Acta Manilana 59 (2011), pp.

19–30
Printed in the Philippines
ISSN: 0065–1370

Improvement of textural properties of yogurt made

from reduced-fat carabao milk by whey protein content
adjustment and heat treatment of the milk
Janice C. Mariano1, John Hennessy M. Merced1, Abbie June F. Miranda1,
and Philipina A. Marcelo1,2*
1
Department of Chemical Engineering, 2Research Center for the Natural and Applied Sciences
University of Santo Tomas, España Boulevard, 1015 Manila, Philippines

If caseins and whey proteins were present in appropriate proportion in milk, and the milk
was heat-treated at temperatures well above the usual milk pasteurization temperatures,
these components have the ability to form firm gel of uniform porosity through heat-induced
protein-protein interactions. In this study, the whey protein-to-casein ratio (by mass) in
reduced-fat carabao milk was adjusted from its natural ratio to 0.25 and 0.35 by the addition
of whey protein isolate (WPI). The milk samples were then heat-treated at 80°C for 25 min.
The heat-treated milk samples were used in yogurt-making, and the viscosity evolution
during acidification with starter culture inoculation was monitored to determine and compare
gel setting times. Compression, puncture and syneresis tests were done on the yogurt
samples to determine the effects of proteins ratio adjustment and heat treatment on the
texture of the yogurt samples. It was found that the milk with 0.25 whey protein-to-casein
ratio exhibited the most satisfactory results in all the tests. This suggests that the combination
of proteins ratio adjustment and heat treatment used led to textural improvement of the
yogurt, and may enable the production of set yogurt from reduced-fat carabao milk without
using carbohydrates-based stabilizers and texture enhancers.

Keywords: yogurt, carabao milk, whey protein, whey protein isolate, casein, viscosity

INTRODUCTION microorganisms. Subjecting milk to heat

Protein enrichment and heat treatment are
important in controlling the consistency
and textural properties of yogurt [1]. Heat
treatment is done to improve the keeping
quality of milk by deactivating
*To whom correspondence should be addressed
Email: pamarcelo@mnl.ust.edu.ph /
philip.marcelo@gmail.com
treatment primarily affects the behavior
of milk proteins — caseins and serum
proteins — in terms of conformational
changes they undergo. During heat
treatment, no noticeable heat induced
effects can be observed on the structure of
casein micelles in the temperature range
of 70–100°C [2]. However, heating has a
definite effect on the serum, or whey,
protein fraction of milk, which is primarily

Acta Manilana • Volume 59 (2011)

 Mariano JC et al. / Acta Manilana 59 (2011)
composed of β-Lactoglobulin (β-Lg) and α-
Lactalbumin (α-La). During heating, the
reactive thiol group is exposed in β-Lg due
to conformational changes of the molecule
[3]. This reactive thiol group can form links
with other proteins having a reactive thiol
group, or through thiol group-disulfide
exchange. The reaction between thiol
groups makes the denaturation process
irreversible [4]. The process of denaturation
and subsequent aggregation of β-Lg
constitutes the polymerization process, in
which the unfolding of the globular protein
represents the initiation step [5]. β-
Lactoglobulin also exhibits interactions
with casein micelle thiol group-disulfide
bridge exchange reaction through κ-
caseins, which are located at the casein
micelle surface. The initial step of this
process is believed to be physical in nature,
but the final interaction is covalent (i.e.,
disulfide linked) [6]. As a result, the “hairy
strands” on the casein micelle surface
contain casein associated with whey
proteins after heating. Incorporating heat
treatment in milk preparation, therefore,
results in a complex mixture of native whey
proteins, whey protein aggregates and
casein micelle covered with whey protein
[7].
Yogurt is made by the acidification of milk
through lactic acid bacteria (LAB). During
fermentation, LAB transforms milk sugar
(lactose) into lactic acid. The lowering of pH
due to milk acidification promotes the
process of casein coagulation [8]. Casein
micelle begins to destabilize at pH 5.3
where colloidal calcium phosphate (CCP),
which supports the casein micelle structure
by binding the casein submicelles together,
leaches into the serum phase, setting a
condition that favors micellar aggregation
[9, 10]. Complete precipitation of casein
occurs at its isoelectric point of pH 4.6. The
precipitated casein complex constitutes the
yogurt curd. The yogurt matrix formed was
20
believed to exhibit much favorable
consistency and textural properties if the
matrix was primarily made up of κ-casein-
β-Lg aggregate [11].
This study aims to exploit the increased
denaturation of whey proteins by elevating
both the heat treatment temperature and
the whey protein to casein ratio to improve
yogurt texture. Carabao (Bubalus bubalis
carabanesis) milk with reduced fat content
was used in the study. The whey protein-
to-casein ratio in the milk was adjusted
from the natural ratio in milk samples to
0.25 and 0.35 by the addition of whey
protein isolate (WPI). The milk samples
were then heat-treated to a temperature
moderately higher than the pasteurization
temperature of 80°C, for 25 min. According
to Dannenberg and Kessler [12] more
extensive whey protein denaturation
occurs at higher temperatures. However,
cases of prolonged heating at high
temperature and excessive protein addition
in yogurts result in a grainy consistency of
the product [1]. This kind of yogurt texture
is a result of vigorous and uncontrolled
protein-protein interactions where large
protein aggregates that separate from the
yogurt gel network are formed. The
challenge, therefore, is to find the suitable
proteins ratio and heat treatment
conditions.
The results of this study could lead to
textural properties improvement of yogurt
made from carabao’s milk. Texture is one of
the factors that consumers consider in
buying food products, thus improvement
of the texture of a nutritious product, such
as yogurt, is a step in making the product a
more attractive and commercially viable
vehicle to deliver nutrients through the
human diet. Buffalo milk, which was found
to contain more caseins and minerals than
cow’s milk is potentially a good raw
material for this purpose [13]. The
 Improvement of textural properties of yogurt
utilization of carabao’s milk, which is the
milk from Philippine buffalo, for dairy
product development and manufacture
will not only result in a wide range of
nutritious choices for consumers but will
also help in achieving high and sustained
productivity in the local dairy industry.
EXPERIMENTAL
Materials
Pasteurized whole carabao milk (DVF Dairy
Farm, Talavera, Nueva Ecija, Philippines)
was purchased from a local retailer and the
WPI was NZMP, Alacen 894, with 2% (w/
w) moisture and 90% (w/w, dry basis)
protein, which was a gift from Fonterra
Philippines (Pasig City, Philippines). The
starter culture was bought from Danasia
Philippines (Makati, Philippines).
Skimming of milk
The pasteurized carabao milk was
skimmed using the Ultra 8V-2 Centrifuge
(LW Scientific, GA, U.S.A.) at 3300 rpm for
30 min, following the method developed by
Hendrix and Reyes [14]. After
centrifugation, the fat accumulated on the
surface was removed using a small plastic
spatula. For more effective skimming, the
milk was centrifuged for another 10 min
and the remaining fat skimmed off. Three
batches of milk were skimmed to produce
reduced-fat samples, which were used in
the experiments. Representatives of the
reduced-fat milk, together with the whole
milk samples were sent to a private
laboratory for chemical analyses.
Compositional analysis of milk
Compositional analyses, which included
the total solids, fat, lactose, total nitrogen,
non-protein and non-casein nitrogen
contents, were determined for both the
pasteurized whole and skimmed carabao
milk samples by the First Analytical
Services & Technical Cooperative (F.A.S.T.)
Laboratories (Cubao, Quezon City,
Philippines).
Preparation of milk samples
Based on the compositional analysis of the
reduced-fat carabao milk, the amount of
WPI powder to be added that corresponded
to the desired whey protein-casein ratio in
the milk was computed. The following
equations were used:
WP from milk = weight of milk × [NCN – NPN) × 6.38]
Whey protein to casein ratio = (WP from milk +
WP from WPI) ÷ (Weight of milk × [(TN – NCN)]
× 6.38])
where: WP = whey protein
TN = total nitrogen
CN = non-casein nitrogen
NPN = non-protein nitrogen
6.38 = constant used in converting
%nitrogen to %protein
The WPI powder was dispersed in the milk
by gentle stirring for 90 min using PC-620
hotplate/magnetic stirrer (Corning Inc., NY,
U.S.A.). The mixture was then allowed to
equilibrate by storing overnight in the
refrigerator at 4°C. The equilibrated milk
solutions were used in the subsequent parts
of the experiment.
Standardization of milk pH
The pH of the milk was measured before
incubation and adjusted to 6.8 ± 0.02 as
necessary for pH standardization. This
ensured that the rate of pH decline due to
the addition of LAB was approximately
uniform in the milk samples.
Preparation of yogurt
A batch of pasteurized whole milk (blank)
and reduced-fat carabao milk with whey
protein-to-casein ratios adjusted to 0.25
21
 Mariano JC et al. / Acta Manilana 59 (2011)
and 0.35 were prepared as previously
described. Following the method developed
by Mesina et al. [15], the milk samples were
then heated using a water bath at 80°C with
constant stirring for 25 min. After heating,
the milk samples were immediately cooled
using a cold bath to the starter culture
inoculation temperature of 45°C. The
freeze-dried starter culture was then added
to the milk with an approximate weight of
0.04 g per batch. The product was stirred
gently for 10 min to ensure uniform
distribution of the culture. The milk sample
mixtures were distributed in 20 mL
sterilized containers and placed in an
incubator (Binder Inc., Tuttlingen,
Germany) for about 12 h at a temperature
of 45°C. After fermentation, the yogurts
produced were stored in a cooler at about
4°C. They were then used in the subsequent
parts of the experiments. Three batches of
milk samples were studied.
Yogurt characterization
The ability of heat-denatured whey protein
to improve yogurt texture was verified by
comparing the viscosity evolution of milk
samples as they set to yogurt gels, the
strength and firmness of the yogurt gels
that are related to sensory quality, and the
percent syneresis from the yogurt at
specific time intervals.
Viscosity test. The viscosity test was done
to determine the time duration of the milk
samples to set to yogurt gels, and to
investigate the effects of heat treatment
parameters and whey protein-to-casein
ratio on the gel setting behavior of the milk.
The whey proteins in the milk solution was
polymerized and allowed to interact with
the casein micelles by heat treatment and
stored at 4°C for 12 h. LAB was then added
at 45°C. The milk was set up in a water bath
with the temperature maintained at 45°C.
Continuous viscosity measurement after
22
the addition of LAB bacteria was carried
out using the Brookfield LVT viscometer
(Brookfield, MA, U.S.A.) and the viscosity
reading was recorded until a constant
reading was achieved, which was deemed
the onset of milk gelation.
Compressibility test. The compressibility
test was done to estimate and compare the
strength of the yogurt gel samples. A flat
round disk was placed on top of the yogurt
sample with the same cross-sectional area
as the disk. Pressure was applied to
compress the yogurt by adding weights in
small increments on the disk until the
yogurt collapsed. The total mass of the disk
and the weight added were recorded, and
the pressure computed by:
P= F
A
where: P = pressure applied on the yogurt
sample, Pa
F = (mass of the disk + mass
of added weights) × acceleration
due to gravity, N
A = cross-sectional area of the disk
or the yogurt sample, m2
The pressure necessary to cause the
collapse of the yogurt network was
recorded. The pressure was related to the
firmness of the yogurt gel network as well
as the porosity of the gel. Void spaces are
present in a gel network and they are filled
with water. During the compression test
the void spaces, or pores, were compressed
and water was expelled from the yogurt
network. The test was performed on all
samples in duplicate.
Puncture test. An improvised glass
instrument, which consisted of a calibrated
glass needle of measured mass, was used
for the puncture test. The instrument was
elevated to different heights which are 8,
10 , and 12 cm, and was released directly
on the yogurt surface and allowed to
 Improvement of textural properties of yogurt
puncture the yogurt within a fixed time
interval. For each elevation the length of
the glass imbedded in the yogurt was
recorded at a fixed time interval. This test
was done to measure and compare the
cohesiveness of the yogurt samples.
This test also allowed the comparison of
the water holding capacity of the samples.
When the instrument was dropped at a
certain height, the initial fraction of glass
length submerged in the yogurt sample
was recorded. The total glass length
imbedded into the yogurt was noted after
30 min. The space that was occupied by the
puncture instrument was the void space
in the yogurt gel that held water. The water
rose to the surface as the puncture went
deeper, and more water was expelled. A
large amount of water expelled would
mean weak and less cohesive network. The
variation of the drop height was done to
see the impact of force applied to the
samples.
Syneresis index
Time-dependent syneresis. The liquid
expelled from each yogurt samples
produced was recorded at the end of the
first, fifth and seventh day of storage. It was
done by carefully removing and weighing
the liquid that separated from the yogurt
gel. This corresponded to the initially
entrapped liquid in the yogurt network
and slowly separated due to diminishing
Table 1. Compositional analysis of whole and reduced-fat carabao milk
Parameters, Whole
% (w/w) Carabao Milk
Total Solids 18.2
Fat 8.0
Lactose 4.7
Total Nitrogen 0.5
Non-Protein Nitrogen 0.03
Non-Casein Nitrogen 0.04
water holding capacity of the yogurt gel
during storage [16]. The percent liquid
expelled was computed using the equation
below [17].
Amount of liquid removed
%Syneresis = ________________________ × 100
Amount of water in the yogurt use
Forced syneresis. After 7 days of storage,
samples were centrifuged for 30 min at
3300 rpm. Empty test tubes were initially
weighed, and then weighed once more after
adding the samples. After centrifugation,
the liquid expelled from the yogurt gel using
pipette and the test tubes containing the
samples were weighed again. The syneresis
percentage was calculated by dividing the
mass of the liquid separated from the yogurt
after 7 days by the amount of liquid
expelled after centrifugation and then
multiplying by 100.
RESULTS AND DISCUSSION
Compositional analysis of the milk
The compositional analyses of the milk
samples used in the experiments are shown
in Table 1. It is interesting to note that while
the protein contents are about the same,
the whole carabao milk contained more
than double of cow’s milk fat content, which
is 3.1% (w/w) on average. This is consistent
with the findings of Ahmad et al. [13] for
raw buffalo milk from the Murrah breed of
B. bubalis. However, for this study, it was
Reduced-fat
Test Method
Carabao Milk
12.6 AOAC 990.20, Air Oven
0.31 Mojonnier Extraction
4.8 AOAC 930.28, Gravimetry
0.3 Kjeldahl
0.03 Kjeldahl
0.06 Kjeldahl
23
 Mariano JC et al. / Acta Manilana 59 (2011)

Table 2. pH of samples casein micelle surface more effectively,

with different whey protein-to- thereby causing an increase in viscosity of
casein ratios the milk samples continuously as
acidification ensues until they set to yogurt
Test Samples pH
gels [22]. This behavior was quantified by
Blank 6.80  0.00
monitoring how the apparent viscosities
0.25 Ratio 6.79  0.01
0.35 Ratio 6.78  0.02 of the milk samples evolved with time.
 

The milk samples slowly thickened until

about 30 min. The viscosity reading then
started to increase more considerably, and
necessary to reduce the fat content of the
then rapidly, until they reached the limit of
carabao milk to ascertain that textural
the viscometer, which indicate that the
characteristics were due mainly to the
samples then started to set into gels.
adjusted protein ratio and heat treatment.
Therefore, the fat content of the milk
Figure 1 shows the viscosity curves for the
samples were reduced by skimming up to
different milk samples considered in this
the limit of available means.
study. The milk sample of 0.35 whey
protein-to-casein ratio exhibited a rapid
Textural characterization tests pH of milk.
increase in viscosity after 40 min. This
The pH of the different milk samples with
indicates high protein aggregation rate and
and without adjusted whey protein-to-
extensive interaction between caseins and
casein ratios are shown in Table 2. The
whey proteins within a short period of
variation in pH prior to heat treatment can
incubation. Excess amount of whey protein
affect the mechanism of denaturation
present in the milk sample must have led
because the molecular conformation of the
to interaction between whey proteins other
whey proteins is pH-dependent [18]. At
than just κ-casein-β-Lg interactions (κ-Cβ-
higher pH values, heat treatment will result
L) [23]. On the other hand, the blank showed
in the formation of more whey protein
a considerable increase in viscosity only
aggregates while lower pH will result in
after 100 min, which was slower compared
more association of whey proteins with
with both milk samples with 0.25 and 0.35
casein micelle [7, 19, 20]. These emphasize
whey protein-to-casein ratios. This slow
the importance of keeping the pH of milk
samples the same prior to heat treatment.
It was to ascertain that the variation in
1100
yogurt characteristics was due to the heat 1000
treatment and the adjustment of whey
Apparent Viscosity (mPa.s)

900
800
protein-to-casein ratio. 700
600
Viscosity. Heat treatment denatures the 500
400
globular whey proteins, and high-heat 300
Blank

treatment leads to high degree of whey 200 0.25 Ratio

100 0.35 Ratio

protein denaturation and polymerization 0
of the denatured proteins [21]. Aside from 0 20 40 60 80 100 120 140 160 180 200

being more effective water binder Time (min)

compared with those in their native form, Figure 1. Change in the apparent viscosity with time
the denatured proteins interact with the of the blank and the milk samples with whey
casein micelles through the k-caseins on the protein-to-casein ratios adjusted to 0.25 and
0.35.

24
 Improvement of textural properties of yogurt
viscosity increase was a result of limited
interactions between whey proteins and
caseins. The milk sample with 0.25 whey
protein-to-casein ratio exhibited a
moderate viscosity build-up, which started
to increase considerably after about 75 min.
It was deemed that the shorter the time it
took for the milk sample to set to yogurt
gel, the stronger the gel network formed by
the milk sample, and that the result
indicated that as the whey protein-to-
casein ratio increased in the heat-treated
milk samples, the shorter the time for the
onset of gelation. This is consistent with the
findings of Beaulieu et al. [24]. However, it
was unclear how this supposed gel
strength could be related with textural
properties relevant to sensory quality. This
was further investigated in the subsequent
tests.
Compression test. In the compression test,
compressive pressure was applied on the
gels and the corresponding change in their
volume measured against the applied
pressure until the gel collapsed. The
compressive pressures at which the gels
collapsed were recorded as indication of
their strength. These were then compared
and analyzed.
The yogurt gel produced from the blank
required the lowest compressive pressure
of only about 220 Pa before a change of
0.2 cm became apparent, followed by its
subsequent collapse (Fig. 2). Moreover,
water was expelled as the yogurt gel was
compressed and the pores of the gel network
where water was held collapsed upon
compression. This behavior must be due to
weak gel network brought about by
insufficient protein interactions that hold
the gel network together [25].
The yogurt gel made from the milk sample
with 0.25 whey protein-to-casein ratio
required greater pressure of about 1400 Pa
7000
6000
Blank Ratio C
Pressure Applied (Pa)
5000 0.25 Ratio
0.35 Ratio
4000
3000
2000
B
1000
A
0
Yogurt Samples
Figure 2. Compressive pressure measured for yogurt
made using: (A) blank, and the milk samples with
(B) 0.25, and (C) 0.35 whey protein-to-casein
ratios at which structure collapse was observed.
before structural collapse became apparent.
The yogurt gel also expelled water as
compression ensued, which means that the
formed yogurt was also porous and had a
satisfactory water holding capacity, but the
gel network was stronger compared with
that produced from the blank. According
to Tamime and Robinson [26], an increased
protein interaction and protein-protein
bond increases the elastic character of the
gel, making yogurt less susceptible to
rupture. It was deemed that the addition of
whey protein lead to a more extensive whey
protein-casein interaction, which
strengthened the gel network, while
maintaining a porosity that allowed
satisfactory water holding capacity. A
strong gel network with satisfactory water
holding capacity is desirable because the
water held in the network acts as a
plasticizer [27]. This will render the yogurt
to be firm but not rigid. Also, this will allow
the yogurt to retain water-soluble
nutrients, such as minerals and the
remaining native whey proteins, making
the yogurt more nutritious.
The yogurt gel made from the milk sample
with 0.35 whey protein-to-casein ratio
required the largest pressure of more than
5000 Pa for apparent collapse, and expelled
very little liquid compared to the other two
samples (Fig. 2). It was deemed that at a
25
 Mariano JC et al. / Acta Manilana 59 (2011)

whey protein-to-casein ratio of 0.35, the network without making the gel rigid and
amount of whey protein must have been dense. This is relevant to sensory and
excessive so that a massive protein-protein overall quality related to mouthfeel and
interactions as well as casein cross-linking spooning the yogurt.
via denatured whey proteins took place
during heating and acidification, making a The yogurt gel from the milk sample with
strong network with small pores [28]. This 0.35 whey protein-to-casein ratio allowed
made the yogurt gel very dense and, a shorter length of the puncture glass to be
therefore, rigid and with small water imbedded in it. This must be due to the
holding capacity. dense and highly cohesive texture of the
yogurt, which must have been rendered by
Puncture test. The experimental results (Fig. a more massive protein-protein interaction
3) show that the yogurt gel samples from and casein cross-linking due to excessive
the blank and the milk with 0.25 whey amount of whey proteins [28]. Once again,
protein-to-casein ratio allowed the same this is consistent with the results of the
length of the puncture glass to be imbedded compression test.
in them within the predetermined time
interval. However, close observation The combined results of the compression
indicated that the yogurt gel from the blank and puncture tests indicate that the
milk sample expelled more liquid. This addition of whey proteins improved the
observation is in agreement with the strength and stability of the yogurt gel
compression test results based on the water network, which lead to the improvement
expelled upon puncture. of water holding capacity of the gel.
However, an excessive amount of whey
It is interesting to note that, while the proteins may also render the gel to be rigid,
yogurt from the milk with 0.25 whey dense and with reduced water holding
protein-to-casein ratio exhibited greater capacity, which are characteristics of
network strength in the compression test, overly stabilized gel and are, generally, not
its response to the puncture test was desirable [1]. While samples made from the
similar to that of the less strong yogurt from blank and 0.25 whey protein-to-casein milk
the blank. This indicates that the addition exhibited similar behavior during
of whey protein strengthened the gel puncture test, the latter exhibited greater

3.6
3.2
2.8 A
14 12 12 Blank Ratio
2.4
Length of Imbedded Rod (mm.)

10.5 10.5 0.25 Ratio

% Liquid Expelled

12 2 0.35 Ratio
9 Drop Height = 8 cm.
10 8
1.6
Blank Ratio
8 0.25 Ratio 1.2 B
0.35 Ratio
6 0.8
4 0.4 C
2 A A B B C C 0
0
Initial Final Initial Final Initial Final –0.4
Yogurt Samples –0.8

Figure 3. Imbedded length of the glass puncture Figure 4. % Liquid expelled from yogurt made using:
instrument in the yogurt made using: (A) blank, (A) blank, and milk with (B) 0.25, and (C) 0.35
and in the yogurt samples with (B) 0.25, and (C) whey protein-to-casein ratios.
0.35 whey protein-to-casein ratios.

26
 Improvement of textural properties of yogurt
gel strength in the compression test.
However, whether this translates to
greater stability against syneresis during
storage is unclear. Therefore, syneresis tests
were subsequently performed.
Syneresis. Syneresis is the extraction or
expulsion of liquid from a gel [29]. Figure 4
shows the percent liquid expelled from
yogurt made from blank and milk samples
with 0.25 and 0.35 whey protein-casein
ratios. The data shows that yogurt samples
from milk with no added whey protein had
the highest percentage of expelled water
compared with yogurts from the milk
samples with added whey proteins.
The blank exhibited high percentage of
liquid expulsion because of weak protein
interactions due to limited amount of whey
proteins, which lead to weak gel network.
Samples with added whey proteins
exhibited relatively smaller amount of
liquid expelled compared with the yogurt
from blank. This can be attributed to
sufficient amounts of whey proteins for k-
casein-b-Lg interactions [30]. In addition,
cross-linking of caseins by polymerized
whey proteins enabled the forming of a
stable gel network to hold more water
within a prolonged period. On the other
hand, excessive amount of whey protein
may lead to the formation of whey protein
aggregates in addition to whey protein-
linked casein aggregates, which result in
pore tightening and cause lowering of water
holding capacity [1].
Time dependent syneresis. The effect of the
increased whey protein-to-casein ratio
was studied by Lorenzen et al. [31] where
it was found that cross-linking of protein
chains can stabilize the three-dimensional
network of yogurt gel and prevent liquid
expulsion as a result of a decrease in gel
porosity, thus reducing syneresis.
The combined results of the compression
and puncture tests indicate that, compared
with the other two samples, the yogurt gel
prepared from milk with 0.35 whey
protein-to-casein ratio was denser and,
therefore, must have been less porous [32].
Therefore, its water holding capacity can
be expected to be smaller compared with
the other samples. Results of the syneresis
test (Fig. 4), showing the 0.35 sample to have
the lowest % liquid expelled confirmed this
hypothesis.
Forced syneresis. Syneresis during
prolonged storage was simulated by
subjecting the samples to centrifugation.
The %liquid expelled from the different
yogurt samples made from milk samples
of different whey protein-to-casein ratios
were tested (Fig. 5). The experimental results
can verify the water holding capacity and,
therefore, the porosity of the yogurt samples
as well. The amount of liquid separated
from the sample affirmed that the yogurt
with more whey proteins was more
resistant against syneresis.
Based on the results (Fig. 5), the 0.35 whey
protein-to-casein ratio had the smallest
amount of water expelled, which is
consistent with the results gathered from
the time-dependent syneresis. This
34.8
34.4
A
34.0
33.6 Blank Ratio
33.2 0.25 Ratio
% Liquid Expelled
32.8 B 0.35 Ratio
32.4
32.0
31.6
31.2
30.8
30.4
30.0
29.6 C
29.2
28.8
28.4
28.0
Figure 5. Estimation of the water holding capacities
of yogurt samples made using: (A) blank, and
milk with (B) 0.25, and (C) 0.35 whey protein-
to-casein ratios.
27
 Mariano JC et al. / Acta Manilana 59 (2011)

15
A The overall results of the syneresis tests are
14
13 consistent with the results of the previous
12 tests, which indicated that the yogurt gel
10
11 Blank Ratio made from milk sample with 0.25 whey
0.25 Ratio
9 0.35 Ratio protein-to-casein ratio had the most
% Syneresis

8
satisfactory gel strength and water holding
7
6
B capacity. This may potentially lead to
5 satisfactory sensory quality.
4
3
2
C CONCLUSION
1
0 The adjustment of whey protein-to-casein
Yogurt Samples
ratio in reduced-fat carabao milk to 0.25,
Figure 6. % Syneresis from yogurt made using (A) and the heat treatment of this milk at a
blank, and milk with (B) 0.25, and (C) 0.35
whey protein-to-casein ratios.
temperature higher than pasteurization
temperature of 80°C for 25 min, resulted in
significant improvements of yogurt texture.
indicates that this sample had the smallest The yogurt gel exhibited relatively high
water holding capacity while the blank had water holding capacity and low
the highest water holding capacity. This is %syneresis, moderate gel setting time as
indicative of the lesser porosity of the 0.35 shown by the moderate increase in
yogurt sample, which must have been viscosity during acidification, and
brought about by the highly stabilized gel moderate gel strength compared with those
network from the excessive amount of samples made from the milk with whey
whey proteins added [22]. protein-to-casein ratio adjusted to 0.35 and
that with unadjusted ratio. These
Figure 6 shows the %syneresis based on the
characteristics are deemed ideal in
total liquid expelled on the seventh day of
producing reduced-fat, high-protein, set
test and the liquid expelled by
yogurt without using carbohydrates-
centrifugation. While the yogurt sample
based stabilizers and texture enhancers.
made from the blank had the highest water
holding capacity, Fig. 6 indicates that it also
ACKNOWLEDGEMENT
gave the highest %syneresis while the
yogurt from the milk with 0.35 whey The authors thank the University of Santo
protein to casein ratio had the smallest Tomas Research Center for the Natural and
%syneresis. The yogurt sample from the Applied Sciences for the research fund; and
milk with 0.25 whey protein-to-casein ratio Fonterra-Philippines, through Ms. Theresa
had a water holding capacity close to that Jusay, for providing the WPI used in the
of the blank (Fig. 5), but showed an average experiments as sponsorship in kind.
%syneresis that was more than twice lower
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