Food Chemistry 151 (2014) 343–351

Contents lists available at ScienceDirect

Food Chemistry
journal homepage: www.elsevier.com/locate/foodchem

Biocomposite cellulose-alginate films: Promising packaging materials

Juho Antti Sirviö a, Aleksi Kolehmainen a, Henrikki Liimatainen b, Jouko Niinimäki b, Osmo E.O. Hormi a,⇑
a
Department of Chemistry, University of Oulu, P.O. Box 3000, FI-90014, Finland
b
Fibre and Particle Engineering Laboratory, University of Oulu, P.O. Box 4300, FI-90014, Finland

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

Article history: Biocomposite films based on cellulose and alginate were produced using unmodified birch pulp,
Received 19 April 2013 microfibrillated cellulose (MFC), nanofibrillated cellulose (NFC) and birch pulp derivate, nanofibrillated
Received in revised form 19 September 2013 anionic dicarboxylic acid cellulose (DCC), having widths of fibres ranging from 19.0 lm to 25 nm as cel-
Accepted 6 November 2013
lulose fibre materials. Ionically cross-linked biocomposites were produced using Ca2+ cross-linking. Addi-
Available online 16 November 2013
tion of micro- and nanocelluloses as a reinforcement increased the mechanical properties of the alginate
films remarkably, e.g. addition of 15% of NFC increased a tensile strength of the film from 70.02 to
Keywords:
97.97 MPa. After ionic cross-linking, the tensile strength of the film containing 10% of DCC was increased
Cellulose
Nanocellulose
from 69.63 to 125.31 MPa. The biocomposite films showed excellent grease barrier properties and
Reinforcement reduced water vapour permeability (WVP) after the addition of cellulose fibres, except when unmodified
Alginate birch pulp was used.
Biocomposite Ó 2013 Elsevier Ltd. All rights reserved.
Ionic cross-linking

1. Introduction
Composite materials derived from biobased sources have drawn
lots of attention during the recent years (Hansen & Plackett, 2008).
One of the most studied applications of biocomposites is in environ-
mentally friendly food packaging materials (Kristo, Biliaderis, &
Zampraka, 2007). Natural polymers, such as polysaccharides, are
ideal starting materials for these kinds of composites due to their
biodegradability, biocompatibility, low toxicity and renewability.
Cellulose is a linear polysaccharide of (1–4)-linked b-D-glucose,
produced by a variety of plants and algae, along with some bacte-
ria. In nature, cellulose is produced as fibrous material which has
long been used as a main component in paper, and as a starting
material e.g. for edible packages (Ayranci & Tunc, 2001; Cheng,
Abd Karim, & Seow, 2008). Additionally, the use of cellulose micro-
and nanofibres in composite materials has been a scientific interest
in recent years (Abdul Khalil, Bhat, & Ireana Yusra, 2012). Cellulose
micro- and nanofibres have been studied as reinforcement agents
in synthetic (Zoppe, Peresin, Habibi, Venditti, & Rojas, 2009) and
natural polymers (Johnson, Zink-Sharp, Renneckar, & Glasser,
2009).
Alginate is another type of linear polysaccharide, consisting of
(1–4)-linked b-D-mannuronate and a-L-guluronate units. The rela-
tive proportions of mannuronate and gulunorate vary within the
source of the alginate (Johnson, Craig, & Mercer, 1997). The most
⇑ Corresponding author. Tel.: +358 294 48 1631; fax: +358 8 553 1593.
E-mail address: osmo.hormi@oulu.fi (O.E.O. Hormi).
common source of alginate is the cell wall of brown algae, and
for commercial purposes alginate is extracted from seaweed. Algi-
nate and alginate-based biocomposites have been used as, for
example, adsorbents for ionic dyes (Sui et al., 2012), antibacterial
films (Benavides, Villalobos-Carvajal, & Reyes, 2012), wound heal-
ing materials (Sikareepaisan, Ruktanonchai, & Supaphol, 2011), and
stimulus response drug releasing materials (Mouriño et al., 2011).
Although alginate based biocomposites have strong potential to
be used for example in packaging applications, they usually lack
the desired mechanical properties, such as high strength. To
increase the mechanical properties of alginate based biocompos-
ites, cellulose nanocrystals have been studied as reinforcement
agents in alginate films (Cheng, Lu, Zhang, Shi, & Cao, 2012). How-
ever, there is only a scarcity of information on the mechanical
properties of biocomposite materials with an alginate matrix rein-
forced with different types of wood cellulose fibres. In this paper,
four different wood cellulose fibres, with widths ranging from
19.0 lm to 25 nm, were used as reinforcement agents for alginate
biocomposites films to be utilised in application such as packaging.
Effects of the different fibres on the film properties and on the
compatibility between cellulose fibres and alginate were studied.
Moreover, an ionic cross-linking of the biocomposite films with
Ca2+ ion, and grease and water vapour barrier properties were also
studied. Properties of films, and the compatibility between cellu-
lose fibres and alginate were studied by Fourier transfer infrared
spectroscopy (FTIR), field emission scanning electron microscopy
(FESEM), energy-dispersive X-ray spectroscopy (EDS) and wide-
angle X-ray diffraction (WAXD).

0308-8146/$ - see front matter Ó 2013 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.foodchem.2013.11.037
344 J.A. Sirviö et al. / Food Chemistry 151 (2014) 343–351
2. Experimental section
2.1. Materials
Bleached birch pulp (Betula verrucosa and pendula) was ob-
tained as dry sheets and used as a fibre material after being disin-
tegrated in deionised water. The properties of cellulose are
presented elsewhere (Liimatainen, Sirviö, Haapala, Hormi, & Nii-
nimäki, 2011). The commercial grade microfibrillated wood cellu-
lose (MFC, KY-100G) was obtained from Daicel Industries Ltd.
(Japan).
The combined treatment of Valley beating and high-pressure
homogenisation was used to prepare nanofibrillated celluloses
(NFC) from the birch pulp. Prior to homogenising, the bleached
birch was ground in a Valley beater to a dry weight of 2.0% over
50 h, yielding 5503 g of weight. The samples where then diluted
to a dry weight of 0.7% and homogenised with a homogeniser
(Invensys APV 2000, Denmark) at a pressure of 350–1200 bar to
create NFC. NFC had same charge density as original pulp
(0.08 mmol/g). Charge density is mainly due to the presence of
hemicelluloses.
Nanofibrillated dicarboxylic acid cellulose (DCC) with a charge
density of 0.75 mmol/g was prepared as reported earlier
(Liimatainen, Visanko, Sirvio, Hormi, & Niinimaki, 2012).
Sodium alginate (Scogin XL, low viscosity) was kindly donated
by FMC Biopolymer (Ireland). Oil Red O (1-([4-(xylylazo)
xylyl]azo)-2-naphthol), used in grease barrier tests, was obtained
from Sigma–Aldrich (Germany) and Ottawa-grade sand (particle
size 20–30 mesh) from Fisher Scientific (UK) and Turpentine (Oulu,
A1 Wood turpentine, Finland) from Kiilto Ltd. All of other
chemicals used in the periodate and chlorite oxidations (NaIO4,
NaClO2 and CH3COOH), aldehyde and carboxyl content analyses
(NH2OHHCl, CH3COONa2H2O, NaCl, and NaOH) and ionical
crosslinking (CaCl22H2O) were obtained as p.a. grade from
Sigma–Aldrich (Germany) and were used without further purifica-
tion. Deionised water was used throughout the experiments.
2.2. Preparation of biocomposite films
The biocomposite films based on cellulose and alginate were
prepared using the solvent-casting method. At first, alginate was
slowly dissolved in deionised water. Then cellulose (unmodified
birch pulp, MFC, NFC or DCC) dispersion was added to this solution,
whilst varying the amount of cellulose from 0% to 50% of the film
dry weight. Total volume of the alginate and cellulose dispersions
was 135 ml and a dry weight of films was 1.515 g, resulting in films
with a grammage of 30 g/m2. After addition of the cellulose disper-
sion, the resulting alginate/cellulose mixture was dispersed using
an UltraTurrax disperser (Germany) (11,000 rpm) for ten minutes
to obtain a homogeneous dispersion after which a water circula-
tion vacuum was used to remove air bubbles from the dispersion.
Finally, the dispersion was cast in a polystyrene tray of 500 cm2
area. Films were allowed to dry from 3 to 5 days at room temper-
ature to obtain 13 biocomposite films with 1%, 10%, 15%, 20% and
50% of MFC, 1%, 5%, 10% and 15% of NFC, 1% and 10% of DCC, and
50% of birch pulp of the film dry weight were, designated as
MFC1, MFC5, MFC10, MFC15, MFC20, MFC50, NFC1, NFC5, NFC10,
NFC15, DCC1, DCC10 and BIRCH50, respectively. Pristine alginate
film was also produced as a reference.
2.3. Preparation of ionically cross-linked biocomposite films
In order to ionically cross-link a film, it was immersed in 400 ml
of 10% CaCl22H2O solution for 30 min. The film was then the care-
fully removed from the solution and rinsed thoroughly with
deionised water. The film was placed on a polycarbonate plate
and small weights were placed on the edges of the film to prevent
the shrinkage of the film during air-drying (24 h). Ionic cross-
linking was conducted with a pristine alginate, BIRCH50, MFC50,
NFC10 and DCC10 films and they were designated as ICBICH50,
ICMFC50, ICMFC10, ICDCC10, respectively.
2.4. Mechanical properties
The films were cut into 15  150 mm stripes and conditioned at
23 °C and 50% humidity for 48 h before testing. Specific tensile
strength (r/q), elongation at break and specific modulus (E/q)
were determined according to ISO Standard, 1995 and Lorentzen
& Wettre tensile strength tester (Sweden). A test speed of
50 mm/min was used. Initial distance between grips was set to
100 mm. Each test was replicated five times and the results are
presented as averages of these measurements. Tensile strength
(r) and modulus (E) was calculated using the density (q) from r/
q and E/q, respectively.
Film thicknesses were determined as averages of five random
positions of the film using Lorentzen & Wettre thickness metre
(Sweden).
2.5. Field emission scanning electron microscope and energy-
dispersive X-ray spectroscopy
FESEM (Zeiss Ultra Plus, Carl Zeiss SMT AG, Germany) was used
to examine morphologies of used cellulosics and the surface of the
films. Fibres were sputter-coated with Pd, and an accelerating volt-
age of 5 kV was used whilst the films were sputter-coated with car-
bon, and an accelerating voltage of 15 kV was used. EDS was used
to obtain surface elemental analysis of the films.
2.6. Fourier transform infrared spectrum
FTIR spectra of films were recorded using a Bruker FT-IR spec-
trometer (USA) using spectral width ranging from 4000 to
400 cm1 with 2 cm1 resolution and an accumulation of 32 scans.
2.7. X-ray diffraction
For X-ray diffraction (XRD) analyses, samples were analyzed be-
tween 2h = 5° and 114° with angle step size 2h = 0.02° in a D8 Dis-
cover X-ray Diffractometer (Bruker AXS Inc., Madison, MI, USA)
equipped with a Co Ka (40 kV/35 mA) source. Pure birch pulp,
MFC, NFC and DCC films were prepared by casting a 1% solution
of cellulose dispersion on a Teflon plate to obtain a film with a
grammage of 30 g/m2. The degree of crystallinity of pure cellulose
films in terms of the crystallinity index (CrI) was calculated from
the peak intensity of the main crystalline plane (0 0 2) diffraction
(I002) at 22.8° and from the peak intensity at 18.0° associated with
the amorphous fraction of cellulose (Iam) according to following
equation (Segal, Creely, Martin, & Conrad, 1959)
 
I002  Iam
CrI ¼  100%
I002
2.8. Grease barrier properties
The grease resistance of the films was determined according to
TAPPI Standard, 2010. Each test was replicated four times: two
times for the bottom layers and two times for the upper layers of
the 50  50 mm2 film specimens.
The method involves the use of turpentine colored with an oil-
soluble red dye to measure the penetration of the grease through a
 J.A. Sirviö et al. / Food Chemistry 151 (2014) 343–351
greaseproof material. Moisture-free colored turpentine was
prepared by mixing 100 ml of turpentine with 1.0 g of Oil Red O
red dye and 5 g of anhydrous CaCl2 in a flask. The flask was closed
with stopper, and the mixture was shaken well and allowed to
stand for 10 h. Finally, the mixture was filtrated and stored in an
airtight bottle.
For grease barrier tests, a film was placed on a white printing
paper, and 5 g of sand was placed on top of the film. A volume of
1.1 ml of colored turpentine was added to the sand pile. The film
was carefully slid to a new unexposed portion of the paper, and
the time when colored-turpentine-stained paper was observed
was recorded. During the first minute, the film was slid every
15 s, then at 1 min intervals for the next 5 min and finally at
5 min intervals until a maximum time of 30 min was reached.
2.9. Water vapour permeability
Water vapour permeability (WVP) was determined according to
ASTM Standard, 1995. All films were conditioned at 23 °C and 50%
humidity for 48 h before being tested. Schott DuranÒ laboratory
glass bottles with a hole drilled in their caps were filled to one-
fourth their volume with deionised water. The test specimen was
placed between the bottle and cap, and a rubber seal was used to en-
sure a tight seal. The test assembly was weighed and placed in a con-
dition-controlled chamber (23 °C and 50% humidity, equipped with
a fan to ensure good circulation of air inside the chamber) and the
weight lost was recorded in one hour intervals for 8 h. A steady state
was reached with all samples during the first hour. The slope of the
strain line of weight lost vs. time was calculated, and the water va-
pour transmission rate (WVTR) was determined by dividing the
slope by the exposed film area. WVP was calculated as follows:
WVTR
WVP ¼
SðR1  R2 Þ
where S is the saturation vapour pressure at the test temperature
(28  102 Pa at 23 °C), R1 is the relative humidity in the dish ex-
pressed as a fraction (1), and R2 is the relative humidity at the va-
pour sink expressed as a fraction (0.5).
3. Results and discussion
3.1. Cellulose and alginate biocomposite films
3.1.1. Preparation of biocomposite films
FESEM images of the cellulose fibres used to obtain biocompos-
ite films are presented in Supplementary data (Fig. S1). The un-
treated birch pulp consisted mainly of cellulose fibres (width
19.0 lm) with 3.4% of fines, according to FibreMaster image ana-
lyzer. MFC consisted of long fibrils, but also included larger mi-
cro-sized particles such as pieces of cell walls. NFC consisted
mainly of nanofibril bundles with widths smaller than 100 nm,
whilst DCC contained single nanofibrils and nanofibril bundles
with widths of below 25 nm. However, it is possible that the fibrils
existing in the suspension are actually thinner compared to the fi-
brils observed in FESEM images due to the agglomeration of nano-
fibres during the sample preparation (Liimatainen et al., 2012).
The appearance of dried films depended on the amount and the
type of cellulose used in the film. Alginate films with no added cel-
lulose were fully transparent, and the transparency decreased as
the amount of cellulose increased: MFC50 and BIRCH50 were tur-
bid with low transparency. Due to the small fibre sizes, films with
NFC and DCC were only slightly less transparent compare to pris-
tine alginate film. All the films were flexible and could be easily
handled. Films with cellulose had higher apparent roughness than
the pure alginate film. The surface facing towards the tray during
345
the drying was significantly smoother compared to the surface
facing towards the air during the drying. Nano-sized celluloses
(NFC and DCC) provided smoothest surfaces, whereas MFC and
especially unmodified birch pulp reinforced films were signifi-
cantly rougher. The visual appearance of the films obtained from
FESEM is shown in a later chapter.
3.1.2. Preparation of ionically cross-linked biocomposite films
Cross-linking can be used to increase the strength of materials
(Bhattacharya, Rawlins, & Ray, 2009). It can also be used to
decrease water solubility of polymers, such as polysaccharides
(Ramesh, Hasegawa, Sugimoto, Maki, & Ueda, 2008). To obtain
cross-linked biocomposite films, Ca2+ ions were used as an ionic
crosslinker. The mechanism of the ionic cross-linking of alginate
is well described in literature (Sikorski, Mo, Skjåk-Bræk, & Stokke,
2007). During the drying, fractures were observed in the pristine
alginate film and intact ionically cross-linked alginate film could
not be produced. Fractures may be due to the high tension caused
by film shrinking. However, intact ionically cross-linked films from
BIRCH50, MFC50, NFC10 and DCC10 were obtained.
The appearances of ICBIRCH50 and ICMFC50 were smooth and
they were only slightly wrinkled on the edges. ICNFC10 in turn
was more wrinkled and bent, whilst ICDCC10 was smooth and only
slightly bent on the edges of the film. Minor fractures were ob-
served at the edges of ICDCC10. Due to the higher amount of algi-
nate, ICNFC10 and ICDCC10 were more prone to shrinking, and
hence they were more wrinkled compared to ICBIRCH50 and
ICMFC50.
All films dissolved or disintegrated in water without cross-link-
ing treatment. After cross-linking the films still wetted easily, but
they did not dissolve in water. It is known that cross-linking of
alginate with multivalent ions decreases dramatically the water
solubility of the alginate (Nakamura, Nishimura, Hatakeyama, &
Hatakeyama, 1995).
3.2. Fourier transform infrared spectrum
FTIR spectra of the pristine alginate film, MFC50, NFC10, DCC10
and BIRCH50 before and after crosslinking are presented in Fig. 1
and assignments of the most characteristic vibration modes of all
of the biocomposite films determined by literature (Fuks, Filipiuk,
& Majdan, 2006; Papageorgiou et al., 2010) are presented in
Supplementary data (Table S1). In the spectrum of the pristine algi-
nate film, characteristic vibration bands of m(COO)asym and
m(COO)sym are observed at 1613.66 and 1414.54 cm1, respectively,
and the band of –C–O stretching vibrations is observed at
1038.02 cm1.
In the FTIR spectra of the biocomposite films m(COO)asym peak is
shifted to a lower wavenumber as the amount of cellulose is in-
creased. The band of m(COO)asym of MFC1 is at 1614.42 cm1 and
it is shifted to 1607.97 cm1 when the amount of MFC is increased
to 50%. The lowest m(COO)asym wavenumber is observed with
BIRCH50 (1603.19 cm1). This may be due to the presence of large
amount of hemicelluloses in birch pulp, which may have better
interaction with alginate than cellulose. The –C–O stretching vibra-
tion bands in MFC50, BIRCH50, and NFC10 are also shifted to
slightly lower wavenumber and are at 1031.51, 1027.27 and
1026.18 cm1, respectively. Wavenumber of the peak of –C–O of
DCC10 is close to pristine alginate film, 1035.50 cm1, which indi-
cates that, compared to other cellulose materials, anionic DCC has
lowest interaction with alginate.
After the crosslinking, the m(COO)asym band of the pristine algi-
nate film is shifted to a lower wavenumber (1602.78 cm1),
whereas the m(COO)sym band has a slightly higher wavenumber
(1420.94 cm1). This is due to the different interactions between
monovalent sodium and divalent calcium ions with alginate
346 J.A. Sirviö et al. / Food Chemistry 151 (2014) 343–351

Fig. 1. FTIR spectra of alginate and biocomposite films (a) before and (b) after cross-linking.

(Papageorgiou et al., 2010). From the Table 1 it can be observed
that Dm(COO) (m(COO)asym  m(COO)sym) of the pristine alginate is
decreased from 199.12 to 181.84 cm1 after ionic cross-linking,
which indicates that a more uniform, structured, carboxylate
groups are formed during the cross-linking (Fuks et al., 2006). It
has been proposed that there is ‘pseudo bridged’ unidentate
complex between alginate and multivalent metal cations
(Papageorgiou et al., 2010). However, recent density functional
calculations have shown that there is only an ionic interaction
between alkaline metals and alginate (Agulhon, Markova, Robitzer,
Quinard, & Mineva, 2012).
Values of Dm(COO)Ca of the ICMFC50 and ICBIRCH50 have also
been shifted from 194.12 to 181.01 cm1 and from 188.90 to
175.12 cm1, respectively. However, values of Dm(COO) of films
containing nano-sized cellulose are virtually the same before and
after cross-linking. Nano-sized celluloses may interact with cal-
cium alginate which leads to the change of the bonding between
alginate and the calcium ion.
It has been previously proposed that anionically charged
nanocelluloses can take part in the formation of highly cross-linked
alginate after calcium cross-linking (Lin, Bruzzese, & Dufresne,
2012). Nanocelluloses used in the present study had anionic
charges of 0.08 mmol/g (NFC) and 0.75 mmol/g (DCC) and thus
NFC and DCC may form ionic cross-linking network with calcium
alginate. Even though birch pulp has same anionic charge as NFC,
it has significantly larger fibre size and lower surface area, due to
which birch pulp has fewer ionic groups available to interact with
calcium alginate than NFC.
3.3. Field emission scanning electron microscope and energy-
dispersive X-ray spectroscopy
FESEM images of biocomposite films are presented in the Fig. 2.
Images are from the surface faced towards the tray during the dry-
ing. The films were observed to be very sensitive and especially the
films with high amount of alginate were easily destroyed during
the zooming and thus only large scale images were obtained. The
pristine alginate film (Fig. 2a) has a smooth and even surface
(white spots on the surface are holes formed on the surface due
to the electron beams of the microscope). The surface of MFC50
(Fig. 2b) was also relative smooth and even, however, some cellu-
lose fibrils are buried under alginate matrix. Compared to MFC50,
the surface of BIRCH50 (Fig. 2c) is significantly less even and large
pores can be observed on the surface. This is due to the much larger
fibre size of birch pulp compared to MFC. Due to the nano-sized fi-
bres, the surfaces of NFC10 (Fig. 2d) and DCC10 (Fig. 2e) are
smooth and no individual fibres can be observed in the FESEM
images.
After ionic cross-linking, the surfaces of pristine alginate, NFC10
and DCC10 films were smooth and identical to non-cross-linked
films (images not shown). However, the surface of ICBIRCH50
(Fig. 2f) was much more even and fewer pores were observed com-
pared to BIRCH50. It is possible that shrinkage of alginate during
the cross-linking compresses the BIRCH50 film, which in turn re-
duces the pore sizes of the film. ICBIRCH50 (Fig. 2g) surface faced
towards the air during the drying was also examined and it is ob-
served to be rougher having individual cellulose fibres clearly vis-
ible. In a comparison, the surface of ICMFC50 (Fig. 2h) was also
observed to be rough, but due to the smaller fibre size, no individ-
ual fibres or visible pores can be observed on its surface.
Results from EDS-elemental analysis are presented in Supple-
mentary data (Table S2). All surface sodium ions were changed
to calcium ions during the cross-linking. In case of ICNFC10, 0.6%
of chlorine is still present in the surface, which indicates that small
amount of unreacted CaCl2 remains on the surface of film after
washing. Interestingly, all films except ICMFC50 have similar
amounts of calcium ions on their surfaces (17.0–20.0%). ICMFC50,
which has only 3.9% of calcium in its surface, may have higher
amount of cellulose fibres on the surface compared to other
biocomposite films, which reduces the amount of calcium bound
to the surface of film.

Table 1
3.4. Wide-angle X-ray diffraction
Asymmetric and symmetric carboxyl vibration frequencies of alginate.

Sample m(COO)asym m(COO)sym Dm(COO) X-ray diffractograms of the pristine alginate, BIRCH50 MFC50,
Alg 1613.66 1414.54 199.12 NFC10 and DCC10 films before and after ionic cross-linking are
MFC50 1607.97 1413.85 194.12 presented in Fig. 3, along with the diffractograms of the pure birch
NFC10 1609.31 1420.79 188.52 pulp, MFC, NFC, DCC films. All celluloses showed characteristic
DCC10 1609.13 1414.78 194.35
BIRCH50 1603.19 1414.29 188.9
peaks of cellulose I with the main 2h diffraction angles close to
ICAlg 1602.78 1420.94 181.84 14.5, 16.0, and 22.2° associated with the 101, 101 and 0 0 2 crystal-
ICMFC50 1606.32 1425.31 181.01 line planes, respectively. CrI’s of the birch pulp, MFC, NFC and DCC
ICNFC10 1609.31 1420.79 188.52 were 56.6%, 63.1%, 48.8% and 44.9%, respectively, which are consis-
ICDCC10 1614.54 1421.09 193.45
tent with the literature (Iwamoto, Nakagaito, & Yano, 2007). On the
ICBIRCH50 1598.18 1422.48 175.7
other hand, the diffractogram of the pristine alginate film shows
 J.A. Sirviö et al. / Food Chemistry 151 (2014) 343–351 347

Fig. 2. FESEM images of the surface of the films faced towards the tray during the drying: (a) pristine alginate film, (b) MFC50, (c) BIRCH50, (d) NFC10, (e) DCC10, (f)
ICBIRCH50 and images of the surfaces of the biocomposite films faced towards the air during the drying: (g) ICBIRCH50 and (h) ICMFC50.

only a broad halo which indicates amorphous nature (Huq et al.,
2012).
The characteristic cellulose peaks are undetectable in the dif-
fractograms of NFC10 and DCC10 due to the large amorphous peak
of alginate, but they are easily observed in the diffractograms of
MFC50 and BIRCH50. The CrI of cellulose in BIRCH50 is observed
to decrease to 48.3%, making it 17% less crystalline than the pure
birch pulp film. The CrI of MFC50 is decreased even more, and it
was observed to be 47.2%, making it 34% less crystalline than the
pure MFC film. This is an unusual phenomenon; however, it has
been proposed that the hydrogen bonding interaction between cel-
lulose crystals and additives can lead to decrease of the crystallin-
ity of cellulose (Ul-Islam, Khan, & Park, 2012). Additives, such as
chitosan, can penetrate cellulose fibres and disrupt the hydrogen
bonding of cellulose leading to decrease in crystallinity (Ul-Islam,
Shah, Ha, & Park, 2011). In this context, alginate may penetrate cel-
lulose fibres during the film preparation and interact with crystal-
line parts of the cellulose, which leads to the decrease of the
crystalline order of cellulose. Furthermore, it is possible the larger
surface area of MFC fibres than birch pulp fibres allows more effi-
cient penetration of alginate into MFC than birch pulp. This is
shown in the higher decrease of the crystallinity of cellulose in
MFC50 than in BIRCH50.
After ionic cross-linking, there is no change in the X-ray
diffractograms of the pristine alginate films, NFC10 and DCC10.
However, the crystallinity of the films containing birch pulp and

(a) DCC10
(b) ICDCC10

DCC
ICNFC10
NFC10
Intensity

NFC ICBRICH50
Intensity

BRICH50

BIRCH

MFC50 ICMFC50

MFC
ICAlg
Alg

20 40 20 40

2θ (degree) 2θ (degree)

Fig. 3. XRD patterns for the biocomposite films (a) before and (b) after the cross-linking. XRD patterns pristine alginate and pure birch pulp (BIRCH), MFC, NFC and DCC films
are also shown.
348 J.A. Sirviö et al. / Food Chemistry 151 (2014) 343–351

MFC are increased, ICMFC50 and ICBIRCH50 having CrI of 55.2% (a) 130
and 54.2%, respectively. This phenomenon has previously been ob-

Specific strength (kNm/kg) and

served with the blend cotton linter cellulose and alginate mem- 110
brane after ionic cross-linking with Ca2+ (Yang, Zhang, Peng, &
90

strength (MPa)
Zhong, 2000).
70
kNm
50 /kg
3.5. Mechanical properties
30 MPa
3.5.1. Specific tensile strength and tensile strength
10
Thicknesses of films are presented in Supplementary data
(Fig. S2). The thicknesses of the films increased with the amount
of added cellulose due to the fibrous structure of cellulose. The dif-
ference between the thicknesses of the films containing the nano- (b) 20
and microfibrillated and unfibrillated cellulose was enormous, e.g.

Specific modulus (MNm/kg) and

18
the BIRCH50 was 88% thicker than MFC50. The addition of a small
16
amount of NFC to alginate does not have a significant impact on

modulus (GPa)
14
the thickness of films; however, NFC15 has a 23% higher thickness
compared to the pristine alginate film. Thicknesses of films con- 12
MNm
taining DCC were slightly higher compared to the pristine alginate 10
/kg
films and the films containing NFC. This suggests that some DCC 8 GPa
agglomerates are formed during the preparation of films contain- 6
ing DCC and alginate. 4
After the cross-linking, the thickness of the alginate films with
birch pulp and MFC is decreased whilst thickness of NFC10 is in-
creased 17% during ionic cross-linking. On the other hand, there
is no significant change in the thickness of the DCC10 after cross- (c) 3.5
linking. 3.25
An increase of the thickness in the alginate films during the 3
cross-linking has been reported previously (Rhim, 2004). The in- 2.75
Strains (%)

crease is due to the changes in the morphology of the alginate dur-
ing the cross-linking in the aqueous calcium chloride solution
(Russo, Malinconico, & Santagate, 2007). However, in the case of
ICMFC50 and ICBIRCH50, the decreases in the thicknesses are
probably due to the shrinkage of the alginate which in turn com-
presses the voids in the biocomposites and makes these films
denser.
Tensile strengths of the films are presented in Fig. 4a. To take
into account high differences in the densities of films, r/q values
have also been reported. The pristine alginate film had an r/q va-
lue of 49.0 kN m/kg and films with 1% to 15% of MFC have statisti-
cally the same r/q values, whilst MFC20 has a 17% and MFC50 a
38% greater r/q values than the pristine alginate film. On the other
hand, BIRCH50 had practically the same r/q value as the pristine
alginate film.
Compared to MFC, nano-sized NFC has a clearly better rein-
forcement effect on the alginate film. An r/q value of
80.10 kN m/kg was obtained when 10% of NFC was used, which
is 63% and 19% greater than those of MFC10 and MFC50. However,
the increase of NFC content from 10% to 15% did not further en-
hance r/q of the films.
Interestingly, DCC1 and DCC10 have practically the same r/q
values as pristine alginate film. This could be explained by the elec-
trostatic repulsion between the anionic carboxyl groups of alginate
and DCC, which prevents effective interaction between alginate
and DCC, previously observed in FTIR study.
Values of r showed slightly different pattern compared to r/q
values due to the large variations on the densities of films.
Fig. 4a shows that when the amount of MFC in the composite
was increased, r values of the films decreased compared to pristine
alginate films, and MFC10 has a 29% lower r value than pristine
alginate film. The r value of BRICH50 was also significantly lower
than the r values of the other films. As previously noted, based on
the FESEM images, surface of BIRCH50 was uneven, and the pores
on the surface may serve as the breaking points in the film during
tension.
2.5
2.25
2
1.75
1.5
1.25
1
Fig. 4. (a) Values of r/q (kN m/kg) and r (MPa) b) E/q (MNm/kg) and E (GPa) and
(c) strains (%) of films (error bar represents standard deviations).
On the other hand, NFC1 has a 15% higher r value compared to
the pristine alginate film and by increasing NFC content in the film
to 15%, the r value is increased 22%. NFC15 had an r value of
97.97 MPa, which is 40% greater that of the pristine alginate film.
Compared to the MFC films, NFC films have significantly higher r
values, e.g. NFC10 had 63% greater r value than MFC10. It has pre-
viously been reported that 8% nanosized cellulose, i.e. nanocrystal-
line cellulose (NCC) increases the r of alginate film from 57 to
75.24 MPa (Huq et al., 2012). This and our present studies suggest
that NCC and NFC have quite similar reinforcement effect on algi-
nate films.
The r value of DCC1 was 13% greater than that of the pristine
alginate film. However, when the amount of DCC is increased from
1% to 10%, the r value of the film is decreased from 79.37 to
69.63 MPa. The observation further indicates poor interaction be-
tween alginate and DCC.
Mechanical properties of the biocomposite films were also stud-
ied after ionic cross-linking. The slides of ICNFC10 were wrinkled
and uneven which affected on their properties as minor fractures
were formed during the preparation of the slides, and the accurate
results of ICNFC10 was not obtained. On the other hand, ICMFC50,
BIRCH50 and ICDCC10 were smooth and easily processed and they
showed higher r/q values compared to their non-cross-linked
 J.A. Sirviö et al. / Food Chemistry 151 (2014) 343–351
counterparts. Highest improvement in r/q was observed with
ICDCC10 which had 54% greater r/q value than DCC10. The r/q va-
lue of MFC50 and BIRCH50 increased 13% and 10% after ionic cross-
linking, respectively.
The r values of films were also remarkably increased during
cross-linking and ICMFC50 had a 49% greater r value than
MFC50, and ICDCC10 has a r value of 125.31 MPa which is 80%
greater r compared to DCC10. To the best of our knowledge this
is the highest r of alginate biocomposite films reported so far
and comparable with cellophane, a commonly used cellulose-
based packaging material as r value of cellophane is reported to
be as high as 120 MPa (Spence, Venditti, Habibi, Rojas, & Pawlak,
2010). As previously noted, based on the FTIR studies, the anionic
groups can take part in the formation of cross-linked network dur-
ing the ionic cross-linking, which results in enhancement of the
mechanical properties of the film.
Interestingly, the r value of BIRCH50 increased 93% during the
ionically cross-linking. As previously noted, based on the FESEM
images, cross-linking made film containing birch pulp more even
and reduced the amount of pores on the surface, which reduced
the amount of breaking points in the film. However, r value of
ICBIRCH50 is still 67% lower that of pristine alginate film and three
fold lower than the r value of ICDCC10.
3.5.2. Specific modulus and modulus
The E/q values of the films are presented in Fig. 4b. The pristine
alginate film had an E/q value of 9.38 M Nm/kg. An increase of the
MFC amount from 1% to 15% does not increase the E/q value of the
biocomposite film; however, MFC20 and MFC50 had 25% and 47%
greater E/q value than the pristine alginate film, respectively.
Interestingly, BIRCH50 showed 19% greater E/q value that of the
pristine alginate film. However, MFC50 had a 24% greater E/q value
than BIRCH50.
The E/q values of the films containing NFC are consistent with
the r/q values. When amount of NFC was increased, the E/q value
of the biocomposite film was also increased, and NFC10 had 55%
and 31% higher E/q value than the pristine alginate film and
MFC10, respectively. No significant increase in the E/q values can
be observed when amount of NFC is increased from 10% to 15%.
Furthermore, the addition of DCC to alginate film does not increase
the E/q of film.
E values of the films shows slight decrease when the amount of
MFC was increased. However, statistically all of the MFC films
showed almost same E that of the pristine alginate film. Similarly
to the r values, the E value of BIRCH50 is also significantly lower
than the E values of the other films.
Addition of the NFC to the alginate film increased E values of the
film and NFC1 had 13% and NFC5 27% higher E values than the pris-
tine alginate film. In addition, NFC10 had 51% higher E value than
MFC10. Statistically there was no difference in E values between
the pristine alginate film and films containing DCC fibres.
The E/q value of MFC50 was not affected by ionic crosslinking;
however, the E value of ICMFC50 was 30% lower than the E value of
MFC50. The E/q value of ICBIRCH50 was also almost same as the
E/q value of BIRCH50 and the E value of BIRCH50 was actually
increased 69% after cross-linking. In the biocomposite films with
10% of DCC, the E/q and the E values were increased 21% and
41%, respectively, after ionic cross-linking.
3.5.3. Elongation at break
Values of the strain of the films are presented in Fig. 4c. It is
known that incorporation of nanocellulose fibres to other polysac-
charide matrix’s decreases strain of the films (Fernandes et al.,
2010). Films with 10% and 15% NFC showed significant decrease
in strain and they had 66% and 61% lower strains than the pristine
349
alginate film, respectively. Statistically, other films showed similar
strain values than the pristine alginate film.
The strain values of the biocomposite films were slightly de-
creased during the cross-linking, and strains of the ICMFC50,
ICBIRCH50 and ICDCC10 were 28%, 42% and 7% lower compared
to their non-cross-linked counterparts.
Results from mechanical studies showed that addition of cellu-
lose, especially NFC to the alginate films increased their strengths
and moduli values, but decreased strain and flexibility of the film.
The improvements in the mechanical properties of composites
containing cellulose fibres are due to the formation of tightly
hydrogen bonded networks between fibres and matrix.
It was also observed that the chemical nature of the cellulose fi-
bre plays important part in the formation of strong biocomposites.
Before cross-linking, the anionic DCC did not have reinforcement
effect on the alginate, due to the electrostatic repulsion between fi-
bres and alginate. Even though there are anionic groups in NFC, the
charge density is significantly lower that of DCC and consequently
NFC had good compatibility with alginate. On the other hand, an-
ionic groups of the DCC were observed to take part in the ionic
cross-linking network after Ca2+ cross-linking, resulting in remark-
able increase in the mechanical properties of the film.
3.6. Barrier properties
3.6.1. Grease barrier properties
All of the studied cellulose-containing alginate films except
BIRCH50 had excellent grease barrier properties, i.e. the dyed tur-
pentine did not penetrate films within the 30 min testing period.
However, the dyed turpentine penetrated BIRCH50 immediately
which was probably due to porous structure of the film previously
noted, based on the FESEM images. On the other hand, surfaces of
the other films were smooth and visible pores were not observed
based on FESEM images. Moreover, the good grease barrier proper-
ties of alginate explain partly the excellent grease barrier proper-
ties of the biocomposite film (Ham-Pichavant, Sèbe, Pardon, &
Coma, 2005).
After ionic cross-linking, films retained their grease barrier
properties and no penetration of the dyed turpentine was observed
in ICNFC10, ICDCC10 and in ICMFC50. However, penetration of the
dyed turpentine occurred immediately with ICBIRCH50.
3.6.2. Water vapour permeability
The WVP of the pristine alginate film, BIRCH50, MFC50, NFC10
and DCC10 was measured before and after ionic crosslinking
(Fig. 5a). The WVP of pristine alginate film was 8.4  107 g/
(Pa s m2). Due to the high hydrophilicity, alginate has low water
vapour barrier properties (Olivas & Barbosa-Cánovas, 2008). The
addition of micro- and nanocellulose fibres to the alginate film de-
creased the WVP, e.g. the WVP of the MFC50 was a 23% lower than
that of the pristine alginate film. BIRCH50 had in turn a 6% higher
WVP that of the pristine alginate film.
It is known that barrier properties of films are improved when
additives have lower permeability and materials are well dispersed
(Khan et al., 2010). Furthermore, addition of micro- and nano-sized
cellulose fibres to the alginate matrix makes alginate films more
tortuous, and thus decreases the pathways to water molecules to
pass through the film (Huq et al., 2012). This could explain the im-
proved water vapour barrier properties of alginate films after addi-
tion of MFC, DCC and NFC fibres. The large pores, observed in
FESEM images, may create direct pathways for water vapour to
penetrate through BIRCH50, resulting in decreased water vapour
barrier properties compared to the pristine alginate film.
It is known that ionic cross-linking improves water vapour bar-
rier properties of alginate and composites containing alginate
(Rhim, 2004). Here water vapour barrier properties were also
350 J.A. Sirviö et al. / Food Chemistry 151 (2014) 343–351
Fig. 5. (a) WVP (g/(Pa s m2)) and (b) SWVP (kg m/(Pa s m2)) of films (error bar
represents standard deviations).
observed to improve after ionic cross-linking and both ICNFC10
and ICDDC10 have a 13% lower WVP than the pristine alginate film.
Furthermore, ICNFC10 has a 6% lower WVP than NFC10 and
ICDD10 has a 9% lower WVP than DCC10.
WVP of MFC50 was increased after cross-linking, and ICMFC50
has a 14% higher WVP than MFC50. However, the WVP of ICMFC50
is still a 6% lower than the pristine alginate film. On the other hand,
the water vapour barrier properties of BIRCH50 was decreased
after cross-linking and ICBIRCH50 has 11% and 5% lower WVP com-
pared to BIRCH50 and the pristine alginate film, respectively. As
previously noted, based on the FESEM images, the number of pores
in the surface of BIRCH50 was reduced during the cross-linking.
This in turn reduced the direct pathways of water vapour to pene-
trate through the film.
To take into account the differences in the thicknesses of films,
SWVPs are presented in Fig. 5b. There were no significant differ-
ences in the SWVPs of the films, expect in the films containing
birch pulp, i.e. SWVP of BRICH50 is more than threefold
higher than that of the pristine alginate film. After cross-linking,
SWVP of the BIRCH50 decreased 32%, but was still 91% higher
compared to pristine alginate films. Compared to common
packaging materials, such as cellophane, all of the biocomposite
films had lower SWVP than cellophane whose SWVP is reported
to be 6.9  1014 kg m/(Pa s m2) (Péroval, Debeaufort, Despre, &
Voilley, 2002).
4. Conclusion
In this work, biocomposite films based on cellulose and alginate
were produced using four different wood cellulose fibres with
different fibre sizes, chemical nature and proportions. Films were
also further cross-linked using Ca2+. Celluloses showed good inter-
action with alginate matrix and the mechanical properties of films
were increased by increasing the amount of the fibre in the
biocomposite and by decreasing the fibre size of cellulose. How-
ever, despite the nanosized fibre structure, anionic charges pre-
vented an efficient interaction between DCC and alginate without
a cross-linking. On the other hand, anionic carboxylate groups
created a highly intense cross-linked network between alginate
and DCC, when the film was ionically cross-linked with Ca2+. A bio-
composite film with remarkably high mechanical strength was
produced by ionically cross-linking the alginate film containing
10% of DCC. All of the biocomposite films except the film with
unmodified birch pulp, showed excellent grease barrier properties.
Furthermore, the WVP of alginate film was decreased by the addi-
tion of MFC and nano-sized cellulose fibres, and by ionic cross-link-
ing. This work demonstrates that biocomposite films based on
cellulose fibres and alginate have high potential to be used as high
strength packaging materials.
Acknowledgments
The authors acknowledge the support from the Future
Biorefinery Programme of Forestcluster Ltd. V.T.T. is acknowledged
for the pulp analysis.
Appendix A. Supplementary data
Supplementary data associated with this article can be found, in
the online version, at http://dx.doi.org/10.1016/j.foodchem.2013.
11.037.
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