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Carbohydrate Polymers
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Article history: Nanocellulose was successfully isolated from Gelidium elegans red algae marine biomass. The red algae
Received 17 April 2016 ber was treated in three stages namely alkalization, bleaching treatment and acid hydrolysis treat-
Received in revised form 13 June 2016 ment. Morphological analysis was performed by eld emission scanning electron microscopy (FESEM)
Accepted 19 June 2016
and transmission electron microscopy (TEM). TEM results revealed that the isolated nanocellulose had
Available online 20 June 2016
the average diameter and length of 21.8 11.1 nm and of 547.3 23.7 nm, respectively. Fourier transform
infrared (FTIR) spectroscopy proved that the non-cellulosic polysaccharides components were progres-
Keywords:
sively removed during the chemically treatment, and the nal derived materials composed of cellulose
Red algae
Pulp
parent molecular structure. X-ray diffraction (XRD) study showed that the crystallinity of yielded product
Fiber had been improved after each successive treatments subjected to the treated ber. The prepared nano-
Nanocellulose dimensional cellulose demonstrated a network-like structure with higher crystallinity (73%) than that
Polysaccharides of untreated ber (33%), and possessed of good thermal stability which is suitable for nanocomposite
material.
2016 Elsevier Ltd. All rights reserved.
1. Introduction strength and modulus, high surface area, high aspect ratio, high
tensile strength and stiffness, cost effectiveness, light weight and
During the past decade, several reports on the preparation environmental benets due to recyclable and biodegradable, the
of nanostructured cellulose materials from vary lignocellulosic market demand for this natural ber has increased exponentially
biomass resources have been extensively reported such as soft- (Lam, Male, Chong, Leung, & Luong, 2012). In recent year, a new
wood kraft pulp, hardwood elemental chlorine free pulp, cotton nanomaterial source (solid nanocellulose ber) was successfully
ber, recycled pulp, ax ber, sisal ber, ramie ber, bamboo isolated from marine biomass such as Posidonia oceanica ball and
residue, wheat straw, grass ber and microcrystalline cellulose leaves (Bettaieb, Khiari, Dufresne, Mhenni, Putaux, & Bou, 2015;
using top-down technologies (Bettaieb et al., 2015). Cellulose Bettaieb, Khiari, Dufresne, Mhenni, & Belgacem, 2015; Bettaieb
is one of the most abundant natural biopolymer on the Earth, et al., 2015). Compared to land plant, marine biomass has become
and widely used for many industry areas such as papermaking, more popular as a promising source for nanocellulose production
food packaging, pharmaceutical applications and green compos- near future. The advantages of using marine biomass over ligno-
ites innovative applications (Bettaieb, Khiari, Dufresne, Mhenni, & cellulosic biomass for the biopolymer production are: (i) low or
Belgacem, 2015). Due to its unique characteristics such as high zero content of natural physicochemical barriers in algae biomass,
thus no harsh chemical treatment is required to remove their native
recalcitrant structure in order to increase the cellulose accessibility
(Korzen, Pulidindi, Israel, Abelson, & Gedanken, 2015); (ii) marine
Corresponding author at: Nanotechnology & Catalysis Research Centre
algae contains of higher yields of stored carbohydrates, and capa-
(NANOCAT), Institute of Postgraduate Studies, University of Malaya, 50603 Kuala
ble to grow under simple growth requirements by utilizing carbon
Lumpur, Malaysia. Tel.: +603 79676960.
Corresponding author at: Nanotechnology & Catalysis Research Centre dioxide, light and inorganic nutrients efciently, which no cost are
(NANOCAT), Institute of Postgraduate Studies, University of Malaya, 50603 Kuala required for providing (Jeong, Choi, Lee, & Oh, 2012); (iii) rapid
Lumpur, Malaysia. Tel.: +603 79676954. growth rate of algae plant and can be harvested within short period
E-mail addresses: leehweivoon@um.edu.my (H.V. Lee), jcjuan@um.edu.my of time compare to others terrestrial plants, and thus capable to
(J.C. Juan).
http://dx.doi.org/10.1016/j.carbpol.2016.06.083
0144-8617/ 2016 Elsevier Ltd. All rights reserved.
Y.W. Chen et al. / Carbohydrate Polymers 151 (2016) 12101219 1211
increasing the demand of feedstocks for nanocellulose production disintegration (Li et al., 2012) and enzyme-assisted hydrolysis
(Kim, Wi, Jung, Song, & Bae, 2015). (Janardhnan & Sain, 2007). Up to date, acid hydrolysis is still consid-
Marine seaweeds can be classied into three different groups ered the most effective and highly effective in the role of cellulose
including red algae, green algae and brown algae. However, the depolymerization process for conversion of cellulose into its nano-
study showed that red algae such as Gelidium is a highly potential dimensional structure. Cellulose bers are subjected to strong acid
biomass sources for various application due to its high carbo- hydrolysis such as sulfuric (H2 SO4 ), hydrochloric (HCl), and phos-
hydrate content and can be hydrolyzed into smaller molecular phoric acid (H3 PO4 ) to break down the long cellulose polymeric
products such as glucose and galactose (Jeong et al., 2012; Kim et al., chain into smaller dimensions. Under the controlled hydrolysis
2015). In the previous studies, this algae biomass species had been conditions of temperature, time, agitation and acid concentra-
used for production of useful chemicals such as bioethanol (Kim tion, the amorphous domains of cellulose matrix are preferentially
et al., 2015; Korzen et al., 2015) and soluble sugars (mainly glucose, hydrolyzed by the hydronium ions (H3 O+ ) and leave the highly
5-hydroxymethylfurfural (HMF), formic acid, and levulinic acid) crystalline segments unaltered (Yahya, Lee, & Hamid, 2015). Tra-
(Jeong et al., 2012). Many researchers treated the algae biomass ditionally, highly crystalline nanocellulose was prepared by acid
under harsh treatment conditions (i.e. high temperature or pres- hydrolysis as compared with others hydrolysis techniques. H2 SO4
sure) in order to yield different types of organic products as a hydrolysis technique has been widely used for isolating highly
liquid fraction, rather than solid cellulosic materials. The red algae crystalline cellulose from various sources, such as sisal ber (De
biomass such as Gelidium has a rich content of carbohydrates, Rodriguez, Thielemans, & Dufresne, 2006), ax ber (Cao, Dong,
which is mainly composed of cellulose and agar (Jeong et al., & Li, 2007), ramie ber (Zoppe, Peresin, Habibi, Venditti, & Rojas,
2012; Yoon et al., 2010). Therefore, it is possible to obtain solid 2009), grass ber (Pandey et al., 2009), cotton ber (Wang, Tian, &
crystalline nanoparticles (nanocellulose) rather than water soluble Zhang, 2010), bamboo pulp (Liu, Zhong, Chang, Li, & Wu, 2010),
fermentable sugars monomers under the controllable hydrolysis sugarcane bagasse (Mandal & Chakrabarty, 2011), China cotton,
conditions. South Africa cotton, waste tissue paper (Maiti et al., 2013), rice
The combination of alkaline treatment, oxidative bleaching and straw (Jiang & Hsieh, 2013), borer powder (Hu et al., 2014), recycled
acid hydrolysis are the most common and effective multiple-step newspaper (Mohamed, Salleh, Jaafar, Asri, & Ismail, 2015), empty
purication process. It has been widely used to isolate cellulosic fruit bunch (Ching and Ng, 2014; Goh et al., 2016), forest residues
material from typical terrestrial biomass plants, such as coconut (Moriana, Vilaplana, & Ek, 2016), and arecanut husk (Chandra,
husk (Rosa et al., 2010), rice husk (Johar, Ahmad, & Dufresne, George, & Narayanankutty, 2016). Recently, Jiang and Hsieh (2013)
2012), mengkuang leaves (Sheltami, Abdullah, Ahmad, Dufresne, & and Oun and Rhim (2016) investigated the effect of different
Kargarzadeh, 2012), Hibiscus cannabinus (Zaini, 2013), and empty hydrolysis treatments on the crystalline index of nanocellulose
fruit bunch (Ching & Ng, 2014; Goh, Ching, Chuah, Abdullah, derived from rice straw cellulose, and the results showed that the
& Liou, 2016). In general, the main components of red algae H2 SO4 -treated nanocellulose rendered higher crystallinity (90.7%)
consists of mucilaginous materials (agar) and energy dense sub- than that of mechanical blending (82.5%) and TEMPO-mediated
strate such as cellulose material (Seo, Lee, Lee, & You, 2010). oxidation (64.4%). It is important to produce high crystallinity
This cellulosic material can be served as a potential feedstock nanocellulose as this is the most signicant properties that have
for bioconversion to nanocellulose. Like others cellulosic biomass greater inuence on the reinforcing mechanical capabilities than
originated from land plants, the long polymeric chains cellulose other physicochemical properties such as aspect ratio, thermal sta-
extracted from algae can also be hydrolyzed using strong min- bility and surface area (Moriana et al., 2016; Silvrio, Neto, Dantas,
eral acid (i.e. sulfuric acid and hydrochloric acid) into smaller & Pasquini, 2013).
solid particles in nano-dimension (i.e. nanocellulose). In order to As abovementioned that, a combination of alkalization, bleach-
produce high quality cellulose pulp from algae more effectively, ing process and acid hydrolysis was effective in isolation of
non-cellulosic components must be removed from the algae during nanocellulose from numerous land biomass sources, therefore we
the extraction process. Arvizu-Higueras research group (Arvizu- believe that same strategy could be applied to marine biomass to
Higuera, Rodrguez-Montesinos, Murillo-lvarez, Munoz-Ochoa, & produce high crystallinity nanocellulose. Gelidium marine biomass
Hernndez-Carmona, 2008) reported that 7% NaOH alkalization is available enormously around the world, especially in the shal-
treatment had a signicant and positive effect in solubilizes the agar low coast with the 310 m depth below the water of many East
from the marine algae plant (Gracilaria vermiculophylla) at 85 C Asian countries, such as Malaysia, Singapore, Japan, China, and
for 2 h. Besides that, color of yielded nanocellulose is also play a Korea. The production of red algae has increasing exponentially
very important factor because it can affect the consumer accep- from 5.3 million tons in 2006 to 10.8 million tons in 2011 (Kim
tance in various applications (Saelee, Yingkamhaeng, Nimchua, et al., 2015). Typically, the exploitation of Gelidium marine biomass
& Sukyai, 2016). The red algae initially was exhibited in reddish as the main source of commercial agar production and food indus-
color due to the presence of rhizoidal laments components in try will generates huge amount of solid residues after extracting the
the ber which contributed to its color (Seo et al., 2010). There- agar component, which are currently not valorized and normally
fore, a successive bleaching treatment is necessary to remove the indiscriminately discharged into environment (Ennouali, Ouhssine,
natural pigment and chlorophyll for production of highly puried, Ouhssine, & Elyachioui, 2006; Seo et al., 2010). In fact, this solid
whiteness extracted cellulose pulp form the red algae biomass. residue can be fully utilized for the production of nanocellulose
Basically, two main oxidizing bleaching agents namely sodium because it consists of large amount of cellulose. Therefore, the
chlorite (NaClO2 ) and hydrogen peroxide (H2 O2 ) were used to Gelidium red algae appear to be a new promising candidate for
bleach the algae biomass ber to produce bleached red algae pulp nanocellulose production due to their abundance and availability
(Bettaieb, Khiari, Dufresne, Mhenni, Putaux et al., 2015; Bettaieb than terrestrial biomass. In 2010, Seo et al. (2010) was rst reported
et al., 2015; Saelee et al., 2016). Due to environmental concern, on the utilization of two different species of red algae, namely Gelid-
chlorite-free hydrogen peroxide was selected as oxidizing bleach- ium amansii and Gelidium corneum for the production of bleached
ing agent for pulping process in this study. pulp in papermaking industry. The results revealed that the paper
The most common choices of isolating nanocellulose are acid made by using the isolated red algae pulp possessed of particularly
hydrolysis (Ching & Ng, 2014; Ching, Rahman, Ching, Sukiman, & high smoothness and opacity than typical wood pulp. However,
Cheng, 2015; Liew, Thielemans, & Hewakandamby, 2016), TEMPO- they did not examine the physicochemical characterizations (i.e.
mediated oxidation (Tanaka, Saito, & Isogai, 2012), mechanical
1212 Y.W. Chen et al. / Carbohydrate Polymers 151 (2016) 12101219
surface morphological studies, crystallinity structure and thermal -cellulose content was extracted from the resultant sample
properties) of isolated cellulose pulp. after determining their holocellulose content by further treating
In this study, Gelidium elegans was used to produce nanocel- the bers with 50 mL of 17.5% NaOH solution according to the
lulose. In addition, we propose for the rst time the isolation of ASTM D1103-55T. Then, the suspension was ltered and dried in a
nanocellulose from this marine biomass via a three-stage puri- weighed crucible until constant weight achieved.
cation process, namely alkalization, bleaching process and acid Lignin content was analyzed according to ASTM D1106-56. The
hydrolysis treatment. We have replaced traditional bleaching ground ber was treated with 3 mL 72% H2 SO4 at 20 C for 2 h under
agent namely (NaClO2 ) with greener and more environmental- constant stirring. The mixture was then diluted to a 3% concentra-
friendly hydrogen peroxide (H2 O2 ) during pulping process. The tion of H2 SO4 and boiled for another 4 h under reux condenser.
physicochemical properties and structural changes of prepared Upon completion, the suspension was washed and ltered with hot
nanocellulose from Gelidium red algae was examined in terms of water and dried in an oven until a constant weight was achieved.
chemical functional groups (FTIR), crystallinity (XRD), surface mor- The nanocellulose yield was calculated gravimetrically by dry-
phology studies (FESEM and TEM), and thermal stability (TGA) ing the resultant precipitate after the repeated centrifugation
systematically. (8000 rpm, 10 min) of the aliquot (10 g) at 40 C under vacuum (Kos,
Anzlovar, Kunaver, Huskic, & Z agar, 2014). On the other hand, the
2. Experimental yield of isolated cellulose from the untreated ber was determined
according to the following equation,
2.1. Materials
Celluloseyield(%) = wo /wt 100
The Gelidium elegans Ktzing red algae biomass was collected where wo refers to the dry weight of the whiteness cellulose sample
from an algae culture lab. The biomass was washed repeatedly and wt refers to the initial weight of the untreated ber prior to
with distilled water in order to remove dirt and contaminations purication process.
on the bers surface, and dried at 50 C in an oven. The dried
bers were grounded into the powder form by using a commer- 2.3.2. Fourier transform-infrared (FTIR) spectroscopy
cial miller. All chemicals and reagents utilized in this study were The FTIR spectra of bers were examined using a Fourier
purchased from Merck Chemicals (Malaysia), and used as received transform infrared spectrometer (Bruker IFS 66/S) from 4000 to
form without any further purication. Commercial alpha cellu- 400 cm1 at a resolution of 4 cm1 . The samples were ground and
lose (-cellulose, purity > 98%) and microcrystalline cellulose (MCC, mixed with KBr with a ratio of 1: 100 (w/w), and then pressed into
purity > 98%) was purchased from Sigma-Aldrich (USA). ultra-thin pellets before the analysis.
Table 1
Chemical composition analysis of untreated ber, alkali-treated, extracted cellulose and acid-treated nanocellulose.
Material -cellulose (%) Hemicellulose (%) Lignin (%) Ash (%)a Extractive (%)
Untreated 17.2 2.3 29.5 1.4 4.5 0.3 30.4 0.5 17.6 0.9
Alkali-treated 55.7 1.1 8.2 0.6 3.4 0.6 27.4 0.9
Bleached 88.6 0.6 2.0 0.2 0.7 0.3 8.3 0.2
Acid hydrolyzed 90.8 0.8 1.3 0.1 <0.5 8.2 0.1
a
The ash content was determined by thermogravimetry based on ASTM D5360 method.
of nanocellulose was summarized in Table 1. The mass fraction of treated with acid hydrolysis. The nal yielded nanocellulose com-
cellulose in the untreated ber was found to be 17.2% and hemicel- prised of 90.8% of -cellulose, 1.3% of hemicellulose and <0.5% of
lulose content of 29.5%, which comparable to the values reported lignin, resulting in production of highly puried cellulose ber.
previously (Gao et al., 2015; Yoon et al., 2010). This red alga nor- The cellulosic material was successfully extracted from the dried
mally consists of higher hemicellulose content than that in green Gelidium marine biomass at a 15.5% yield after alkalization and
and brown algae biomasses. Hemicellulose content in the red algae bleaching process. On the other hand, the yield of isolated nanocel-
biomass including Gelidium amansii and Porphyra umbilicalis were lulose, with respect to the initial amount of extracted cellulose,
24.37 and 47.16%, respectively (Gao et al., 2015). The differential on was 52.1%. The yield of isolated Gelidium nanocellulose was higher
the hemicellulose content of untreated ber observed in this study than that of nanocellulose isolated from rice straw cellulose (16.9%)
might be due to the different origins and environmental conditions (Jiang & Hsieh, 2013), microcrystalline cellulose (28.6%) (Zhou,
(Li et al., 2009). 2012), bamboo bleached ber (30%) (Brito, Pereira, Putaux, & Jean,
Obviously, when the dewaxed ber was subjected to the 2012) following a similar acid hydrolysis procedure.
alkalization treatment with NaOH, the hemicellulose content of
alkali-treated ber was reduced signicantly from 29.5 to 8.2%.
However, the primary removal of lignin in the treated ber does 3.2. Fourier transform-infrared spectroscopy (FTIR) analysis
not occur effectively during the alkalization treatment as the decre-
ment of lignin was low (4.53.4%). In order to efcient remove the Fig. 2(a) and (b) shows the FTIR spectroscopic analysis of the
lignin content, H2 O2 bleaching process was conducted. As shown untreated, alkali-treated, bleached and acid hydrolyzed Gelidium
in Table 1, most of the lignin was successfully degraded during the elegans ber. Generally, a broad peak at 3400 cm1 was observed
oxidative bleaching process, as the percentage of lignin decreased in all the samples spectra mainly due to hydrogen bond of O H
from 3.4 to 0.7%. Besides that, it was found that hemicellulose stretching vibration, which is indicates the hydrophilic tendency
content was also decreased to a certain extent from 8.2 to 2.0% of the bers (Chirayil et al., 2014). In addition, at the 2900 cm1
during the bleaching process. These nding reected that oxida- and 1640 cm1 was attributed to the sp3 C H stretching vibra-
tive bleaching process has capability of removing most of the lignin tion and the OH bending of the absorbed water, respectively (Tan,
and hemicellulose. It is not surprising because similar observa- Abd Hamid, & Lai, 2015). The peak near 1160 cm1 , which appears
tion has been reported by Chings group (Ching and Ng, 2014). The in all the spectra, associated with the C O C asymmetric stretch-
mass fraction of -cellulose in Gelidium increased exponentially ing of the cellulose, and this can be explained the intensity of this
(17.288.6%) due to the successive hydrolyzation of hemicellulose particular peak was gradually increased from untreated ber to
and lignin during the pretreatment. The hemicellulose and lignin nanocellulose as the cellulose content of the treated ber getting
fraction of the ber were further reduced when the bleached ber increased during the pretreatment (Saelee et al., 2016). Moreover,
the skeletal vibration of C O C pyranose ring skeleton (Yahya et al.,
1214 Y.W. Chen et al. / Carbohydrate Polymers 151 (2016) 12101219
Fig. 2. (a) FTIR spectra of untreated and chemically-treated Gelidium ber at different preparation stages, and (b) zoom of the 1900700 cm1 region.
2015) in cellulose ber was clearly indicated in the FTIR peak at The peak at 1315 cm1 (Saelee et al., 2016) and 1428 cm1
1050 cm1 for all the ber. (Fortunati et al., 2013; Saelee et al., 2016) was associated with
Others than similarity, several notable changes can be found in C H bending of cellulose and CH2 symmetric bending of cellu-
the spectra of bleached and acid hydrolyzed bers compared to lose, respectively. Meanwhile the peak at 1371 cm1 represented
untreated Gelidium ber. The absorbance at 1530 cm1 indicated the C H asymmetric deformation and C O symmetric stretching
the existence of the stretching vibration of N H band of the pro- within the polysaccharide aromatic rings of cellulose (Tan et al.,
tein amide II structure (Dmytryk, Saeid, & Chojnacka, 2014), and 2015). It is interesting to observe that untreated seaweed did not
this peak was also assigned to the C C stretching from aromatic exhibit peaks at 1315 cm1 and 1371 cm1 , as shown in Fig. 2b. This
hydrocarbons of lignin (Ching & Ng, 2014). The absorption bands at is due to the majority of the cellulose content was embedded by
1245 cm1 was mainly due to the stretching vibration mode of the the agar as well as others non-cellulosic materials in the untreated
acyl-oxygen CO OR associated with the hemicellulose (Mohamed ber. Besides that, the cellulose extracted (bleached ber) from the
et al., 2015), and this band was also related to the presence of the Gelidium ber was comparable with the commercial available alpha
-l-galactose 6-sulfate units that primarily found in the agar com- cellulose (-cellulose) and microcrystalline cellulose (MCC). Based
position (Guerrero, Etxabide, Leceta, Penalba, & de la Caba, 2014). A on the FTIR characteristics, Gelidium extracted cellulose pulp pat-
peak located at approximately 872 cm1 was assigned to the pres- terns exactly matched with that of -cellulose and MCC (Fig. 2).
ence d-galactose-4-sulfate of the carrageenan in the ber (Pereira, This suggested that extracted cellulose and nanocellulose has been
Sousa, Coelho, Amado, & Ribeiro-Claro, 2003). These three peaks successfully produced from red seaweed.
were clearly observed in the untreated ber; however, the intensity This results were concordance with other literatures (Chen et al.,
of these peaks was greatly reduced in alkali-treated ber and com- 2016a; Chirayil et al., 2014; Hamid et al., 2015; Tan et al., 2015;
pletely disappeared in bleached and acid treated bers. This result Yahya et al., 2015), in which the extracted cellulose and nanocel-
clearly demonstrated that non-cellulosic polysaccharides such as lulose showed a similar FTIR patterns. It is important to note that
lignin and hemicellulose were partially removed after the alkaliza- the extracted nanocellulose shows absorbance signal at 1428 cm1 ,
tion and then thoroughly removed after bleaching treatment. 1160 cm1 , 1110 cm1 , and 898 cm1 (Fortunati et al., 2013) indi-
A peak at 1110 cm1 indicated that presence of -glucosidic cated that nanocellulose produced was primarily in the form of
ether linkages (C O C) resulted in the vibration of wagging, cellulose I structure.
deformation and twisting modes of anhydro-glucopyranose ring
skeleton (Tan et al., 2015). In addition, a small peak at 898 cm1 was
owing to the -glycosidic linkages between the anhydroglucose 3.3. Field emission scanning electron microscopy (FESEM)
rings in the cellulose, and also represents the glycosidic 4 C1 ring analysis
conformation deformation (Chirayil et al., 2014; Hamid, Zain, Das,
& Centi, 2015). In the case of bleached and acid hydrolyzed bers, Fig. 3 illustrates the FESEM images of the Gelidium elegans ber
the strong absorption of the two prominent peaks at 1110 cm1 at different pretreatment stages. Fig. 3(a) shows the untreated ber
and 898 cm1 indicated that more cellulose content was exposed displayed a smooth bers surface due to the presence of oil and
in the treated ber after the successive pretreatments to remove waxes. The presence of oil and waxes in the raw ber is com-
the non-cellulosic polysaccharides, whereas the absorption peak mon and vastly reported (Saelee et al., 2016). Alkalization removed
at 898 cm1 remained unaltered for the acid hydrolyzed ber indi- majority of the agar and non-cellulosic components from the inner
cated that the typical structure of -d-glucopyranosyl structure parts of the Gelidium ber via debrillation and depolymerization.
in cellulose parent chain can be retained after the acid hydroly- Depolymerization and debrillation by alkalization is shown in
sis treatment (Hamid et al., 2015; Saelee et al., 2016). In addition, Fig. 3(b). It was observed that alkalization has loosened the biomass
the absorbance peaks at 1428 cm1 , 1371 cm1 and 1315 cm1 in structure, which resulted in swelling and solubilizing agar materi-
bleached and acid hydrolyzed bers were mainly contributed by als of the treated ber. This eventually facilitating the destruction
the presence of cellulose (Chen, Lee & Abd Hamid, 2016a; Chen, of the bers structure, partial cracking of the bers surface layer
Lee & Hamid, 2016b). and dissolves the agar compound. Thus, bleaching agents will react
Y.W. Chen et al. / Carbohydrate Polymers 151 (2016) 12101219 1215
Fig. 3. FESEM images of (a) untreated, (b) alkali-treated, (c) bleached and (d) acid hydrolyzed Gelidium ber.
more readily into the alkali-treated ber by penetrating deeper into was found in the range 501600 nm. Thus, the aspect ratio of the
the inner parts through the cracks of the bers. resultant nanocellulose was relatively high, which is about 25. The
Fig. 3(c) clearly indicated that more ber bundles were sep- isolated nanocellulose seems to be ner and possesses a porous net-
arated into individualized brils structure due to removal of work, which implied that hydronium (H3 O+ ) ions has successfully
cementing materials after alkalization and bleaching treatment. diffused into the compact and rigid structure of extracted cellu-
Each of this cellulose ber appeared to be composed of several to lose. This has resulted selective elimination of amorphous regions
hundreds microbrils, and leads to the formation of cellulose com- of cellulose long brils into smaller sizes. This results appeared to be
pact structure. The average diameter of untreated seaweed ber similar to previous study (Yahya et al., 2015), in which nanocellu-
and extracted cellulose were 124 3.8 m and 5 1.5 m, respec- lose was also found in ber network arrangements (spider web-like
tively. The diameter reduced was mainly due to the destruction of structure) with variable width between 10 and 60 nm.
the primary cell wall of the ber during removal of non-cellulosic Table 2 summarizes the values of geometrical characteris-
materials. Similar observation on the destruction of the bers bun- tics for the nanocellulose derived from various biomass sources
dles structure after alkali-treatment and bleaching process was also and collected from the literature. The aspect ratio of nanocellu-
reported by others (Chirayil et al., 2014; Sheltami et al., 2012; Zaini, lose derived from Gelidium (L/d = 25) in this study was similar
2013). to that reported for Eucalyptus wood (L/d = 24) (Beck-Candanedo,
The average diameter of the ber further reduced to Roman, & Gray, 2005), which under the similar hydrolysis pro-
1.5 0.2 m after the acid treatment, as shown in Fig. 3(d). The cess. However, the aspect ratio of yielded nanocellulose was larger
aggregation of extracted cellulose was broken down in great extent than the value reported for the nanocellulose isolated from rich
during acid hydrolysis treatment. The hydronium ions (H3 O+ ) from husk (L/d = 1015) (Johar et al., 2012), cotton (L/d = 10.7) (Martins,
the sulfuric acid (H2 SO4 ) responsible for the penetrating amor- Teixeira, Corra, Ferreira, & Mattoso, 2011), Curaua (L/d = 1317)
phous structure at higher rate compared to crystalline areas during (Correa, de Morais Teixeira, Pessan, & Mattoso, 2010) and kenaf
acid hydrolysis. This will lead to hydrolytic cleavage of the gly- bast (L/d = 13) (Kargarzadeh et al., 2012) via H2 SO4 hydrolysis under
cosidic bonds in amorphous structure which exists as loose and similar hydrolysis conditions. In order to conrm the effective-
low ordered orientation parts. As a result, the micro-sized cellu- ness of acid hydrolysis, the dimensions of isolated nanocellulose
lose bers were separated from the bril bundles and gave rise to was compared with the nanomaterial isolated via different hydrol-
a fragmentary breakdown in brillar structure. This ndings are ysis approach such as ionic liquid BmimHSO4 (Tan et al., 2015),
in accordance with previously study (Tan et al., 2015), in which Ni(NO3 )2 transition metal salt (Yahya et al., 2015) and glycols com-
nanocellulose has been extracted from cotton linter. bined with methane sulfonic acid (Kunaver, Anzlovar, & Z agar,
2016). The results revealed that the acid-hydrolyzed nanocellulose
3.4. Transmission electron microscopy (TEM) analysis had larger aspect ratio than other promising hydrolysis approach
(Table 2).
Based on TEM micrographs, it was evidenced that nano-sized Moriana et al. (2016) reported that the cellulosic structures with
brils were successfully isolated from the Gelidium elegans ber an aspect ratio values above 10, the minimum value for a good
(Fig. 4). The results revealed that the particle diameter of nanocellu- stress transfer within the interaction between the bers and the
lose was in the range of 565 nm with a calculated average diameter matrix, able to provide a good reinforcing capability and improve-
of 21.8 11.1 nm, and had the highest content of 49% at the interval ment in the thermal and mechanical properties when adding into
of 1120 nm. In addition, the nanocellulose exhibited an aver- the polymer matrices, even in low concentrations. Therefore, the
age length of 547.3 23.7 nm, in which most of the bers length high aspect ratio of nanocellulose isolated from Gelidium could pro-
1216 Y.W. Chen et al. / Carbohydrate Polymers 151 (2016) 12101219
Fig. 4. (a) TEM micrograph of nanocellulose, (b) length and (c) diameter distribution of isolated Gelidium nanocellulose.
Table 2
Geometrical properties of cellulose nanomaterial derived from different crude cellulose starting materials.
Starting materials Reaction conditions Width (nm) Length (nm) Aspect ratio Hydrolyzing catalyst Reference
0 -0.2 0 0
0 100 200 300 400 500 600 700 800 0 100 200 300 400 500 600 700 800
(a) Temperature (oC) (b) Temperature (oC)
80 80
1.9
60 0.9 60
1.4
40 40
0.9
0.4 Char yield:
20 Char yield: 20 8.2% 0.4
8.3%
0 -0.1 0 -0.1
0 100 200 300 400 500 600 700 800 0 100 200 300 400 500 600 700 800
(c) Temperature (oC) (d) Temperature (oC)
Fig. 6. Thermogravimetric analysis of (a) untreated, (b) alkali-treated, (c) bleached and (d) acid hydrolyzed ber.
phous regions of the samples was characterized at the 2 value of biomass because high crystallinity will eventually provide high ten-
around 18.5 (Segal et al., 1959). The presence of cellulose I was also sile strength as well as better reinforcement for natural composite
aligned with FTIR results. There are other peaks from untreated raw materials (Yahya et al., 2015).
seaweed which mainly corresponded to two mineral compounds,
namely silica (SiO2 ) and weddellite (CaC2 O4 2H2 O). The presence
of these impurities is generally found in the sea biomass (Bettaieb, 3.6. Thermogravimetric analysis (TGA)
Khiari, Dufresne, Mhenni, Putaux et al., 2015; Bettaieb et al., 2015).
After the pulping process, the intensity of reection peaks pre- Fig. 6 shows the thermogravimetric (TG) and its derivative
sented the mineral salts was gradually reduced, and the cellulose thermogravimetric (DTG) curves of the untreated, alkali-treated,
I form was well preserved. The XRD characterization revealed the bleaching-treated and acid treated Gelidium ber. Weight loss was
presence of main intensity peaks of cellulose at 2 values of 16.5 observed in all the samples at the temperature of <100 C which was
(110), 22.5 (200) and 34.5 (004) for bleached and acid hydrolyzed corresponded to evaporation of moisture bounded loosely on the
bers, corresponding to a typical cellulose I structure (Chen et al., ber surface and the chemisorbed water bounded inside the ber
2016a). This nding suggested that the cellulose was successfully (Chandra et al., 2016). The presence of absorbed water was also
extracted from red seaweed pulp after the alkalization, bleaching detected by FTIR characteristic peak at 1640 cm1 which represent
treatment, and acid hydrolysis process. All these results were in bending vibration of water intermolecular H-bonding.
good agreement with the FTIR result, in which the non-cellulosic In this study, untreated ber started to degrade at 225 C, and
materials were removed. the DTG curve revealed a major decomposition peak at 308 C.
The crystallinity indexes for the samples are also listed in Fig. 5. Besides that, a broad peak was also observed at ca. 640 C which
The calculated crystallinity index for untreated algae biomass, corresponded to decomposition of inorganic compounds such as
alkali-treated, bleached and acid hydrolyzed ber was found to weddellite into CaO that mainly occurred at 650 C (Frost and
be 33, 51, 63 and 73%, respectively. These results clearly demon- Weier, 2003). For the alkali treated ber, the onset thermal decom-
strated that the crystallinity of the treated material progressively position (Ton ) shifted to higher temperature of 243 C with the
improved during the chemical extraction process. This is because maximum rate of degradation temperature (Tmax ) at 317 C. This
of non-cellulosic polysaccharides components, and amorphous was due to the agar component in treated ber started to solubilize
regions in the ber were removed. Besides that, it was observed during alkalization process. This is not surprising because similar
that acid hydrolyzed ber had the maximum crystallinity index thermal degradation trend has also been reported for the sea algae
(73%). As explained earlier, hydronium ions (H3 O+ ) has success- biomass samples (Muradov et al., 2015).
fully penetrated more readily into the amorphous regions of the It is noteworthy that decomposition of bleached ber was seen
extracted cellulose, and then hydrolytic cleaved of the -1,4- at 246 C with a Tmax value of 319 C. This nding indicated that
glycosidic linkages within the cellulose chains to releases the H2 O2 -bleaching process could eventually enhance the thermal
individual crystallites segments (Tan et al., 2015). It is interesting stability of the bleached ber due to the successive removal of non-
to obtain high crystallinity of nanocellulose from Gelidium algae cellulosic impurities and the inorganic mineral salts. These results
are well agreed with XRD result (Fig. 5) in which the disappear-
1218 Y.W. Chen et al. / Carbohydrate Polymers 151 (2016) 12101219
Kargarzadeh, H., Ahmad, I., Abdullah, I., Dufresne, A., Zainudin, S. Y., & Sheltami, R. Pereira, L., Sousa, A., Coelho, H., Amado, A. M., & Ribeiro-Claro, P. J. (2003). Use of
M. (2012). Effects of hydrolysis conditions on the morphology, crystallinity, FTIR, FT-Raman and 13 C-NMR spectroscopy for identication of some
and thermal stability of cellulose nanocrystals extracted from kenaf bast bers. seaweed phycocolloids. Biomolecular Engineering, 20(4), 223228.
Cellulose, 19(3), 855866. Reddy, J. P., & Rhim, J.-W. (2014). Isolation and characterization of cellulose
Kim, H. M., Wi, S. G., Jung, S., Song, Y., & Bae, H.-J. (2015). Efcient approach for nanocrystals from garlic skin. Materials Letters, 129, 2023.
bioethanol production from red seaweed Gelidium amansii. Bioresource Rosa, M. F., Medeiros, E. S., Malmonge, J. A., Gregorski, K. S., Wood, D. F., Mattoso, L.
Technology, 175, 128134. H. C., et al. (2010). Cellulose nanowhiskers from coconut husk bers: effect of
Korzen, L., Pulidindi, I. N., Israel, A., Abelson, A., & Gedanken, A. (2015). Single step preparation conditions on their thermal and morphological behavior.
production of bioethanol from the seaweed Ulva rigida using sonication. RSC Carbohydrate Polymers, 81(1), 8392.
Advances, 5(21), 1622316229. Roohani, M., Habibi, Y., Belgacem, N. M., Ebrahim, G., Karimi, A. N., & Dufresne, A.
Kos, T., Anzlovar, A., Kunaver, M., Huskic, M., & Z agar, E. (2014). Fast preparation of (2008). Cellulose whiskers reinforced polyvinyl alcohol copolymers
nanocrystalline cellulose by microwave-assisted hydrolysis. Cellulose, 21(4), nanocomposites. European Polymer Journal, 44(8), 24892498.
25792585. Saelee, K., Yingkamhaeng, N., Nimchua, T., & Sukyai, P. (2016). An environmentally
Kunaver, M., Anzlovar, A., & Z agar, E. (2016). The fast and effective isolation of friendly xylanase-assisted pretreatment for cellulose nanobrils isolation from
nanocellulose from selected cellulosic feedstocks. Carbohydrate Polymers, 148, sugarcane bagasse by high-pressure homogenization. Industrial Crops and
251258. Products, 82, 149160.
Lam, E., Male, K. B., Chong, J. H., Leung, A. C., & Luong, J. H. (2012). Applications of Segal, L., Creely, J., Martin, A., & Conrad, C. (1959). An empirical method for
functionalized and nanoparticle-modied nanocrystalline cellulose. Trends in estimating the degree of crystallinity of native cellulose using the X-ray
Biotechnology, 30(5), 283290. diffractometer. Textile Research Journal, 29(10), 786794.
Li, R., Fei, J., Cai, Y., Li, Y., Feng, J., & Yao, J. (2009). Cellulose whiskers extracted from Seo, Y.-B., Lee, Y.-W., Lee, C.-H., & You, H.-C. (2010). Red algae and their use in
mulberry: a novel biomass production. Carbohydrate Polymers, 76(1), 9499. papermaking. Bioresource Technology, 101(7), 25492553.
Li, J., Wei, X., Wang, Q., Chen, J., Chang, G., Kong, L., et al. (2012). Homogeneous Sheltami, R. M., Abdullah, I., Ahmad, I., Dufresne, A., & Kargarzadeh, H. (2012).
isolation of nanocellulose from sugarcane bagasse by high pressure Extraction of cellulose nanocrystals from mengkuang leaves (Pandanus
homogenization. Carbohydrate Polymers, 90(4), 16091613. tectorius). Carbohydrate Polymers, 88(2), 772779.
Liew, S. Y., Thielemans, W., & Hewakandamby, B. (2016). Separation of sulphuric Silvrio, H. A., Neto, W. P. F., Dantas, N. O., & Pasquini, D. (2013). Extraction and
acid from an acid suspension of cellulose nanocrystals by manual shaking. characterization of cellulose nanocrystals from corncob for application as
Journal of Nano Research, 38, 5872. reinforcing agent in nanocomposites. Industrial Crops and Products, 44,
Liu, D., Zhong, T., Chang, P. R., Li, K., & Wu, Q. (2010). Starch composites reinforced 427436.
by bamboo cellulosic crystals. Bioresource Technology, 101(7), 25292536. Soni, B., & Mahmoud, B. (2015). Chemical isolation and characterization of different
Maiti, S., Jayaramudu, J., Das, K., Reddy, S. M., Sadiku, R., Ray, S. S., et al. (2013). cellulose nanobers from cotton stalks. Carbohydrate Polymers, 134, 581589.
Preparation and characterization of nano-cellulose with new shape from Tan, X. Y., Abd Hamid, S. B., & Lai, C. W. (2015). Preparation of high crystallinity
different precursor. Carbohydrate Polymers, 98(1), 562567. cellulose nanocrystals (CNCs) by ionic liquid solvolysis. Biomass and Bioenergy,
Mandal, A., & Chakrabarty, D. (2011). Isolation of nanocellulose from waste 81, 584591.
sugarcane bagasse (SCB) and its characterization. Carbohydrate Polymers, 86(3), Tanaka, R., Saito, T., & Isogai, A. (2012). Cellulose nanobrils prepared from
12911299. softwood cellulose by TEMPO/NaClO/NaClO2 systems in water at pH 4.8 or 6.8.
Martins, M. A., Teixeira, E. M., Corra, A. C., Ferreira, M., & Mattoso, L. H. (2011). International Journal of Biological Macromolecules, 51(3), 228234.
Extraction and characterization of cellulose whiskers from commercial cotton Wang, Y., Tian, H., & Zhang, L. (2010). Role of starch nanocrystals and cellulose
bers. Journal of Materials Science, 46(24), 78587864. whiskers in synergistic reinforcement of waterborne polyurethane.
Mohamed, M., Salleh, W., Jaafar, J., Asri, S., & Ismail, A. (2015). Physicochemical Carbohydrate Polymers, 80(3), 665671.
properties of green nanocrystalline cellulose isolated from recycled Yahya, M. B., Lee, H. V., & Hamid, S. B. A. (2015). Preparation of nanocellulose via
newspaper. RSC Advances, 5(38), 2984229849. transition metal salt-catalyzed hydrolysis pathway. BioResources, 10(4),
Moriana, R., Vilaplana, F., & Ek, M. (2016). Cellulose nanocrystals from forest 76277639.
residues as reinforcing agents for composites: a study from macro- to Yoon, J. J., Kim, Y. J., Kim, S. H., Ryu, H. J., Choi, J. Y., Kim, G. S., et al. (2010).
nano-dimensions. Carbohydrate Polymers, 139, 139149. Production of polysaccharides and corresponding sugars from red seaweed.
Muradov, N., Taha, M., Miranda, A. F., Wrede, D., Kadali, K., Gujar, A., et al. (2015). Advanced Materials Research, 93, 463466. Trans Tech Publ.
Fungal-assisted algal occulation: application in wastewater treatment and Zaini, L. H. (2013). Isolation and characterization of cellulose whiskers from kenaf
biofuel production. Biotechnology for Biofuels, 8(1), 1. (Hibiscus cannabinus L.) bast bers. Journal of Biomaterials and
Oun, A. A., & Rhim, J.-W. (2016). Characterization of nanocelluloses isolated from Nanobiotechnology, 4(1), 3744.
Ushar (Calotropis procera) seed ber: effect of isolation method. Materials Zhou, Y. M. (2012). Effect of nanocellulose isolation techniques on the formation of
Letters, 168, 146150. reinforced poly(vinyl alcohol) nanocomposite lms. Express Polymer Letters,
Pandey, J. K., Kim, C.-S., Chu, W.-S., Lee, C. S., Jang, D.-Y., & Ahn, S.-H. (2009). 6(10), 794804.
Evaluation of morphological architecture of cellulose chains in grass during Zoppe, J. O., Peresin, M. S., Habibi, Y., Venditti, R. A., & Rojas, O. J. (2009).
conversion from macro to nano dimensions. e-Polymers, 9(1), 12211235. Reinforcing poly(-caprolactone) nanobers with cellulose nanocrystals. ACS
Applied Materials & Interfaces, 1(9), 19962004.