Eur Food Res Technol (2009) 229:281286
DOI 10.1007/s00217-009-1056-6
ORIGINAL PAPER
Rheological properties of whey protein isolate stabilized emulsions
with pectin and guar gum
Emine Alben Erelebi Esra Ibanoflu
Received: 7 January 2009 / Revised: 6 March 2009 / Accepted: 9 March 2009 / Published online: 28 March 2009
Springer-Verlag 2009
Abstract Dynamic oscillatory and steady-shear rheologi-
cal tests were carried out to evaluate the rheological proper-
ties of whey protein isolate (WPI) stabilized emulsions with
and without hydrocolloids (pectin and guar gum) at pH 7.0.
Viscosity and also consistency index of emulsions
increased with hydrocolloid concentration. At
= 20 s1,
the value of viscosity of the emulsion with 0.5% (w/v) pec-
tin was about Wvefold higher than that of the emulsion with-
out pectin. Flow curves were analyzed using power law
model through a Wtting procedure. Flow behaviour index of
all emulsions except for containing 0.5% (w/v) guar gum
was approximately in the range of 0.91.0, which corre-
sponds to near-Newtonian behaviour. The shear thinning
behaviour of emulsions containing 0.5% (w/w) guar gum
was conWrmed by Xow behaviour index, n, of 0.396. Both
storage (G) and loss modulus (G) increased with an
increase in frequency. Emulsions behaved like a liquid with
G > G at lower frequencies; and like an elastic solid with
G > G at higher frequencies. EVect of guar gum was more
pronounced on dynamic properties. Phase angle values
decreased from 89 to <10 with increasing frequency and
indicated the viscoelasticity of WPI-stabilized emulsions
with and without pectin/guar gum.
Keywords Whey protein isolate Pectin Guar gum
Rheology Power law model
E. A. Erelebi (&) E. Ibanoflu
Department of Food Engineering,
Gaziantep University, 27310 Gaziantep, Turkey
e-mail: alben@gantep.edu.tr
Introduction
Whey proteins are widely used in food formulation to con-
trol texture and improve nutritional and functional proper-
ties [13]. The principal components of whey protein are
-lactoglobulin, -lactalbumin, bovine serum albumin (BSA)
and immunoglobulins [4]. Whey protein dispersions are
generally used to stabilize oil-in-water (O/W) emulsions.
O/W emulsions occur in many industrial processes and are
the basis of many food products [5]. Kinetically stable
emulsions can be formed by using emulsiWers and/or thick-
ening agents. EmulsiWers are mainly surface-active proteins
which can adsorbed on the droplet surfaces lowering sur-
face tension and preventing the droplets from coming close
enough together to aggregate. Thickening agents are mainly
polysaccharides used to increase viscosity of the continu-
ous phase and enhance the emulsion stability by retarding
the droplets movement. The main stabilizers used are xan-
than gum, galactomannans, intact or modiWed starches, pro-
pylene glycol alginate, pectin and carboxymethyl cellulose
[6]. The combination of protein and polysaccharide delivers
a range of properties to emulsions: physicochemical stabil-
ity, storage stability, rheology, texture and mouthfeel.
In recent years, the rheological properties of emulsions
have been investigated from both the theoretical and practi-
cal standpoint [68]. The rheological properties of Xuid food
should be carefully taken into account for designing and
modelling purposes [9], food product development, quality
control, sensory evaluation [10], consumer acceptability and
long-term stability purposes [11]. The huge number of struc-
tural parameters that inXuence emulsion stability also aVect
rheology. Thus, during the last years, a large number of
researchers have addressed a wide range of topics related to
emulsion rheology [7, 12, 13], but relatively limited studies
have been performed on comparison of the inXuence of
123
282
ionic and neutral hydrocolloids on emulsion rheology. Gen-
erally, steady state Xow measurements and oscillatory
(dynamic) measurements are used for the characterization of
food emulsions [7]. Steady state Xow measurements are
used for characterizing the shear thinning nature of food sta-
biliser [14]. Oscillatory experiments are used to obtain infor-
mation on the viscoelastic and gel characteristics of foods.
Concerning emulsion rheological behaviour, it ranges
between Newtonian and pseudoplastic [13]. Emulsion
formed of milk protein and soya oil showed Newtonian
behaviour, but kappa carrageenan addition resulted in vis-
cosity increase and clear pseudoplastic behaviour, indicating
reversible Xocculation of oil droplets. Protein emulsions
containing xanthan gum and gum arabic showed pseudo-
plastic behaviour [15], which approached Newtonian char-
acter as the concentration of gum increased [16]. Generally,
gums and stabilizers have non-Newtonian rheology and they
impart non-Newtonian character to the emulsion even when
the amount of the dispersed phase is low [13]. Tavares and
Lopes de Silva [3] reported that the presence of galactoman-
nan had a negative eVect on the network development of
galactomannan-WPI systems, increasing the temperature of
gelation and the viscous character of the system, and
decreasing the storage modulus (G).
The objectives of the present work were (1) to investi-
gate the eVects of pectin and guar gum on rheological prop-
erties of whey protein isolate (WPI)-stabilized emulsions
(2) to determine rheological parameters k and n, which are
related to consistency and Xow index, respectively, by
using power law model.
Materials and methods
Materials
Whey protein isolate was kindly provided from Davisco
Foods International (BIPRO, Lot No. JE 030-3-420, Le
Sueur, MN). The manufacturers analysis of WPI was
98.0% protein (dry weight basis), 0.3% fat, 1.7% ash and
4.4% moisture. Low-methoxyl pectin (methoxy content
8.9%; P-9135) and guar gum (G-4129) were purchased
from Sigma Chemical Company. Commercially available
reWned soybean oil was obtained from the local supermar-
ket and used without further puriWcation. BuVer salts,
potassium dihydrogen orthophosphate and disodium hydro-
gen orthophosphate, were purchased from Analar analytical
reagent; BDH Chemical Ltd.
Preparation of protein solutions
Potassium dihydrogen orthophosphate and disodium hydro-
gen orthophosphate were dissolved in deionized water to
123
Eur Food Res Technol (2009) 229:281286
prepare phosphate buVer solution (pH 7.0, 0.05 M). Stock
solutions of WPI were prepared at room temperature by
dissolving the powders in phosphate buVer while stirring
with magnetic stirrer for 1 h.
Preparation of polysaccharide solutions
Pectin (low methoxyl) and guar gum stock solutions were
prepared in phosphate buVer by stirring the dispersions vig-
orously for 30 min at room temperature, followed by heat-
ing at 50 C until the solution became clear.
Emulsion preparation
Oil-in-water emulsions (16 ml) were prepared with 25.0%
(v/v) soybean oil and 75.0% (v/v) of WPI solutions. The
two phases were homogenized using lab scale homogenizer
(Art-Miccra D-8, Germany) at 33,000 rpm for 5 min to
obtain a Wnal protein concentration of 0.5% (w/v). Emul-
sions were also prepared in the presence of 0.1 or 0.5% (w/
v) pectin/guar gum. All emulsions were prepared immedi-
ately before being loaded into the rheometer.
Rheological measurements
Rheological measurements were performed at 25
0.01 C, with a RheoStress RS-1 controlled stress rheome-
ter (HAAKE, Karlsruhe, Germany), using a cone and plate
geometry (cone diameter 35 mm, angle 2, gap 0.108 mm).
The plate was equipped with a Peltier temperature control
that allows rapid temperature control. The slippage at the
sample/plate interface was negligible. For each measure-
ment, 2.0 ml of emulsion were carefully deposited over the
plateau of the rheometer. After the plateau has been contact
with the cone, the exposed surface of sample was covered
with a thin layer of silicone oil to prevent evaporation dur-
ing the measurement. All samples were allowed at rest for
5 min after loading to allow temperature equilibration and
induced stress to relax. Stress sweep tests (1 Hz at 25 C)
were made to determine the linear viscoelastic region of all
samples; a stress value of 1 Pa was chosen for all the
frequency tests. Oscillatory (dynamic) tests were performed
inside the linear viscoelastic region and the storage modu-
lus (G) and loss (G) modulus were recorded versus
frequency. Frequency sweeps were done from 0.001 to
10 Hz at 1 Pa and at 25 C, but in most cases some results
had to be discarded due to equipment limitation at low
frequencies. Data analysis software (Reowin Pro Data
Manager Version 2.64) was used to obtain the experimental
data (G, G, tan , etc.).
Steady state Xow measurements were carried out in the
range of 0200 s1 during 5 min and rheological parame-
ters (shear stress, shear rate, apparent viscosity) were
Eur Food Res Technol (2009) 229:281286 283

obtained from the software. Experimental Xow curves were 10

Wtted to the power law model ( = k
n) using Sigma Plot 0.0%
0.1%
2000 software, where k (Pa sn) is the consistency index and 0.5%
n (dimensionless) is the Xow behaviour index. 1

Viscosity (Pa.s)
Statistical analyses
0.1

Each experiment was replicated twice using freshly pre-

pared samples and the values reported are means of these
0.01
repetitions. A one-way analysis of variance (ANOVA) and
general linear model with LSD (least signiWcant diVerence)
test were used to establish the signiWcance diVerences
0.001
among the mean values at the 0.05 signiWcance level. The 1 10 100
statistical analyses were performed using SPSS 8.0 system Shear rate (s-1)
software. Fig. 1 EVect of pectin concentration on the viscosity of 0.5% (w/v)
WPI-stabilized emulsions with 25% (v/v) soybean oil at 25 C, pH 7.0

Results and discussion

100

Steady-state measurements 0.0%

0.1%
0.5%
Power law model
Figure 1 shows the eVect of pectin concentration on viscos-
Shear stress (Pa)

ity of WPI-stabilized emulsions at 25 C, pH 7.0. The vis-
cosity of the emulsions increased with the addition of
pectin. A signiWcant increase (P < 0.05) in the viscosity
was observed when the amount of pectin was increased. At

= 20 s1, the value of viscosity of the emulsion with 0.5%
(w/v) pectin was about Wvefold higher than that of the
emulsion without pectin. This increase in the viscosity can
be explained by the occurrence of phase separation at
microscopic level. In fact, at pH 7.0 both pectin and WPI
molecules carry net negative charges and there is no com-
plexation between these macromolecules. When the pectin
concentration in the mixture is above a critical concentra-
tion, a depletion eVect leads to an attractive force between
the protein aggregates and promotes the formation of large
microgel particles of protein, thereby causing phase separa-
tion, and increasing the viscosity of the system [17]. As
shown from Fig. 1, viscosity of emulsions decreased with
increasing shear rate. As the shear rate suYciently increases
to overcome the Brownian motion, the emulsion droplets
become more ordered along the Xow Weld and oVer less
resistance to Xow and hence the lower viscosity [5].
Figure 2 shows the Xow curves of WPI-stabilized emul-
sions and the Wts of power law model at diVerent pectin
concentrations. The shear stress showed a practically linear
dependence on shear rate. Flow curves were modelled with
the power law and the parameters were given in Table 1.
n < 1.0 corresponds to shear thinning behaviour, n > 1.0
corresponds to shear thickening behaviour and n = 1.0 cor-
responds to Newtonian behaviour [18]. The emulsions with
and without pectin behaved like a near-Newtonian Xuid.
Addition of pectin had no signiWcant eVect on Xow type.
10
1
0.1
1 10 100
Shear rate (s-1)
Fig. 2 Flow curves of 0.5% (w/v) WPI-stabilized emulsions with 25%
(v/v) soybean oil at diVerent pectin concentrations
Flow behaviour index of emulsions was approximately in
the range of 0.91.0 both in the absence and presence of
pectin, which corresponds to near-Newtonian behaviour.
The consistency index is an indicator of the viscous nature
of the emulsion and was observed to be increasing with
pectin concentration from 0.035 to 0.147 Pa sn (R2 > 0.980)
and its trend is in agreement with that of viscosity versus
shear rate data in Fig. 1.
EVect of guar gum concentration on viscosity of WPI-
stabilized emulsions was shown on Fig. 3. In the presence
of low amount of guar gum (0.1% w/v), the major observa-
tion was similar to that of the WPI-stabilized emulsions in
the presence of pectin, whereby the viscosity of the emul-
sions gradually increased with guar gum addition from
0.033 to 0.053 Pa s at
= 22 s1. A signiWcant increase in
the viscosity was observed when guar gum concentration
was 0.5% (w/v), and the viscosity of the emulsion with

123
284 Eur Food Res Technol (2009) 229:281286

Table 1 Rheological parameters from power law model for WPI- 100
pectin/guar gum stabilized emulsions
Hydrocolloid Pectin Guar gum
conc. (w/v %) 10
n 2
n k (Pa s ) R n k (Pa sn) R2

Shear stress (Pa)

0.0 0.876 0.035 0.980 0.876 0.035 0.980
0.1 0.973 0.067 0.999 0.984 0.047 0.998 1

0.5 1.000 0.147 0.999 0.396 5.382 0.994 0.0%

0.1%
0.1 0.5%
Power law model
10

0.0%
0.1% 0.01
1 10 100
0.5%
Shear rate (s-1)
Viscosity (Pa.s)

1
Fig. 4 Flow curves of 0.5% (w/v) WPI-stabilized emulsions 25%
(v/v) soybean oil at diVerent guar gum concentrations

0.1
general tendency of both the consistency index and the Xow
behaviour index was mainly determined by the hydrocol-
loid concentration: increasing guar gum concentrations
largely increased the viscosity and hence the consistency
0.01
1 10 100 index of emulsions.
Shear rate (s-1)

Fig. 3 EVect of guar gum concentration on the viscosity of 0.5%
(w/v) WPI-stabilized emulsions with 25% (v/v) soybean oil at 25 C,
pH 7.0
0.5% (w/v) was 15-fold higher than that of the emulsion
with 0.1% (w/v) guar gum.
Flow curves of WPI-stabilized emulsions at diVerent
guar gum concentrations were also Wtted to power law
model (Fig. 4) and power law parameters were shown on
Table 1. Power law parameters showed that the emulsions
without guar gum and with 0.1% (w/v) guar gum behaved
like a near-Newtonian Xuid since Xow behaviour index was
approximately 0.91.0. In the presence of 0.5% (w/v) guar
gum, shear stress versus shear rate plot deviated from line-
arity and these emulsions showed shear-thinning behaviour.
Guar gum is known to exhibit pseudoplastic behaviour and
hence induces a shear-thinning behaviour, as evidenced by
the values of the Xow behaviour index, n [0.396 in the pres-
ence of 0.5% (w/v) guar gum]: whereas this parameter
approximated unity in the absence of guar, a progressively
smaller value was obtained at increasing guar gum concen-
trations [19]. The lower the value of n the more shear thin-
ning behaviour the emulsion exhibits. Shear thinning
character is associated with the Xocculation of fat droplets.
This is usually the case with weakly Xocculated emulsions
or those to which a thickener is added [20]. Flocculation
increases the apparent dispersed phase volume and leads to
the formation of nonspherical aggregates, both factors
contributing to increase the emulsion consistency [1]. The
Oscillatory measurements
Dynamic frequency sweep tests were performed in the lin-
ear viscoelastic range to determine the frequency depen-
dence of the storage modulus (G) and loss modulus (G).
Oscillatory rheological measurements of storage and loss
modulus indicate whether the emulsion system is strongly
or weakly Xocculated [11]. In this later category the system
is essentially liquid-like. Figure 5 illustrates the mechanical
spectra describing the viscoelastic behaviour of WPI-stabi-
lized emulsions at diVerent pectin concentrations. Magni-
tudes of both G and G increased with frequency and they
were almost frequency dependent. At all pectin concentra-
tions studied, emulsions exhibited a liquid-like behaviour at
low frequencies where loss modulus (G) was higher than
storage modulus (G). This behaviour corresponds to the
characteristics of viscoelastic Xuids according SteVe [21].
When G is much higher than G at low frequencies, the
energy used to deform the material is dissipated viscously
and the materials behaviour is liquid-like [22]. At higher
frequencies G and G crossed-over and at frequencies
approximately above 7.0 Hz, G became greater than G
and the system showed a solid-like behaviour. This was
also conWrmed by phase angle (tan ) values. Phase angle
value (tan  = G/G) can provide information about the
nature of the viscoelastic response of the emulsion system
and measures the energy loss compared to energy stored in
cyclic deformation. A phase angle of 90 indicates the material
is fully viscous while an elastic material is characterized by

123
Eur Food Res Technol (2009) 229:281286
100
0.0%
0.1%
10 0.5%
G', G'' (Pa)
1 1
0.1
0.01
0.01 0.1 1 10
Frequency (Hz)
Fig. 5 Mechanical spectra of WPI-stabilized emulsions at diVerent
pectin concentrations (G solid symbols, G open symbols)
phase angle value approaches to 0 [23]. At lower frequen-
cies, phase angle values of WPI-stabilized emulsions were
around 89 at all pectin concentrations and the system
showed a liquid-like response. This value reduced with
increasing frequency and approached to less 10 and sup-
ported the viscoelasticity of these WPI-stabilized emul-
sions.
Dynamic rheological tests of WPI-stabilized emulsions
at diVerent guar gum concentrations showed liquid-like
character at lower frequencies and G and G crossing each
other at high frequencies and showed solid-like character
(Fig. 6). Both G and G were frequency dependent and
increased with the addition of guar gum. A gradual change
on the relative values of G and G was observed as the
guar gum concentration was increased. This eVect was
more pronounced in the presence of 0.5% (w/v) guar gum.
The addition of guar gum caused the formation of much
stronger (more elastic) structure. This stronger structure in
the presence of polysaccharide can be attributed either to
the formation of a highly Xocculated droplet network struc-
ture that gives this pronounced viscoelastic behaviour or to
the formation of a gel-like structure in the continuous phase
by the polysaccharide itself [7]. It could also be resulted
from changes in the viscoelastic proWle which mean that the
macromolecular mobility within the network was decreased
due to the presence of the neutral polysaccharide [3]. Phase
angle values of WPI-stabilized emulsions in the absence
and presence guar gum were decreased with increasing fre-
quency and found to changing between 89 and 10 within
studied frequency range. This results indicated a viscous
behaviour at low frequencies and a viscoelastic behaviour
with increasing frequencies. We concluded that WPI-stabi-
lized emulsions with and without guar gum had a viscoelas-
tic property and it could be characterized as weak gels as a
typical behaviour of dressings and emulsions as reported
previously [13].
285
100
0.0%
0.1%
10 0.5%
G', G'' (Pa)
0.1
0.01
0.01 0.1 1 10
Frequency (Hz)
Fig. 6 Mechanical spectra of WPI-stabilized emulsions at diVerent
guar gum concentrations (G solid symbols, G open symbols)
Conclusion
Dynamic rheological properties and Xow behaviour of
WPI-stabilized emulsions with and without pectin/guar
gum were investigated. All emulsions except for containing
0.5% (w/v) guar gum showed near-Newtonian Xuid behav-
iour. With the addition of 0.5% (w/v) guar gum, emulsions
showed shear thinning behaviour. Increasing hydrocolloid
concentration increased both viscosity and consistency
index. So, Xow properties of the emulsions could be con-
trolled by appropriate manipulation of composition. Power
law model provided a good Wtting (R2 > 0.980) for experi-
mental data. Both storage modulus and loss modulus were
frequency dependent. Dynamic rheological tests also dem-
onstrated that both WPI-pectin and WPI-guar gum stabi-
lized emulsions behaved like a liquid at low frequencies
and like a solid at higher frequencies. So we concluded that
emulsions showed a viscoelastic behaviour. This was also
conWrmed by phase angle values.
Acknowledgments We thank The ScientiWc and Technological Re-
search Council of Turkey (TUBITAK) for Wnancial support. We also
thank Davisco Foods International for providing the WPI sample.
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