Food Hydrocolloids 41 (2014) 44e52

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Food Hydrocolloids
journal homepage: www.elsevier.com/locate/foodhyd

Impact of sucrose and trehalose on different agarose-hydrocolloid

systems
Natalie Russ*, Birgitta I. Zielbauer, Thomas A. Vilgis
Max Planck Institute for Polymer Research, Ackermannweg 10, 55128 Mainz, Germany

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

Article history: Adding high amounts of sugar influences the physical properties of hydrocolloid systems enormously,
Received 31 October 2013 due to their mutual interactions with each other as well as with water molecules. The impact of the two
Accepted 18 March 2014 disaccharides sucrose and trehalose on the thermo-mechanical properties of agarose based hydrogels is
Available online 29 March 2014
investigated. Agarose, a typical gelling agent, is combined with the nongelling agents sodium-alginate
and xanthan, two polyelectrolytes which differ significantly in their flexibility. Material properties,
Keywords:
such as viscoelasticity, water holding capacity, gelling temperature and thermal stability can be adapted
Agarose
by the controlled addition of sugar. These properties are investigated by strain and temperature
Alginate
Xanthan
dependent oscillatory rheological measurements as well as by controlled drying experiments. The ex-
Disaccharide periments clearly indicate the competition of the different carbohydrates for water. The differences in
Viscoelasticity water binding of sucrose and trehalose are clearly visible. A phenomenological model is presented and
Water holding capacity illustrates the different effects of the sugar molecules on the hydrocolloid systems. It is suggested that
hydrate shells and the differences in local water binding on atomistic and nano scales have strong effects
on macroscopic properties.
Ó 2014 Elsevier Ltd. All rights reserved.

1. Introduction gelling properties of agarose and the composite gels agarosee

Hydrocolloids are widely used in daily life and especially in the
food industry to improve the quality and to adjust the texture of
different food products. They are well established as thickening
additives, as gelling agents or to stabilize dispersions and solutions.
Additionally, their applications in different food products are often
affected by the presence of high sugar concentrations, which are
used as sweeteners or to change the texture of the resulting
product to produce an overall acceptable mouth feel. For many
products such as confectionery, candies and bakery products, high
sugar concentrations are essential since they define the local
physical interactions of proteins, starches and hydrocolloids, which
may have been added as stabilizers and texturizers. In this study
the impact of the disaccharides sucrose and trehalose on the
thickening properties of well-defined model systems, rather than
on real foods, which are multicomponent materials with many
competing interactions, is investigated. Nongelling hydrocolloids
such as alginate (in the absence of bivalent calcium ions) and
xanthan have been chosen, and the impact of sugar addition on the
* Corresponding author. Tel.: þ49 6131 379149.
E-mail addresses: russ@mpip-mainz.mpg.de (N. Russ), zielbauer@mpip-mainz.
mpg.de (B.I. Zielbauer), vilgis@mpip-mainz.mpg.de (T.A. Vilgis).
alginate and agaroseexanthan is studied.
For some basic information about the chemical and physical
properties of the hydrocolloids agarose (Imeson, 2010; Stanley,
2006), xanthan (Sworn, 2010; Urlacher & Noble, 1999) and algi-
nate (Clare, 1993; Onsøyen, 1999) the reader is referred to the
literature. The gelation mechanism and final gel properties of
agarose and its modification by additions of non-gelling hydrocol-
loids with different flexibility have been the main topic of previous
work (Russ, Zielbauer, Koynov, & Vilgis, 2013). The gelation of
agarose consists of a two-step mechanism with an initial helix
formation and subsequent aggregation of the helices (Nordqvist &
Vilgis, 2011; Normand, Lootens, Amici, Plucknett, & Aymard,
2000). The addition of the nongelling agents xanthan and algi-
nate influences this gelation mechanism in different ways. Xanthan
molecules in solution behave as charged and very stiff, almost rigid
rods which undergo above a critical concentration, a so-called non-
equilibrium jamming transition (Edwards & Vilgis, 1986), resulting
in an immobilization of the rods at random positions with random
orientations. This rigid and randomly oriented structure of the
ensemble of xanthan rods reduces the mobility and diffusion of the
dissolved and homogeneously distributed agarose chains. Conse-
quently, the gelation of agarose under spatial restrictions limits the
elasticity and stability of the network. Alginate in contrast, consists

http://dx.doi.org/10.1016/j.foodhyd.2014.03.020
0268-005X/Ó 2014 Elsevier Ltd. All rights reserved.
 N. Russ et al. / Food Hydrocolloids 41 (2014) 44e52
as well as agarose of entangled and flexible polymer coils, which
hardly hinder the diffusion of the agarose chains and the gel for-
mation. Additionally the flexible alginate coils get incorporated into
the meshes of the agarose network where they act as reinforcing
fillers, resulting in a more stable and elastic network. Most of these
effects are ruled by large scale conformational chain properties,
long range interactions between charges and short range dipolar
interactions, in contrast to other properties induced by low mo-
lecular weight sugars.
The addition of low molecular weight sugars to hydrocolloid
solutions or gels influences the physical properties of those by
competing effects on local length scales. It will be shown below,
that local interactions (hydrate shells, additional hydrogen
bonding) have a strong effect in macroscopic quantities like moduli
and viscosity (Fioretto et al., 2013). Adding sugar to hydrocolloid
systems at constant polymer concentrations leads to a “replace-
ment” of free water by sugar molecules, leading to an effectively
increased hydrocolloid concentration with respect to the amount of
solvent. Therefore the distance between the polymer chains de-
creases, and the mean intermolecular polymerepolymer in-
teractions increase. Moreover, the sugar molecules compete with
the hydrocolloids for the available water to form hydrate shells. On
the other hand, the sugar molecules can interact with the polymer
chains of the hydrocolloids via hydrogen bonds or minimalize the
electrostatic repulsion in polyelectrolytes.
In this study, two disaccharides, sucrose and trehalose, are
investigated. Sucrose consists of glucose and fructose combined by
a (1, 2)-linkage, whereas trehalose is composed of two glucose rings
with a (1, 1)-bond. These two disaccharides strongly differ in their
interactions with water molecules, due to differences in their
chemical structure regarding the positions and orientation of their
hydroxyl groups (Gharsallaoui, Rogé, & Mathlouthi, 2008). The
dynamic state of water molecules around the sugar molecules in
solution depends on the polar interaction between water and sugar
molecules, which is affected by the number of equatorial hydroxyl
groups (n(e-OH)). Uedaira, Ikura, and Uedaira (1989) proposed that
the sugar molecules, possessing a higher number of e-OH groups
per molecule have a stronger stabilizing effect on the water struc-
ture. Sucrose has a high solubility in water, whereas the trehalose
molecules with their higher n(e-OH) interact very strongly with the
water molecules via hydrogen bonds, resulting in a higher water
holding capacity (Branca et al., 2001). The differences between
mono- and disaccharides in affecting hydrocolloid properties have
been addressed elsewhere (Maurer, Junghans, & Vilgis, 2012).
In order to evaluate the impact of sucrose and trehalose on the
mechanical and thermal properties of the different hydrocolloid
systems, amplitude and temperature dependent oscillatory
rheology measurements have been performed. Additionally, the
water holding capacity of the different gels was examined by drying
them under defined conditions. Based on these results, simple
models are proposed to describe the impact of sucrose and treha-
lose on the gelling and thickening properties of agarose, xanthan
and alginate as well as on their mixtures agaroseexanthan and
agaroseealginate.
2. Materials and methods
2.1. Materials
Agarose (CAS No. 9012-36-6) was purchased as fine white
powder from Fisher Scientific GmbH with a specific gelling tem-
perature (Tgel) between 34 and 45  C and a gel strength of >1000 g/
cm2 as given by the producer. Na-alginate (CAS No. 9005-38-3) was
purchased from SigmaeAldrich Chemie GmbH. Xanthan gum
(reinst, E-415, CAS-No. 11138-66-22) was from Carl Roth GmbH &
45
Co. KG. Sucrose (CAS No. 57-50-1) and trehalose (CAS No. 99-20-7)
were purchased from Theka Analytical and Acros Organics (Thermo
Fisher Scientific Inc.), respectively. Sucrose had a purity of 99.0%
and D-trehalose was about 99% anhydrous.
2.2. Sample preparation
1%, 1.25% and 1.7% w/w of agarose was dispersed in distilled
water or in the appropriate sugar (sucrose or trehalose) solutions
with 20% and 40% w/w. For the moisture content analysis, sugar
solutions with 10 and 30% w/w were used additionally. The
dispersion was heated up to 90  C and kept there for 5 min while
stirring (200 rpm). By using a contact thermometer and a snap-cap
vial, a constant temperature of 90  C and minimal water evapora-
tion could be assured. After 5 min the hot clear solution was poured
into moulds with a defined geometry. For the amplitude-sweep
tests, moulds with a diameter of 25 mm and a height of 3 mm
and for the moisture content analysis moulds with 50 mm diameter
and a height of 1 mm were used. Excess solution was wiped off with
a broad razor blade. The solution was allowed to cool down to room
temperature for 15 min, then the moulds were removed and the set
gels hardened in the refrigerator at 4  C for 24 h. To produce the 1-1
mixtures of agaroseealginate and agaroseexanthan, the pure
xanthan or alginate dispersions have been prepared first. For this,
1%, 1.25% and 1.7% w/w of the xanthan and alginate powder were
dispersed in distilled water or sugar solutions and stirred for 24 h at
room temperature at 200 rpm. Then 1% w/w agarose was dispersed
in distilled water or sugar solution like before and mixed in a 1-1
ratio with the hydrocolloid dispersions. The mixture was shaken by
hand and heated up again to 90  C for 5 min while stirring at
200 rpm. Afterward the gels hardened as described above, first at
room temperature and then in the refrigerator.
2.3. Dynamic viscoelastic measurements
The measurements of the viscoelastic properties as well as the
solegel transition of the hydrocolloids were carried out with a
stress controlled rheometer (Bohlin Gemini 200, Malvern, UK). Two
different tests were performed: amplitude-sweep test and
temperature-sweep test.
Amplitude tests were performed for g ¼ 0.001-1 at a constant
frequency f ¼ 1 Hz and temperature T ¼ 25  C. Five periods per
point were measured with a delay time of 0.5 s and an integration
time of 5 s. Measurement of five different samples were averaged
and the standard deviation taken as error. The plateau modulus G0
has been extracted from these averaged curves at a strain well in
the linear viscoelastic region. Alginate solutions and xanthan dis-
persions were measured with a stainless steel coaxial cylinder (C25,
Malvern, UK) while for gelled agarose samples a plateeplate ge-
ometry of stainless steel with a diameter of 25 mm was used. For
the latter, an adequate contact between samples and measuring
system without significant compression of the sample had to be
ensured. Therefore the gap size for each sample was set manually.
After inserting the sample, the upper plate was set to a gap of
3000 mm and lowered in steps of 100 mm until the contact between
sample and plate was distinct and visible. To avoid slippage of the
samples at high deformations, sandpaper with a grain size 80 was
glued onto both plates.
The temperature-sweep test yields information about the sole
gel transition and the thermal stability of the hydrocolloid gels.
Storage G0 and loss G00 modulus are measured as a function of
temperature with f ¼ 1 Hz and strain g ¼ 0.001 or 0.01. Integration
and delay time were the same as for the amplitude-sweep. Here
three different samples for each system investigated were
measured and the results were averaged. 1350 mL of the hot and
46 N. Russ et al. / Food Hydrocolloids 41 (2014) 44e52
liquid sample (sample preparation see 2.2 without gel setting) were
deposited onto the preheated measuring system (PP 40, stainless
steel) of 80  C and the upper plate was lowered slowly to a gap of
1000 mm. The liquid sample was cooled down from 80  C to 20  C at
a rate of 1 K/min and the moduli were determined as a function of
temperature. Temperature control was enabled by a coaxial peltier
cylinder system. To avoid water evaporation during the measure-
ment, a thin film of paraffin oil (viscosity h ¼ 25 mPas, Merck KGaA)
protected the sample. To guarantee a continuous contact between
sample and plates during gelling and melting an auto-tension
control was employed. This function allows maintaining a con-
stant normal force on the sample during the solid phase such that
expansion or contraction of the gels due to thermal variations can
be compensated.
2.4. Moisture content analysis
To investigate the water holding capacity and the syneresis
behavior, the hydrogels were dried with a halogen moisture
analyzer (HR83, Mettler Toledo, Germany). The samples (sample
preparation see 2.2) were dried at two different temperatures:
50  C, a temperature far from the melting point of the gels, and
80  C, a temperature near the melting point. While drying, the
weight loss is detected in dependence of time and the moisture
content can be directly determined in percent of the total sample
mass. The measurement was stopped, within 5 s no weight changes
bigger than 1 mg could be detected. The following results are
averaged over the measurements of three different samples for
each investigated system.
3. Results and discussion
3.1. Viscoelastic properties
High concentrations of disaccharides, such as sucrose and
trehalose, affect the viscoelastic properties of hydrocolloids by
different mechanisms. If hydrocolloid/sugar systems are prepared
by simply substituting the water by the same amount of sugar so-
lution as it has been done here, an effectively increased hydrocol-
loid concentration results due to replacement of water molecules
by sugar molecules. This leads to an increase in the elastic (G0 ) as
well as viscous (G00 ) modulus in all hydrocolloid systems (1%
agarose, 1% alginate, 1% xanthan, 1% agaroseealginate and 1%
agaroseexanthan). To determine the effect of sugar correctly, sys-
tems with the same amount of water have to be compared.
Dispersing 1% w/w of hydrocolloid in a 20% or 40% w/w sugar so-
lution provides an effective concentration of 1.25% or 1.7% hydro-
colloid, with respect to the water present in the system. Therefore, a
1.25% and 1.7% hydrocolloid system without sugar was used for
comparison in order to eliminate the effect of increasing moduli
due to water shortage. Fig. 1 shows the average of the plateau
modulus G0 , obtained from the amplitude-sweep test, in the linear
viscoelastic region for the different hydrocolloid systems. Only
single frequency measurements were performed here, because
although agarose gels exhibit viscoelastic behavior, they behave
almost like a pure elastic material and show only a slight frequency
dependence in the solid state (Labropoulos, Niesz, Danforth, &
Kevrekidis, 2002; Ross, Pyrak-Nolte, & Campanella, 2006).
3.1.1. Agarose þ sucrose or trehalose
The increase in the elastic modulus of the 1% w/w agarose gel by
adding 20% or 40% sugar (Fig. 1a), as already observed by
Deszczynski, Kasapis, MacNaughton, and Mitchell (2003);
Deszczynski, Kasapis, and Mitchell (2003) can be explained by
the reduced amount of water and the thus increased effective
concentration of agarose. Kasapis, Al-Marhoobi, Deszczynski,
Mitchell, and Abeysekera (2003); Kasapis, Al-Marhoobi, Deszc-
zynski, Mitchell, and Abeysekera (2004) described the stabilizing
effect of sugar molecules in hydrocolloid systems. They assumed
that the hydrogen bonds between the polymer chains are
strengthened by adding sugar molecules, and more stable and
stronger junctions in the hydrogel networks develop. Nishinari
et al. (1992) also observed an increase of the elastic modulus of
agarose gels by the addition of sucrose. Based on a “zipper model”
(Watase, Kohyama, & Nishinari, 1992) these authors explained this
observation by an increase of the number of junction zones, a
decrease of the size of each junction zone and consequently, a
decrease of the rotational freedom of a link in a molecular zipper.
Here we were interested in a comparison of gels with equal poly-
mer to water ratios with and without sugar molecules as co-solutes.
This comparison with an agarose gel of a concentration corre-
sponding to the effective concentration of agarose in the sugar
containing gel (1.25% or 1.7%) shows a decrease in elasticity with
the addition of sucrose as well as trehalose (Fig. 1a).
One reason for this decrease can be that the presence of the
sucrose or trehalose molecules disturbs the mobility and diffusion
of the agarose chains (Fig. 2a), and causes a steric hindrance. Thus,
during cooling, less agarose polymer chains, or chain portions along
their contour, are able to associate to double helices, and conse-
quently, less double helices can aggregate to the three dimensional
network, resulting in a lower elasticity and stability. Kasapis et al.
demonstrated as well a drop in elasticity with sugar concentrations
above 40% w/w, indicating a disaggregation of the polysaccharide.
They explain this by the increasing shortage of water molecules,
which additionally form stable hydrogen bonds with sugar mole-
cules, and thus gradually deprive the polysaccharides of a hydration
layer, which is required for the thermodynamic stability of inter-
molecular helices and the subsequent building of a network
(Kasapis et al., 2003).
3.1.2. Alginate þ sucrose or trehalose
For alginate solutions on the contrary, the addition of sucrose or
trehalose increases the elastic modulus, even above that of the
aqueous reference system with correspondingly reduced amount of
water (1.25% or 1.7%) (Fig. 1b). Thus the added sugar molecules
seem to stabilize the system of intersecting and entangled polymer
coils additionally. Both sugar types as well as alginate possess hy-
droxyl groups and are able to form hydrogen bonds, which, ac-
cording to recent computations, contribute energy of about 20 kJ/
mol respectively 8 to 10 kBT (Wendler, Thar, Zahn, & Kirchner,
2010). Thus, sucrose or trehalose molecules may act as “linker”
via hydrogen bonds between distant alginate polymer chains and
new junction zones can be formed, which increase the moduli (and
viscosity). In alginate solutions the sucrose or trehalose molecules
attach between the flexible alginate polymer chains and form
hydrogen bonds with the hydroxyl groups of the alginate mole-
cules. Thereby the average distance between the chains is reduced
and the density of entanglements in the solution increases (Fig. 2b).
The elastic character of the alginate solution increases and more
energy can be stored during deformation.
3.1.3. Xanthan þ sucrose or trehalose
The addition of sucrose or trehalose to the 1% xanthan disper-
sion increases the elastic modulus of the hydrocolloid system
slightly due to the increased effective polysaccharide concentra-
tion. But the comparison with the 1.25% or 1.7% concentrated so-
lutions shows that the ability to store energy during deformation is
strongly reduced by the addition of sucrose or trehalose (Fig. 1c).
This behavior can be explained by a hindrance of the jamming
transition in the xanthan dispersion. The hydrated sucrose or
 N. Russ et al. / Food Hydrocolloids 41 (2014) 44e52 47

Fig. 1. Average of the elastic plateau modulus G0 at a constant frequency f ¼ 1 Hz, temperature T ¼ 25  C and strain g for a) agarose g ¼ 0.01, b) alginate g ¼ 0.1, c) xanthan g ¼ 0.01,
d) agaroseealginate g ¼ 0.01 and e) agaroseexanthan g ¼ 0.01. In each case we compared the aqueous systems with 1.25% or 1.7% w/w and 1% w/w hydrocolloid concentration to
the 1% hydrocolloid system including 20% or 40% w/w sugar. The measurements of the hydrocolloid mixtures with 40% w/w trehalose content could not be interpreted, because
after gelation, trehalose was crystalized.

trehalose molecules accumulate between the hydrocolloids and
enlarge the distance between the rod-like and stiff xanthan mole-
cules. Thereby the electrostatic repulsion of the single rods is
reduced and the mobility of the molecules increases (Fig. 2c). The
system of ordered and rigid rods seems to be destabilized by the
addition of sucrose or trehalose; in consequence the elastic
modulus decreases. The dynamics of the xanthan molecules are
slow due to their high molecular weight; in addition the random
orientation of the stiff xanthan chains in the log jam phase has very
slow dynamics. Compared to the diffusion of (free) sugar molecules
and water, they can be considered as quasi fixed, which defines a
clear separation of time scales. Low molecular weight sugars may
arrange with single xanthan rods, but, in contrast to alginate, they
will not be able to act as intermolecular linkers at arbitrary
48 N. Russ et al. / Food Hydrocolloids 41 (2014) 44e52

Fig. 2. Schematic illustration to show the impact of the sugar molecules in the hydrocolloid solution of a) agarose, b) alginate, c) xanthan d) agaroseealginate mixture and e)
agaroseexanthan mixture. Hexagonal symbols represent the disaccharide molecules (sucrose or trehalose), thin lines and helices the agarose and thick lines the alginate polymers.
In the agarose solution, the sugar molecules hinder the diffusion of polymer chains and double helices. In the alginate solution, the sugar molecules act as linker between the
polymer chains, and in the xanthan solution the sugar molecules reduce the electrostatic repulsion. In the agaroseealginate mixture, the mobility of the agarose polymers is limited
by the less flexible alginate coils additionally. For agaroseexanthan mixtures, free sugar molecules as well as xanthan rods hinder the agarose network formation.

positions between the xanthan rods. The energy gain provided by
hydrogen bonding appears much lower than the electrostatic en-
ergy of the randomly oriented log jammed xanthan ensemble and
the quasi-free motion of the low molecular components.
3.1.4. Agaroseealginate þ sucrose
The mixtures agaroseealginate and agaroseexanthan with the
addition of 40% trehalose could not be measured reasonably,
because trehalose was crystallized after the mixture gelled. Here,
the competition for water molecules is strongly enlarged and the
solubility limit of trehalose is exceeded. In the agaroseealginate
mixture the elastic modulus of the higher concentrated gel is
considerably higher than that of the system with added sugar
(Fig. 1d). That means the addition of 20% or 40% sugar decreases the
elasticity of the resulting gel. The elasticity of the pure agarose
system decreases only about 43% by the addition of 40% sucrose,
whereas in the mixture with alginate the elasticity decreases about
more than 56%. Fig. 1b already demonstrated that the elasticity of
the alginate solution increases by the addition of sugar. The added
sucrose molecules enhance the interactions between the single
alginate polymer chains, resulting in more elastic but less flexible
interpenetrating polymer coils. Thus, the addition of high amounts
of sugar to the agaroseealginate mixture leads to the presence of
less flexible alginate coils, and therefore an increased hindrance of
the mobility and diffusion of the agarose polymer chains compared
to the pure agaroseealginate system. Consequently, a less elastic
and stable agarose network forms (Fig. 2d).
3.1.5. Agaroseexanthan þ sucrose
The addition of 20% sucrose or trehalose increases the elastic
modulus of the system containing agarose and xanthan signifi-
cantly, whereas the addition of 40% sucrose shows only a very slight
increase. The elastic modulus with the gels prepared with 20 or 40%
sugar solutions is significantly reduced compared to the corre-
sponding system with higher polymer concentration. Compared to
the 1.7% concentrated composite gel, G0 of the 40% sucrose con-
taining gel decreases over 83% (Fig. 1e), for 20% sucrose almost 60%.
Thus, the addition of sucrose molecules strongly decreases the
elastic modulus of the agaroseexanthan mixture. The xanthan
molecules as well as the sucrose molecules hinder the mobility and
diffusion of the agarose polymer chains and the association to the
double helices, followed by the aggregation to the three-
dimensional network, is strongly restricted (Fig. 2e). Thus, the
resulting agarose network consists of less and weaker junction
zones, resulting in a less elastic and stable composite gel. The value
of G0 of the system with 40% sucrose falls even below the value of
the gel with 20% sucrose. It is not fully clear if this result is caused
only by the hindering effect of the sugar molecules described
above, or if inhomogeneities due to the very low water content in
the composite gel play a role additionally.
Comparing the different disaccharides sucrose and trehalose,
trehalose exhibits a stronger impact on the viscoelastic properties.
This effect is due to the better ability of trehalose to bind water
molecules in its hydration shell compared to sucrose. Branca et al.
(2001) identified a stronger interaction between trehalose and
water by viscosity measurements. Dissolved trehalose molecules
have a distinctive tendency to build hydrogen bonds due to their
higher number of e-OH groups. According to Kawai et al. n(e-OH)
per saccharide is 6.3 for sucrose and 8.0 for trehalose (Kawai,
Sakurai, Inoue, Chûjô, & Kobayashi, 1992). In hydrocolloid solu-
tions containing sugar molecules, the water molecules favor the
bonding with sugar molecules from an entropic point of view,
especially with trehalose. Thus the influence of trehalose is always
larger due to entropically profitable hydrogen bonds. Fig. 3 shows
schematically the different interactions of sucrose and trehalose
with the surrounding water molecules. Regarding the molecular
structure of the two disaccharides in Fig. 3, it gets obvious that
trehalose possesses more equatorial orientated OH-groups than
sucrose. The number of water molecules in the co-sphere is in
proportion to the number of OH groups in the sugar molecules, and
the e-OH groups are able to interact with water in a manner which
forms a long-lived hydration structure, since they match the un-
perturbed water lattice (Uedaira et al., 1989). The special linkage of
the two glucose rings in the trehalose molecule provides solely
equatorial arranged OH groups, which facilitates the accommoda-
tion of solute molecules within the H-bond network due to the
small perturbation induced on surrounding water molecules and
leads to a higher hydration number for trehalose (Branca et al.,
2001; Fioretto et al., 2013), resulting in bigger hydrate shells.
Nishinari, Kohyama and Watase (Nishinari & Watase, 1992; Watase
 N. Russ et al. / Food Hydrocolloids 41 (2014) 44e52 49

Fig. 3. Schematic comparison of the hydrate shells of sucrose and trehalose. The
number of e-OH groups determines the interactions between water molecules and
sugar molecules. Trehalose provides more e-OH groups than sucrose, resulting in a
bigger hydrate shell. Equatorial and axial OH groups are marked by adjacent letters e
and a, respectively.

et al., 1992) investigated the effects of different polyols and sugars

on the solegel transition of kappa-carrageenan or agarose gels.
They suggested as well different influences related to the number of
equatorial attached OH groups in the sugar molecules.

3.2. Solegel transition

The following section discusses the temperature dependent

oscillation measurements at constant frequency and strain. Fre-
quency dependent measurements of G0 and G00 would be the most
accurate method to determine the gel point, however they are time
consuming and difficult to perform in the vicinity of the gel point
due to the time dependent nature of the gelation process. Here the
gelling temperature is determined as the crossover of G0 and G00 . For
agaroseexanthan, the system exhibits elastic character even in the
sol state, since xanthan is a weak gel itself at the applied low
amplitude and dominates the modulus in the mixed system in the
sol state with G0 > G00 . Therefore, no crossover of G0 and G00 is ex-
pected and the exact gelling point of agarose is masked. However,
tan d (Russ et al., 2013), which shows a slow decrease during
cooling, starts dropping at a much faster rate at a temperature,
where G0 and G00 show a steep increase. This temperature is
therefore assumed to indicate the gelation of the agarose network
and was taken as Tgel for the system agaroseexanthan. Fig. 4aec
show the dependence of the moduli on temperature and the cor-
responding gelling temperatures for the 1.7% agarose gel, 1.7%
agaroseealginate and 1.7% agaroseexanthan as well as for the
corresponding 1% gels with 40% sugar content.
In Table 1 the gelling temperatures for the different hydrocolloid
systems are given. Here it gets obvious that the gelling tempera-
tures of the composite gels without sugar addition are much lower
than the gelling temperature of the pure agarose gel. Due to the
lower agarose concentration in the mixtures the probability that
two agarose helices meet and aggregate is reduced and the for-
mation of the three dimensional network is decelerated
(Labropoulos et al., 2002). On the contrary, the temperature of the
solegel transition is shifted to higher values in all three systems by
the addition of sucrose and even stronger of trehalose. In the
agarose gel the addition of 40% trehalose provokes a shift of about
8.1%, in the agaroseealginate mixture of about 9.1%, and in the
mixture with xanthan even a shift of about 11.3% towards higher
temperatures is demonstrated. This increase of the gelling point Tgel Fig. 4. Logarithmic plot of G0 (black) and G00 (grey) versus temperature during cooling
for high trehalose content results from the entropic preference of at f ¼ 1 Hz and constant strain g for a) agarose/sugar g ¼ 0.01, b) agaroseealginate/
the water molecules to bind sugar, especially trehalose molecules, sugar g ¼ 0.001 and c) agaroseexanthan/sugar g ¼ 0.001.

due to their high number of equatorial OH groups. Due to the water-

trehalose bonds, at high temperatures, the surrounding free water,
50 N. Russ et al. / Food Hydrocolloids 41 (2014) 44e52

Table 1
Gelling temperatures for 1.7% agarose, 1% agarose in 40% sugar solution, 1.7%
agaroseealginate/-xanthan and 1% agaroseealginate/-xanthan in 40% sugar
solution.

Tgel ( C) 1.7% þ40% sucrose þ40% trehalose

Agarose 37.3  0.3 38.5  0.1 40.6  0.1

AgaroseeAlginate 33.7  1.0 34.2  0.1 37.1  0.1
AgaroseeXanthan 34.4  0.6 36.0  1.1 38.8  1.2

in which the agarose helices diffuse, is reduced. Thus the proba-

bility, that two or more agarose helices interact by direct aggre-
gation, is enlarged and the three-dimensional network building is
accelerated resulting in higher gelling temperatures. In the com-
posite gel with xanthan, the surrounding space of the agarose he-
lices is reduced by the rigid and ordered structure of the stiff
xanthan rods in addition, resulting in a solegel transition at higher
temperatures. Other studies (Nishinari et al., 1992; Nishinari,
Watase, Miyoshi, Takaya, & Oakenfull, 1995; Watase et al., 1992)
also demonstrated a shift in Tgel of agarose gels to higher temper-
atures with the addition of sugar. They observed a shift to higher
temperatures with increasing number of equatorial OH groups,
confirming the stronger impact of trehalose shown here.

3.3. Water holding capacity

Hydrogels are susceptible to syneresis and tend to release water

spontaneously or upon energy addition. Heat exposition forces the
bound water molecules to transfer into the gaseous state by
migrating to the surface and leaving the gels or dispersions while
drying out. The ability to retard this effect of syneresis depends on
the water holding capacity of the different hydrocolloids. To study
this, the moisture content as a function of time was measured at
two different temperatures (50  C and 80  C). The diffusion of water
molecules to the surface is a transport property and depends on the
mobility and velocity of the molecules. Two parameters were
analyzed, the residual moisture content and the kinetics of losing
water.
The residual moisture content results from the difference be-
tween the theoretical water content and the maximum value of the
measured moisture content. This residual moisture content corre-
lates with the water holding capacity of the hydrocolloids. The
higher the percentage of residual moisture, the stronger is the
binding between water molecules and disaccharides or hydrocol-
loids. Fig. 5 shows the residual moisture content in dependence of
the sugar concentration for the agarose/sugar gels, alginate/sugar
solutions, xanthan/sugar dispersions and the composite gels at an
exposition temperature of 50  C. The results of the measurements
with 80  C demonstrated the same trend and are not shown here.
All hydrocolloid systems show increasing residual moisture con-
tents with increasing sugar concentrations. This consistent rise
results from the increasing number of hydrogen bonds between
water and disaccharide molecules. In the presence of sugar, the
water molecules are bound stronger, and during heat exposition
the transition of the water molecules from liquid to gas is retarded.
Thus, the water binding is elevated and the removal of water from
the system gets more and more difficult, resulting in a higher
amount of residual moisture content after drying at a given tem-
perature. For higher sugar concentrations, 40 and 50% w/w, an
obvious discrepancy between sucrose and trehalose is observable.
As mentioned earlier, due to the sterical position of its hydroxyl
groups, trehalose has a distinctive ability to bind water molecules
(Fig. 3). Therefore trehalose forms larger hydration shells and the
release of water molecules requires more energy. Interestingly
alginate and xanthan show a nearly similar exponential-like
Fig. 5. Residual moisture content as a function of the sugar concentration at constant
heat exposition T ¼ 50  C for a) agarose, alginate and xanthan and b) agaroseealginate
and agaroseexanthan. The filled symbols mark the addition of sucrose and the open
symbols that of trehalose.
increase in the residual moisture content with increasing sugar
concentration. Up to 30% sugar content the residual moisture
content increases only slowly but reaches values comparable to
agarose for 40% and 50% trehalose. To improve the water holding
capacity of the thickeners significantly, at least a concentration of
40% w/w is needed. In contrast, agarose shows an improvement in
water holding, even for smaller sugar concentrations. In the agarose
system, the sugar molecules as well as the water molecules are
confined in the network, in comparison to the nongelling agents
where the sugar molecules are more mobile. In the agarose
network, the water molecules are strongly fixed at the sugar mol-
ecules enclosed in the network and during evaporation, the agarose
network is shrinking, and thereby confining the sugar even more. A
diffusion of the water molecules bound to the sugar is thus hin-
dered, leading to a higher residual moisture content for the agarose
systems even for low concentrations of sugar. On the contrary, for
the nongelling hydrocolloids, these are diffusing freely and thus the
water molecules can reach the surface and evaporate easier. Thus,
in the agarose gel the sucrose and trehalose molecules can
contribute more to the water binding and the addition of only 10%
sucrose or trehalose increases the water holding capacity of the
 N. Russ et al. / Food Hydrocolloids 41 (2014) 44e52
agarose gels. The mixtures of gelling and thickening agents show a
similar trend as the pure agarose system, because there the agarose
network has the same restricting effect as in pure agarose. The
residual moisture content increases nearly linear with increasing
sugar concentration. No significant differences between the thick-
eners alginate and xanthan whether alone or in mixtures, can be
recognized. However, at high sugar contents an obvious difference
between the disaccharides gets visible, which can again be attrib-
uted to the different water binding abilities of sucrose and
trehalose.
To analyze the kinetics of the water loss, the measured water
content was plotted against time and an empirical exponential fit
function (y ¼ A1 expðt=t1 Þ þ y0 ) was applied to the beginning of
the measurement curve. To compare the fits, the slope at t ¼ 0 (A1/
t1) was calculated. This initial slope describes the velocity of water
evaporation at start of exposition, i.e. at a short time scale. It can
thus be associated with the free (unbound) water, which evapo-
rates already at short times. The higher values the slope reaches,
the faster moisture is released and the worse is the water holding
capacity of the hydrocolloid system. Fig. 6 demonstrates the
dependence of this slope on the sugar concentration at T ¼ 50  C
and T ¼ 80  C. A distinct difference between the hydrogels and the
hydrocolloid dispersions or pure disaccharide solutions in Fig. 6a
can be observed. The agarose containing gels consist of a three-
dimensional network with meshes filled with water molecules.
Due to this confinement, the mobility of the water molecules is
additionally hindered and the diffusion of the molecules to the
surface is retarded. The interactions in the alginate and xanthan
Fig. 6. Initial slope of the drying curve versus sugar concentration at constant tem-
perature a) T ¼ 50  C and b) T ¼ 80  C. The filled and open symbols mark the addition
of sucrose and trehalose, respectively.
51
dispersions are mainly based on hydrogen bonds and reach much
lower strength than in the agarose gels as deducible from the lower
values of the slopes. This low water holding capacity is additionally
demonstrated by the comparison with pure sugar solutions, which
show only slightly lower values. The difference between alginate
and xanthan gets more distinctive at the measuring temperature of
80  C. The alginate/sugar dispersions reach smaller slopes than the
xanthan/sugar dispersions, indicating a better water holding ca-
pacity of alginate compared to xanthan. The formation of hydrogen
bonds between hydrocolloids and water molecules is always
accompanied by a loss of entropy. The water molecules entropically
prefer the binding with the flexible alginate chains (see above).
Here, the hydroxyl groups are randomly distributed along the chain
and the water molecules are not forced into a certain orientation.
Regarding the stiff and rigid xanthan rods, the positions of the
hydroxyl groups are fixed due to the ordered structure in the sys-
tem. The orientation of hydrogen bonds is strongly forced into the
given direction of the xanthan rods in the jammed configuration
which demands a larger entropy loss. The binding of water mole-
cules with alginate molecules is more favored than the binding
with xanthan molecules, and results in a better water holding
ability of alginate. Remarkable is the independence of the slope
from the sugar concentration. All hydrocolloid systems as well as
the disaccharide solutions show an almost equal slope. At the
beginning of the measurement (short times), first the water mol-
ecules, which are bound via longerange interactions to the hy-
drocolloids, are evaporated. This does not depend on the amount of
sugar molecules. At later stages of the measurement, the amount of
short-range bounded water molecules depends strongly on the
local interactions, such as polarity, ionic strength and specific sugar
concentration. Thus, the value of the residual moisture content
varies with increasing sugar concentration at a long measurement
time.
4. Conclusion
This study examines the impact of the two disaccharides su-
crose and trehalose on the thermal and mechanical properties of
the non-gelling hydrocolloids alginate and xanthan as well as of
the gelling agent agarose and their mixtures agaroseealginate
and agaroseexanthan. Amplitude and temperature dependent
rheological measurements yield information about the visco-
elastic properties and the different changes in the hydrocolloid
systems by the addition of high amounts of sugar. For all
hydrogels, pure agarose gels as well as the mixtures, the gelling
point is shifted to higher temperatures by the addition of sugar.
This results from the competition of hydrocolloids and sugar for
free water leading to an effectively increased hydrocolloid con-
centration, and thus an increased probability for gel formation.
Trehalose always shows a stronger effect, due to its higher ability
to form hydrogen bonds and bind water. The water holding ca-
pacity at long and short time scales is influenced by the different
natures of the hydrocolloids and the molecular structure of the
disaccharides. Agarose has a better water holding capacity at
short-time scales due to its ability to confine the water molecules
in its network and thus hinder their diffusion. The residual
moisture content of agarose containing systems is increased by
the addition of small amounts of sugar molecules, which get
confined in the network during drying and hampering the
diffusion of water to the surface. In general, this study demon-
strates that hydrogels consisting of an agarose network can be
modified by controlled addition of sugars. Material specific
properties like viscoelasticity, texture, water holding capacity,
gelling temperatures and thermal stability can be adapted and
the scope of application enlarged.
52 N. Russ et al. / Food Hydrocolloids 41 (2014) 44e52
Acknowledgment
The authors would like to thank the MPIP soft matter food sci-
ence group, especially Sania Maurer and Gustav Waschatko, for
always fruitful and helpful discussions. Special thanks to Andreas
Hanewald for the technical support with the rheometer and Doris
Kirsch for the linguistic refinement.
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