Cellulose (2014) 21:1143–1148
DOI 10.1007/s10570-014-0165-y
ORIGINAL PAPER
The dynamic development of exclusion zones on cellulosic
surfaces
Belkis Sulbarán • Guillermo Toriz •
G. Graham Allan • Gerald H. Pollack •
Ezequiel Delgado
Received: 27 August 2013 / Accepted: 6 January 2014 / Published online: 17 January 2014
Ó Springer Science+Business Media Dordrecht 2014
Abstract The dynamics of the development of solute
exclusion zones produced by water in contact with
various cellulosic surfaces are reported. The term
‘‘exclusion zone’’ (EZ) refers to the ordered water
volume present immediately contiguous to hydrophilic
surfaces. As such, three examples of cellulose-based
films, i.e., cellophane, cellulose acetate with 0.94
degree of substitution (CA0.94) and cellulose acetate
with 2.51 degree of substitution (CA2.51) were com-
pared by exposure to sulfated-polystyrene microspheres
(2 lm diameter) suspended in water. Zones that were
not penetrated by the microspheres were observed in
each case and measured by means of an optical
microscope. The thickness of these exclusion zones
adjacent to the cellulosic surfaces increased progres-
sively with time and reached a maximum value of
190 lm after 1 h of exposure. Zone formation was
influenced by roughness, crystallinity, and the degree of
acetylation of the surfaces. For the cellulosic surfaces,
B. Sulbarán  G. Toriz  E. Delgado (&)
Departamento de Madera, Celulosa y Papel, Universidad
de Guadalajara, C.P. 45110 Guadalajara, Mexico
e-mail: ezedelfor@gmail.com
G. G. Allan
Department of Chemical Engineering and College of the
Environment, University of Washington, Seattle, WA,
USA
G. H. Pollack
Department of Bioengineering, University of Washington,
Seattle, WA, USA
the rougher the surface, the thicker was the EZ formed.
Both the accessibility and availability of the hydroxyl
groups on the cellulosic surfaces also influenced the
thickness of the resultant EZ in the sequence, cellophane
[ CA0.94 [ CA2.51. The surface of cellulose acetate
with the greater substitution degree (CA2.51) produced
the thinnest EZ. The diminution of the capacity of these
cellulose derivatives to instigate water-molecule order-
ing is considered to be due to the hydrophobicity
imparted by the acetyl group content.
Keywords Water-ordering  Exclusion zone 
Cellulosic surfaces  Surface roughness 
Crystallinity
Introduction
Water, which is involved in a wide variety of
biological, physical, and chemical processes, has
dipole–dipole interactions that make water molecules
self-associate by hydrogen bonding, forming three-
dimensional networks (Vogler 1998; Lombardo et al.
2009). Extensive ordering of water molecules occurs
near hydrophilic surfaces and is known as the ‘‘fourth
phase’’ of water (Vogler 1998; Zheng and Pollack
2003; Chai et al. 2008; Pollack 2013). Ordered water
has a hexagonal crystalline structure similar to ice, and
its viscosity, freezing point and permittivity are
different from bulk water. Biological tissues, gels,
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and natural as well as synthetic polymers have
structured water adjacent to their surfaces (Zheng
et al. 2006; Chai et al. 2008; Pollack, 2013).
The ordering of the molecules in water by a surface
can create a zone from which solutes of different types
are excluded (Zheng and Pollack 2003; Zheng et al.
2006; Pollack et al. 2009). The thickness of this
exclusion zone (EZ) can extend to hundreds of
micrometers from the surface (Zheng and Pollack
2003; Zheng et al. 2006; Chai et al. 2009; Das and
Pollack 2013). Such long-range water ordering has
been confirmed on hydrophilic surfaces as diverse as
mica, quartz, clay, sand and glass by the use of a
variety of spectroscopy methods and novel instru-
mentation (Das and Pollack 2013). Several of these
studies have also shown organized water extending
into the bulk for at least 50–100 nm (*200–500 water
layers) (Zheng et al. 2006). Furthermore, a comparison
of bulk and the water in the EZ illustrates that the latter
has a reduced adsorption of infrared radiation (infrared
images), negative electrical potential gradients, and
shorter relaxation times in nuclear magnetic resonance
experiments (NMR-T1) (Zheng and Pollack 2003;
Zheng et al. 2006; Chai et al. 2009; Das and Pollack
2013). The decrease of relaxation times in the EZ
implies that the water therein has a more stable
structure than free water, is similar to a liquid crystal,
and solutes are excluded therefrom because free water
is more easily available for interaction (Zheng et al.
2006; Vyalikh et al. 2007; Chai et al. 2009). In
addition, water within the EZ absorbs UV light at the
wavelength of 270 nm and this is interpreted as a
‘‘signature’’ of the ordered nature of water in the
vicinity of charged or hydrophilic surfaces (Chai et al.
2008). From all of these observations, it is concluded
that water in the EZ is physically and chemically
different from ordinary bulk water.
In the formation of a paper web from an aqueous
suspension of cellulose fibers and additives, water
serves as a transport vehicle for the accommodation of
the fibers and the development of adhesion between
cellulose surfaces (Hubbe 2006). Adhesion between
cellulose fibers is essential for the development of both
the dry and wet strength properties of paper and
paperboard (Hubbe 2006; Delgado et al. 2011). It has
long been accepted that the main basis of bonding
between fibers is the formation of hydrogen bonds
between the hydroxyl groups on the cellulose surfaces
(Hubbe 2006). However, the surface roughness of
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Cellulose (2014) 21:1143–1148
cellulose fibers can also affect bonding. Initially, there
are long-range interactions between water and fibers,
which create forces that compact fibers as water leaves
the web, forcing the fibers into intimate contact. When
water drains off by the application of vacuum and
pressure, more water–air interfaces appear, thus
increasing surface tension. Then, the capillary and
surface tension forces, together known as Campbell
forces, bring fiber surfaces into more intimate contact,
facilitating short-range interactions. However, the
nature of the short-range mechanisms that lead to the
extensive development of networks of hydrogen bonds
between fibers as the water content is reduced down to
8–10 % is not fully understood. Questions therefore
arise as to how much fiber surface is really in close
contact in order to favor the formation of hydrogen
bonds, and whether this kind of intermolecular force is
sufficient to account for the exhibited paper strength.
Moreover, the location and role of the remaining water
in ‘‘dry’’ paper (7–8 % moisture content) remains
unknown (Delgado et al. 2011). For these reasons, the
existence of ordered water near hydrophilic surfaces
may influence the bonding of cellulose fibers, espe-
cially in the consolidation stage of a sheet of paper
(Delgado et al. 2011).
It can therefore now be hypothesized that the water
adjacent to the surface of the cellulose fibers may
possess certain ordering that favors interfiber bonding.
As a result, the bonds between ordered water mole-
cules could act as bridges to join surfaces hundreds or
thousands of nanometers apart. This sort of water bond
bridging may act in concert with other short-range
interactions such as electrostatic attractions, polar
interactions, hydrogen bonding and van der Waals
forces (Delgado et al. 2011). It is also possible that this
bridging phenomenon has an effect on the exclusion or
repulsion of a variety of polymeric additives, organic
or inorganic fine particles, and other particles sus-
pended or dissolved in the fiber slurry used to form the
sheet.
The general objective of the present study was to
understand the nature of the EZ produced by water
molecules adjacent to cellulosic surfaces. To pursue
this objective, the following studies were undertaken:
(a) characterization of the solute EZ (size and
development with time) next to cellulosic surfaces;
and (b) establishment of the effects of surface
topography and surface hydrophilicity on the size of
the EZ. Surface hydrophilicity was calculated by
Cellulose (2014) 21:1143–1148
Fig. 1 A schematic illustration of the chamber used to place
microspheres in contact with the polymeric films
measuring surface crystallinity and relating it to the
accessibility and occurrence of surface hydroxyl
groups. Due to the difficulty of studying this phenom-
enon with individual cellulose fibers or fibrous
networks (e.g., paper), regenerated cellulose film
(cellophane) and cellulose acetate films were selected
as examples of cellulosic surfaces.
Experimental
Materials
Four types of hydrophilic surfaces were used: (a) Naf-
ion 117 (Sigma Aldrich, Toluca, Mexico) was
employed as a reference; (b) cellulose acetate
(CA0.94) with a 0.94 degree of substitution (DS)
(Fluka, Toluca, Mexico); (c) cellulose acetate with
2.51 DS (CA2.51) from Sigma Aldrich (Toluca,
Mexico); and (d) cellophane films from Artefactos de
Papel, S.A. de C.V. (Mexico City, Mexico). Monodis-
perse, sulfate-functionalized polystyrene micro-
spheres of 2 lm diameter (Polysciences, Warrenton,
PA) were suspended in deionized water (1 mg/mL).
Exclusion zone observations
A strip of each polymer film was placed along the
length of a circular cavity carved in a glass plate as
shown in Fig. 1. The suspension of microspheres was
then added. The concentration of the microspheres
was 1 mg/mL.
Observations of the formation of exclusion zones
were carried out with an inverted microscope (Zeiss
Axiovert I35) equipped with a 109 objective. Video
recordings were taken at specific intervals during a
total of 60 min of exposure, and recorded with a
digital camera (Scion Corporation, Inc., color digital
camera, Model #CFW-1312C). The EZ thickness was
determined from the images using a software program
1145
(Image J). The program measured the distance
between the film surface and the near edge of the
excluded microspheres in the media.
Topography of the polymer surfaces
Surface topography was measured by atomic force
microscopy (AFM) and a Veeco Dimension 3100
scanning probe microscope (SPM) in the Nanotech
User Facility at the University of Washington, USA.
The system was operated by a NanoScope IVa
Controller, integrated with a Dimension Hybrid
XYZ Closed-Loop Scanner. The cantilevers (tapping
mode, silicon, from Nanosensors) had a typical
resonance frequency of 250–300 kHz. All measure-
ments were performed in the wet state. Roughness
was determined from records obtained in the tapping
mode (height image) (Yuliwati et al. 2011) and is
defined as:
ZLy ZLx
1
Ra ¼ jf ðx; yÞjdxdy
LxLy
0 0
where: Ra is the mean roughness. This is the mean
value of the surface relative to the center plane; f(x, y)
is the surface relative to the center plane; Lx and Ly
are the dimensions of the surface.
Cellulosic film crystallinity
Measurements of crystallinity were carried out in a
powder X-Ray Diffractometer (XRD SIEMENS D500
Model Kristallograph) with a Cu Ka radiation
(0.1542 nm wavelength) at the University of Guad-
alajara, Mexico. High angle XRD data were collected
from 5°–50° at 0.02° increments and 1 s count times
using films placed on a quartz sample holder. For
cellulosic films, the crystallinity (%) was calculated as
described by Segal et al. (1959):
 
ðI002  IamÞ
IC ¼  100
I002
where IC crystallinity (%); I002 is the crystalline peak
of the maximum intensity at 2h between 22° and 23°
for cellulose I; Iam is the crystalline peak of the
minimum intensity at 2h between 18° and 19° for
cellulose I.
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Computation of the number of available surface
hydroxyl groups
In order to calculate the number of surface hydroxyl
groups per lm2 a crystal lattice model was used for
cellulose I in a wet state as previously reported by
Houtman and Atalla (1995). The lattice had dimen-
sions of 37 9 29 Å2 and included five cellulose chains
with seven anhydroglucose units per chain. This
model would represent the minimum surface area for
an idealized cellulose surface since the cellulose
molecules are then in the most compact state. This
calculation indicated that 9.78 9 106 hydroxyl groups
per lm2 are available for interaction with free water.
Accessibility and degree of substitution were
considered in the estimation of the actual number of
available hydroxyl groups on each of the cellulosic
surfaces. Accessibility was related to the amorphous
part of cellulose, defined as (1—the crystallinity
index). Finally, the amount of free hydroxyl groups
was calculated from:
 
# available OH IC
¼ 1  ð3  DSÞ
lm2 100
 9:78  106
where IC crystallinity (%); DS degree of substitution
(there are three available hydroxyl groups per anhy-
droglucose unit); 9.78 9 106 is the amount of
hydroxyl groups per lm2 in a wet cellulose I lattice.
Results and discussion
The aim of this work was to define the exclusion zones
for cellulosic surfaces and compare it with that of the
extensively studied Nafion (a chain-end sulfated
perfluorohydrocarbon). The establishment of the
exclusion zones took longer times with cellulosic
surfaces than with Nafion, on which the EZ began to
develop instantaneously and grew rapidly (Fig. 2). It is
also shown that the more acetylated the cellulose
surface, the lower the thickness of the resultant EZ,
and the longer the time needed for development.
Nafion showed the largest EZ when compared to the
cellulose acetate and cellophane films. The latter
started to form a measurable EZ after 10 min of
exposure. The EZ for CA0.94 films reached 50 lm in
thickness after a 20 min exposure and a little less
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Cellulose (2014) 21:1143–1148
Fig. 2 The time course of the EZ thickness generation by
different film surfaces in contact with a water suspension of
2 lm-sulfated microspheres (lines are drawn only to guide the
eye)
(40 lm) for CA2.51 films at a 30 min exposure. It can
also be noted that EZ of cellophane, CA0.94 and
Nafion seemed to converge after 60 min. In longer
experiments (90 min) with all the surfaces (data not
shown) the zones became separated again, denoting
their fluctuating nature.
These observations can be explained as follows:
Nafion has surface ionic sulfate groups that can
interact instantaneously with water, which allows for
the early development of an EZ. In contrast, the
cellulose hydroxyl groups on the surface take more
time to interact with the water molecules in the
aqueous phase. The more substituted the cellulose
derivative, the longer it took for the hydroxyl groups
of the water to interact with the nonionic surface and to
develop an EZ. The fact that exclusion zones seem to
converge at longer times is ascribed to the ability of
the less substituted cellulose acetate to rearrange its
surface hydroxyl groups, and also to the fluctuating
nature of the zones. Figure 3 shows the optical
micrographs for each film tested after 1 h of exposure.
The EZ is clearly thicker for the less substituted
cellulose derivative, and even thicker for the Nafion
film.
The topography of the cellulosic and Nafion films
was measured in order to determine whether a change
in roughness was related to the extent of EZ formation.
The effect of surface roughness on the EZ thickness is
Cellulose (2014) 21:1143–1148
a b
Fig. 3 The exclusion zones generated by different film surfaces in contact with a water suspension of 2 lm sulfated microspheres at a
60 min exposure time for a Nafion, b cellophane, c CA0.94 and d CA 2.51
Fig. 4 The effect of cellulosic surfaces roughness (horizontal
top axis) and surface hydroxyl groups (horizontal bottom axis)
on the EZ thickness (error bars represent SD of 10 EZ
measurements)
noteworthy since the EZ extends for many microns
even though the surface roughness of films is within
the nanometer scale. Moreover, for the cellulosic
surfaces, the roughness decreased with increased
degree of substitution. Also, for the cellulosic surfaces
under study, larger EZ values corresponded to a
greater roughness because the layers of structured
water fluctuated more and created a wider and more
diffuse EZ (Fig. 4).
On the other hand, the Nafion film has a low surface
roughness (46 nm) and generates the thickest EZ
(220 lm) although the swelling in such films is
negligible. Thus, the effect of swelling of the cellu-
losic surfaces needs to be considered and may prove to
be a more significant parameter, since it is implied that
the appearance of an EZ is a consequence of the
1147
c d
interactions between the hydrophilic groups present on
the surface and the water molecules of the media. In
fact, in our assays it was observed that a swelling of the
surfaces occurred before an EZ could be detected.
Certainly, more data on this phenomenon needs to be
acquired before any further conclusions on this issue
can be drawn.
The thickness of the EZ as a function of available
hydroxyl groups on the cellulosic surfaces is also
presented in Fig. 4. The actual surface availability of
hydroxyl groups depends on the number present and
those left after acetylation and their accessibility. It
has been demonstrated that surface accessibility is
higher for cellophane than for cotton and that was
attributed to the lower crystallinity index of cello-
phane (Östenson et al. 2006). Consequently, the values
of the crystallinity index were utilized to calculate the
degree to which hydroxyl groups in amorphous
regions are available for interaction with water. On
the other hand, according to Houtman and Atalla
(1995), a crystal lattice for cellulose in the wet state
with dimensions of 37 9 29 Å2, which includes five
cellulose chains, has sufficient space to accommodate
692 water molecules and has 9.78 9 106 hydroxyl
groups per nm2 available for interaction with free
water. This case would be for the minimum surface
area, i.e., for the best packing of cellulose, and this was
the value selected for calculations of the OH avail-
ability per lm2. For the cellulosic materials studied
here, the cellophane formed a thicker EZ than any of
the cellulose acetate films, followed by CA0.94 and
then by the more substituted CA2.51. The large
fluctuation of EZ size exhibited by cellophane stems
from multiple factors that may include roughness and
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a rather low crystallinity. It is to be noted that the
roughness and OH availability of cellulosic surfaces
follow the same trend regarding the formation of the
EZ. Thus, the thickness of the EZ depends primarily
on the surface hydrophilicity, which was the greatest
in the non-substituted cellulose (cellophane). Also, the
amount of free hydroxyl groups depends on the
available surface area, which increased with higher
roughness (Fig. 4).
Conclusions
Cellulosic surfaces create observable and measurable
exclusion zones when in contact with micron-sized
polymeric spheres suspended in water. The exclusion
zones extend to thicknesses between 40 and 200 lm,
depending on the exposure time, the crystalline
character of the surface and the content of hydropho-
bic groups, i.e., the availability of surface hydroxyl
groups. This behavior is in accord with current data
reported in the literature for other hydrophilic mate-
rials. Furthermore, it is established that surface
roughness affects the extent of exclusion zone forma-
tion thereon.
Acknowledgments Financial support for this work was
provided by the National Council of Science and Technology
of Mexico (CONACYT) and by an NIH Transformative grant
awarded to Professor Pollack. The authors acknowledge the
Nanotech User Facility at the University of Washington in
Seattle, USA, for experimental support during the AFM
measurements.
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