Industrial Crops and Products 50 (2013) 707714

Contents lists available at ScienceDirect

Industrial Crops and Products

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

Cellulose nanocrystals from pineapple leaf, a new approach for the

reuse of this agro-waste
Roni Marcos dos Santos a , Wilson Pires Flauzino Neto a , Hudson Alves Silvrio a ,
Douglas Ferreira Martins a , Nolio Oliveira Dantas b , Daniel Pasquini a,
a
Instituto de Qumica, Universidade Federal de Uberlndia, Campus Santa Mnica, Av. Joo Naves de vila, 2121, 38400-902, Uberlndia, Minas Gerais,
Brazil
b
Instituto de Fsica, Universidade Federal de Uberlndia, Campus Santa Mnica, Av. Joo Naves de vila, 2121, 38400-902, Uberlndia, Minas Gerais, Brazil

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

Article history: Pineapple leaf (PL) is an annually renewable agricultural residue, available in abundance, which is used
Received 7 April 2013 very rarely and is of limited value at present. Therefore, this agro-waste deserves to be better and/or
Received in revised form 8 August 2013 properly used. The aim of this study was to explore PL as a source of raw material for the production of
Accepted 16 August 2013
cellulose nanocrystals (CN). The CN were extracted by acid hydrolysis at 45 C for 5, 30 or 60 min, using
20 mL of H2 SO4 (9.17 M) for each gram of material. The resulting CN were characterized by crystallinity
Keywords:
index, FTIR, morphology (shape and size) and thermal stability. Among the hydrolysis conditions carried
Pineapple leaf
out, the best extraction time was 30 min. At this extraction time, the CN presented a needle-shaped
Cellulose nanocrystals
Agricultural residue
nature, high thermal stability (225 C), high crystallinity (73%), an average length of 249.7 51.5 nm and
Reuse a diameter of 4.45 1.41 nm, giving an aspect ratio (L/D) of around 60. Therefore, CN obtained from PL
has great potential as reinforcement in the manufacture of nanocomposites. The production of CN from
this underutilized agro-waste has commercial application potential that can add value to the pineapple
cultivation, generate extra income for farmers and also help in agribusiness diversication. In addition,
the reuse of these residues allows a signicant reduction in both the volume of waste accumulated in the
environment and in the extraction of raw materials.
2013 Elsevier B.V. All rights reserved.

1. Introduction residues in industrial processes for the generation of value-added

In recent years, the quest for sustainable development has moti-
vated efforts toward maximizing the efciency of the use of raw
materials and minimizing the creation of waste (Ashori, 2008). In
this context, the use of biomass residues as feedstock for the pro-
duction of energy and materials has been the object of intensive
academic and industrial research (Mishra et al., 2004; Reddy and
Yang, 2005; Schievano et al., 2009; Silva et al., 2009). The reuse of
these residues allows a signicant reduction both in the volume of
waste accumulated in the environment and in the extraction of raw
materials. Thus, an efcient reuse of these wastes is of great impor-
tance, not only for minimizing the environmental impact, but also
for obtaining a higher prot.
Agriculture is an important sector in the Brazilian economy
(Rahman, 2011). Diversication of the industry is crucial in encour-
aging economic stability and growth. The utilization of these crop
Corresponding author. Tel.: +55 34 3239 4143; fax: +55 34 3239 4208.
E-mail addresses: pasquini@iqufu.ufu.br, danielpasquini2005@yahoo.com.br
(D. Pasquini).
products, such as in the production of high performance materi-
als, could be an additional source of revenue for farmers and also
help in agro-industry diversication by providing a non-food-based
market for agro-wastes (Alemdar and Sain, 2008; Flauzino Neto
et al., 2013; Rahman, 2011; Reddy and Yang, 2005; Silvrio et al.,
2013).
Pineapple is one of the most popular tropical fruits in the world
and their crop occupies a prominent position in the Brazilian
agricultural sector. Brazil is one of the main producers of this
crop, accounting for approximately 10.9% of the world production
(http://www.cnpmf.embrapa.br/planilhas/Abacaxi Mundo 2010.
pdf). Currently, the main focus of the pineapple industry in this
country is the fruits and related foodstuffs and consequently the
other plant parts (stems, roots and especially leaves) are consid-
ered agricultural residues of pineapple cultivation (dos Santos
et al., 2001; Fagundes and Fagundes, 2010). The post-harvest
residue comprises mainly pineapple leaves, which are mostly
burned to eliminate fungi and other parasites, composted or just
crammed to rot (de Aquino, 2006; Maniruzzaman et al., 2011). This
is due to the lack of adequate technology for this purpose, as well
as the ignorance of the farmers about the existence of commercial

0926-6690/$ see front matter 2013 Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.indcrop.2013.08.049
708 R.M.d. Santos et al. / Industrial Crops and Products 50 (2013) 707714
uses for leaves that can generate extra income for these farmers
(de Aquino, 2006; Mishra et al., 2004).
The practices of decomposing and burning the pineapple leaf
(PL) in situ do not contribute to the improvement of plantation yield,
as reported in previous literature (Ahmed et al., 2002; Mohamed
et al., 2009).
PL is an annually renewable agricultural residue, that is
biodegradable, available in abundance (inexpensive), used very
rarely and of limited value at present. In addition, after harvest-
ing, PL waste remains, causing various problems for farmers to deal
with. There is a great demand to nd other end uses for these agri-
cultural cellulosic wastes (Cherian et al., 2010, 2011; Kengkhetkit
and Amornsakchai, 2012; Maniruzzaman et al., 2011). Hence, with-
out any additional cost input, PL bers can be obtained for industrial
purposes.
Several processes and products have been reported that uti-
lize PL as a raw material. These include the extraction of cellulose
ber and nanober, the production of paper, textiles and com-
posites (Banik et al., 2011; Cherian et al., 2010; Chollakup et al.,
2011; Kengkhetkit and Amornsakchai, 2012; Mishra et al., 2004;
Threepopnatkul et al., 2009). However, there is not yet any pub-
lished work on the extraction of cellulose nanocrystals (CN) from
PL.
CN have attracted immense interest as a novel nanostruc-
tured material during recent years. CN are very high crystallinity
nanoparticles derived from cellulosics bers. CN are a very high-
value material, since they can transform the performance of
existing products as well as helping to create new, unique and
improved products. The unique combination of amazing physico-
chemical properties and environmental benets allows that the CN
offer a wide range of potential applications. At present, the main
application of CN is as a reinforcing agent in the nanocomposite
eld. Others elds of potential applications are packaging, paints,
coatings, special papers, cosmetics, pharmaceuticals, biomedical
materials, textiles, the automotive industry, aerospace, building
materials, the electronic and electrical industry, and many others
(Moon et al., 2011; Peng et al., 2011; Podsiadlo et al., 2005; Silvrio
et al., 2013). One specic example of the application of CN is the
solidication of liquid crystals for optical applications, e.g. security
paper (Beck et al., 2011; Revol et al., 1998).
It is known that the morphology and properties of the CN
depend mainly on the source of the original cellulose, of the extrac-
tion process and their parameters (Beck-Candanedo et al., 2005;
Elazzouzi-Hafraoui et al., 2008). Therefore, the isolation and further
analysis of the characteristics of CN from many kinds of cellulosic
resources is necessary and relevant for the efcient comparison and
exploitation of these resources (Chen et al., 2011; Flauzino Neto
et al., 2013; Silvrio et al., 2013).
CN have been isolated from different vegetable sources, such
as cotton and wood pulp (Beck-Candanedo et al., 2005; Teixeira
et al., 2010), and from animal sources such as tunicates (Berg et al.,
2007). In addition, there are only a few papers which describe the
isolation of whiskers from agricultural byproducts, such as soy hulls
(Flauzino Neto et al., 2013), corncob (Silvrio et al., 2013), rice husk
(Rosa et al., 2012) and sesame husk (Purkait et al., 2011).
In this work, CN were extracted from PL under different condi-
tions of sulfuric acid hydrolysis in order to obtain a material with
a high crystallinity index, thermal stability, aspect ratio and yield.
Different techniques were employed to characterize the PL at dif-
ferent stages of treatment. The characteristics investigated were
the chemical composition, crystallinity index, thermal stability,
surface charge and morphology (shape and size). The aim in this
study was to investigate the viability of this agricultural residue as
a simple and low-cost source of CN and the possibility of adding
value to the pineapple cultivation through a new approach to their
utilization.
2. Experimental
2.1. Materials and methods
The PL used in this study was obtained from residues after har-
vesting pineapple in the So Mateus farm (Comendador Gomes,
Minas Gerais, Brazil). The plant species used was Ananus coso-
mus belonging to the Bromeliaceae family. The other reagents
employed in this study were: sulfuric acid (95.098.0 wt.%, Vetec,
P.A.), sodium hydroxide (Vetec), potassium hydroxide (Vetec),
sodium chlorite (NaClO2 , technical grade, 80%, SigmaAldrich),
glacial acetic acid (Synth), and cellulose membrane (D9402,
SigmaAldrich).
2.2. Preparation of cellulose nanocrystals
2.2.1. Purication
The raw PL was ground in a mill. After that, the PL was treated
with a sodium hydroxide aqueous solution of 2% (w/w) for 4 h at
100 C under mechanical stirring, washed several times with dis-
tilled water until the alkali was completely removed, and nally
dried at 50 C for 12 h in an air-circulating oven. After this treat-
ment, the material was bleached with a solution made up of equal
parts (v:v) of acetate buffer (27 g NaOH and 75 ml glacial acetic
acid, diluted to 1 L of distilled water) and aqueous sodium chlorite
(1.7 wt% NaClO2 in water). This bleaching treatment was performed
at 80 C for 4 h. The bleached material was washed repeatedly in
distilled water until the pH of the material became neutral and
subsequently dried at 50 C for 12 h in an air-circulating oven. The
material content throughout these chemical treatments was about
46% (w/w). The material which resulted after the purication was
the treated pineapple leaf (TPL). The bleaching process utilized has
been used/adapted by other authors (de Rodriguez et al., 2006;
Siqueira et al., 2010a; Flauzino Neto et al., 2013; Silvrio et al.,
2013).
2.2.2. Isolation of cellulose nanocrystals
After the chemical treatment was completed, the TPL was milled
with a blender and then used for the extraction of CN by acid
hydrolysis. The hydrolysis was performed at 45 C for 5 min, 30 min
or 60 min under vigorous and constant stirring. For each gram of
TPL we used 20 mL of H2 SO4 64% (w/w) (9.17 M). Immediately fol-
lowing the hydrolysis, the suspension was diluted 10-fold with
cold water to stop the hydrolysis reaction, and centrifuged twice
for 10 min at 7000 rpm to remove the excess acid. The precipitate
was then dialyzed with tap water to remove non-reactive sulfate
groups, salts and soluble sugars, until the neutral pH (4 days) was
reached. Subsequently, the resulting suspension of dialysis process
was ultrasonicated for 10 min and stored in a refrigerator at 4 C.
Some drops of chloroform were added in the CN suspension to avoid
any bacterial growth. The cellulose nanocrystals from pineapple
leaf were labeled CNPL5 or CNPL30 or CNPL60 , depending on the
time of extraction.
2.3. Characterizations and measurements
2.3.1. Birefringence analysis
Aliquots of CN suspensions at same concentration
(7.103 g mL1 ) were placed in a glass bottle, and then these
bottles were placed in front of a source of polarized light
and photographed by a camera equipped with a polarized
light lter while being agitated with the aid of a magnetic
stirrer.
 R.M.d. Santos et al. / Industrial Crops and Products 50 (2013) 707714
Fig. 1. Photographs of (a) the pineapple cultivation, (b) untreated pineapple leaves, (c) ground pineapple leaves, and (d) treated pineapple leaves.
2.4. Gravimetric analysis
The hydrolysis yields were calculated by drying aliquots of the
CN suspensions with a known volume at 105 C for 12 h in an air-
circulating oven.
2.5. Chemical composition
The chemical composition of PL and TPL was investigated as
follows: the lignin content was measured according to a standard
method of the Technical Association of Pulp and Paper Indus-
try TAPPI T13M-54; the holocellulose (-cellulose + hemicellulose)
content was estimated by the acid chlorite method (Browning,
1967); the -cellulose content was determined treating the holo-
cellulose with potassium hydroxide solutions of 5 and 24% (w/w),
according Browning (Browning, 1967); the hemicellulose content
was found by subtracting the -cellulose part from the holocellu-
lose content; and the ash content was measured by considering the
percentage difference between the initial weight of the dried ber
of the sample and that after calcination for 4 h at 800 C (Trindade
et al., 2005). An average of three replicates was calculated for each
sample.
2.5.1. Fourier transform infrared spectroscopy (FTIR)
A Shimadzu IRPrestige-21 Infrared spectrophotometer was used
to obtain spectra for the PL, TPL, CNPL5 , CNPL30 and CNPL60 . The KBr
disk (ultra-thin pellets) method was used in taking the IR spectra.
Samples were ground and mixed with KBr (sample/KBr ratio, 1/100)
to prepare pastilles. The experiments were carried out in the range
of 5004000 cm1 with a resolution of 4 cm1 and a total of 32 scans
for each sample.
2.6. X-ray diffraction (XRD)
The X-ray diffractograms of PL, TPL, CNPL5 , CNPL30 and CNPL60
were obtained at room temperature within a 2 range from 5 to 40
and a scan rate of 2 min1 . The equipment used was a diffractome-
ter Shimadzu LabX XRD-6000, operating at a power of 40 kV with a
current of 30 mA and Cu K radiation (1.5406 A). Before performing
the XRD, all samples were dried at 50 C for 12 h in an air-circulating
oven.
The diffractograms were deconvoluted into peaks and halos
referring to the crystalline and amorphous regions, respectively.
709
These deconvolutions were obtained using the Pseudo-Voigt 2 peak
function from Origin 7.0, which is shown in Eq. (1), and these
deconvolutions were evaluated according to the two-phase model
(Cerqueira et al., 2006).

2 wL
y = y0 + A mu
 4(x xc )2 + w2
L

4 ln 2 (4 ln 2/w2 )(xxc )2
+ (1 mu )  e G (1)
WG
where wL and wG are the width at half maximum for Lorentz and
Gauss components of the above equation, respectively, A is the area
and mu is the prole shape factor.
Considering this model, the crystallinity indexes (CrI) of the
samples were calculated using Eq. (2):
 Ac

CrI = 100 (2)
Ac + Aa
where Ac and Aa are the areas under the crystalline peaks and the
amorphous halos, respectively determined by the deconvolutions.
2.6.1. Atomic force microscopy (AFM)
AFM measurements were performed with Shimadzu SPM-9600
equipment for evaluating the morphology of CNPL5 , CNPL30 and
CNPL60 . A drop of a diluted nanocrystals aqueous suspension (to
about 5.105 g mL1 ) was deposited onto a freshly cleaved mica
surface and air-dried. AFM images were obtained at room temper-
ature in the dynamic mode with a scan rate of 1 Hz and using Si
tips with a curvature radius of less than 10 nm and a spring con-
stant of 42 N m1 . The dimensions of nanocrystals were determined
using VectorScan software (software for Shimadzus SPM-9600).
To eliminate the effect of tip radius on width measurements, we
measured the heights of the nanocrystals, which are not subject
to peak broadening artifacts, and assumed the nanocrystals to be
cylindrical in shape (Beck-Candanedo et al., 2005). Seventy-ve
nanocrystals were randomly selected to determine the average
length, width and aspect ratio. For each nanocrystal, one measure-
ment of the length and two measurements of the diameter were
performed and the aspect ratio was calculated.
2.6.2. Thermal characterization (TG)
Thermal stabilities of PL, TPL, CNPL5 , CNPL30 and CNPL60 were
evaluated using Shimadzu DTG-60H equipment. The analysis
710 R.M.d. Santos et al. / Industrial Crops and Products 50 (2013) 707714
Fig. 2. FTIR spectra of PL, TPL, CNPL5 , CNPL30 and CNPL60 .
conditions were: a nitrogen atmosphere with ow 30 mL min1 ,
heating rate of 10 C min1 , temperature range from 25 to 600 C,
sample mass between 5 and 7 mg and aluminum pans.
3. Results and discussion
3.1. Purication, chemical composition and FTIR
A diluted alkali treatment was performed to remove the lignin,
hemicelluloses, waxes, pectins, proteins, soluble mineral salts, sil-
ica and ash, while bleaching was applied to remove the lignin
residues (Dufresne et al., 1997; Flauzino Neto et al., 2013; Sheltami
et al., 2012). The yield of purication was 40% (by dry mass). Fig. 1
shows the physical aspect of the PL before and after purication
(TPL). Visually, the white color of the material after purication
indicates that a great part of the initial non-cellulosic components
was removed. The contents of the cellulose were 36.3 3.8% and
74.5 4.2%, of the hemicelluloses were 22.9 2.0% and 20.4 2.6%,
of the lignin were 27.53 1.94% and 8.72 1.63%, and ash were
2.85 0.24% and 2.28 0.11%, for PL and TPL respectively (the val-
ues are based on dry basis).
It was veried that the percentage of mass consumption of the
main components of the PL due to the purication process was 18.0,
64.4 and 87.4% for -cellulose, hemicelluloses and lignin, respec-
tively. The mass loss of cellulose during the purication process
can be related to two factors: (i) the breakdown of cellulose chains
during bleaching and (ii) the loss of material inherent in various
ltering processes and mass transfer (Flauzino Neto et al., 2013).
Taking into account that purication was performed to remove
non-cellulose components and mainly lignin, which acts as a natu-
ral agglutinative hindering acid hydrolysis, the purication process
has reached its goal because the composition of TPL is suitable for
the extraction of cellulose nanocrystals (low content of lignin and
high content of cellulose).
Fig. 2 shows the FTIR spectra of PL, TPL, CNPL5 , CNPL30 and
CNPL60 . The prominent peak at 1742 cm1 in the spectrum of PL
is attributed to the acetyl and uronic ester groups of the hemi-
celluloses or the ester linkage of carboxylic group of ferulic and
p-coumaric acids of lignin and/or hemicelluloses (Kejun et al., 2011;
Sun et al., 2005). In the same spectrum, the peak at 1514 cm1 is
associated with the C C aromatic skeletal vibration of lignin (Kejun
et al., 2011; Sun et al., 2005; Vargas et al., 2011; Xiao et al., 2001),
and the band near 1254 cm1 corresponds to the axial asymmetric
strain of C O C, which is commonly observed when C O e.g.
Fig. 3. The resulting colloidal suspensions for CNPL5 , CNPL30 and CNPL60 after a few
hours of rest.
in ether, ester, and phenol groups are present (Siqueira et al.,
2010b). Comparing the data shown in the FTIR spectra for PL and
TPL, it could be noted that the lack of peaks at 1742 cm1 , 1514 cm1
and 1254 cm1 in the spectrum of TPL is due to the signicant
removal of hemicelluloses, and mainly lignin, by the purication
process (alkali and bleaching treatments).
The peak at 1061 cm1 is assigned to the C O stretching and the
C H rock vibrations of the cellulose (Alemdar and Sain, 2008). The
small increase in this peak for TPL in relation to PL indicates that the
TPL have higher cellulose content. Similar behavior was observed
when comparing the spectra of TPL with CNPL5 , CNPL30 and CNPL60 .
This peak appeared in all of the spectra and the differences pre-
sented suggest that the CNPL5 , CNPL30 and CNPL60 samples has a
very high content of cellulose.
3.2. Isolation of Cellulose nanocrystals, gravimetric and
birefringence analysis
The yields of the CNPL, with respect to the initial amount of dried
TPL bers, for CNPL5 , CNPL30 and CNPL60 were 77, 65 and 55 wt%
respectively; these values are consistent with the literature data
(Silvrio et al., 2013; Teixeira et al., 2011, 2010).
Among the several methods for preparing CN, acid hydroly-
sis is the most well-known and widely used (Peng et al., 2011).
This process breaks down the disordered and amorphous parts of
the cellulose, releasing single and well-dened crystals. Thus, this
process is based on the quicker hydrolysis kinetics presented by
amorphous regions, as compared to crystalline ones (Habibi et al.,
2010; Peng et al., 2011; Teixeira et al., 2011).
During the hydrolysis process with sulfuric acid, sulfate groups
are introduced on the surface of the CN by esterication of the
hydroxyl groups from cellulose. This allows an anionic stabilization
by repulsive forces, leading to the achievement of stable aque-
ous dispersions of CN (Beck-Candanedo et al., 2005; Lima and
Borsali, 2004; Silva and DAlmeida, 2009). Also, it is known that
the increased extraction time results in higher sulfate content in
the CN (Flauzino Neto et al., 2013; Roman and Winter, 2004). Only
the hydrolysis times of 30 and 60 min led to stable aqueous suspen-
sions being obtained (as shown in Fig. 3). The suspension obtained
with 5 min of hydrolysis tended to aggregate after a few hours (as
seen in Fig. 3), probably this particle agglomeration is related to the
large size and small amount of surface charges of these particles.
Birefringence was used to conrm the presence of isolated
nanowhiskers in a suspension and is considered by some authors as
 R.M.d. Santos et al. / Industrial Crops and Products 50 (2013) 707714
Fig. 4. Photograph of aqueous suspension of CNPL5 , CNPL30 and CNPL60 observed
between cross nicols showing the formation of birefringent domains.
a good dispersibility criterion in suspension (Silva and DAlmeida,
2009; Silvrio et al., 2013). This birefringence results from two ori-
gins: (1) a structural anisotropy form of cellulose and (2) a ow
anisotropy resulting from the alignment of the nanocrystals under
ow, generally operated before observation. Fig. 4 shows a bire-
fringence of different cellulose suspensions in water, indicating
that CN were isolated in all three isolation conditions. The birefrin-
gence was somewhat weaker for CNPL5 suspension compared to
the CNPL30 and CNPL60 suspensions. It is possible that the hydroly-
sis time was not sufcient to separate single CN from TPL bers
as effectively as the others hydrolysis times and, therefore, the
birefringence is not as strong (Oksman et al., 2011).
3.3. X-ray diffraction
The XRD patterns of PL, TPL, CNPL5 , CNPL30 and CNPL60 are
shown in Fig. 5. These patterns are typical of semi-crystalline mate-
rials with an amorphous broad hump and crystalline peaks. In all
Fig. 5. X-ray diffractograms of PL, TPL, CNPL5 , CNPL30 and CNPL60 .
711
of the diffractograms proles (Fig. 5), there is a predominance of
type I cellulose, veried by the presence of peaks at 2 = 15 (plane
1 0 1), 17 (plane 101), 21 (plane 0 2 1), 23 (plane 0 0 2) and 34
(plane 0 0 4) (Borysiak and Garbarczyk, 2003; Flauzino Neto et al.,
2013). The peak at 2 = 26.5 present in all the spectra is related to
the sample holder.
The CrI was found to be about 49, 64, 69, 73 and 68% for the
PL, TPL, CNPL5 , CNPL30 and CNPL60 , respectively. The higher CrI
value of TPL compared to PL can be well understood by the reduc-
tion and removal of amorphous non-cellulosic compounds induced
by the alkali and bleaching treatments performed in the puri-
cation process. The increase in the CrI value of CNPL5 in relation
to TPL was also observed, due to the partial removal of the para-
crystalline domains during the acid hydrolysis. A similar behavior
was observed when comparing the diffraction patterns of CNPL5
and CNPL30 samples.
The CNPL60 sample presented a decrease in crystallinity with
respect to CNPL30 . A similar effect of hydrolysis time in excess was
observed by some authors for cellulose nanocrystals from sugar-
cane bagasse bers and pea hull bers (Chen et al., 2009; Teixeira
et al., 2011), although the rod-like structures were maintained,
supported by AFM images.
3.4. Atomic force microscopy (AFM)
An accurate morphological examination of the CN is essential
not only for the promoters of the manufacturing process of the CN,
but also for developers of functional applications. In this way, AFM
topography measurements were performed in order to have a pre-
cise characterization of the dimensions of the individual crystallites
(Flauzino Neto et al., 2013).
Determining the exact dimensions of CN is complicated by the
specic limitations of the different analytical methods used. In the
case of AFM, tip/sample broadening represents the main limitation,
resulting in an overestimation of CN dimensions. Since the CN are
assumed to be cylindrical in shape, the height of the CN was taken
to be equivalent to the diameter, to compensate for image widening
due to the convolution of the tip and the particle (Beck-Candanedo
et al., 2005; Flauzino Neto et al., 2013; Kvien et al., 2005). However,
the tip broadening effects causes an error in the length measure-
ments, but this is unavoidable (Beck-Candanedo et al., 2005).
Fig. 6 shows AFM images of CNPL5 , CNPL30 and CNPL60 . For the
samples CNPL30 and CNPL60 , the AFM images presented needle-
like nanoparticles throughout, conrming that the extraction of
CN from PL was successful. However, the AFM images of sample
CNPL5 showed micro-sized bers and some needle-like nanoparti-
cles. Therefore, it is clear that the hydrolysis conditions employed
for this sample (CNPL5 ) were not sufcient to completely isolate CN
from TPL bers. This is in agreement with the weak birefringence
and particle agglomeration observed for this suspension sample
(CNPL5 ).
Fig. 7 shows the length (L), width (D) and aspect ratio (L/D) his-
tograms of CNPL30 and CNPL60 obtained by several AFM images, as
described in the Experimental Section. The statistics of the length,
width and aspect ratio are shown in Table 1.
The increase in extraction time resulted in a slightly shorter
length for CNPL60 when compared with CNPL30 . This was expected,
since the long extraction time (60 min) partially destroyed areas of
the crystalline domains, as seen by XRD analysis.
Table 1
Length, width, aspect ratio, for CNPL30 and CNPL60 obtained by AFM pictures.
Length (nm) Diameter (nm) Aspect ratio (nm)
CNPL30 249.7 51.5 4.45 1.41 60.1 19.5
CNPL60 190.2 36.5 4.18 1.44 50.4 20.7
712 R.M.d. Santos et al. / Industrial Crops and Products 50 (2013) 707714

Fig. 6. AFM images of CNPL5 , CNPL30 and CNPL60 .

By studying histograms it is clear that the increase in hydrolysis
time resulted in a reduction in particle size, but no signicant dif-
ference in width among the CNPL30 and CNPL60 could be detected,
when the standard deviation of each value was taken into account.
As a consequence of the whiskers preparation conditions,
a decrease in the aspect ratio (L/D) of CNPL60 compared with
CNPL30 was observed. This suggests that CNPL30 can give a bet-
ter reinforcing effect than CNPL60 at the same ller loading levels
(Eichhorn et al., 2010). The average aspect ratio for the CNPL found
in this work is close to the largest values ever reported in the lit-
erature; therefore, these particles have great potential to be used
as reinforcing agents in nanocomposites (Kalia et al., 2011; Silvrio
et al., 2013).
The results of the morphological investigation by AFM are
consistent with other reports in the literature, where CN were
extracted from different sources (Bai et al., 2009; Beck-Candanedo
et al., 2005; de Rodriguez et al., 2006; Elazzouzi-Hafraoui et al.,
2008; Kvien et al., 2005; Rosa et al., 2010; Siqueira et al., 2010a;
Teixeira et al., 2011).
3.5. Thermogravimetric analysis (TGA)
At present, the main application of CN is as a reinforcing agent
in the nanocomposite eld, and typical processing temperatures
for thermoplastics rise above 200 C, thus the thermal stability of
these crystals is a key factor in order for them to be used as effective
reinforcing materials (Roman and Winter, 2004). The thermogravi-
metric (TG) results of the PL, TPL, CNPL5 , CNPL30 and CNPL60 are
shown in Fig. 8. In all cases, a small weight loss was found in the
range of 25150 C, due to the evaporation of the water of the
materials or low molecular weight compounds (about 7%). The ini-
tial degradation temperatures were found to be around 216, 244,
245, 225 and 220 C for the PL, TPL, CNPL5 , CNPL30 and CNPL60 ,
respectively.
The initial degradation temperature of the TPL (244 C) was
appreciably increased compared to that of the original PL (216 C).
Due to the low initial decomposition temperatures of hemicellu-
loses, lignin and pectin, the higher initial temperature of thermal
decomposition of the TPL is related to the partial removal of hemi-
celluloses, lignin, and pectins by purication processes (alkali and
bleaching treatments) (Alemdar and Sain, 2008; Chen et al., 2011;
Flauzino Neto et al., 2013).
As reported in previous studies, treatment with sulfuric acid
leads to a remarkable decrease in thermal stability of CN. This
occurs because the incorporation of sulfate groups on the surface
of the cellulose after hydrolysis has a catalytic effect in its reactions
of thermal degradation (Roman and Winter, 2004).
Taking into account that acid hydrolysis leads to a remarkable
decrease in thermal stability of CN, it was expected that the initial
degradation temperature of CNPL5 were smaller than for TPL, but

Fig. 7. Length (L), width (D) and aspect ratio (L/D) histograms of CNPL30 and CNPL60 obtained by several AFM images.
 R.M.d. Santos et al. / Industrial Crops and Products 50 (2013) 707714
Fig. 8. Thermogravimetric (TG) curves of the PL, TPL, CNPL5 , CNPL30 and CNPL60 .
it was actually almost the same (245 vs. 244 C). This can be clearly
explained by the higher cellulose content of CNPL5 compared to
TPL and the small hydrolysis time (small amount of sulfate groups)
of sample CNPL5 .
The CNPL30 sample presented a decrease in initial degradation
temperature with respect to CNPL5 , as expected since the CNPL30
sample had a higher sulfate content than CNPL5 . Similar behavior
was observed when comparing the initial degradation tempera-
ture of samples CNPL30 and CNPL60 . These results are consistent
with results obtained from the Chemical Composition, XRD and
FTIR measurements.
4. Conclusions
The present work shows that PL is an interesting source of
raw material for the production of CN, due to the characteristics
of the obtained nanocrystals. Chemical treatment performed with
sodium chlorite and alkali removed the non-cellulosic components
resulting in bers with a low content of lignin and a high con-
tent of cellulose, which were therefore suitable for extracting CN.
Through the AFM images it was observed that there was an incom-
plete isolation of CN after 5 min of hydrolysis by the employed
conditions (sample CNPL5 ). However, above 30 min of hydroly-
sis it was possible to obtain stable aqueous suspensions of CNPL
which are negatively charged, due the presence of sulfate groups.
The yields of acid hydrolysis, with respect to the initial amount of
dried TPL bers, for CNPL5 , CNPL30 and CNPL60 were 77, 65 and
55 wt%, respectively. The increase in the hydrolysis time resulted
in a decrease in the dimensions and also in the aspect ratio (L/D) of
the CNPL.
For an extraction time of 30 min, the CN presented a needle-
shaped nature, high thermal stability (225 C), high crystallinity
(73%), an average length of 249.7 51.5 nm and a diameter of
4.45 1.41 nm, giving an aspect ratio of around 60 (which is among
the largest values reported in the literature). It can be concluded
from these results that the CN obtained from PL has great poten-
tial to be used as reinforcement agents for the manufacture of
nanocomposites and also for diversied applications.
The production of CN from this underutilized agro-waste has
commercial application potential that can add value to the pineap-
ple cultivation, generate extra income for farmers and also help
in agribusiness diversication. In addition, the reuse of these
residues allows a signicant reduction both in the volume of
waste accumulated in the environment, as in the extraction of raw
materials.
713
Acknowledgements
The authors thank CAPES/PROAP, CNPq and FAPEMIG for nan-
cial support.
References
Ahmed, O.H., Husni, M.H., Anuar, A.R., Hana, M.M., 2002. Effect of residue man-
agement practices on yield and economic viability of Malaysian pineapple
production. Journal of Sustainable Agriculture 20 (4), 8394.
Alemdar, A., Sain, M., 2008. Isolation and characterization of nanobers from agri-
cultural residues wheat straw and soy hulls. Bioresource Technology 99,
16641671.
Ashori, A., 2008. Wood-plastic composites as promising green-composites for auto-
motive industries. Bioresource Technology 99, 46614667.
Bai, W., Holbery, J., Li, K.C., 2009. A technique for production of nanocrystalline
cellulose with a narrow size distribution. Cellulose 16, 455465.
Banik, S., Nag, D., Debnath, S., 2011. Utilization of pineapple leaf agro-waste for
extraction of bre and the residual biomass for vermicomposting. Indian Journal
of Fibre & Textile Research 36, 172177.
Beck, S., Bouchard, J., Berry, R., 2011. Controlling the reection wavelength of irides-
cent solid lms of nanocrystalline cellulose. Biomacromolecules 12, 167172.
Beck-Candanedo, S., Roman, M., Gray, D.G., 2005. Effect of reaction conditions on the
properties and behavior of wood cellulose nanocrystal suspensions. Biomacro-
molecules 6, 10481054.
Berg, O., Capadona, J.R., Weder, C., 2007. Preparation of homogeneous disper-
sions of tunicate cellulose whiskers in organic solvents. Biomacromolecules 8,
13531357.
Borysiak, S., Garbarczyk, J., 2003. Applying the WAXS method to etimate the super-
molecular structure of cellulose bres after mercerization. Fibres & Textiles in
Eastern Europe 11, 104106.
Browning, B.L., 1967. Methods of Wood Chemistry, 1st ed. Interscience Publishers,
New York.
Cerqueira, D.A., Rodrigues Filho, G., Assunco, R.M.N., 2006. A new value for the
heat of fusion of a perfect crystal of cellulose acetate. Polymer Bulletin 56,
475484.
Chen, Y., Liu, C., Chang, P.R., Cao, X., Anderson, D.P., 2009. Bionanocomposites based
on pea starch and cellulose nanowhiskers hydrolyzed from pea hull bre: effect
of hydrolysis time. Carbohydratre Polymers 76, 607615.
Chen, W., Yu, H., Liu, Y., Hai, Y., Zhang, M., Chen, P., 2011. Isolation and char-
acterization of cellulose nanobers from four plant cellulose bers using a
chemical-ultrasonic process. Cellulose 18, 433442.
Cherian, B.M., Leo, A.L., de Souza, S.F., Thomas, S., Pothan, L.A., Kottaisamy, M.,
2010. Isolation of nanocellulose from pineapple leaf bres by steam explosion.
Carbohydrate Polymers 81, 720725.
Cherian, B.M., Leo, A.L., de Souza, S.F., Costa, L.M.M., de Oliveira, G.M., Kottaisamy,
M., Nagarajan, E.R., Thomas, S., 2011. Cellulose nanocomposites with nano-
bres isolated from pineapple leaf bers for medical applications. Carbohydrate
Polymers 86, 17901798.
Chollakup, R., Tantatherdtam, R., Ujjin, S., Sriroth, K., 2011. Pineapple leaf
ber reinforced thermoplastic composites: effect of ber length and ber
content on their characteristics. Journal of Applied Polymer Science 119,
19521960.
de Aquino, M.S., 2006. Desenvolvimento de uma Desbradeira para Obtenco da
Fibra da Folha do Abacaxi. Master Thesis. Federal University of Rio Grande do
Norte.
de Rodriguez, N.L.G., Thielemans, W., Dufresne, A., 2006. Sisal cellulose whiskers
reinforced polyvinyl acetate nanocomposites. Cellulose 13, 261270.
dos Santos, M.A.T., Nepomuceno, I.A.S., de Abreu, C.M.P., de Carvalho, V.D., 2001.
Teores de polifenis de caule e folha de quatro cultivares de abacaxizeiro. Revista
Brasileira de Fruticultura 23 (2), 274276.
Dufresne, A., Caville, J., Vignon, M., 1997. Mechanical behavior of sheets prepared
from sugar beet cellulose microbrils. Journal of Applied Polymer Science 64,
11851194.
Eichhorn, S.J., Dufresne, A., Aranguren, M., Marcovich, N.E., Capadona, J.R., Rowan,
S.J., 2010. Review: current international research into cellulose nanobres and
nanocomposites. Journal of Materials Science 45, 133.
Elazzouzi-Hafraoui, S., Nishiyama, Y., Putaux, J.L., Heux, L., Dubreuil, F., Rochas, C.,
2008. The shape and size distribution of crystalline nanoparticles prepared by
acid hydrolysis of native cellulose. Biomacromolecules 9, 5765.
Fagundes, N.S., Fagundes, N.S., 2010. Restos culturais do abacaxizeiro na alimentaco
de ruminantes. Revista Eletrnica Nutritime 7 (3), 12431247.
Flauzino Neto, W.P., Silvrio, H.A., Dantas, N.O., Pasquini, D., 2013. Extraction and
characterization of cellulose nanocrystals from agro-industrial residue soy
hulls. Industrial Crops and Products 42, 480488.
Habibi, Y., Lucia, L.A., Rojas, O.J., 2010. Cellulose nanocrystals: chemistry, self-
assembly, and applications. Chemical Reviews 110, 34793500.
Kalia, S., Dufresne, A., Cherian, B.M., Kaith, B.S., Avrous, L., Njuguna, J., Nassiopou-
los, E., 2011. Cellulose-based bio- and nanocomposites: a review. International
Journal of Polymer Science 2011, 135.
Kejun, S., Juntao, Z., Ying, C., Zongwen, L., Lin, R., Cong, L., 2011. Accelerating the
degradation of green plant waste with chemical decomposition agents. Journal
of Environmental Management 92, 27082713.
714 R.M.d. Santos et al. / Industrial Crops and Products 50 (2013) 707714
Kengkhetkit, N., Amornsakchai, T., 2012. Utilisation of pineapple leaf waste for plas-
tic reinforcement: 1. A novel extraction method for short pineapple leaf ber.
Industrial Crops and Products 40, 5561.
Kvien, I., Tanem, B.S., Oksman, K., 2005. Characterization of cellulose whiskers
and their nanocomposites by atomic force and electron microscopy. Biomacro-
molecules 6, 31603165.
Lima, M.M.S., Borsali, R., 2004. Rodlike cellulose microcrystals: structure, properties,
and applications. Macromolecular Rapid Communications 25, 771787.
Maniruzzaman, M., Rahman, M.A., Gafur, M.A., Fabritius, H., Raabe, D., 2011. Modi-
cation of pineapple leaf bers and graft copolymerization of acrylonitrile onto
modied bers. Journal of Composite Materials 46, 7990.
Mishra, S., Mohanty, A.K., Drzal, L.T., Misra, M., Hinrichsen, G., 2004. A review on
pineapple leaf bers, sisal bers and their biocomposites. Macromolecular Mate-
rials and Engineering 289, 955974.
Mohamed, A.R., Sapuan, S.M., Shahjahan, M., Khalina, A., 2009. Characterization of
pineapple leaf bers from selected Malaysian cultivars. Journal of Food, Agricul-
ture & Environment 7 (1), 235240.
Moon, R.J., Martini, A., Nairn, J., Simonsen, J., Yougblood, J., 2011. Cellulose nano-
materials review: structure, properties and nanocomposites. Chemical Society
Reviews 40, 39413994.
Oksman, K., Etang, J.A., Mathew, A.P., Jonoobi, M., 2011. Cellulose nanowhiskers
separated from a bio-residue from wood bioethanol production. Biomass &
Bioenergy 35, 146152.
Peng, B.L., Dhar, N., Liu, H.L., Tam, K.C., 2011. Chemistry and applications of nanocrys-
talline cellulose and its derivatives: a nanotechnology perspective. The Canadian
Journal of Chemical Engineering 9999, 116.
Podsiadlo, P., Choi, S., Shim, B., Lee, J., Cuddihy, M., Kotov, N.A., 2005. Molecularly
engineered nanocomposites: layer-by-layer assembly of cellulose nanocrystals.
Biomacromolecules 6, 29142918.
Purkait, B.S., Ray, D., Sengupta, S., Kar, T., Mohanty, A., Misra, M., 2011. Isolation
of cellulose nanoparticles from sesame husk. Industrial Engineering Chemistry
Research 50, 871876.
Rahman, M.A., 2011. Study on modied pineapple leaf ber, journal of textile and
apparel. Technology and Management 7 (2), 116.
Reddy, N., Yang, Y., 2005. Biobers from agricultural byproducts for industrial appli-
cations. Trends in Biotechnology 23, 2227.
Revol, J.-F., Godbout, L., Gray, D.G., 1998. Solid self-assembled lms of cellulose with
chiral nematic order and optically variable properties. Journal of Pulp and Paper
Science 24, 146149.
Roman, M., Winter, W.T., 2004. Effect of sulfate groups from sulfuric acid hydrolysis
on the thermal degradation behavior of bacterial cellulose. Biomacromolecules
5, 16711677.
Rosa, M.F., Medeiros, E.S., Malmonge, J.A., Gregorski, K.S., Wood, D.F., Mattoso, L.H.C.,
Glenn, G., Orts, W.J., Imam, S.H., 2010. Cellulose nanowhiskers from coconut
husk bers: effect of preparation conditions on their thermal and morphological
behavior. Carbohydrate Polymers 81, 8392.
Rosa, S.M.L., Rehman, N., de Miranda, M.I.G., Nachtigall, S.M.B., Bica, C.I.D., 2012.
Chlorine-free extraction of cellulose from rice husk and whisker isolation. Car-
bohydrate Polymers 87, 11311138.
Schievano, A., DImporzano, G., Adani, F., 2009. Substituting energy crops with
organic wastes and agro-industrial residues for biogas production. Journal of
Environmental Management 90, 25372541.
Sheltami, R.M., Abdullah, I., Ahmad, I., Dufresne, A., Kargarzadeh, H., 2012. Extraction
of cellulose nanocrystals from mengkuang leaves (Pandanus tectorius). Carbohy-
drate Polymers 88, 772779.
Silva, R., Haraguchi, S.K., Muniz, E.C., Rubira, A.F., 2009. Aplicaces de bras lig-
nocelulsicas na qumica de polmeros e em compsitos. Qumica Nova 32,
661671.
Silva, D.J., DAlmeida, M.L.O., 2009. Cellulose whiskers. O Papel 70, 3452.
Silvrio, H.A., Flauzino Neto, W.P., Dantas, N.O., Pasquini, D., 2013. Extraction
and characterization of cellulose nanocrystals from corncob for application as
reinforcing agent in nanocomposites. Industrial Crops and Products 44, 427436.
Siqueira, G., Abdillahi, H., Bras, J., Dufresne, A., 2010a. High reinforcing capability
cellulose nanocrystals extracted from Syngonanthus nitens (Capim Dourado).
Cellulose 17, 289298.
Siqueira, G., Bras, J., Dufresne, A., 2010b. Luffa cylindrical as lignocellulosic source of
ber, microbrillated cellulose, and cellulose nanocrystals. Bioresources 5 (2),
727740.
Sun, X.F., Xu, F., Sun, R.C., Fowler, P., Baird, M.S., 2005. Characteristics of degraded
cellulose obtained from steam-exploded wheat straw. Carbohydrate Research
340, 97106.
Teixeira, E.M., Corra, A.C., Manzoli, A., Leite, F.L., Oliveira, C.R., Mattoso, L.H.C., 2010.
Cellulose nanobers from white and naturally colored cotton bers. Cellulose
17, 595606.
Teixeira, E.M., Bondancia, T.J., Teodoro, K.B.R., Corra, A.C., Marconcini, J.M., Mat-
toso, L.H.C., 2011. Sugarcane bagasse whiskers: extraction and characterizations.
Industrial Crops and Products 33, 6366.
Threepopnatkul, P., Kaerkitcha, N., Athipongarporn, N., 2009. Effect of surface
treatment on performance of pineapple leaf berpolycarbonate composites.
Composites Part B 40, 628632.
Trindade, W.G., Hoareau, W., Megiatto, J.D., Razera, A.T., Castellan, A., Frollini, E.,
2005. Thermoset phenolic matrices reinforced with unmodied and surface-
grafted furfuryl alcohol sugar cane bagasse and curaua bers: properties of bers
and composites. Biomacromolecules 6, 24852496.
Vargas, A.M.M., Cazetta, A.L., Garcia, C.A., Moraes, J.C.G., Nogami, E.M., Lenzi, E., Costa,
W.F., Almeida, V.C., 2011. Preparation and characterization of activated carbon
from a new raw lignocellulosic material: amboyant (Delonix regia) pods. Journal
of Environmental Management 92, 178184.
Xiao, B., Sun, X.F., Sun, R.C., 2001. Chemical, structural, and thermal characteri-
zation of alkali-soluble lignins and hemicelluloses, and cellulose from maize
stems, rye straw, and rice straw. Polymer Degradation and Stability 74,
307319.