doi: 10.1111/1471-0307.

12238

ORIGINAL
RESEARCH Approaches to minimise yoghurt syneresis in simulated
tzatziki sauce preparation
PATROKLOS VARELTZIS,1 KONSTANTINOS ADAMOPOULOS,2*
E F S T R A T I O S S T A V R A K A K I S , 2 A T H A N A S I O S S T E F A N A K I S 2 and
ATHANASIA M. GOULA3
1
Department of Food Technology, Alexandrian Technological Institute of Thessaloniki, Thessaloniki GR – 574 00,
2
Department of Chemical Engineering, Faculty of Engineering, Aristotle University of Thessaloniki, Thessaloniki, and
3
Department of Food Science and Technology, Faculty of Agriculture, Aristotle University of Thessaloniki,
Thessaloniki, Greece

The phenomenon of syneresis becomes more profound in increased moisture yoghurt-based prod-
ucts. Such a product is the traditional Greek appetizer tzatziki that contains cucumber, a vegetable
with a high moisture content. During the manufacture of tzatziki, the addition of cucumber causes
the protein network to break up, provoking an increase in syneresis. The aim of this study was to
investigate a tzatziki manufacturing procedure that will lead to significantly decreased syneresis.
Two different manufacturing procedures were compared: the extra moisture coming from cucumber
was introduced before or after fermentation. The effect of adding whey protein concentrate (WPC),
albumin, sodium caseinate or a mixture of these was also studied. The results show that the addi-
tion of extra moisture before yoghurt fermentation leads to a significantly lower syneresis (7.5%)
and higher consistency (2000 cp) than those obtained in the case of addition after fermentation
(25% and 1500 cp, respectively). The use of albumin, WPC or a mixture of albumin, WPC and
sodium caseinate further decreased the phenomenon of syneresis to below 5%, without altering the
colour of the product (DΕ* < 2.3).
Keywords Syneresis, Stirred yoghurt, Tzatziki, Yoghurt, Casein, Whey protein concentrate.

maintained for 4–6 h. The acid development of

INTRODUCTION
Yoghurt is defined as a fermented milk product
produced with thermophilic lactic bacteria, usu-
ally Streptococcus thermophilus and Lactobacil-
lus delbrueckii ssp. bulgaricus (Mottar et al.
1989; Lucey et al. 1999; Ginovart et al. 2002).
All yoghurt-manufacturing procedures are basi-
cally the same. The milk is clarified and sepa-
rated into cream and skim milk and then
standardised to achieve the desired fat and pro-
tein content. The mixture is homogenised using
high pressures (10–20 MPa) at temperatures of
55–70 °C (Tamime and Robinson 1999). The
milk is pasteurised by heating for 30 min at
*Author for 85 °C or 10 min at 95 °C. Next, the milk is
correspondence. E-mail: cooled to approximately 43 °C, which is an
costadam@eng.auth.gr
optimum growth temperature for the yoghurt
© 2015 Society of starter culture that is added to the milk in a
Dairy Technology fermentation tank. A temperature of 43 °C is
yoghurt is carefully monitored until the pH
reaches 4.0–4.7; then, the fermentation is
stopped by rapid cooling (Connolly 1978).
During milk fermentation, the casein becomes
unstable and coagulates to form a firm gel, com-
posed of strands of casein micelles, with whey
entrapped within this matrix, which is inter-
locked via hydrogen bonds, forming a protein
matrix. Yoghurt structure is the result of disul-
phide bonding between k-casein and denatured
whey proteins and by aggregation of casein as
the pH drops to the isoelectric point of the
casein proteins during fermentation (Damin
et al. 2009).
An important aspect of a milk gel is whey
separation, which refers to the appearance of a
liquid on the surface of milk gel. It is a common
defect in fermented milk products such as
yoghurt (Lucey et al. 1998). Syneresis is defined

Vol 69, No 2 May 2016 International Journal of Dairy Technology 191

Vol 69, No 2 May 2016
as the shrinkage of gel, and this occurs concomitantly with
expulsion of liquid or whey separation and is related to
instability of the gel network resulting in the loss of the
ability to entrap all the serum phase (Walstra 1993).
According to Lucey et al. (1998), some possible causes of
wheying-off in acid gels are very high incubation tempera-
tures, excessive treatment of the mix, low total solids con-
tent (protein and/or fat) of the mix, movement or agitation
during or just after gel formation and very low acid produc-
tion (pH > 4.8) (Magenis et al. 2006; Donato and Guyom-
arc’h 2009). Magenis et al. (2006) reported that factors
influencing yoghurt texture and syneresis include total solids
content, milk composition (proteins, salts), homogenisation,
type of culture, acidity resulting from the growth of bacte-
rial cultures and heat pretreatment of milk.
Traditionally, the solid content of milk is increased for
yoghurt production. The three main systems available nowa-
days are good options to achieve desired protein and solids
contents: (i) addition of protein ingredient powders (skimmed
milk, whey protein concentrates, caseinates); (ii) evaporation
of water from milk under vacuum; or (iii) removal of water
by membrane filtration (Tamime et al. 2001). Fortification
with skim milk powder (SMP) is the common practice to
increase the solid content in conventional yoghurt manufac-
ture, but when enrichmenting the protein content is the main
target, the amount of SMP that can be added to provide extra
protein content becomes limited, since too high levels of
SMP can lead to a powdery taste and high lactose content,
which ultimately results in a highly acidic product (Abd El-
Khair 2009; Supavititpatana et al. 2009; Marafon et al.
2011a,b). An alternative approach of fortification of the milk
for yoghurt manufacture is by ultrafiltration. Protein concen-
trates produced by ultrafiltration have better nutritional value
than that produced by traditional methods. Another advan-
tage of UF milk is that it contains a higher level of protein
with a lower level of lactose than normal milk (Karlsson
et al. 2005). However, UF concentration of milk for yoghurt
manufacture leads to an excessively firm coagulum and a
more viscous product than conventional SMP fortification
(Schkoda et al. 2001).
The phenomenon of syneresis becomes more profound in
increased-moisture yoghurt-based products. Such a product
is the traditional Greek appetizer tzatziki. Tzatziki is made
of strained yoghurt mixed with cucumbers (in a ratio of
4:1), garlic, salt, olive oil and sometimes lemon juice, and
dill or mint or parsley. Strained or Greek-style yoghurt is
traditionally made by straining fermented yoghurt curd in a
cloth bag to reach the desired solids level by removing acid
whey. This step is achieved by mechanically separating the
whey from the curd using either a centrifugal separator or
membrane filtration (Nsabimana et al. 2005) or by fortifica-
tion of milk with milk protein concentrates (Bong and
Moraru 2014). Cucumber is a vegetable with a high mois-
ture content (~96% w/w), which practically increases the
192
moisture content of tzatziki. During the manufacture of tzat-
ziki, migration of water from the high moisture content
ingredient (cucumber) to the low moisture content ingredient
(yoghurt) occurs. Thus, the gel matrix of the yoghurt is bro-
ken and cannot hold the extra water, resulting in increased
syneresis.
The aim of this study was to investigate a tzatziki manu-
facturing procedure that will lead to significantly decreased
syneresis. For experimental purposes, a simulated tzatziki
sauce preparation was used. This preparation was strained
yoghurt with added moisture where, instead of adding
whole cucumber, garlic and dill, only the extra moisture
coming from cucumber was added. As stirring of yoghurt
breaks up the protein network leading to an increased syner-
esis, we tested the hypothesis that extra moisture can be bet-
ter retained by introducing it before the formation of
yoghurt. To this end, two different ways of manufacturing
were compared: the extra water coming from cucumber was
added (a) before and (b) after fermentation and compared to
tzatziki made from a commercial readymade strained
yoghurt. The effect of adding whey protein concentrate,
albumin, sodium caseinate or mixtures of these protein
ingredients on syneresis was also studied.
MATERIALS AND METHODS
Materials
Homogenised, pasteurised fresh cow’s milk (72 °C for 15 s)
(Mevgal S.A., Thessaloniki, Greece) was used as a stock of
raw material throughout the experimental work to avoid var-
iation in milk composition.
Globulal 70 N whey protein concentrate (70% protein
content) and Emulac sodium caseinate (50% protein con-
tent) were purchased from Meggle AG (Wasserburg, Ger-
many), whereas hen egg albumin powder (78% protein
content) was obtained from Kallbergs Industri AB (T€ ore-
boda, Sweden).
Streptococcus thermophilus and Lactobacillus delbrueckii
ssp. bulgaricus were obtained from Aristomenis D. Phikas
& Co (Thessaloniki, Greece). The cultures were stored at

20 °C in a concentrated form before use.
Soft potable water was used for tzatziki manufacture.
Yoghurt–Tzatziki manufacture
Tzatziki is made of strained yoghurt mixed with cucumbers
in a ratio of 4:1. The moisture contents of strained yoghurt
and cucumber are about 82 and 96% w/w, respectively
(Trichopoulou and Georga 2004). For experimental pur-
poses, a simulated tzatziki sauce preparation was used. This
preparation was strained yoghurt with added moisture,
where instead of adding whole cucumber, garlic and dill,
only the extra moisture coming from cucumber was added.
Thus, during tzatziki manufacture, 1.2 kg of water was
added per 5 kg of yoghurt/yoghurt formulation.
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Two different yoghurt-manufacturing procedures were fol-
lowed as depicted in Figure 1. In the first case (Figure 1a),
water was introduced after the incubation of condensed milk
with the micro-organisms, whereas in the second one (Fig-
ure 1b), the water was added before the incubation. In both
cases, different mixes of protein ingredients were added as
agents for improving texture and reducing syneresis. The
protein ingredients used were (w/w) whey protein concen-
trate 1% and 5%, sodium caseinate 1% and 5%, and albu-
min 1% and 5%, as well as a protein mixture 5% consisting
of 2% albumin, 2% whey protein concentrate and 1%
sodium caseinate. The percentages were based on prelimin-
ary experiments. Both preparations were compared to a con-
trol sample, which was manufactured using a commercially
ready strained yoghurt, with the same total solids and fat
contents.
The milk was concentrated form an initial solid content of
about 11.2% to a final concentration of approximately
21.5% in a rotary vacuum evaporator (Model R 114, Buchi
Laboratoriums-Technik, Flawil, Switzerland) at 45 °C. A
quantity (500 mL) of the milk samples was prewarmed to
45–50 °C in a microwave oven and heated in a water-boil-
ing bath to 80 °C. When this temperature was reached, the
samples were kept for 30 min in the bath and protein ingre-
dients were added. In the case presented in Figure 1b,
where the water was added before the incubation, soft pota-
ble water in a quantity that accounts for the extra moisture
content of the tzatziki was also added. After cooling to

Fresh milk (3.5% fat)

Condensation by boiling (continuous stirring)

Heating at 80 ºC for 30 min

Protein ingredients addition

(a) (b)

Addition of water corresponding to

Cooling to 42 ºC (temperature of incubation)
the cucumber of tzatziki

Inoculation and incubation for 5 h Cooling to 42 ºC (temperature of incubation)

(till pH reached 4.5)

Cooling of yogurt at room temperature (1.5 h) Inoculation and incubation for 5 h

(till pH reached 4.5)

Refrigerated storage (4 ºC) Cooling of yogurt at room temperature (1.5 h)

Addition of water corresponding to Refrigerated storage (4 ºC)

the cucumber of tzatziki

Refrigerated storage (4 ºC)

Figure 1 Tzatziki manufacturing procedure (a) water added after fermentation and (b) water added before fermentation.

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42 °C, the mixes were inoculated with the active starter
(9 g/53.5 kg), divided into three equal portions in plastic
cups and incubated at 42 °C in a fermentation store until a
4.7 pH was reached in about 5 h. The yoghurt was then
cooled to room temperature for 1.5 h by placing the cups in
a bath of cold water and stored at 4 °C in the cold refrigera-
tor. In the case presented in Figure 1a, where the water was
added after the incubation, the quantity of water that
accounts for the extra moisture content of the tzatziki was
added during storage. All samples were prepared in tripli-
cates. Formulations used for tzatziki manufacture are listed
in Table 1.
Tzatziki samples were drawn at different cold storage
intervals (2, 9, 14, 16, 18, 22, and 25 days) and character-
ised for viscosity, syneresis, pH and colour.
Physicochemical characteristics
Solubility
Milled protein ingredient powders were dispersed (1% w/w)
in deionised water and the pH adjusted with 2 N NaOH to
7.0. Dispersions were centrifuged at 20000 g for 15 min.
Protein concentrations were determined according to Lowry
et al. (1951). Solubility was obtained from the concentration
ratio of the supernatant and the dispersion before centrifuga-
tion.
at 100 rpm. The sample temperature was 4 °C. For a rela-
tive comparison between treatments, viscosity reading was
taken at the point of the 30th s and torque was maintained
at all times between 10% and 100%.
Moisture content
Moisture was determined by an Ohaus moisture analyzer
(Model MB35 Halogen, Ohaus Co., Pine Brook, NJ, USA).
Colour
Colour was determined using a Hunter Lab colorimeter. L*,
a* and b* values were determined, while the total difference
in colour (DΕ) was calculated using the following equation:
qffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
DE ¼ ðL0
Ls Þ2 þ ða0
as Þ2 þ ðb0
bs Þ2 , where Lo*,
ao* and bo* are the values for the control sample, and Ls*,
as* and bs* are the values for the sample. DΕ ~ 2.3 corre-
sponds to just noticeable difference.
Statistical analysis
Data were analysed by a general linear model procedure of
the Fisher’s protected-least-significant-difference test using
SAS (SAS Inst., Cary, NC, USA). This test combines analy-
sis of variance (ANOVA) with comparison of differences
between the means of the treatments at the significance level
of P < 0.05.

Syneresis RESULTS AND DISCUSSION

Syneresis was measured according to Keogh and O’Ken-
nedy (1998). Briefly, yoghurt (30–40 g) was centrifuged (T
52.1, VEB MLW Zentrifugenwerk Engelsdorf, Germany) at
222 g for 10 min at 4 °C. The clear supernatant was poured
off, weighed and recorded as syneresis (%).
Viscosity
Viscosity measurements were obtained using a Brookfield
viscometer (Model DV-II+; Brookfield Engineering Labs,
Inc., Middleboro, MA, USA) with a No 4 spindle rotating
Effect of protein ingredients addition on syneresis rate
of tzatziki
The protein ingredients used as agents for improving water-
holding capacity and decreasing syneresis were whey pro-
tein concentrate (WPC), albumin and sodium caseinate.
Their measured properties are presented in Table 2. The
choice of protein ingredients was based on their ability to
decrease syneresis in tzatziki manufactured using commer-
cial strained yoghurt. Figure 2a,b show the effect of the

Table 1 Formulations used for tzatziki manufacture

Ingredients (%)
Treatment Yoghurt Milk Water Whey protein concentrate Albumin Sodium caseinate Active starter
Control 80 – 20 – – – –
1% whey protein concentrate – 77.6 21.4 1.0 – – 0.02
1% albumin – 77.6 21.4 – 1.0 – 0.02
1% sodium caseinate – 77.6 21.4 – – 1.0 0.02
5% whey protein concentrate – 74.5 20.5 5.0 – – 0.02
5% albumin – 74.5 20.5 – 5.0 – 0.02
5% sodium caseinate – 74.5 20.5 – – 5.0 0.02
5% protein mixture – 74.5 20.5 2.0 2.0 1.0 0.02

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syneresis, while a more profound effect on syneresis was

Table 2 Protein solubility and pH of solutions at 25 °C (avg  SD,
exhibited by increasing albumin concentration from 1 to
n = 4)
5%.
Protein (% w/w) Solubility (%) pH The effects of protein and total solids (TS) are difficult to
5% whey protein concentrate 33.0  1.5b 6.70  0.08c study separately, as the two variables cannot be modified
5% albumin 37.9  1.7a,b 7.24  0.02b independently. Increasing TS improved yoghurt texture,
5% sodium caseinate 31.7  0.7b 6.91  0.01a,c based on sensory and instrumental evaluations. Generally,
Means in the same column followed by the different superscript let-
higher TS caused an increase in density and reduced pore
ters are significantly different (P < 0.05).
size in the protein matrix of the yoghurt gel. This led to a
reduction in syneresis and improvement of the water-hold-
(a) 30 ing capacity of the gel. Keogh and O’Kennedy (1998)
explained that casein and b-lactoglobulin interact chemically
25 on heating. This effectively increases the concentration of
Syneresis (% w/w)

gel-forming protein in the yoghurt matrix and reduces syner-

20
A
esis through increased entrapment of serum within the inter-
15 B stices of the whey protein molecules attached to the surface
C of the casein. The effects of different levels of TS were
10 D
studied by Tamime and Robinson (1999), and they found
that the consistency was greatly improved as solids
5
increased from 12 to 20 g/100 g. The greatest change was
0 observed from 12 to 14 g/100 g, whereas levels above
9 14
16 g/100 g resulted in less pronounced change. According
16
18 to Prentice (1992), increase in protein levels is the principal
22
Time (days) 25
factor influencing texture and enrichment of milk with milk
powder results in the development of chains and aggregates
(b) 30 of casein micelles. Wu et al. (2001) demonstrated that
25
water-holding capacity was related to the ability of the pro-
teins to retain water within the yoghurt structure. These
Syneresis (%w/w)

20 researchers further suggested that the fat globules in the

15
10
5 because the high whey protein-to-casein ratio induced
0
9
14 16
18
22
Time (days) 25
Figure 2 Effect of protein addition on syneresis rate during storage of
tzatziki-simulated samples. Addition level of (a) 1% (w/w) and (b) 5%
(w/w). (A) control sample, (B) whey protein concentrate, (C) albumin
and (D) sodium caseinate.
level of protein addition on the syneresis of tzatziki during
cold storage at 4 °C. Each data point in the figure represents
average values of three replications. The repeatability for
syneresis expressed as the average standard deviation of the
three replications was 0.3%. All three protein ingredients at
both levels (1 and 5%) significantly decreased syneresis
compared to the control sample. However, comparing the
effect of the different concentrations of the same protein
ingredient, it was concluded that increasing the concentra-
tion of WPC and sodium caseinate did not further decrease
A milk may also play an important role in retaining water.
B
C Tzatziki samples containing albumin had lower syneresis
D rates than the others, whereas syneresis was reduced with
increased proportions of sodium caseinate. This may be
shrinkage of the gel, which led to increased whey drainage
or whey separation (Lucey 2004). Keogh and O’Kennedy
(1998) explained that casein and b-lactoglobulin interact
chemically on heating. This effectively increases the con-
centration of gel-forming protein in the yoghurt matrix and
reduces syneresis through increased entrapment of serum
within the interstices of the whey protein molecules attached
to the surface of the casein. Saint-Eve et al. (2006)
observed that sodium caseinate provides a gel with a hetero-
geneous structure with large pores. When yoghurts were
enriched with whey protein, the protein network was more
uniform and the pores of the gel were smaller than those of
yoghurts to which sodium caseinate had been added.
Effect of way of added moisture on syneresis rate of
tzatziki
Although the phenomena occurring during syneresis are not
fully understood, it is agreed that increased syneresis with
storage time is usually associated with severe casein net-
work rearrangements (van Vliet et al. 1997) that promote

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whey expulsion. As stirring the formed gel would add an
extra destabilising effect, we tested the hypothesis that water
would be better retained in stirred yoghurt products if it was
added before fermentation.
Syneresis was significantly lower (P < 0.05) for all tzat-
ziki samples in which water was added before fermentation
(Figure 3), except for the sample with 5% added albumin,
in which there was no difference in syneresis. This was also
true for the control sample, which exhibited an almost con-
stant syneresis around 7%, throughout the storage period.
However, the control sample in which the water was added
after fermentation had a much higher syneresis, at a level
around 25%. Samples with 5% albumin and 5% of the pro-
tein mixture exhibited the least syneresis.
Rheological properties of yoghurt have been well studied.
Yoghurts have flow properties that are characteristic of a
non-Newtonian and weakly viscoelastic fluid (Ramaswamy
and Basak 1991, 1993; Benezech and Maingonnat 1993;
Skriver et al. 1993). Keogh and O’Kennedy (1998) studied
A B C D
(a) 35
30
Syneresis (% w/w)
the role of milk fat, protein, gelatin and hydrocolloids
(starch, locust bean gum/xanthan mixture) on the rheology
of yoghurt. These authors showed that the consistency index
(K) and syneresis were more frequently influenced by the
composition than the behaviour index (n) and the critical
strain (cc). Lee and Lucey (2006) investigated the structural
breakdown of the original (intact) yoghurt gels that were
prepared in situ in a rheometer, as well as the rheological
properties of stirred yoghurts made from these gels. The
same authors found that the rheological properties of
yoghurts were greatly influenced by the physical properties
of the original intact (set) yoghurt gels. Water in milk gels
is physically trapped within the gel network, meaning that
the tendency for whey separation is primarily linked to
dynamics of the casein network rather than mobility of the
water molecules (van Vliet and Walstra 1994).
Effect of protein addition and way of moisture addition
on viscosity, pH and colour of tzatziki
Protein addition increased yoghurt viscosity. Similar results
were found by Abu-Jdayil (2003), who found greater vis-
cosity in yoghurts of the labneh type, with higher protein
content. Yoghurts containing WPC had lower viscosity val-
ues than the others. These results are in agreement with

25 those reported by Modler et al. (1983) and Guzman-Gonz-

alez et al. (1999), who found that casein-based products
20
tended to produce firmer gels with less syneresis than
15 yoghurts fortified with whey protein. Modler and Kalab
10
(1983), using electron microscopy studies, showed that
yoghurts prepared with casein, SMP and MPC exhibited a
5 high degree of fused micelles when compared to yoghurts
0 stabilised with WPCs.
2 4 6 8 10 12 14 16 18 20 Adding the extra moisture to the system before fermenta-
tion and setting probably allows casein and whey proteins
Time (days)
to arrange themselves in a network and trapping the extra
moisture, rendering yoghurts less susceptible to syneresis
A B C D
(b) 35 compared to samples in which the extra moisture is added
after fermentation. This is further strengthened by the obser-
30 vation that the viscosity of tzatziki samples with water
added before fermentation was also found to be higher than
Syneresis (% w/w)

25
that of all the respective samples in which the water was
20
added after fermentation (Figure 4). For yoghurt products,
15 steps such as mixing result in a viscosity reduction that is
only partially restored after shearing is stopped. Recovery of
10
structure is called ‘rebodying’ and is a time-dependent phe-
5 nomenon. Structural recovery also affects the apparent vis-
cosity of yoghurts (Lee and Lucey 2010). Arshad et al.
0
(1993) reported that glucono-d-lactone (GDL)-induced gels
2 4 6 8 10 12 14 16 18 20
had only 30% recovery of the original value of the dynamic
Time (days) moduli even after allowing 20 h for recovery after shearing.
Figure 3 Syneresis during storage of tzatziki-simulated samples with As far as the effect of protein ingredients on pH changes,
added water (a) after and (b) before incubation. (A) control sample, (B) there was a small decrease in the pH from the time the fer-
1% whey protein concentrate, (C) 5% albumin and (D) 5% protein mix- mentation was stopped till 48 h after storing the product at
ture. 4 °C (postacidification) and slowly continued till the 10th

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(a) A B C D Control 5%WPC

8000 4.9
5%Albumen 5%Caseinate
7000
4.7
6000
Viscosity(cp)

5000 4.5
4000
4.3
3000

pH
2000 4.1

1000
3.9
0
1 3 5 7 9 11 13 15 17 19 21 3.7
Time (days)
3.5
(b) 8000 A B C D 0 5 10 15 20 25 30
Time (days)
7000
Figure 5 Change of pH in tzatziki-simulated samples with 5% w/w
6000 addition of different protein ingredients.
Viscosity(cp)

5000
4000
(a) 3.5 A B C D
3000
2000 3.0

1000 2.5

0 2.0
ΔΕ*

1 3 5 7 9 11 13 15 17 19 21
1.5
Time (days)
1.0
Figure 4 Viscosity of tzatziki-simulated samples with added water (a)
after and (b) before incubation. (A) control sample, (B) 1% whey protein 0.5
concentrate, (C) 5% albumin and (D) 5% protein mixture.
0.0
0 5 10 15 20
day of storage. A decrease in pH during storage is expected Time (days)
as a result of the activity of LB (Tamime and Robinson
1999); this has been observed in lactic beverages (Oliveira A B C D
et al. 2001). Postacidification occurred independently of the (b) 2.5
ingredient or the concentration used and was 0.20 pH units
on average. A similar trend was observed by Damin et al. 2.0
(2009). There was no significant pH difference between the
control and the samples throughout the storage period for 1.5
ΔΕ*

the 1% level of addition. On the other hand, at the 5% level

of addition, a significant (P < 0.05) difference was observed 1.0
(Figure 5). This can be attributed to the higher buffering
capacity brought by the added protein ingredients. There 0.5
was a high correlation coefficient between the pH through-
out the storage period and the respective syneresis, that is 0.0
0.90–0.95. 0 5 10 15 20
Furthermore, there was no significant effect of the way of Time (days)
extra water addition on the pH. There was, however, a ten- Figure 6 Total colour difference of tzatziki-simulated samples with
dency for a slightly higher pH during the first 2 weeks of added water (a) after and (b) before incubation in relation to the control
storage for the samples with added water after fermentation sample. (A) control sample, (B) 1% whey protein concentrate, (C) 5%
(~0.1–0.15 pH units, results not shown). albumin and (D) 5% protein mixture.

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Protein addition significantly affected the colour of
yoghurt (P < 0.05). An increase in the lightness and a
decrease in yellow colour of yoghurts were observed. The
results were in agreement with the observation of Gonzalez-
Martınez et al. (2002), who studied the change in colour as
affected by addition of casein. The higher milk protein con-
tent gave better protein coagulation. The coagulation of pro-
tein affected the structure and surface properties of yoghurt.
Mor-Mur and Yuste (2003) reported that the increased pro-
tein coagulation enhanced the light absorption that resulted
in the lighter tons.
Addition of water before fermentation also led to better
colour retention of the tzatziki samples (Figure 6) for all
treatments. In tzatziki samples in which water was added
before fermentation, DE* is below 2.3, that is the value that
corresponds to just noticeable difference. Albumin, even
though significantly improving syneresis, led to a greater
colour change and, therefore, it might not be acceptable for
product formation.
CONCLUSIONS
The hypothesis that extra moisture can be better retained in
stirred yoghurt-based products (e.g. tzatziki) by introducing
the extra moisture before the formation of yoghurt was suc-
cessfully tested. Results clearly show that, even without the
addition of protein ingredients, in tzatziki samples with
water added before fermentation, syneresis was lower and
consistency was higher than in those where the water was
added during the stirring of yoghurt. The use of albumin,
WPC or a mixture of albumin, WPC and sodium caseinate
further decreased the phenomenon of syneresis.
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