Industrial Crops and Products 35 (2012) 274–279

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Industrial Crops and Products

journal homepage: www.elsevier.com/locate/indcrop

Steam explosion pretreatment reproduction and alkaline delignification reactions

performed on a pilot scale with sugarcane bagasse for bioethanol production
G.J.M. Rocha a,∗ , A.R. Gonçalves b , B.R. Oliveira a , E.G. Olivares a , C.E.V. Rossell a
a
Bioethanol Science and Technology National Laboratory - CTBE, P.O. Box 6170, 13083-970 Campinas, SP, Brazil
b
Department of Biotechnology, Lorena Engineering School, São Paulo University, P.O. Box 116, 12600-970 Lorena, SP, Brazil

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

Article history: This work was focused on the steam explosion pretreatment reproduction and alkaline delignification
Received 25 April 2011 reactions on a pilot scale for the ethanol production, through different varieties of natural sugar-
Received in revised form 6 July 2011 cane bagasse, pretreated bagasse and delignified pretreated bagasse (cellulosic pulp). The possible
Accepted 8 July 2011
chemical composition differences of the various types of bagasse, as well as the chemical composi-
Available online 30 July 2011
tion variations of the materials in the 20 processes of pretreatment and delignification on the pilot
scale were verified. The analytical results of the 20 samples of most diverse varieties and origins of
Keywords:
natural sugarcane bagasse considering planting soils, planting periods and weather; show no signif-
Sugarcane bagasse
Reproducibility
icant chemical differences. It is evident that only with the chemical composition it is not possible
Pretreatment to verify the differences between the varieties of sugarcane bagasses. The research results may offer
Steam explosion some evidences of these varieties, but it is not a reliable parameter. The pilot process of steam explo-
Delignification sion pretreatment and the alkaline delignification process of pretreated material showed through
Pilot scale analytical results a good capacity of reproduction, as the standard differences were below 2.7. The aver-
age allowed in the pretreatment and alkaline delignification processes were 66.1 ± 0.8 and 51.5 ± 2.6
respectively, ensuring an excellent reproduction capacity of the processes obtained through chemical
characterizations.
© 2011 Elsevier B.V. All rights reserved.

1. Introduction of sugarcane bagasse components is desirable for both economical

Sugarcane bagasse is the solid residue remaining after crushing
the sugarcane to obtain its juice which is used for sugar or ethanol
production. The enormous sugar and ethanol production in Brazil
generates huge amounts of bagasse. During the 2010/2011 har-
vest, more than 625 million tons of sugarcane were crushed, which
generated around 208 million tons of sugarcane bagasse (CONAB,
2011). The sugarcane bagasse is currently used as the main source
of the energy required by the sugar and ethanol mills, as well as
for generating electricity to be sold. However, an important part of
the produced bagasse is belittled. It is sure that with the necessary
technological improvements of the boilers and the processes; it is
possible to supply the energy needs of the plants with only half of
the produced bagasse (Rocha et al., 2011). The sugarcane bagasse
surplus could be used in more than forty different applications, such
as production of ethanol, pulp and paper, boards, animal feed and
furfural (Gálvez, 2000; Mussatto et al., 2006). The integral usage
∗ Corresponding author. Tel.: +55 19 3518 3179; fax: +55 19 3518 3164.
E-mail address: george.rocha@bioetanol.org.br (G.J.M. Rocha).
and environmental reasons.
The sugarcane bagasse as well as any other type of plant biomass
is composed by cellulose, hemicelluloses, lignin, and small amounts
of extractives and mineral salts. The structural components are
distributed in a lamellar structure (Fengel and Wegener, 1989).
The hydrolysis is crucial for the conversion of bagasse polysaccha-
rides, mainly the cellulose, into valuable products. However, the
strongest cellulose crystalline arrangement, and the lignin protec-
tive effects as well as the hemicelulloses hard access to the enzymes
and the catalytic acids to the ␤1 → 4 glycosidic bonds, constitute a
serious obstacle to the hydrolysis (Canettieri et al., 2007).
In order to make the cellulose hydrolysis easier, different kinds
of physical, chemical and physic-chemical pretreatment methods
have been proposed. Pretreatment and delignification processes
are aimed to disrupt the cellulose–hemicelluloses–lignin complex,
and they are important technological steps for fractioning the lig-
nocellulosic materials into their main components, so they can be
used according to a biorefinery philosophy (Chandra et al., 2007;
Martín et al., 2007; Mussatto et al., 2006; Rocha, 2000; Saad et al.,
2008).
In order to establish pretreatment processes strong enough to
separate the cell wall arrangement and mild avoiding a significant

0926-6690/$ – see front matter © 2011 Elsevier B.V. All rights reserved.
doi:10.1016/j.indcrop.2011.07.010
 G.J.M. Rocha et al. / Industrial Crops and Products 35 (2012) 274–279
Fig. 1. Flowsheet of procedures for the separation of major components of sugarcane bagasse.
chemical degradation of the biomass components, through easily
removable chemical elements causing a low environmental impact,
low energy demand using low-cost equipment is the challenge for
today’s chemical industry (Canettieri et al., 2007).
The traditional pretreatment processes are severe, destructive
and not efficient enough. Recently, innovative methods were able
to separate the three main polymeric constituents of the lignocel-
lulosic biomass and to decrystallinize the cellulose with minimal
chemical alteration of the hemicellulose and lignin (Kumar et al.,
2009; Yang and Wyman, 2008).
Different approaches of dilute acid pre-hydrolysis have been
shown to be effective in pretreatment processes (Kumar et al.,
2009; Taherzadeh and Karimi, 2008). Sulfuric acid is the most com-
monly used acid in pretreatment of sugarcane bagasse nowadays
(Lavarack and Griffin, 2002), but other reagents, such as hydrochlo-
ric, nitric and phosphoric acids can also be used for this purpose
(Gámez et al., 2006; Rodríguez-Chong et al., 2004). Currently,
the usage of acetic acid along with peroxide hydrogen has been
reported as a way to remove lignin prior to bagasse enzymatic
hydrolysis (Tan et al., 2010).
1.1. Bagasse chemical composition
The bagasse is composed by cellulose, hemicellulose and lignin,
as most of the natural macromolecules, even though it presents
little amounts of other low molecular weight compounds.
Rocha et al. (2011) reported the following values for the
sugarcane bagasse chemical composition: cellulose 45.5 ± 1.1,
hemicellulose 27.0 ± 0.8, total lignin 21.1 ± 0.9, mineral com-
pounds 2.2 ± 0.1 and extractives 4.6 ± 0.3, showing that this plant
biomass contains a considerable amount of cellulose and other
components of interest for the Biorefinery Industry.
In general, the study of hydrolysis conditions to obtain fer-
mentable hemicellulosic hydrolysate, as well as the delignification
processes to obtain cellulose in order to produce ethanol was suc-
cessfully concluded in laboratory scale. Different reactor designs for
275
the hydrolysis and delignification processes scale-up were studied
(Rodrigues et al., 2010).
The present study evaluated the use of sugar cane bagasse in its
full heterogeneous nature. This aims at exploring the variability of
this resource, regarding its chemical composition and yields after
steam explosion and alkaline delignification. Hence, the results
reported in the present work are representative of the reality that
will be faced by initiatives to scale up the processes for bioethanol
production.
2. Methodology
Twenty sugarcane bagasse samples of different varieties consid-
ering places where they have been planted and types of soils, as well
as the year period when they have been cultivated and the weather;
have been demonstrated according to their chemical composition.
The bagasses in their majority were collected in sugar and ethanol
mills from São Paulo State and Brazilian Northeast.
The natural bagasse was pretreated using steam explosion
processes. These procedures are designed to separate, by hydrol-
ysis, the hemicelluloses present in the biomass. The pretreated
bagasse, which basically contains cellulose and lignin, was delig-
nified through the usage of an alkaline solution in order to separate
the lignin present in biomass. These processes are shown in Fig. 1.
2.1. Steam explosion pretreatment
Twenty different samples of natural bagasse were pretreated
by steam explosion on the pilot scale and they were analyzed.
These procedures are designed to separate, by auto hydrolysis, the
hemicellulose present in the biomass. Sugarcane bagasse contain-
ing approximately 50% of moisture was transferred to a stainless
steel reactor with capacity of 200L and sealed. Steam was injected
in the reactor up to a pressure of almost 1.3 MPa equivalent to
13 kgf cm−2 (190 ◦ C) and kept there for a period of 15 min. After this
276 G.J.M. Rocha et al. / Industrial Crops and Products 35 (2012) 274–279
time, the reactor was suddenly depressurized. The steam explosion
pretreated bagasse was transferred to a cyclone with capacity of
1000L and collected in a plastic barrel. This material was washed
into a semi-industrial centrifuge of 100L at 1100 rpm until the
complete removal of the sugar-rich liquid fraction (hemicellulosic
hydrolysate).
2.2. Delignification
For the cellulignin fractions obtained in the pretreatment it
was applied a pilot scale alkaline delignification. The steam explo-
sion pretreated bagasse was reacted with a NaOH solution 1.0%
(w/v). The delignification reactions were made in a stainless steel
316L reactor with a 350L capacity, fitted with mixing and heating
systems, using a solid–liquid ratio 1:10 (w/v). The operation was
carried out at 100 ◦ C for 1 h.
After the delignification reaction, the reactor was unloaded and
the soluble fraction (containing liquor lignin) along with the cel-
lulosic pulp was collected and separated through centrifugation.
The liquor, rich in lignin, was stored for future chemical or biologi-
cal reactions of this material. The cellulosic pulp was then washed
with water until it no longer produced a yellow-colored fluid due
to the presence of lignin (neutral pH) and stored in cold at 4 ◦ C.
Twenty reactions of this process in pilot scale, using the same
steam explosion pretreated bagasse, were analyzed.
2.3. Sugarcane bagasse fractions physical and chemical
characterization
Natural bagasses, pretreated bagasses as well as pretreated
delignified bagasses (cellulosic pulp) were chemically analyzed in
the laboratories of Engineering School of Lorena (EEL-USP).
The analytical methodology used in this study was developed by
Rocha et al. (1997) and validated by Gouveia et al. (2009). Analyses
were performed in triplicate.
2.4. Insoluble and soluble lignin determination
All samples have been ground to 16/60 mesh, although only
the natural bagasse sample was removed from the extractives with
organic solvent (ethanol/cycle-hexane, 2:1, v/v) and washed with
water at 70 ◦ C.
A sample of about 2 g without extractives, dry weight, was
transferred to a Becker of 100 mL and treated with 10 mL of 72%
H2 SO4 under vigorous mixing for 7 min in a bath maintained at
45.0 ± 0.5 ◦ C. The reaction was interrupted through the addition
of 50 mL of distilled water. The sample was immediately trans-
ferred to a 500 mL Erlenmeyer, where the acid was diluted by water
addition up to a final volume of 275 mL.
The bottle was sealed with aluminum paper and autoclaved
for 15 min at 1.05 atm to complete the hydrolysis of the oligomer
remaining. After decompression, the bottle was removed from the
autoclave and cooled to room temperature.
The hydrolysed material was separated from solids through
filtration using a paper filter (Nalgon, 18.5 cm diameter, ash
0.00020 g) previously weighed. The hydrolysate was collected in
a volumetric bottle of 500 mL and the remaining solid in the paper
filter was washed with 50 mL of distilled water portions until reach-
ing the bottle volume, was stored for further analysis.
The lignin retained in paper filter was washed with distilled
water until exemption from sulphates (approximately 1500 mL)
and dried in oven at constant temperature of 105 ◦ C. Its content
was obtained by gravimetry. Soluble lignin in the hydrolyzate
was determined by UV-spectroscopy in a 5-mL aliquot of the
hydrolyzate, which was adjusted to pH 12 with 6 M NaOH and 10-
fold diluted. The absorbance of the solution was read at 280 nm
(Perkin Elmer LAMBDA 25). The concentration of soluble lignin was
determined using the following equation:
Clig. = 4.187 × 10−2 (Alig280 − Apd280) − 3.279 × 10−4 g/L (1)
where Clig. = lignin concentration, in g/L; Alig280 = lignin solu-
tion extinction in 280 nm; Apd280 = A1 + A2 e2 = e1 extinction in
280 nm, sugar decomposition products (furfuraldehyde and HMF),
whose A1 A2 concentrations were determined in advance through
HPLC and e1 and e2 through the UV spectroscopy.
2.5. Ash content determination in insoluble lignin
After determining the insoluble lignin in sulfuric acid 72% it was
transferred to a porcelain crucible previously weighed. The sam-
ple was slowly calcined to 300 ◦ C for 2 additional hours at 800 ◦ C
in a muffle. After cooling the crucible in a dissector, the ash mass
was determined in analytical balance (adquated from the standard
ASTM, 1966).
2.6. Carbohydrates determination
The levels of celobiose, glucose, xylose, arabinose and acetic acid
present in the filtrated solution on sulfuric acid hydrolysis 72% of
lignocellulosic materials were analyzed through HPLC. A small sam-
ple of the solution was filtered in a Sep Pak C18 cartridge and then
examined for its contents of celobiose, glucose, xylose, arabinose
and acetic acid through high performance liquid chromatogra-
phy (HPLC), a chromatograph Shimadzu applying a model CR 7A
refraction index detector Shimadzu model RID-6A, for a column in
which was used Aminex HPX 87H (300 mm × 7.8 mm, BIO-RAD. The
mobile phase was H2 SO4 0.005 mol/L with a flow of 0.6 mL/min, at
45 ◦ C. The concentrations of glucose and celobiose were converted
into cellulosic pulp. The concentrations of xylose and arabinose
were converted to hemicelluloses, and the concentration of acetic
acid was converted to acetyl groups. The masses were divided due
to their dry weight of the original material and multiplied by the
hydrolysis factor. The conversion factors from glucose and celo-
biose to cellulosic pulp are 0.90 and 0.95, respectively. Similarly,
xylose and arabinose were converted to hemicellulose using the
factor 0.88. The conversion factor of acetic acid to acetyl group is
0.72.
The concentrations of the compounds were determined due to
the calibration curves drawn for each component.
2.7. Furfuraldehyde and hydroxymethylfurfural determination
(of sugars decomposition products)
The furfuraldehyde and hydroxymethylfurfural in the fil-
tered material for the sulfuric acid hydrolysis with 72% of
lignocellulosic materials were analyzed through HPLC using a chro-
matograph Shimadzu model C-R7A detector with an UV visible
Shimadzu SPD model – applying a column RP 10A-18 (C-18) of
125 mm × 4 mm (Hawlett-Packard), and in the mobile phase a solu-
tion of acetonitrile–water 1:8 (v/v) with 1% of acid acetic, at a flow
of 0.8 mL/min to 25 ◦ C.
The middle areas of chromatograms for furfuraldehyde and
hydroxymethylfurfural were converted into cellulose and hemi-
cellulose using the conversion factors 1.37 and 1.29, respectively
(experimental values).
 G.J.M. Rocha et al. / Industrial Crops and Products 35 (2012) 274–279 277

Table 1
Analytical results of the chemical composition of natural bagasse.

Samples Cellulose Hemicellulose Lignin Ash Extractives

1 42.4 22.9 21.5 3.4 9.3

2 42.7 23.7 21.9 2.0 3.9
3 43.5 25.3 22.1 2.1 4.0
4 41.8 18.9 23.0 2.5 3.7
5 43.2 26.9 22.5 3.5 4.0
6 44.8 26.7 22.6 2.6 4.0
7 45.7 26.5 21.5 2.9 3.7
8 44.9 26.3 22.1 2.7 3.9
9 45.0 26.4 23.4 2.5 2.7
10 43.1 25.7 23.9 3.7 2.9
11 41.5 26.8 24.7 5.5 2.4
12 43.9 26.1 23.3 4.6 2.5
13 40.6 26.7 24.6 4.6 3.6
14 40.9 26.9 23.9 5.1 3.2
15 42.3 25.3 22.5 1.0 5.8
16 45.2 24.7 21.8 1.0 5.7
17 42.3 24.1 21.9 1.5 6.0
18 42.8 24.3 22.4 1.5 4.9
19 42.9 24.2 22.5 1.5 5.3
20 43.2 26.0 25.4 1.4 4.0

Average 43.1 25.2 22.9 2.8 4.3

Standard deviation 1.4 1.9 1.1 1.4 1.6
Confidence interval 0.6 0.8 0.5 0.6 0.7

3. Results and discussion
3.1. The chemical composition of sugarcane bagasse and its
derivates
Production and analytical results of the natural bagasse chemi-
cal composition from the 20 samples of several sugarcane varieties
are shown in Table 1. The confidence intervals values were obtained
using a confidence level of 95%.
The results found for natural bagasse are similar to those found
in the literature (Agnihotri et al., 2010; Rocha et al., 2011; Rocha and
Silva, 2005). Although the analyzed samples have different origins
and varieties, planting soils, planting periods as well as weather;
Chemically, it has not been observed any significant differences, as
shown in Table 1.
The samples of 20 reactions of steam explosion pretreated sug-
arcane bagasse of a wide range of varieties as well as the results
from the pretreatment reactions were properly analyzed. The val-
ues of the chemical composition of these pretreated bagasses are
shown in Table 2. The confidence interval values were obtained
using confidence level of 95%.
The average results obtained from the 20 reaction samples of
the pretreated bagasse show that even using a pilot scale pro-
cess it was reproduced as it can be seen analyzing the deviation
standards at Table 2. It shows an average mass production of
66.1% with a deviation standard of 0.8. The major differences
are observed in the amount of residual lignin due to the lack
of a more efficient control of the pilot process variables, mainly
the temperature. Recent studies developed at EEL/USP show that
a 5 ◦ C increase in the pretreatment processes results in signif-
icant changes in the pretreated material chemical composition
(Silva, 2010; Silva et al., 2010). Sugarcane bagasse major compo-
nents solubilization after the pretreatment process is shown in
Fig. 2.

Table 2
Chemical composition of pretreated sugarcane bagasses.

Samples Cellulose Hemicellulose Lignin Ash Mass yield%

1 55.9 4.4 37.1 2.5 66.2

2 55.5 4.1 37.6 2.8 65.7
3 57.6 4.5 34.5 1.8 67.0
4 56.0 8.5 30.1 5.2 66.6
5 61.4 7.4 29.9 1.3 64.2
6 56.8 7.7 33.8 1.5 65.0
7 57.2 6.6 34.4 1.6 65.5
8 56.5 7.3 34.4 1.6 65.8
9 58.0 7.5 33.6 1.3 66.0
10 55.8 9.2 33.6 1.4 67.0
11 56.9 7.8 34.0 1.9 66.8
12 59.4 7.8 30.0 2.2 64.7
13 57.0 7.8 32.2 2.4 65.5
14 57.2 7.2 32.4 2.0 66.3
15 56.7 6.0 30.0 4.4 67.0
16 56.3 6.4 30.3 4.4 66.4
17 58.8 5.3 30.5 4.7 66.0
18 58.5 5.3 30.5 4.7 66.1
19 57.6 5.7 30.4 4.5 67.2
20 60.9 5.0 31.2 3.2 66.5

Average 57.5 6.6 32.5 2.8 66.1

Standard deviation 1.6 1.5 2.4 1.4 0.8
Confidence interval 0.7 0.6 1.0 0.6 0.4
278 G.J.M. Rocha et al. / Industrial Crops and Products 35 (2012) 274–279

Table 3
Chemical composition of cellulosic pulps.

Samples Cellulose Hemicellulose Lignin Ash Mass yield%

1 82.2 4.0 10.5 3.5 55.8

2 82.9 4.1 9.6 3.6 55.4
3 88.0 3.9 4.5 3.5 51.0
4 88.0 3.6 4.8 3.6 50.9
5 88.6 4.2 5.0 2.5 50.0
6 86.5 4.3 5.5 2.9 52.5
7 89.4 4.0 4.6 2.0 48.1
8 90.9 4.3 4.8 1.1 47.9
9 88.6 4.1 4.2 3.5 50.5
10 89.4 4.1 4.5 3.4 49.9
11 86.0 4.5 5.3 3.5 52.0
12 87.6 2.8 5.8 5.7 53.2
13 84.5 3.1 6.5 5.5 54.0
14 83.0 3.2 8.3 5.1 53.8
15 82.3 3.2 9.4 5.1 55.5
16 89.6 3.4 5.5 1.4 48.0
17 84.6 3.6 9.4 2.0 54.4
18 88.5 4.9 4.3 2.5 49.8
19 88.1 5.0 4.3 2.4 48.8
20 88.0 4.9 4.5 2.6 49.2

Average 86.8 4.0 6.1 3.3 51.5

Standard deviation 2.7 0.6 2.1 1.3 2.6
Confidence interval 1.2 0.3 0.9 0.6 1.2

From the values demonstrated by the respective productions
we can observe an excellent hemicelluloses hydrolysis. It solubi-
lized an average of 82.7% with a standard deviation of 4.3. The
process hydrolyzed 11.8% of cellulose with a deviation of 3.7. Prob-
ably, the hydrolyzed cellulose is the polymer amorphous region.
The lignin solubilized 7.9% on average with a standard deviation of
9.1. It was observed in the experiments 1, 2. 3, 7 and 8 there was an
increase in lignin content compared with the values of lignin from
the natural bagasse, indicating that probably occurred aromatics
and non-aromatics extractives condensation.
The samples of 20 delignification reactions in the pilot scale of
the steam explosion pretreated bagasse of different varieties were
analyzed. The chemical composition values of these cellulosic pulps
are shown in Table 3. The confidence interval values were obtained
using a confidence level of 95%.
Alkaline delignification step for the steam-exploded bagasse
was performed aimed to decrease inhibitory effects showed by
cellulolytic enzymes due to the presence of lignin.
The values reported in Table 3 show that the amount of cellulose
increased to an average of approximately 87% and the removal of
hemicellulose and lignin exceeded 90% in this study.
The reproduction of the process was very good as well as the
deviation standard for its production, so was the content for each
component analyzed, none of them exceed 2.7 with confidence
interval value of 1.2.
Sugarcane bagasse major components solubilization after the
pretreatment and delignification processes are shown in Fig. 3.
From the values obtained through the respective production we
can observe an excellent removal of lignin from the biomass. It
solubilized a 92.7% average with a deviation standard of 3.9. The
hemicellulose hydrolysis came to 94.7% with deviation standard of
0.9. The process hydrolyzed 31.1% of cellulose with a deviation of
3.5. It evidences that even in milder conditions, the steam explo-
sion pretreatment and alkaline delignification processes cause a
substantial cellulose loss, which for the ethanol production from
this component is required a final product cost analysis.
The average results of the delignified pretreated bagasse sam-
ples of twenty reactions show that, even applying a pilot scale, a
good reproduction of the process can be observed, as it can be
seen through the deviation standards. It shows an average mass
production of 51% with a deviation standard of 2.6. The greater devi-
ations are observed in the amounts of cellulose and residual lignin

Fig. 2. Solubilization of the major components of sugarcane bagasse after steam Fig. 3. Graph of solubilization of the components in pretreatment and delignifica-
explosion pretreatment. tion processes.
 G.J.M. Rocha et al. / Industrial Crops and Products 35 (2012) 274–279
in the cellulosic pulp. A better control of the differences on the pilot
process, temperature mainly, would decrease this variation.
4. Conclusion
The analytical results of the 20 natural sugarcane bagasse sam-
ples of the most diverse varieties and origins considering planting
soils, periods of the year as well as weather; show no significant
chemical composition differences.
It is evident that with only the chemical composition it is not
possible to verify the differences between the sugarcane bagasses
varieties. The extract amount may offer some evidences of this
varieties, but they are not a reliable parameter.
In order to differentiate these varieties it is necessary the use
of other analytical techniques such as (X-ray fluorescence), or
any other spectrometric method or physical method to observe
nanomorphological changes.
The steam explosion pretreatment pilot process and the
alkaline delignification process of pretreated material proved
through the analytical results to have a good reproduction,
once the deviation standards were below 2.7. The produc-
tion average for the pretreatment and delignification processes
were 66.1 ± 0.8 and 51.5 ± 2.6 respectively, confirming the excel-
lent reproduction of the processes observed in the chemical
characterizations.
References
Agnihotri, S., Dutt, D., Tyagi, C.H., 2010. Complete characterization of bagasse of
Saccharum officinerum early species – co 89003 for pulp and paper making.
Bioresource 5 (2), 1197–1214.
ASTM Methods, 1966. Standard Test Method for Lignin in wood. D.110656, pp.
396–398.
Canettieri, E., Rocha, G.J.M., de Carvalho, J.R., Silva, J.B.A., 2007. Optimization of acid
hydrolysis from the hemicellulosic fraction of the Eucalyptus grandis residue
using response surface methodology. Bioresour. Technol. 98, 422–428.
Chandra, R.P., Bura, R., Mabee, W.E., Berlin, A., Pan, X., Saddler, J.N., 2007. Substrate
pretreatment: the key to effective enzymatic hydrolysis of lignocellulosics? Adv.
Biochem. Eng. Biotechnol. 108, 67–93.
CONAB (National Supply Company), 2011. The Sugarcane Production in Brazil will
Reach a New Record (accessed 12.1.11.) http://www.conab.gov.br.
Fengel, D., Wegener, G., 1989. Wood Chemistry, Ultrastructure Reactions. Walter de
Gruyter, Berlin/New York.
Gálvez, L.O., 2000. Diversified productions in sugarcane agro-industry. In: Gálvez,
L.O. (Ed.), Handbook of Sugarcane Derivatives. , 3rd ed. ICIDCA, Havana/Cuba,
pp. 3–17.
279
Gámez, S., González, J.J., Ramírez, J.A., Garrote, G., Vázquez, M., 2006. Study of the
sugar cane bagasse hydrolysis by using phosphoric acid. J. Food Eng. 74, 78–88.
Gouveia, E.R., Nascimento, R.T., Souto-Maior, A.M., Rocha, G.J.M., 2009. Validation
of the methodology for the chemical characterization of the sugarcane bagasse.
Quím. Nova 32, 1500–1503.
Kumar, R., Mago, G., Balan, V., Wyman, C.E., 2009. Physical and chemical character-
izations of corn stover and poplar solids resulting from leading pretreatment
technologies. Bioresour. Technol. 100, 3948–3962.
Lavarack, B.P., Griffin, G.J., 2002. The acid hydrolysis of sugarcane bagasse hemi-
cellulose to produce xylose, arabinose, glucose and other products. Biomass
Bioenergy 23, 367–380.
Martín, C., Rocha, G.J.M., Pérez, M., López, Y., Hernández, E., Plasencia, Y., 2007. Acid
prehydrolysis, alkaline delignification and enzymatic hydrolysis of rice hulls.
Cell. Chem. Technol. 41, 129–135.
Mussatto, S., Dragone, G., Rocha, G.J.M., Roberto, I., 2006. Optimum operating con-
ditions for brewer’s spent grain soda pulping. Carbohydr. Polym. 64, 22–28.
Rocha, G.J.M., Silva, F.T., Araújo, G.T., Curvelo, A.A.S., 1997. A fast and accurate method
for determining the cellulose and polyoses through HPLC. In: V Brazilian Sym-
posium Proceedings on the Chemistry of Lignin and Other Wood Components,
vol. 5, Curitiba, PR, Brazil, pp. 113–115.
Rocha, G.J.M., 2000. Sugarcane bagasse oxygen-assisted delignification. Ph.D. Thesis.
Institute of Chemistry, University of São Paulo.
Rocha, G.J.M., Silva, J.S., 2005. Comparative study of cellulose pulps obtained from
sugarcane bagasse pretreated by an acid solution and through steam explosion.
In: XVI Brazilian Congress Proceedings of Chemical Engineering-COBEQ, Santos,
SP, Brazil, pp. 255–257.
Rocha, G.J.M., Martin, C., Soares, I.B., Souto Maior, A.M., Baudel, H.M., Moraes, C.A.,
2011. Dilute mixed-acid pretreatment of sugarcane bagasse for the ethanol pro-
duction. Biomass Bioenergy 35, 663–670.
Rodrigues, R.C.L.B., Rocha, G.J.M., Rodrigues Jr., D., Filho, H.J.I., Felipe, M.G.A., Pessoa
Jr., A., 2010. Scale-up of diluted sulfuric acid hydrolysis for producing sug-
arcane bagasse hemicellulosic hydrolysate (SBHH). Bioresour. Technol. 101,
1247–1253.
Rodríguez-Chong, A., Ramírez, J.A., Garrote, G., Vázquez, M., 2004. Hydrolysis of
sugar cane bagasse using nitric acid: a kinetic assessment. J. Food Eng. 61,
143–152.
Saad, M.B., Oliveira, L., Candido, R.G., Quintana, G., Rocha, G.J.M., Gonçalves, A.R.,
2008. Preliminary studies on sugarcane straw fungal treatment for organosolv
pulping. Enzyme Microb. Technol. 43, 220–225.
Silva, V.F.N., 2010. Studies of pretreatment and enzymatic saccharification of agroin-
dustrial wastes, as stages in the process for obtaining the cellulosic ethanol.
Master’s Degree Dissertation. University of São Paulo - Lorena Engineering
School, Lorena, SP, Brazil.
Silva, V.F.N., Arruda, P.V., Felipe, M.G.A., Gonçalves, A.R., Rocha, G.J.M., 2010.
Cellulosic hydrolysates fermentation obtained through sugarcane bagasse enzy-
matic saccharification pretreated by hydrothermal processing. J. Ind. Microbiol.
Biotechnol. 37, 1476–5535.
Taherzadeh, M.J., Karimi, K., 2008. Pretreatment of lignocellulosic wastes to improve
ethanol and biogas production: a review. Int. J. Mol. Sci. 9, 1621–1651.
Tan, H., Yang, R., Sun, W., Wang, S., 2010. Peroxide-acetic acid pretreatment to
remove bagasse lignin prior to enzymatic hydrolysis. Ind. Eng. Chem. Res. 49,
1473–1479.
Yang, B., Wyman, C.E., 2008. Pretreatment: the key for unlocking low-cost cellulosic
ethanol. Biofuels Bioprod. Bioref. 2, 26–40.