Ultrasonics Sonochemistry 21 (2014) 216–225

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Ultrasonics Sonochemistry
journal homepage: www.elsevier.com/locate/ultson

Alkaline and ultrasound assisted alkaline pretreatment for

intensification of delignification process from sustainable raw-material
Preeti B. Subhedar, Parag R. Gogate ⇑
Chemical Engineering Department, Institute of Chemical Technology, Matunga, Mumbai 400 019, India

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

Article history: Alkaline and ultrasound-assisted alkaline pretreatment under mild operating conditions have been inves-
Received 27 May 2013 tigated for intensification of delignification. The effect of NaOH concentration, biomass loading, temper-
Received in revised form 31 July 2013 ature, ultrasonic power and duty cycle on the delignification has been studied. Most favorable conditions
Accepted 5 August 2013
for only alkaline pretreatment were alkali concentration of 1.75 N, solid loading of 0.8% (w/v), tempera-
Available online 13 August 2013
ture of 353 K and pretreatment time of 6 h and under these conditions, 40.2% delignification was
obtained. In case of ultrasound-assisted alkaline approach, most favorable conditions obtained were
Keywords:
alkali concentration of 1 N, paper loading of 0.5% (w/v), sonication power of 100 W, duty cycle of 80%
Alkali
Ultrasound
and pretreatment time of 70 min and the delignification obtained in ultrasound-assisted alkaline
Pretreatment approach under these conditions was 80%. The material samples were characterized by FTIR, SEM, XRD
Delignification and TGA technique. The lignin was recovered from solution by precipitation method and was character-
Sustainable raw material ized by FTIR, GPC and TGA technique.
Ó 2013 Elsevier B.V. All rights reserved.

1. Introduction matrix and open or moderately break up the recalcitrant structure.

The instability of oil prices and the imminent depletion in the
traditional fossil fuels, have enforced a number of countries to in-
vest a great deal into research and development with an objective
of finding an alternative energy source. Plant biomass has come
forward as one of the important source of alternative energy, as
plant biomass can be utilized in both physico-chemical and biolog-
ical conversion into several valuable products [1]. There is also a
considerable interest in developing renewable energy resources
to replace crude oil. Production of ethanol can be carried out using
inexpensive and plentiful lignocellulosic biomass such as agricul-
tural and forestry residues, wastepaper, a considerable portion of
municipal solid waste, and woody and herbaceous energy crops.
However, even though lignocellulosic material provides an eco-
nomical resource, the method of conversion to ethanol is very com-
plicated [2]. As lignocellulosic biomass is complex and strongly
resistant to enzymatic hydrolysis due to its recalcitrance, pretreat-
ment is needed in order to increase the hydrolysis rates. Various
pretreatment methods have been reported for the delignification
of biomass including steam explosion, acid, and alkaline treatment
[3]. Alkaline pretreatment is one of the most commonly applied
processes among the different proposed pretreatment methods
for delignification of biomass. The intention of pretreatment is to
generate reactive cellulose materials from the lignocellulosic
⇑ Corresponding author. Tel.: +91 22 33612024; fax: +91 22 33611020.
E-mail address: pr.gogate@ictmumbai.edu.in (P.R. Gogate).
Subsequently, biomass can be more readily hydrolyzed by cellulase
enzyme, and converted into monosaccharides, primarily glucose.
Pretreatment is one of the most costly steps in biochemical conver-
sion of lignocellulosic biomass, accounting for up to 40% of the to-
tal processing cost [4]. Furthermore, it strongly affects the
downstream processing and associated costs of operation includ-
ing enzymatic hydrolysis.
Pretreatment of lignocellulosic biomass with alkali such as
NaOH can eradicate or modify the lignin content by rupturing
the ester bonds that form cross links involving xylan and lignin,
thus increasing the porosity of the lignocelluloses [5]. Alkaline pre-
treatments show less sugar degradation and furan derivatives for-
mation. The alkaline pretreatment is very complex involving
numerous reactive and non reactive phenomenons such as dissolu-
tion of non degraded polysaccharides, peeling off reaction (forma-
tion of alkali stable end groups), decomposition of the dissolved
polysaccharides and hydrolysis of glycosidic bonds. Accordingly,
the efficiency of NaOH pretreatment depends considerably on the
operating parameters such as temperature, concentration of NaOH,
treatment time in addition to the inbuilt characteristics of the bio-
mass used [6].
The alkaline pretreatment process can be improved further by
the application of ultrasound [7]. The present study conjugates
alkaline pretreatment with ultrasound irradiation. The ultrasonic
treatment of aqueous media produces cavitation, which generates
conditions of high temperature, pressure and extreme shear forces.
The decomposition of water molecules into free radicals by

1350-4177/$ - see front matter Ó 2013 Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.ultsonch.2013.08.001
 P.B. Subhedar, P.R. Gogate / Ultrasonics Sonochemistry 21 (2014) 216–225
cavitation aids in cleaving the linkages in lignin and xylan networks
[8]. Ultrasound irradiation causes homolysis of lignin–carbohy-
drate bonds releasing lignin [9]. The application of ultrasound for
delignification of lignocellulosic matrix is still in emerging stage.
Ultrasound assisted fractionation of the lignocellulosic biomass im-
proves the effectiveness of the classical treatments such as alkaline
treatment, giving proportionately higher yield and selectivity of the
obtained products [10]. Ultrasound treatment shortens the process-
ing time as well as reduces the requirement of alkali [11]. The main
limitations of the alkaline pretreatment of lignocellulosic biomass
include relatively longer pretreatment time and consumption of
more alkali. Waste paper is an abundant and low cost substrate
for production of glucose by enzymatic hydrolysis. On an average,
waste paper constitutes half a municipal solid waste, and newspa-
per alone 14% of the waste [12]. The extensive use of papers has
produced a huge amount of waste paper, however, it is difficult to
recycle this sustainable resource because of the high processing
cost. Presently, majority of waste paper is land filled or incinerated;
but, both of these methods are of increasing environmental con-
cern. Though European recycling ratio of paper is comparatively
high i.e. 66%, still it has not reached an adequate level yet. Addition-
ally, when paper materials are recycled, they are typically turned
into lower grade paper products. By further recycling of paper, fiber
length in the paper becomes shorter and this shortening of paper fi-
bers decreases the quality of paper. This means that a major fraction
of paper would always be sent to disposal. This fraction contains a
considerable and underutilized source of sugar [13]. Thus, newspa-
per as a substrate has been selected as representative source of
waste lignocellulose. Until now no work has been carried out to
intensify the alkaline pretreatment of sustainable lignocellulosic
biomass such as newspaper. In the present work, ultrasound-as-
sisted alkaline approach has been studied for the delignification
of waste newspaper as a sustainable raw material.
Breakthroughs in pretreatment are still required for both scien-
tific as well as economic reasons. More exhaustive analysis of fac-
tors influencing delignification is required to establish design
related information. The aim of the present study was to investi-
gate most favourable biomass loading, NaOH concentration and
sonication time, sonication power, duty cycle so as to maximize
the ultrasound-assisted alkaline pretreatment with the intention
to improve the delignification. The overall purpose of this research
was to develop an efficient ultrasound-assisted alkaline pretreat-
ment method for the final possible bioconversion of lignocellulo-
sics into bioethanol.
2. Materials and methods
2.1. Materials
Newspaper as a substrate was selected as a lignocellulosic bio-
mass for the pretreatment and was collected from a local supplier.
Its composition based on the dry substrate was: 41.02% cellulose,
24.85% hemicellulose, 23.07% lignin, 5.99% ash and 3.89% moisture.
The composition was determined by using standard method de-
scribed by NREL. The collected newspaper was air dried for 2 h at
105 °C and mechanically grinded using electric mixer grinder in dis-
tilled water. The newspaper used for the experiments was in the
form of slurry. Standard of lignin was obtained as a gift sample from
Waltar Enterprises, Mumbai. Acetyl bromide was procured from
Spectrochem Pvt. Ltd., Mumbai. All other reagents and chemicals
were procured from S.D. Fine chemicals Pvt. Ltd., Mumbai.
2.2. Alkaline pretreatment
A fully baffled 2000 cm3 jacketed glass reactor was used for the
delignification reaction. The temperature of reaction was
217
controlled by circulating water. A uniform suspension of newspa-
per (0.8% w/v) in distilled water was transferred to the reactor fol-
lowed by addition of alkali (NaOH) solution. Immediately after the
addition of the alkali solution, a sample was withdrawn and agita-
tion was started. Samples were frequently withdrawn at regular
time intervals and immediately centrifuged at 10,000g for 20 min
to remove the residual particles. The clear supernatant obtained
was used for measuring the lignin concentration.
2.3. Ultrasound-assisted alkaline pretreatment
A three neck fully baffled, jacketed glass reactor equipped with
mechanical stirrer was used for the reaction. Ultrasonic device was
a probe sonicator obtained from Dakshin Ultrasonics, Mumbai. The
ultrasonic irradiation at a frequency of 20 kHz was transferred
through a titanium cylindrical horn, introduced into the reactor
through the side neck and submerged 1.0 cm into the reaction mix-
ture. The horn had a diameter of 1.1 cm. The experimental setup is
shown in Fig. 1. The procedure followed for the delignification
studies was similar to that mentioned in alkaline pretreatment.
Integral method of analysis was followed in the present work to
calculate the rate constants and it has been observed that first or-
der kinetics explains the delignification kinetics very well. All the
experiments were repeated two times to check the reproducibility
and average values have been reported in the figures. Error bars
have also been shown to illustrate the variation.
2.4. Recovery of lignin
For the recovery of lignin, sample was irradiated with ultra-
sound for delignification using horn at power of 100 W, duty cycle
of 80% and using 1 N NaOH solution at room temperature. The
hemicelluloses were isolated from the sample by precipitation of
the acidified hydrolysate (pH 5.5 adjusted with 10% HCl) with 3
volumes of 95% ethanol. The pellet rich in hemicelluloses was fil-
tered, washed with 70% ethanol and air dried. After evaporation
of ethanol, the alkali soluble lignin was obtained by precipitation
at pH 1.5 adjusted by 10% HCl. The isolated lignin was washed with
acidic water (pH 2.0) and air dried. The residue rich in cellulose
was washed with water and then dried at 105 °C for 6 h [14].
2.5. Analysis
The lignin concentration in solution after reaction was analyzed
by UV Spectrophotometer. Samples were solubilized in a solution
Generator
Horn
Agitator
Reactor
Water out
Drain Water in
Fig. 1. Experimental setup for ultrasound-assisted alkaline pretreatment.
218 P.B. Subhedar, P.R. Gogate / Ultrasonics Sonochemistry 21 (2014) 216–225
of 25% acetyl bromide in glacial acetic acid. Sample tubes were
tightly capped and kept in water bath at 50 °C for 30 min. After
cooling the samples, 2.5 ml glacial acetic acid, 1.5 ml 0.3 M NaOH
and 0.5 ml 0.5 M hydroxylamine hydrochloride solution was added
to all the tubes. Absorbance was measured at 330 nm [15]. The
morphological structure of untreated and pretreated newspaper
was also examined using JSM-7600F Field Emission Gun – Scan-
ning Electron Microscope (JEOL Ltd., Tokyo, Japan) at voltage of
3 kV. A thin layer of the sample was mounted on the copper sample
holder, using a both sided carbon tape and coated with gold of
10 nm thicknesses via an electron beam evaporation system to
make the samples conductive.
FTIR spectra of untreated and pretreated newspaper and recov-
ered lignin were recorded on a Bruker Vertex 80 Vacuum FTIR
spectrometer. Data were collected from 500 to 4000 cm1 wave-
length range with 256 scans for each sample at 4 cm1 resolution.
About 2 mg samples were prepared by mixing with 120 mg of
spectroscopic grade KBr followed by pressing in a standard device
using a pressure of 6000 psi to produce 13 mm diameter pellets.
The background spectrum of pure potassium bromide was sub-
tracted from that of the sample spectrum. X-ray diffraction pat-
terns were collected on a powder X-ray diffractometer (D8
Advance Bruker), with the operating conditions set at 40 mA and
40 mV. It was run over a 2h range of 5–30° at 0.02° step intervals,
with a step time of 0.4 s [16]. Crystallinity index (CrI) is defined as
the percentage of crystallinity material in the biomass, and was
calculated with Eq. (1) [17].
I002  Iam
CrIð%Þ ¼  100%
I002
where I002 is the maximum intensity of 0 0 2 peak at 2h = 22.5° and
Iam is the intensity at 2h = 18.7°.
The molecular weight of acetylated lignin was obtained by gel
permeation chromatography (GPC). The effluent was monitored
with a Waters RI2414 RI-detector. The temperature was set at
35 °C during testing, and the injection volume was 100 ll. The col-
umn was calibrated using monodisperse polystyrene standards in
the range of 370–815,000 g/mol. The tetrahydrofuran (THF) flow
was kept as 1 ml/min and the sample was dissolved in THF at a
concentration of 1 mg/ml. The sample was stored for 24 h at 5 °C
to avoid any variation in the molecular weight. The signal detected
was digitalized at a frequency of 2 Hz, and the molecular weight
distribution (MWD) was calculated from the recorded signal using
normal GPC calculation procedures.
The thermal characterization of newspaper was carried out
using a thermo-gravimetric analyzer (TGA). TGA was used to
examine the thermal decomposition behavior of the untreated
and pretreated newspaper. This analysis records the weight of
sample in dynamic conditions in a programmed temperature range
of 20 and 600 °C at a constant heating rate of 10 °C min1 under an
inert atmosphere of high quality nitrogen (99.5% nitrogen, 0.5%
oxygen content). A sample of 8 ± 1 mg was used for the analysis.
The experimental results were achieved using a Thermax700 Ther-
mo Scientific TGA analyzer. The variation in the mass of the sample
allows drawing the TG (variation in the mass as a function of the
temperature) thermograms.
3. Results and discussion
3.1. Effect of alkali concentration
Generally NaOH pretreatment processes are classified as ‘‘high
concentration and low concentration’’ processes based on the load-
ing used in the process. In low concentration processes, typically
0.1–1 N NaOH at high temperature is used. In this case, reactive
destruction of lignocellulosic biomass occurs. In high concentra-
tion pretreatment, usually 1.5–5 N NaOH at low temperature is
used [18]. In this case, along with lignin, cellulose dissolution in
the alkali solution may also occur. In the present study, newspaper
was treated with different alkali concentrations ranging from 0.25
to 2.25 N NaOH solution. Although many factors contribute to alka-
line pretreatment, favourable alkali concentration should be estab-
lished in order to accomplish effective pretreatment with reduced
load on waste treatment and decreased working capital invest-
ment. The effect of different alkali concentration on delignification
of newspaper has been shown in Fig. 2a. The results from the pres-
ent study showed that as alkali concentration increased, extent of
delignification also increased. The effect of alkali concentration on
the rate of delignification of newspaper is shown in Table 1. It has
been observed that the extent of delignification as well as the first
order kinetic rate constant increased with an increase in the alkali
concentration indicating that higher alkalinity is favorable for the
delignification. The kinetic rate constant increased from 0.0179
to 0.1047 h1 for an increase in alkali concentration from 0.25 to
2.25 N. The change in the extent of delignification was marginal
for alkali concentration change from 1.75 N to 2.25 N and hence
1.75 N alkali concentration has been used for further studies to
get maximum benefit. Lignin degradation during alkaline pretreat-
ment occurs due to the breakage of aryl ether linkages which con-
stitute approximately 50–70% of total linkages found in the lignin.
Higher rate of delignification with an increase in alkali concentra-
tion can be attributed to the fact that hydroxyl ions catalyze the
cleavage of ether linkages in the lignin and thus liberate the soluble
sodium phenolates in the liquid [19]. The breakage of these bonds
ð1Þ
increases the hydrophilicity of lignin in the solution. High alkalin-
ity of the NaOH reagent causes solvation of hydroxyl groups in car-
bohydrates and creates the swelling effect in sugar residue. This
further increases the diffusivity of the reagent through the
capillaries.
The effect of alkali concentration in the presence of ultrasound
has been shown in Fig. 2b. It can be seen from Fig. 2b that lower
alkali concentration is required in ultrasound-assisted alkali pre-
treatment for similar extent of delignification. Significant increase
in delignification was observed as alkali concentration was in-
creased from 0.25 to 1 N, and beyond that only a marginal increase
in delignification was obtained. Thus 1 N alkali concentration has
been selected at which 80% delignification was achieved. Table 2
shows the effect of alkali concentration on rate of delignification
of newspaper in the case of ultrasound-assisted process. The ki-
netic rate constant increased from 0.0034 to 0.0265 min1 as alkali
concentration increased from 0.25 to 1.25 N. Ultrasound has been
found to be useful in increasing the yield and mass transfer [20].
The increase in yield and mass transfer occurs due to acoustic
0.25 N
0.75 N
1.25 N
Delignification (%)
1.75 N
2.25 N
Time (hr)
Fig. 2a. Effect of alkali concentration on delignification of newspaper.
 P.B. Subhedar, P.R. Gogate / Ultrasonics Sonochemistry 21 (2014) 216–225 219

Table 1
Effect of alkali concentration on rate of delignification. 0.2% Loading
0.4% Loading
Rate constant k (h1) R2

Delignification (%)
Alkali concentration (N) Delignification (%)
0.6% Loading
0.25 10.1 0.0179 0.99 0.8% Loading
0.75 21.84 0.0425 0.99 1.0% Loading
1.25 32.98 0.0707 0.98
1.75 40.21 0.0915 0.97
2.25 44.4 0.1047 0.97

0.25 N
0.50 N
0.75 N
Delignification (%)

Time (hours)
1.00 N
1.25 N Fig. 3a. Effect of paper loading on delignification of newspaper.

Time (min)
Fig. 2b. Effect of alkali concentration on delignification of newspaper in ultra-
sound-assisted process.
cavitation. As ultrasonic waves propagate through the liquid med-
ium, expansion and compression cycles are created in liquid med-
ium. During expansion cycles, formed bubbles absorb energy and
grow in size where as in compression cycle, bubbles gets collapsed
which result in an increase in the local temperature and pressure
[21]. Cavitation produces turbulence and liquid circulation (micro
streaming) which results in significant increase in mass transfer
rates resulting in increased delignification extent.
3.2. Effect of loading
Generally, the favourable loading depends on the type of bio-
mass as well as its recalcitrance. The effect of paper loading on del-
ignification reaction was statistically significant. From Fig. 3a, it
can be seen that, on increasing paper loading from 0.2% to 0.8%
(w/v), extent of delignification increased from 9.13% to 40.16%.
However, when the loading was further increased to 1.0% (w/v),
the delignification obtained was marginally reduced to 38.91%.
The obtained results can be attributed to the fact that too high con-
centration of solids in slurry adversely affected the treatment as
the slurry became too difficult to mix. Thus 0.8% (w/v) was found
to be the most favorable paper loading for the present system.
The effect of paper loading on rate of delignification has been
shown in Table 3. Kinetic studies revealed that the first order del-
ignification rate constant increased from 0.0656 to 0.00915 h1
with an increase in the loading from 0.2% to 0.8% (w/v).
In ultrasound-assisted alkaline pretreatment, lower solid load-
ing was required as compared to that required in the absence of
ultrasound. From Fig. 3b, it is clear that, only 0.5% paper loading
was required for 79.8% delignification in the presence of ultra-
sound. In the conventional pretreatment (without ultrasound),
only 40% delignification was obtained using 0.7% loading. The ef-
fect of paper loading on rate of delignification in the case of ultra-
sound-assisted approach has been shown in Table 4. At most
favorable loading of 0.5%, 79.8% delignification was obtained with
kinetic rate constant of 0.0245 min1. Apart from cavitation,
mechanical and thermal energy associated with ultrasound (vibra-
tion and stirring) causes the reduction in particle size and
enhances the mass transfer across the material. The mechanical ef-
fect of ultrasound also increases the contact surface area between
solid and liquid phases due to size reduction of the solid matrix
[22].
3.3. Effect of temperature
The trend observed for delignification with increase in temper-
ature was similar to the previous figures. As temperature of the
reaction increased from 313 to 373 K, delignification extent of
newspaper increased from 16% to 44.02%. But temperature of
353 K was selected as most favorable temperature for the deligni-
fication reaction, because further increase in temperature leads to
the production of inhibitory chemicals and degradation of carbo-
hydrates which ultimately affects the sugar yield. So although tem-
perature plays an important role in deciding the extent of
delignification, it also accelerates polysaccharides degradation
and hence most favorable temperature needs to be selected [23].
The effect of temperature on rate of delignification is shown in Ta-
ble 5. If we assume that the effect of temperature on delignification
follows Arrhenius rule, the activation energy of this delignification
reaction can be determined with the help of following expression
Ea
lnðkÞ ¼ lnðk0 Þ  ð2Þ
RT
where k0 is the pre-exponential factor and R is the molar gas con-
stant, which is equal to 8.314 J/(mol.K).
For determining the kinetic parameters, Arrhenius plot was
made based on rate constants obtained at various temperatures.

Table 2
Effect of alkali concentration on rate of delignification in ultrasound-assisted process. Table 3
1 2
Effect of paper loading on rate of delignification.
Alkali concentration Delignification Rate constant k (min ) R
(N) (%) Paper loading (%) Delignification (%) Rate constant k (h1) R2
0.25 20.78 0.0034 0.99 0.2 31.08 0.0656 0.99
0.50 46.86 0.0095 0.99 0.4 34.2 0.0735 0.99
0.75 72.95 0.0187 0.97 0.6 37.1 0.0806 0.98
1.00 80 0.0249 0.97 0.8 40.16 0.091 0.97
1.25 81.7 0.0265 0.97 1 38.91 0.0885 0.97
220 P.B. Subhedar, P.R. Gogate / Ultrasonics Sonochemistry 21 (2014) 216–225

rate of delignification as the economic aspects are always crucial

0.2% Loading
0.3% Loading making determination of most favorable power dissipation levels
0.4% Loading very important. As power of sonication increased, the extent of del-
Delignification (%)

0.5% Loading
0.6% Loading ignification also increased till 100 W in 70 min of treatment.
0.7% Loading Table 6 shows the effect of ultrasoinc power on the rate of deligni-
fication. As power increased from 60 to 100 W, the delignification
rate constatnt increased from 0.0081 to 0.0239 min1. Beyond
100 W i.e. for 110 W, rate constant increased marginally and hence
100 W was taken as most favourable power with 80.16% delignifi-
cation. Overall a 2-fold increase in the extent of delignification was
obtained with use of ultrasound as compared to only alkaline pre-
treatment. Sun et al. [25] obtained 78.5% delignification of wheat
straw by using ultrasound at 100 W power and 20 kHz frequency,
Time (min) which is in well agreement with the results obtained in the present
study. The ultrasonic power dissipation influences the number of
Fig. 3b. Effect of paper loading on delignification of newspaper in ultrasound-
assisted process. cavitation bubbles formed, their lifetime and the generated cavita-
tion intensity. Cavitational effects may be affected adversely at
higher power level. Number of bubbles may form near the tip of
Table 4 ultrasound transducer at a higher power level, which can obstruct
Effect of paper loading on rate of delignification in ultrasound-assisted process. the transfer of energy from the transducer to liquid medium [26].
Paper loading (%) Delignification (%) Rate constant k (min1) R2 In a specified sonication treatment, the power of sonication and
the processing time required are usually inversely related [27];
0.2 40.21 0.0075 0.99
0.3 55.65 0.0122 0.98 whereas sonication power and yield are directly dependent. Hence,
0.4 72.93 0.0199 0.97 increasing the sonication power till most favourable value would
0.5 79.82 0.0245 0.98 in general cut down the irradiation period required for a specified
0.6 81.73 0.0258 0.98 treatment objective and also increase the yield.
0.7 83.6 0.0281 0.97

3.5. Effect of duty cycle

Table 5 Ultrasonic duty cycle is another important operating parameter

Effect of temperature on rate of delignification.
along with the period of treatment. Duty cycle is the fraction of
Temperature (K) Delignification (%) Rate constant k (h1) R2 time during which an ultrasonic system is operated. A duty cycle
313 16.03 0.0299 0.99 of 10%, for example, is equivalent to sonication for 1 s followed
323 23.69 0.0463 0.99 by a rest period (no sonication) of 9 s [28]. The studies were per-
333 30.76 0.0656 0.98 formed using 40%, 50%, 60%, 70% and 80% duty cycles. A 70% duty
343 36.14 0.08 0.97
cycle appeared to be most favorable. With this cycle, a final delig-
353 40.32 0.0915 0.96
363 42.93 0.1008 0.96
nification of 80.0% was obtained, which was nearly 2.0-fold that
373 44.02 0.1054 0.96 obtained in the absence of ultrasound. Effect of duty cycle on rate
of delignification is shown in Table 6. The rate constant for deligni-
According to the slope of the line presented in Fig. 4, the estimated fication was 0.0242 min1 at most favorable duty cycle of 70%.
activation energy of delignification for newspaper was found to be Ultrasound can be applied in pulse mode and continuous mode.
22 kJ/mol. Huang et al. [24] have reported the activation energy for Application of ultrasound in continuous mode creates problems
delignification reaction of wheat straw with caustic potash-aque- such as erosion of tip as well as it is energy inefficient as compared
ous ammonia solutions as 29.75 kJ/mol, which is in well agreement to pulsed mode. Pan et al. [29] have reported that operating the
with the obtained result in present study. system in both pulsed as well as continuous mode at a specific
intensity gave similar yield. Hence, it is established from this study
that proper ON–OFF timing of ultrasonic system must be deter-
3.4. Effect of ultrasonic power mined for delignification of biomass, as unnecessary increase in
the ON time would lead to undue heating and unnecessary electri-
The ultrasound-assisted experiments were also dedicated to cal energy consumption.
determining the influence of ultrasonic power dissipation on the
3.6. Surface morphology

Since a large fraction of lignin has been removed by ultrasound-

assisted alkaline pretreatment, it became interesting to examine
y = -2731.1x + 5.3479 the physical changes in the biomass. Scanning electron micro-
graphs of untreated and pretreated samples were obtained to
ln k

R² = 0.9462
verify the biomass structural changes caused by the ultrasound-as-
sisted alkaline pretreatment. Representative SEM micrographs of
untreated and pretreated newspaper are shown in Fig. 5, where
significant differences can be easily observed. Untreated newspa-
per had a relatively smooth and intact surface structure which
might be recalcitrant to enzymatic attack and its adsorption onto
the biomass, resulting in lower enzymatic conversion from this
1/T (K-1 )
surface morphology. Significant morphological changes indeed oc-
Fig. 4. The relationship between 1/T and ln(k). curred after pretreatment. Surface structure of newspaper after
 P.B. Subhedar, P.R. Gogate / Ultrasonics Sonochemistry 21 (2014) 216–225 221

Table 6
Effect of ultrasonic operating parameters on rate of delignification.

Ultrasonic operating Delignification Rate constant k R2

parameters (%) (min1)
Power (W)
60 42.77 0.0081 0.99
70 50.21 0.0106 0.98
80 60 0.0138 0.98
90 73.26 0.0197 0.98
100 80.16 0.0239 0.98
110 82.01 0.0259 0.97
Duty cycle (%)
40 63.15 0.0148 0.98
50 69.97 0.0178 0.98
60 75.59 0.0213 0.98
70 79.94 0.0242 0.98
80 80.92 0.0246 0.98

sono-assisted alkaline pretreatment was not smooth, and dis-
rupted surface structure was observed mainly due to the breaking
up the lignin. As a result, some lignin or lignin-carbohydrate com-
plexes were condensed on the surface of the biomass. Furthermore,
the structure of the lignocellulosic biomass was opened up. The
goal of pretreatment is not only to delignify the biomass, which
can inhibit the enzymatic hydrolysis but also to increase the avail-
able surface area of cellulose being exposed to the enzymes. So as
cellulosic structure got opened up because of lignin removal during
pretreatment, the available surface area for enzymatic hydrolysis is
definitely increased.
3.7. Spectral characterization
Infrared spectroscopy is widely used for investigating the struc-
ture of constituents and the chemical changes in lignocellulosic
materials during delignification. The FTIR spectra of untreated
and pretreated newspaper are illustrated in Fig. 6a. The prominent
band around 3414 cm1 and 1000–1200 cm1 indicated the pres-
ence of cellulose in the sample. These bands are attributed to the
O–H stretching vibrations and intermolecular hydrogen bonding
in phenolic and aliphatic structures. The existence of hemicellulose
was confirmed by large bands at 1071, 1372, and 805 cm1. These
bands are assigned for unconjugated C@O stretching in acetyl
group in the hemicelluloses. The bands at 1510 and 1605 cm1
showed the aromatic skeletal vibrations of lignin. These bands
are assigned for C@O stretching in conjugated p-substituted aryl
ketones [30]. These results confirmed the main components of
newspaper.
It can be observed that the intensity of band at 3414 cm1 be-
came strengthened for ultrasound-assisted alkaline pretreatment.
This could be attributed to the increase in the cellulose content
in the pretreated newspaper as compared to the untreated news-
paper. Also in the case of pretreated newspaper, bands for hemicel-
lulose were present but bands for lignin were absent indicating
lignin was removed from the sample to a great extent after
pretreatment.
Fig. 6b shows the infrared spectra of the recovered lignin after
ultrasound-assisted alkaline treatment. The recorded spectra of lig-
nin were analyzed on the basis of assignments suggested by Faix
[31]. The spectra showed a broad band at 3420.62 cm1 which is
attributed to the hydroxyl groups in phenolic and aliphatic struc-
tures. The absorbance of C–H vibrations of the aromatic ring was
detected at 1600.34 cm1 and the band attributed to methoxyl
group appeared at 1416.57 cm1.
3.8. X-ray diffraction (XRD)
X-ray diffraction spectra of untreated and pretreated newspa-
per were studied (Fig. 7). The CrI of newspaper was calculated
based on the Eq. (1). The CrI were 36.70%, 46.90% and 54.20% for
untreated, alkali treated and ultrasound-assisted alkali treated
newspaper, respectively. In case of lignocellulosic biomass, CrI
measures the relative amount of crystalline cellulose in the total
solid. Hence, when lignin which is amorphous in nature was par-
tially removed, CrI would be increased after delignification. This
explanation could be proved by the fact that alkali treated and
ultrasound-assisted alkali treated newspaper had CrI higher as

Untreated Newspaper Pre-treated Newspaper

Fig. 5. SEM images of untreated and pretreated newspaper.
222 P.B. Subhedar, P.R. Gogate / Ultrasonics Sonochemistry 21 (2014) 216–225

100
Pre-treated
newspaper

90
80
Transmittance [%]
70

Untreated
newspaper
60
50
40

2906.18

1737.60

1605.59
1510.89
1426.12

1071.55
1372.33
1266.36

805.81

617.91
576.30
461.52
3500 3000 2500 2000 1500 1000 500
Wavenumber cm-1

Fig. 6a. FT-IR spectra of untreated and pretreated newspaper.

100
90
80
Transmittance [%]

70
60
50

3420.62

1600.34

1416.57

1182.57
1126.79
1045.17

621.78

3500 3000 2500 2000 1500 1000 500

Wavenumber cm-1

Fig. 6b. FT-IR spectra of recovered lignin.

compared to untreated newspaper. It has been reported that, alkali
pretreatment of lignocellulosic biomass causes swelling of cellu-
lose, which results into decreasing crystallinity [32]. Conversely
in the present study, the CrI of alkali and ultrasound-assisted alkali
pretreatment was higher than that untreated one, which could be
explained by the fact that lignin was removed and the fraction of
cellulose in the samples were increased. The maximum value of
CrI for ultrasound assisted treatment also confirms the superiority
of the treatment approach based on ultrasound.
3.9. Molecular weight distribution
Molecular weight distribution of the recovered lignin has been
studied (Fig. 8). The molecular weight average (Mw), number aver-
age (Mn) and polydispersity (Mw/Mn) of the recovered lignin was
found to be 3104 Da, 2755 Da and 1.126 respectively. Lignin
mostly composed of guaiacyl units is expected to show higher
molecular weight than those presenting high contents of syringyl
units [33]. The recovered lignin had Mw of 3104 g/mol, which
 P.B. Subhedar, P.R. Gogate / Ultrasonics Sonochemistry 21 (2014) 216–225 223

Fig. 7. XRD profile of untreated and pre-treated newspaper.

comes under low molecular weight lignin (ranged between 1990
and 3501 g/mol) suggesting higher proportion of syringyl units.
The obtained numbers can be explained because lignin molecular
weight is closely related to the number of C–C bonds between
the structural units, especially those involving C5 of the aromatic
ring [34]. Guaiacyl type units are able to generate this kind of
bonds, and this is not possible in syringyl type of units, as they
have both C3 and C5 positions substituted by methoxy groups.
These C–C bonds are not cleaved due to their higher stability.
The relatively low polydispersity of the lignin indicated the high
fraction of low molecular weight present in the recovered sample.
3.10. Thermogravimetric analysis
TGA is a valuable analytical method to study the physico-chem-
ical properties of macromolecules such as cellulose. Weight change
of a sample is recorded as a function of time or temperature and
portrayed by a TG curve, which conveys information on thermal
behavior of sample. A three stage decomposition weight loss has
been observed as shown in Fig. 9a. In first stage, hemicellulose deg-
radation takes place and in second and third stage the degradation
of cellulose and lignin may take place, respectively [35]. For un-
treated as well as pretreated newspaper, the initial decomposition
i.e. first decomposition occurred between 35 and 120 °C giving al-
most 5–6% weight loss. The first stage (temperature below 120 °C)
corresponds to the drying period where light volatiles, mainly
water are liberated throughout the procedure. Marginal decay of
sample weight was observed (Fig. 9a). The degradation occurs at
a faster rate in second stage compared to first and third stage. Dev-
olatilization is the major step in all thermochemical conversion
process involving biomass and is represented by the second stage
of decomposition. In this stage, significant slope of the TG curves
is observed, corresponding to significant drop in the weight of
samples due to liberation of volatile hydrocarbon from rapid

Fig. 8. Molecular weight distribution of lignin.

224 P.B. Subhedar, P.R. Gogate / Ultrasonics Sonochemistry 21 (2014) 216–225
120
100
Weight percent (%)
80
Untreated
60
Alkali treated
40 Sono-assisted alkali
treated
20
0
0 100 200 300 400
Temp. 0 C
Fig. 9a. TGA results of untreated and pre-treated newspaper.
Fig. 9b. TGA of lignin.
thermal decomposition of hemicellulose, cellulose and some part
of lignin. The second decomposition of the samples occurred be-
tween 200 and 350 °C, representing near about 50–52% weight loss
for the untreated sample and 60–63% weight loss for alkali
pretreated sample. This is primarily due to the fact that 80 wt.%
of biomass is formed by volatile fraction, and only around
20 wt.% is in the form of solid carbonaceous residue [36]. In case
of ultrasound-assisted alkali pretreatment, about 90% weight loss
was observed in the sample. The ultrasound-assisted alkali pre-
treated paper contained mainly cellulose which is a homopolymer
of glucose. The degradation of the cellulose starts around 250 °C. It
is nearly decomposed totally into volatile product at 400 °C. The
third stage decomposition is located between 360 and 500 °C with
near about 40% total weight loss for the untreated sample, 30–32%
weight loss for alkali pretreated and 9–10% weight loss for ultra-
sound-assisted alkali pretreated sample. Untreated sample con-
tained 41.02% cellulose, 24.85% hemicellulose and 23.07% lignin.
After alkaline pretreatment 40% delignification was obtained, thus
13.8% lignin was remained in the sample. Ultrasound-assisted al-
kali pretreated sample retained only 4.6% lignin. Thus owing to less
lignin content, ultrasound-assisted alkaline pretreated sample
decomposed early as compared to the untreated and only alkali
treated sample, again establishing the superiority of the ultrasound
based approach.
Thermogravimetric curve for recovered lignin after ultrasound-
assisted alkaline pretreatment was also studied (Fig. 9b). At the ini-
tial stage of thermal treatment, lignin losses moisture from room
temperature to 120 °C. Degradation of lignin occurred over a broad
temperature interval (120–600 °C), because various oxygen func-
tional groups from its structure have different thermal stabilities,
and their scission occurs at different temperatures. The maximum
weight loss rate occurred in the temperature range of 200–350 °C.
The decomposition of lignin is very complex and mainly depends
on its origin.
4. Conclusions
The current work examined the feasibility of ultrasound-as-
sisted alkaline pretreatment for delignification of sustainable bio-
mass in the form of waste newspaper. Newspaper pretreated
with ultrasound-assisted alkaline pretreatment showed better del-
ignification as compared to only alkaline pretreatment. About
500 600 700
2-fold increase in delignification was achieved by ultrasound-as-
sisted alkaline pretreatment as compared to alkaline pretreatment.
The remaining fibrous material is a potentially superior substrate
for enzymatic hydrolysis and ethanol fermentation owing to its
high cellulose content. Overall, the significant decrease in the pre-
treatment time, temperature and alkali requirement with en-
hanced effectiveness are the most attractive features of
ultrasound-assisted alkaline pretreatment.
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