cm1 corresponds to CeO anti-
and [C4mim][OAc] treatment due to structural transformation from symmetric bridge stretching. A strong peak at 1082 and 1033 cm1
cellulose I to cellulose II. This conrms the reduction in the crys- arises from CeOeC pyranose ring skeletal vibration. Similar
tallinity after ILs treatment of cellulose, hence resulting increase in observation is also reported by Goshadrou et al. (2013) and Kapoor
glucose yield. As discussed above, [C2mim][OAc] and [C4mim][OAc] et al. (2015).
ILs, having higher b value, i.e. 1.27, 1.19 causes higher trans-
formation of cellulose than [C4mim][Cl] and [C4mim][BF4]. The 4. Conclusion
intensities at 3400, 2887, 1328, 1122, 1082, 1033 cm1 peaks get
reduced after treatment resulting to produce 89.8 and 73.9% Cellulose transformation efciencies, for improving the enzyme
glucose yield respectively (Table 1). The intensity of these peaks hydrolysis by ve ILs, i.e. [C2mim][OAc], [C4mim][OAc], [C2mim]
after [C4mim][BF4] treatment remains same as in the case of native, [Cl], [C4mim][Cl] and [C4mim][BF4] were investigated and a corre-
which conrms that [C4mim][BF4] do not causes cellulose structure lation between KeT parameters including hydrogen bond acidity,
hydrogen bond basicity and polarity, viscosity and surface tension
and their glucose yields was established. Among the KeT param-
eters, b value stands as the best predictor and rst parameter needs
100 100 to be considered while checking ILs treatment efciency. [OAc]
anion based ILs with b > 0.72 causes in the maximum reduction of
90
CrI and resulted in 89.7% glucose yield after 72 h of cellulase hy-
80 drolysis and ILs with b 0.72 like [C4mim][BF4] neither induced
80
CrI % of ILs treated cellulose
Glucose yield (%) after 72 h with
100 C/ 5h 130 C/ 2h
Untreated avicel
Untreated avicel
[C mim][BF ]
Absorbance
Absorbance
[C mim][BF ]
[C mim][Cl] [C mim][Cl]
[C mim][Cl] [C mim][Cl]
[C mim][OAc] [C mim][OAc]
[C mim][OAc] [C mim][OAc]
4000 3500 3000 2500 2000 1500 1000 500 4000 3500 3000 2500 2000 1500 1000 500
Wavenumber (cm ) Wavenumber (cm )
a b
Fig. 8. FT-IR spectrum of untreated and regenerated cellulose treated by ve ILs at 100 C/5 h and 130 C/2 h.
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