doi: 10.1111/1471-0307.

12371

ORIGINAL
RESEARCH Effect of whey protein concentrate, pH and salt on
colloidal stability of acid dairy drink (Doogh)
Z A H R A A Z A R Y and A L I N A S I R P O U R *
Department of Food Science and Technology, College of Agriculture, Isfahan University of Technology, Esfahan
84156-83111, Iran

Serum separation of Doogh, an Iranian yoghurt drink, is a major problem. In this study, the effects
of whey protein concentrate (WPC) addition, pH of the product and dissolved salt (sodium chloride)
in milk on the colloidal stability of Doogh were investigated. By increasing the amount of WPC
(from 0.5 to 3%) and salt in milk (from 0 to 1.6%), the serum separation decreased. Increasing pH
values of sample (from 3.5 to 4.5), WPC (from 0.5 to 4%) and salt concentration in milk (from 0 to
2%) also increased the samples’ viscosity. All samples showed Newtonian behaviour except samples
at pH 4.5, containing 4% WPC and 2% dissolved salt in milk.
Keywords Acidified milk drink, Doogh, Colloidal stability, Serum separation, Whey protein
concentrate, Salt.

conformation of j-casein and decreases steric

INTRODUCTION
Doogh is a traditional and nutritious fermented
milk drink produced in Iran and some parts of
the Middle East. This drink is manufactured by
mixing set or stirred yoghurt with water at a
level of 50–60% and salt at a level of 0.2–1%
and it is flavoured with some aqueous extracts
of local herbs (Azarikia and Abbasi 2010; Ghor-
bani Gorji et al. 2011). The pH of this product
is lower than 4.5. Serum separation of Doogh
during shelf life is the main obstacle in the way
of its overall consumption. In unheated milk at
the natural pH (approximately 6.6), j-casein is
on the surface of casein micelles, with the gly-
cosylated C-terminal sequences protruding to
form a negatively charged polyelectrolytic
‘hairy’ layer on the casein micelle surface
(Anema et al. 2004b). These micelles are sta-
bilised by steric and electrostatic repulsion due
to the extended conformation of j-casein
(Tromp et al. 2004; Du et al. 2009). In addition,
colloidal calcium phosphate (CCP), which is
mainly linked to as1, as2 and b-caseins,
improves the stability of casein micelles. It has
*Author for positive charges and crosslinks the negative sites
correspondence. E-mail: of casein micelles. Decreasing the pH of milk
ali.nasirpour@cc.iut.ac.ir
neutralises the carboxylic acid groups and
© 2017 Society of charged side chains of the amino acids on the
Dairy Technology glyco macropeptide hairs, collapses the extended
and electrostatic repulsion (Walstra 1990). Also
colloidal calcium phosphate and the small
amounts of magnesium and citrate associated
with casein micelles are dissolved and caseins
are dissociated into a diffusible fraction of milk.
It is generally admitted that in milk at pH 5.2
calcium that associated with inorganic phosphate
and the total of inorganic phosphate are solu-
bilised. Calcium is totally solubilised only at pH
3.5 (Anema et al. 2004b; Dalgleish et al. 2005;
Gaucheron 2005). Therefore, decreasing the pH
of milk to lower than 4.5 leads to the formation
of casein micelle aggregates.
In the commercial production of yoghurt, the
milk is pasteurised at 90 °C for 10 min. At tem-
peratures above 70 °C, denatured b-lactoglobu-
lin interacts with j-casein via thiol-disulphide
reactions (Donato et al. 2007) and because of
that isoelectric point of b-lactoglobulin is higher
than isoelectric point of the casein micelles, acid
milk gel forms in higher gelation pH compared
with acid-induced gel of unheated milk (Anema
et al. 2004a). Also by adding water to yoghurt
for the production of Doogh causes serum sepa-
ration under gravity. This phenomenon leads to
the separation of this drink into a casein-rich
lower layer and an upper layer of clear ‘serum’,
which can often occupy more than half of the
total volume. (Kiani et al. 2010).

198 Vol 71, No 1 February 2018 International Journal of Dairy Technology

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Hydrocolloids are common additives used to retard separa- addition and dissolved salt in milk on the colloidal stabil-
tion in acidic milk drinks. It was shown that both adsorbing ity of the product.
and nonadsorbing hydrocolloids are capable of stabilising the
system. Adsorbing hydrocolloids such as high methoxy pec-
MATERIALS AND METHODS
tin (Jensen et al. 2010), carboxymethylcellulose (Du et al.
2009) and soya bean soluble polysaccharides (Nobuharaa Materials
et al. 2014) are charged polysaccharides, which can link with Fresh skim milk with pH 6.6 (containing 0.05 wt % fat)
casein micelles via electrostatic interaction and stability of was obtained from the local dairy plant (Pegah Dairy Co,
system is due to electrostatic repulsion, steric repulsion or Esfahan, Iran). Starter culture (PRO-A from Proquiga, La
both. These interactions are highly dependent on the pH and Coru~na, Spain) was used for yoghurt production. WPC
ionic strength of the solution (Azarikia and Abbasi 2010). powder was purchased from Milei (Milei Co., Leutkirch,
‘Nonadsorbing’ hydrocolloids such as guar gum and locust Germany) and stored at 4 °C. Composition of the powder
been gum, by forming a network, can increase the viscosity was measured using standard methods (81.17  0.04% total
of these drinks and entrap water in a network and immobilise protein; 2.34  0.02% minerals; 7.12  0.02% fat).
the casein micelles. Therefore, colloidal stability of the pro- Sodium chloride, lactic acid and other materials used in this
duct increases (Azarikia and Abbasi 2010; Kok 2010). How- study were of analytical grade.
ever, some stabilisers might adversely influence the taste and
mouthfeel of the acidified milk drinks and decrease the Preparation of Doogh samples
acceptability (Koksoy and Kilic 2004). In this research, 18 samples of Doogh were prepared based on
WPC is used for the improvement of physical properties a three-factor D-optimal response surface methodology
of fermented milk products (Anema et al. 2004b) and (RSM) design using Design Expert 6 Software (Stat-Ease
enhances the nutritional value. On the other hand, many fac- Inc., Minneapolis, MN, USA) and are presented in Table 1.
tors affect colloidal stability of acidified milk drink such as Experimental domain was chosen based on preliminary tests.
the amount of total dry matter of product (Kiani et al. Minimum and maximum concentrations of WPC in Doogh
2009), fat content, starter type used for yoghurt production, samples were 0.5 and 4%, respectively. Also three pH levels
time and temperature of incubation (Ozdemir and Kilic 3.5, 4 and 4.5 and 5 concentrations (0%, 0.5%, 1%, 1.33%
2004), homogenisation pressure (Sedlmeyer et al. 2004), and 2%) of dissolved salt in milk were studied.
thermal processing (Sejersen et al. 2007), pH (Du et al. Figure 1 shows a process flow diagram of the production
2009) and ionic strength of the product (Jensen et al. 2010). of Doogh samples. Fresh skim milk was heated at 85 °C for
Adding NaCl to milk at the natural pH decreases the pH 15 min, and then it was cooled down to inoculation temper-
and increases Ca2+ concentrations in the serum phase due to ature (45 °C). Yoghurt starter culture was prepared by
exchanges of divalent cations or protons, which were linked
directly to phosphoseryl residues of micelles by Na+ (Awad
2007); the casein–sodium complex that is more soluble than Table 1 Statistical design for experiment
casein micelle is formed (Strange et al. 1994). In addition, Samples WPC (%) pH of the samples Dissolved salt in milk (%)
sodium ions that associate with the hairy layer of micelles,
1 0.5 4.5 0
at pH lower than 4.5, prevent adsorption of hydrogen ions
2 2.25 3.5 1
on the hairy layer and the repulsive force between micelles 3 2.83 4.5 0
increases the colloidal stability (Schkoda et al. 1999). 4 1.375 4 0.5
The changes in milk with the addition of salt at an 5 0.5 3.5 2
acidic pH are quite different from those created at a neu- 6 0.5 4.5 1.33
tral pH. Previous research shows that increasing the salt 7 4 3.5 0
concentration in the water added to yoghurt to produce 8 4 4.5 2
Ayran results in an increase in protein–protein interactions 9 2.25 4 2
and the serum separation is increased (Koksoy and Kilic 10 2.25 4.5 1
2004). In addition, by adding salt to casein micelles at pH 11 4 3.5 2
4.85 the colloidal calcium phosphate is not exchanged with 12 4 3.5 0
the sodium ion (Gaucheron et al. 2000). Increasing the salt 13 2.25 3.5 0
concentration in casein solution at pH 3–4 leads to a 14 4 4 1
decrease in solubility of caseins (Strange et al. 1994). The 15 0.5 3.5 0
16 4 3.5 2
effect of pH in the range of 3.5–4.5 and the effect of salt
17 0.5 3.5 2
addition to the milk on the colloidal stability of acidified
18 4 4.5 2
milk drinks has not yet been studied. The aim of this
research was to evaluate the effect of pH of Doogh, WPC

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Dissolving WPC in
concentration 0.5 to 4% and
Adding freeze-dried starter to the rest of salt in water
one litre of the sterilized skim
milk
Keeping overnight the WPC
solutions or WPC–salt
Pasteurization of skim milk solutions (5°C)
Incubation of milk (3 hours,
(85°C, 15 min)
45°C) for preparing starter
Denaturation of the solutions
Cooling milk (45°C)
Inoculation of 5% starter culture (85°C, 5 min)

Incubation of milk at 45°C until Cooling the denatured

Adding 0 to 2% salt to milk at
reaching pH near 3.5, 4 and 4.5 solutions (20°C)
pH 6 ± 0.05

Cooling yogurt after Keeping overnight the

fermentation (5°C) solutions (5°C)

Stirring yogurt Adding the solutions to the

stirred yogurt

Mixing the samples Doogh

Adjusting pH of the samples on

3.5, 4 and 4.5 using lactic acid

Homogenisation of the samples

(150 bar, 60°C)

Pasteurisation of the samples

(75°C, 1 min)

Cooling the samples (20°C)

Filling the samples into 500-ml

bottles, sealing and storing

Figure 1 Process flow diagram of the production of the samples.

adding freeze-dried starter to one litre of sterilised skim
milk and incubated for 3 h at 45 °C. The exact amount of
5% w/w of this starter was inoculated to pasteurised skim
milk at 45 °C and it was incubated at 45 °C until the pH
reached near the pH that is proposed in Table 1. In some
samples, the corresponding amount of formulation salt
(based on Table 1) was added to milk after incubation when
pH of milk decreased to about pH 6  0.05. If the salt was
added before inoculation, incubation time was very long to
reach final pH. After fermentation, the yoghurt was cooled
down to 5 °C.
Whey protein solutions or whey protein–salt solutions were
prepared by dissolving WPC (and salt) in water under gentle
stirring at 20 °C for 2 h and kept overnight at 5 °C. WPC
solutions were denatured on a shaking water bath (Memmert,
WNB22, Memmert GmbH, B€uchenbach, Germany) at 85 °C
for 5 min and cooled down to 20 °C and subsequently kept
overnight at 5 °C.
The Doogh samples contained 50% wt yoghurt (dry mat-
ter = 8.4  0.1), 1% wt salt, 0–1% wt lactic acid (to adjust
final pH values of samples), 0.5–4% WPC and 44–48.5%
distilled water.

200 © 2017 Society of Dairy Technology

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Stirred yoghurt was gradually added to whey protein solu-
tion or whey protein–salt solution (depending on the concen-
tration of dissolved salt in milk used for the production of the
yoghurt based on Table 1) and mixed to achieve a homoge-
nous drink. In the next stage, the pH of the samples was
adjusted using lactic acid. Samples were then homogenised at
150 bar and 60 °C (APV lab homogeniser, Albertslund, Den-
mark) and pasteurised at 75 °C for 1 min in water bath (Mem-
mert, WNB22). The samples were cooled down to 20 °C and
filled into 500-mL bottles, sealed and stored at 5 °C.
Serum separation
About 30 mL of each sample were held for 1 day at 5 °C
in graduated glass tubes of diameter 1.4 cm and height
18 cm. The height of the serum phase was measured, and
the results are presented in percentage of serum separation
(divided by the total height of sample in tube and multiplied
by 100) expressed as the serum separation in percentage.
The experiment was performed in triplicate.
Viscosity
The viscosity of Doogh samples was measured at 5 °C by a
rotational viscometer (RVDVII; Brookfield, Stoughton, MA,
USA) using spindle No. 1 at 60, 80, 100, 120, 140 rpm/
min. Before measurement, samples were shaken carefully to
have a homogenous solution.
Measurement of total solid, protein and sodium in
serum and sediment phases
Serum phase was separated from the sediment phase with a
syringe after 7 days. In both phases, total solid (TS) content
was determined by evaporation of 2-g sample in separate
dishes in a boiling bath, followed by oven-drying (UT 5050
E: Heraeus, Hanau, Germany) at 105 °C until constant
weight was attained, and then the samples were cooled in a
desiccator and reweighed. Total nitrogen (TN) and nonpro-
tein nitrogen (NPN) were measured by Kjeldahl method
(UDK 132 VELP Scientifica Kjeldahl, Usmate, Italy). Pro-
tein content of both phases was calculated by the formula
(TN NPN) 9 6.38. Sodium was measured using a flame
photometer (PFP7; Jenway, Dunmow, Essex, UK) based on
the method proposed by Hald (1946).
Statistical analysis
Experimental results were analysed using Design Expert
6.06 Trial software. An appropriate model was selected for
each parameter. The validity of the model was investigated
based on the analysis of variance technique.
RESULTS AND DISCUSSION
Serum separation of Doogh samples
The amount of serum separation of the samples can be pre-
dicted using a quadratic model. Analysis of variance of the
suggested model showed that serum separation was affected
© 2017 Society of Dairy Technology
by concentrating WPC in Doogh samples, concentration of
dissolved salt in milk and the interaction of WPC and dis-
solved salt in milk. Nevertheless, the final pH of the sam-
ples in the studied range had no significant effect
(P < 0.05) on serum separation.
Effect of WPC on serum separation of Doogh samples
Figure 2 (a) shows that with the addition of WPC up to
3.13%, serum separation of the samples decreased. During
thermal treatment of Doogh samples, the added WPC and
casein micelles interacted and whey proteins became cova-
lently bound to casein micelles (Lucey et al. 1999). Accord-
ing to Table 2, the amount of serum separation in sample 1
(the sample containing 4% WPC) was significantly lower
compared with sample 2 (the sample containing 0.5%
WPC). Although the dry material of Doogh sample 1 was
higher than that in Doogh sample 2, dry materials and pro-
tein concentrations were lower in serum phase of Doogh
sample 1. Our results show that samples containing higher
amount of WPC had lower concentration of dry materials
and proteins in their separated serum phase. Moreover, these
samples had lower serum separation. Whey proteins are sol-
uble proteins over a wide range of pH values (Onwulata
and Huth 2008). Therefore, the measured proteins in serum
phase were mainly of whey proteins. Hence, whey proteins
in sample 1 interacted with casein micelles, and therefore,
the protein concentration in serum phase of Doogh sample
was decreased. Previous studies showed that in the presence
of casein-denatured whey protein complexes in fermented
milk, the water binding capacity of the product increases
(Sedlmeyer et al. 2004). In addition, adding whey proteins
to samples may prevent the interactions between casein
micelles and creation of large aggregates at low pH. There-
fore, by increasing the WPC concentration in the samples,
more whey protein–casein interactions occurred and led to
an increase in the dispersibility and solubility of casein
micelles via increasing the soluble part of casein micelles
and increasing the steric hindrance that prevent the creation
of larger casein aggregates.
Effect of dissolved salt in milk on serum separation of
Doogh samples
A comparison of Figure 2 (a,b,c) reveals that with an
increase in salt concentration in milk, serum separation of
the samples decreased. This is because of proton
exchanges of casein micelles. In this case, sodium ions
were exchanged with calcium ions in casein micelles and
sodium–casein complexes were formed. By exchanging
sodium with calcium in casein micelles, the stability and
solubility of casein increased over a wide range of pH.
The results shown in Table 3 confirm that adding salt to
milk decreased the serum separation. On the seventh day
of storage, serum separation in sample 1 (the sample con-
taining 2% salt in milk) was lower than that in sample 2
201
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Figure 2 Effect of WPC concentration and pH on serum separation of Doogh samples (after 24 h at 4 °C) containing 0% (a), 1% (b) and 2%
(c) dissolved salt in milk.
(the sample containing 2% salt in whey protein solution,
i.e. salt was added after fermentation). To confirm the
exchange of sodium with calcium in casein micelles, the
sodium content of serum and sediment phases of samples
were measured. The results show that the sample contain-
ing 2% NaCl dissolved in milk (sample 1) contained
higher and lower sodium in the sediment and serum
phases, respectively, compared with sample 2. According
to these observations, by adding salt to milk, sodium
began to replace calcium on the casein micelles. Therefore,
the solubility of casein increased. This phenomenon could
decrease the amount of sediment in the samples that was
prepared by milk containing salt.
Figure 2 (b,c) show that half and total amount of salt was
dissolved in milk, respectively. In these figures, required
WPC to create the maximum stability in the samples was
lower than that in part A (2.25% WPC in part B and
1.375% WPC in part C). Therefore, whey proteins and dis-
solved salt in milk had a synergistic effect on serum
202
separation of the samples. According to Figure 3 (a,b), the
amount of WPC had more effect on serum separation com-
pared with the amount of dissolved salt content in milk. As
seen in Figure 3 (b) (2.25% WPC), addition of salt to milk
increased the stability of the samples, while its effect was
lower compared with Figure 3 (a) that shows the addition
of 0.5% WPC to samples.
In part C (4% WPC), increasing salt concentration in milk
from 1.5 to 2% led to an increase in serum separation. In
fact, in this condition, the ionic strength of the samples led
to a decrease in the stability of protein particles.
Effect of final pH on serum separation of Doogh
samples
The pH of samples (in the range of our experiments) had
no significant effect on serum separation. However, based
on Figure 2, in the case that samples had their maximum
stability against serum separation, adding more WPC did
not affect serum separation. In this condition, the final pH
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Table 2 Total solid and protein contents of serum and sediment phases in two samples of product

Samples Total solid (%) Protein (%)

WPC (%) pH Dissolved salt in milk (%) Serum separation (%) Serum phase Sediment phase Serum phase Sediment phase
4 3.5 0 7  0b 0.537  0.006b 7.366  0.096a 0.090  0.001b 3.46  0.214a
0.5 3.5 0 61.82  1.82a 2.245  0.105a 2.809  0.11b 0.167  0.014a 1.384  0.096b
The numbers are mean  SD of three replicate. The mean values with different letter(s) in the same column are different at a significant level of
0.05.

Table 3 Amount of total solid and sodium in serum and sediment phases in two samples of product

Samples Total solid (%) Sodium (mg in 100 g)

WPC (%) pH Dissolved salt in milk (%) Serum separation (%) Serum phase Sediment phase Serum phase Sediment phase
0.5 3.5 2 45  0 b
1.833  0.06 b
3.29  0.08 a
137.73  3.32 b
131.27  4.47a
0.5 3.5 0 61.82  1.52a 2.452  0.1a 2.809  0.1b 173.26  4.21a 106.021  3.76b
The numbers are mean  SD of three replicate. The mean values with different letter(s) in the same column are different at a significant level of
0.05.

of the samples had a significant effect on serum separa-
tion. This means by controlling the final pH, maximum
stability could be achieved. For example, in Figure 2 (a),
product stability was not affected by increasing the WPC
concentration from 3.125% to 4%. However, pH of the
samples in this range affected serum separation. In fact, in
this region, added WPC did not react with casein micelles
but they could form soluble and nonsedimentary aggre-
gates or an insoluble aggregate that led to an increase in
serum separation. The pH of the samples was very impor-
tant in the solubility of the aggregates and the casein
micelles. As the pH of the samples was increased from
3.5 to about 4.16, serum separation increased, while further
increase in pH up to 4.5 decreased serum separation. How-
ever, the effect of pH was negligible especially at pH
higher than 4.16.
Also in this condition at pH 4.5 the viscosity of samples
increased. In this case, based on Stock’s law, sedimentation
of aggregated casein micelles was retarded. Figure 3 (c) illus-
trates that at lower than 0.5% and higher than 1.5% of dis-
solved salt in milk, serum separation was not affected by pH
of Doogh samples. Serum separation was not affected by
increasing salt content from 0.5 to 1.5% in milk, whereas in
this condition, pH had an important effect on serum separa-
tion. By increasing the pH from 3.5 to about 4.2, serum sepa-
ration increased while further increase in pH up to 4.5 led to
decrease in the serum separation. However, these changes
were small.
Figure 4 shows that in all three pH values, 3.5, 4 and
4.5, the maximum serum separation corresponded to the
samples containing minimum concentration of WPC and
maximum salt dissolved in the whey protein solution.
Samples with pH 3.5, which contained 3% WPC and 1.6%
dissolved salt in milk, were most stable against serum
separation.
Viscosity of Doogh samples
According to our result, model 2FI was appropriate to deter-
mine the product viscosity. It is a linear model where the
interactions between two factors have a significant effect on
response. Analysis of variance of the suggested model
shows that WPC, dissolved salt in milk, and pH as well as
interactions of pH and WPC, and interactions of pH and
dissolved salt had significant effects on viscosity of the sam-
ples.
Effect of WPC on viscosity of Doogh samples
Figure 5 shows that by increasing the WPC content in the
samples, viscosity was increased, because by increasing the
WPC concentration, the dry material content of Doogh sam-
ples increased (Kiani et al. 2009). Moreover, more interac-
tions occurred between whey proteins and casein micelles;
therefore, the viscosity of product increased (Ozen and Kilic
2009).
Effect of dissolved salt in milk on viscosity of Doogh
samples
As shown in Figure 5, the viscosity of the samples
increased by increasing the dissolved salt in milk as a result
of hydration and swelling of casein micelles (Awad 2007).
Moreover, a comparison of Figure 5 (a,b) reveals that the
effect of added salt into milk on the viscosity of the samples
was lower than that of the WPC content of samples. The
sample containing 0.5% WPC and 2% dissolved salt in milk

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Figure 3 Effect of dissolved salt in milk and pH on serum separation of Doogh samples (after 24 h at 4 °C) containing 0.5% (a), 2.25% (b) and 4%
(c) WPC.
had the lowest viscosity. The sample with 4% WPC and 2%
dissolved salt in milk exhibited the highest viscosity value,
because the size of casein micelles, sample total solids and
interaction between whey proteins and casein micelles were
increased.
Effect of final pH on viscosity of Doogh samples
Figures 5 and 6 reveal that the viscosity of the samples was
increased by increasing the pH value especially at higher
concentrations of dissolved salt in milk (Figure 5) and
WPC (Figure 6). According to the Figure 6, the effect of
WPC on viscosity was lower than the effect of pH. There-
fore, the effect of pH on viscosity of samples was higher
than that of dissolved salt in milk and WPC concentration.
Based on the results of this research, samples with pH 3.5
and contained 0.5% WPC and 0% salt in milk and samples
with pH 4.5 and contained 4% WPC and 2% salt in milk
exhibited the minimum and maximum viscosity values,
respectively.
204
Effect of WPC, pH and dissolved salt in milk on
rheological properties of Doogh samples
Rheological properties play an important role in the product
consistency, mouthfeel and its acceptability. Doogh is
mainly a dilute suspension of protein clusters of milk
including whey proteins and casein acting as colloidal parti-
cles. Intermolecular bonds and thermodynamic properties of
protein and the interactions between particles determine the
rheological characteristic of the product (Kiani et al. 2008).
Previous studies showed that flow behaviour of Doogh
without stabiliser was Newtonian (Foroughinia et al. 2007;
Kiani et al. 2008) because dilution of this product increases
the distance between particles. In addition, because of a low
concentration of whey proteins in this product, crosslinking
between particles decreases. This causes the shear stress, at
high or low shear rates, to have a constant proportionality
with shear rate (Kiani et al. 2008, 2009). In this section, the
effects of WPC, pH of product and dissolved salt in milk
on the rheological properties of the product were
© 2017 Society of Dairy Technology
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Figure 4 Effect of WPC and dissolved salt in milk on serum separation of Doogh samples (after 24 h at 4 °C) with pH 3.5 (a), 4 (b) and 4.5 (c).

Figure 5 Effect of WPC and dissolved salt in milk on viscosity of Doogh samples; samples had pH 4 (a) and 4.5 (b).

investigated. Results of this research showed that in samples the flow behaviour of the samples and the samples showed
with pH 3.5 containing 2% dissolved salt in milk and in nearly Newtonian behaviour. Also, in samples containing
samples with pH 3.5 without dissolved salt in milk, increas- 0.5% WPC and 0% salt in milk, with an increase in pH
ing the WPC concentration from 0.5 to 4% did not affect from 3.5 to 4.5, the flow behaviour of Doogh samples did

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Figure 6 Effect of WPC and pH on viscosity of Doogh samples containing 1% (a) and 2% (b) dissolved salt in milk.
not change; the samples showed nearly Newtonian beha-
viour. Because of the low concentration of WPC in these
samples, the distance between particles was much and they
were less likely to interact. Furthermore, salt was dissolved
in whey protein solution, and the particle size did not
increase. On the other hand, it is revealed that as the pH of
samples was increased from 3.5 to 4.5 in samples contain-
ing 4% WPC and 2% dissolved salt in milk, the sample
behaviour changed from Newtonian to pseudoplastic or
shear-thinning behaviour. One of the most important factors
in the flow behaviour of Doogh is the volume of protein
particles present in the product. In a sample with the highest
pH value, WPC and dissolved salt in milk, the particles had
less space to move and they collided with each other and
created many bonds between them. In fact, the main reason
for the non-Newtonian flow behaviour of Doogh samples
was the overlapping phenomenon of the colloidal particles
in Doogh. These connections were destroyed at higher shear
rates. This caused a shear-thinning flow behaviour in Doogh
samples (Kiani et al. 2008). But, in samples with 0.5%
WPC and pH 3.5, by increasing the dissolved salt in milk
from 0 to 2%, the Newtonian behaviour was evident. In
these samples, WPC concentration was low and these pro-
teins were soluble at this pH value; therefore, the interparti-
cle interactions were low. Also, it is observed that the
increase in dissolved salt in milk from 0 to 2%, in the sam-
ple with pH 3.5 and 4% WPC, flow behaviour of Doogh
samples did not change. Although WPC was relatively high
in these samples, the proteins were soluble due to a low pH
value. Therefore, the particles were apart from each other
and a nearly Newtonian behaviour was evident in the sam-
ples.
CONCLUSION
By increasing the concentration of WPC in Doogh samples
and increasing the salt concentration in milk, the serum
206
separation of the product decreased. Interactions between
WPC and dissolved salt in milk had a significant effect on
serum separation of the samples. The amount of WPC had
more effect on serum separation compared with the dis-
solved salt content in milk. The pH of samples (in the
range of 3.5–4.5) had no significant effect on the stability
of the product. However, when the samples had minimum
serum separation, adding WPC did not increase stability,
whereas in pH values higher and lower than the pH 4.16
the stability of product is increased. Also in this condition
at pH 4.5, the viscosity of the samples increased and phase
separation decreased. The viscosity of the samples
increased by increasing the dissolved salt in milk, concen-
tration of WPC and pH value up to 4.5. The effect of pH
on viscosity of samples was higher than that of dissolved
salt in milk and WPC concentration. Most of Doogh sam-
ples showed Newtonian behaviour, but in samples with pH
4.5, 4% WPC and 2% dissolved salt in milk, pseudoplastic
behaviour was observed.
ACKNOWLEDGEMENT
This research was supported by ‘Iran National Science Founda-
tion, INSF’ which is gratefully acknowledged.
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