Ultrasonics Sonochemistry 38 (2017) 772782

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Ultrasonics Sonochemistry
journal homepage: www.elsevier.com/locate/ultson

High solids emulsions produced by ultrasound as a function of energy

density
Larissa Consoli , Guilherme de Figueiredo Furtado, Rosiane Lopes da Cunha, Mriam Dupas Hubinger
Department of Food Engineering, School of Food Engineering, 80, Monteiro Lobato Street, University of Campinas, 13083-862 Campinas, SP, Brazil

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

Article history: The use of emulsifying methods is frequently required before spray drying food ingredients, where using
Received 29 August 2016 high concentration of solids increases the drying process yield. In this work, we used ultrasound to obtain
Received in revised form 28 November 2016 kinetically stable palm oil-in-water emulsions with 30 g solids/100 g of emulsion. Sodium caseinate,
Accepted 29 November 2016
maltodextrin and dried glucose syrup were used as stabilizing agents. Sonication time of 3, 7 and
Available online 1 December 2016
11 min were evaluated at power of 72, 105 and 148 W (which represents 50%, 75% and 100% of power
amplitude in relation to the nominal power of the equipment). Energy density required for each assay
Keywords:
was calculated. Emulsions were characterized for droplets mean diameter and size distribution, optical
Sonication
Palm oil
microscopy, confocal microscopy, f-potential, creaming index (CI) and rheological behavior. Emulsions
High energy emulsification presented bimodal size distribution, with D[3,2] ranging from 0.7 to 1.4 lm and CI between 5% and
Flocculation 12%, being these parameters inversely proportional to sonication time and power, but with a visual kinet-
ically stabilization after the treatment at 148 W at 7 min sonication. D[3,2] showed to depend of energy
density as a power function. Sonication presented as an effective method to be integrated to spray drying
when emulsification is needed before the drying process.
2016 Elsevier B.V. All rights reserved.

1. Introduction feed characteristics (viscosity, concentration, density) and equip-

Emulsion-based formulations are frequently used in the phar-
maceutical, cosmetics and food fields. More recently, they have
been recognized as potential vehicles for delivering bioactive com-
pounds [1]. The majority of the most known substances with the
ability to bring beneficial health properties, such as polyunsatu-
rated fatty acids, carotenoids, some polyphenolics, and vitamins,
exhibit lipophilic properties. So, oil-in-water emulsions may be
used to protect and deliver formulations loaded with such
compounds.
Spray drying emulsions is an alternative to enhance their shelf
life and also to improve handling properties, transport and storage
issues. This technique is widely used in pharmaceutical, chemical,
cosmetic and food industries [2]. Furthermore, spray drying is the
most used microencapsulation technique of food ingredients. The
process is energy intensive, but cost effective, continuous and flex-
ible [3,4].
Many factors related to the process (e.g., gas inlet temperature,
feed flow rate, drying gas flow rate, atomizing pressure), liquid
Corresponding author.
E-mail addresses: larissa.consoli@gmail.com (L. Consoli), furtado.gf@gmail.com
(G. de Figueiredo Furtado), rosiane@unicamp.br (R.L. da Cunha), mhub@unicamp.br
(M.D. Hubinger).
ment design (co-current or counter-current gas flow, atomizer
type, design) exert influence on the properties of spray-dried
products [2]. Regarding the properties of the powder obtained by
spray drying, it was shown that they are affected by the infeed
emulsions. Feeding liquids with high concentration of solids
increases the drying process yield and, for consequence, reduces
operational costs. It is then common to use infeed solids concentra-
tion as high as possible, being the fluid viscosity the main limiting
factor, because excessive viscosity hinders atomization and the
drying performance [5]. Likewise, characteristics such as stability
and droplet size must also be considered when elaborating formu-
lations for drying. Particularly, the droplet size has a straight rela-
tion to the retention of the materials incorporated as active, since
lower droplet diameter results in higher retention efficiency [6,7].
The ratio between oil phase and encapsulating agents concentra-
tion is also important, because there must be sufficient amount
of carrier for the protection of the oil. Typical total solids concen-
trations usually employed for the production of food emulsions
for spray drying are 20% [8], 30% [911], and 40% [1215], while
the ratio (w/w) oil/total solids ranges from 0.10 [15] up to 0.50
in extreme cases [8].
Ultrasound processors have been successfully employed for the
production of nano and sub-micron oil-in-water emulsions,

http://dx.doi.org/10.1016/j.ultsonch.2016.11.038
1350-4177/ 2016 Elsevier B.V. All rights reserved.
 L. Consoli et al. / Ultrasonics Sonochemistry 38 (2017) 772782
stabilized by a great variety of low molecular weight emulsifiers,
proteins, hydrocolloids and combinations among these materials.
Although emulsification by ultrasound technique has been much
explored in the last years [1618], the integration of this technol-
ogy with spray drying still need investigation, since it perfectly
matches the requirement of producing low diameter oil-in-water
emulsions. Current works concerning the use of ultrasound to pro-
duce oil-in-water emulsions employ moderate to high solids con-
centration, such as 23% [19,20], 3135% [2123] and 42% [22].
However, the oil/total solids ratio used in these works is too high
(> 0.80) and thus not suitable for spray drying.
The choice for natural biopolymers, such as proteins, for emul-
sions stabilization has been preferred in relation to synthetic emul-
sifiers, as a clean label claim [24]. Dairy proteins have been
extensively used as food ingredients due to their excellent emulsify-
ing properties [25]. The caseins, more specifically, have relevant
importance as emulsifiers, because they are capable of rapidly con-
ferring low interfacial tension during droplet formation, due to the
strong amphiphilic characteristics of the major individual caseins
[26]. Sodium caseinate is obtained by the acidification of the whole
casein to pH 4.6, followed by addition of NaOH up to pH 6.7, and fur-
ther pasteurization and spray drying [27]. Because of its outstanding
emulsifying properties, sodium caseinate has been used as ingredi-
ent in many products, and much investigation has been conducted in
order to better clarify its role in emulsion stabilization and to opti-
mize its performance for each system in which it is applied.
Apart from emulsifiers, the use of hydrocolloids as thickening
agents is also common in emulsions formulations, because their
ability to increase viscosity favors stability. Additionally, when
designing emulsions for spray drying, hydrocolloids also con-
tribute for reaching a suitable concentration of solids. Starch
derivatives, such as maltodextrins, and glucose syrups, have been
commonly used with this finality and, beyond functionality, they
represent an economic choice in comparison to other ingredients,
such as some gums and modified starches [5].
This work had as main goal evaluating the effect of the ultra-
sound process parameters of power amplitude and sonication time
on high solid emulsions formation, in order to set the best condi-
tion to produce food emulsions for further spray drying.
2. Material and methods
2.1. Material
Aqueous phase of emulsions was composed of sodium caseinate
(90.7 g protein/100 g on dry weight basis, determined by Kjeldahl
method [28]), which was kindly donated by Alibra Ingredients
(Campinas, SP, Brazil), maltodextrin MOR-REX 1910 (Dextrose
Equivalent (DE) = 912) and Dried Glucose Syrup (DGS) MOR-REX
1930 (DE = 2630), both supplied by Ingredion Brazil Industrial
Ingredients Ltd. (Mogi-Guau, SP, Brazil). The oily phase was com-
posed of palm oil (Cargill Foods Brazil Itumbiara, GO, Brazil) and
ethanol. All other materials used in this work were of analytical
grade.
2.2. Emulsions preparation
2.2.1. Aqueous phase preparation
Emulsions had total solid concentration of 30 g/100 g, where
the oily phase represents 15 g/100 g of the total solids. These con-
centrations of total solids and oil were set as optimum by Frascareli
et al. (2012), and thus the formulation will be used to carry an
active material in our further studies. Sodium caseinate, maltodex-
trin and DGS, at the concentrations of 10.9, 37.9 and 37.9 g/100 g of
solution, respectively, were individually solubilized in Milli-Q
773
water containing 0.01% sodium azide, in order to avoid microbial
growth. The solutions were stirred overnight at room temperature
(
25 C). After this period, they were mixed with equal propor-
tions of maltodextrin and DGS, until it achieved a final protein/car-
bohydrate ratio of 1/5. The pH was then adjusted to 7.5 with NaOH
1 M.
2.2.2. Coarse emulsion
Ethanol (0.4 g/100 g emulsion) was added to palm oil at 50 C
and the mixture was magnetically stirred for 15 s. This oily phase
was added to the aqueous mixture prepared as described in Sec-
tion 2.2.1 and the system was stirred at 4000 rpm for 2 min, using
a rotorstator device (Silverson L5 M-A Laboratory Mixer, Che-
sham, Buckinghamshire, UK). Assays were performed with 300g
(
285 mL) of emulsion.
2.2.3. Ultrasound treatment
Coarse emulsion was completely transferred to a jacketed bea-
ker connected to a thermostatic bath, in order to keep the temper-
ature below 35 C in all assays. An ultrasonic processor Q700
(700 W full power, 20 kHz frequency QSonica Newton, CT,
USA) was used. The probe ( = 12.5 mm) was immersed 24 mm
into the liquid. Sonication time and ultrasound power amplitude
were varied in order to obtain optimum conditions for the produc-
tion of palm oil-in-water emulsions. Emulsions were sonicated for
3, 7 and 11 min at power amplitudes of 50%, 75% and 100%, which
resulted in power values of 72, 105 and 148 W, as given by the
ultrasound equipment. In this document, all other mentions to pro-
cess conditions will be done by means of power and not power
amplitude. Ranges within parameters were evaluated were defined
considering results from preliminary trials, in which we verified
that 3 min was the minimum period of sonication enough to pro-
mote droplet disruption, and intervals lower than 4 min did not
cause significant changes in the system. Since this formulation will
be used in further studies for spray drying, with the addition of a
bioactive material, we considered that the use of excessive sonica-
tion periods could bring negative effects to the bioactive mole-
cules, and then set 11 min as the maximum sonication period. All
assays were performed in duplicate.
2.3. Emulsions characterization
Emulsions obtained both by rotor-stator device and by sonica-
tion were characterized immediately after preparation and 24 h
after this period [9,13,29]. When creaming was observed, the lower
layer of the emulsions was characterized, since this is the phase of
interest when performing spray drying process, due to its higher
concentration of solids.
2.3.1. Emulsions droplet size and size distribution
A laser light scattering equipment (Mastersizer 2000, Malvern
Instruments, Worcestershire, UK) was used to perform size mea-
surements. Droplet mean diameter was expressed as the volume-
surface mean diameter D[3,2] (Sauter mean diameter Eq. (1))
and as the volume-weighed mean diameter D[4,3] (De Brouckere
mean diameter Eq. (2)). The Span, which gives the width of size
distribution, was calculated from Eq. (3). Each sample was ana-
lyzed in triplicate.
X
ni D3i
D3;2 X 1
ni D2i
X
ni D4i
D4;3 X 2
ni D3i
774 L. Consoli et al. / Ultrasonics Sonochemistry 38 (2017) 772782
D0:9  D0:1
Span
D0:5
where ni is the number of droplets with diameter Di , and D0:1 , D0:5
and D0:9 represent the diameter of accumulated distribution of
10%, 50% and 90% of total droplets, respectively.
2.3.2. Optical microscopy
Emulsions microstructure was visualized using an optical
microscope Carl Zeiss Axio Scope A1 (Gottingen, Alemanha), with
a 100x objective. Images were captured by the software Axio
Vision Rel. 4.8.
2.3.3. Confocal laser scanning microscopy (CLSM)
CLSM was used in order to ensure the formation of oil-in-water
emulsions, and also for observing the modification caused by pro-
cess conditions with improved image quality. The oily phase of
emulsions was labeled with the Nile Red dye, in the concentration
of 0.002 g/100 g oil. Samples were examined using a Zeiss LSM
780-NLO confocal on an Axio Observer Z.1 microscope (Carl Zeiss
AG, Germany) using 63x and 100x objectives. Images were col-
lected using 514 nm laser line for excitation and 527728 emission
filters for Nile Red, with pinholes set to 1 airy unit for each channel,
1024  1024 image format, and 1x and 2x optical zoom.
2.3.4. f-potential
The determination of f-potential of oil droplets was made using
a Zetasizer Nano Series (Malvern Instruments, Worcestershire, UK).
The measurement is based on the application of an electrical field
to the electrodes of the cell in which sample is placed. f-potential is
related to the mobility of the charged droplets between the two
electrodes. 100 lL of each emulsion were diluted in 9.9 mL of
Milli-Q water. Measurements were performed in triplicate.
2.3.5. Creaming index (CI)
Immediately after preparation, 100 mL of each emulsion were
transferred to cylindrical glass tubes ( = 26 mm), sealed and kept
at room temperature (
25 C) for 24 h. Phase separation was
expressed as the ratio between the height of the superior phase
(H) and the total height of liquid (HT), as shown in Eq. (4). This
measurement was performed in duplicate.
H
CI
HT
2.3.6. Rheological behavior
Emulsions flow curves were obtained using a cone-plate geom-
etry ( = 60 mm) in a stress-controlled rheometer AR1500ex (TA
Instruments, Elstree, UK). The analysis was done in the range
between 0 and 1000 s1, in 3 min, in a three-step sequence: up-
down-up. The second up curve data was fitted to the Newtonian
model (Eq. (5)) and to the Power Law (Eq. (6)). Rheological mea-
surements were conducted only immediately after sample prepa-
ration, due to the creaming that was observed 24 h later.
r gc_
r k  c_ n
in which r is the shear stress (Pa), g is the viscosity (Pa.s), c_ is the
shear rate (s1), k is the consistency index (Pa.sn) and n is the
behavior index (dimensionless).
2.4. Energy density calculation
The concept of energy density (ED) is applicable for the produc-
tion of emulsions by mechanical devices. It relates the power intro-
duced into the device to the processed volume of the fluid [30]. For
3
ultrasound processors operating in batch, ED is calculated by Eq.
(7) [31], which can also be used for rotor-stator devices, replacing
tson by the stirring time used. The power, in this case, can be
obtained by Eq. (8).
ED P  t son =V 7
where P is the Power delivered to the system (W); tson is the soni-
cation time and V is the volume of the fluid.
P U  i 8
where P is the power delivered to the system (W), U is the electric
tension (V Volts) and i is the electric current (A Ampre).
The ultrasound processor used in this work displayed, during
the process, the power delivered to the fluid and, at the end of
the process, the total amount of energy employed. The energy val-
ues displayed by the equipment were very close to those obtained
using Eq. (7). The energy input of the rotor-stator device was calcu-
lated using Eq. (8), using the electric current measurement given
by the equipment (Silverson L5 M-A Laboratory Mixer, Chesham,
Buckinghamshire, UK). The results for energy density were then
correlated to the volume-surface mean diameter (D[3,2]) of emul-
sions droplets, measured immediately after preparation. Power
and logarithmic functions, Eqs. (9) and (10), were fitted to experi-
mental data for comparison.
D3;2 aEDb 9
D3;2 a ln EDb 10
where D[3,2] is the volume-surface mean diameter, ED is the energy
density and a and b are constants.
2.5. Statistical analysis
The data were statistically analyzed by Tukey test using the
software Microsoft Excel (2016). Differences between means were
considered significant at a 95% confidence level (p 6 0.05).
3. Results and discussion
4 3.1. Emulsions droplet mean diameter, size distribution and
microstructure
Sonication significantly reduced the droplet diameter of emul-
sions, independently of the intensity of the treatment, as shown
by the parameters D[3,2] and D[4,3] in Table 1. The reduction in dro-
plet diameter, promoted by sonication, can also be clearly seen on
the micrographs from Figs. 1 and 3. With respect to the droplets
size distribution, the polydispersity indices (Span) of both coarse
and sonicated emulsions were very similar, although the increase
in sonication time affected it, as will be further discussed in this
section.
5 Regarding the sonicated emulsions, the majority of them pre-
sented bimodal size distribution, independently on the power
6 employed. However, droplet size was strongly affected by such fac-
tor: in Fig. 2, it can be seen that, for the same sonication time, the
curves were shifted to the left when the power was increased. This
fact is also demonstrated by data presented in Table 1: at a fixed
sonication time, the higher the power amplitude, the lower D[3,2]
and D[4,3] mean diameters. Sonication time affected these parame-
ters in the same way. That is, given a fixed power, droplet diame-
ters reduced when the ultrasound treatment was longer, except for
a slight increase in D[4,3] of emulsions sonicated for 11 min, in com-
parison to those sonicated for 7 min at 148 W. These observations
 L. Consoli et al. / Ultrasonics Sonochemistry 38 (2017) 772782 775

Table 1
Mean droplet diameter of palm oil-in-water emulsions obtained by rotor-stator device and ultrasound.

Power (W) Sonication time (min) Immediately after preparation 24 h after preparation
D3;2 D4;3 Span D3;2 D4;3 Span
(lm) (lm) (lm) (lm)
0 4.28 0.29fA 9.55 0.90dA 1.87 0.11aA 7.98 1.15dB 17.87 2.70bB 1.79 0.16abA
eA bcA aA cA aA
72 3 1.46 0.066 2.25 0.06 1.76 0.02 1.47 0.15 2.27 0.32 1.77 0.10aA
7 1.38 0.05deA 2.38 0.11cA 2.37 0.09bdA 1.34 0.15abcA 2.18 0.31aA 2.23 0.16abcA
11 1.18 0.02cdA 2.05 0.08abcA 2.54 0.18bcA 1.11 0.11abcA 1.90 0.25aA 2.47 0.27bcA
105 3 1.36 0.08deA 2.18 0.39bcA 1.87 0.27aA 1.41 0.18bcA 2.17 0.55aA 1.86 0.41abA
7 1.05 0.14bcA 1.99 0.31abcA 2.76 0.32bcB 0.99 0.01abcA 1.69 0.06aA 2.21 0.18abcA
11 0.87 0.01abA 1.69 0.08abcA 2.70 0.28bcA 0.86 0.02abA 1.65 0.14aA 2.61 0.38cA
148 3 1.24 0.04cdeA 2.07 0.10abcA 2.03 0.23adA 1.28 0.00abcA 2.07 0.01aA 2.06 0.02abcA
7 0.73 0.01aA 1.22 0.13aA 1.64 0.14aA 0.73 0.03aA 1.26 0.23aA 1.71 0.19aA
11 0.68 0.05aA 1.45 0.10abA 2.95 0.40cA 0.72 0.01aA 1.40 0.10aA 2.73 0.57cA

Values are averages of two experiments, with each individual sample analyzed in triplicate. Different lowercase letters in each column and different capital letters in each
row, for the same parameter, represent statistically significant difference (p 6 0.05).

confirm that emulsions characteristics are controlled both by son-
ication time and by power when a fixed frequency is used.
Increasing sonication time from 7 to 11 min, at fixed power,
resulted in similar D[3,2] diameters, which suggests that there is a
limit on the time of ultrasound treatment that still promotes
droplet rupture when using the same power. Above this limit,
any amount of energy delivered to the system will be dissipated
as heat. Although the volume-weighed diameter D[4,3] showed
dimension variation due to process conditions, it did not follow
the trends of increasing or decreasing as straight as D[3,2] did,
and its value was statistically similar on most of the assays.
The effect of sonication time was much stronger on the Span of
emulsion droplet sizes than that of power. Long periods of sonica-
tion expose droplets to high amounts of energy, promoting not
only droplet rupture, but also droplet collision and, thus, re-
coalescence and flocculation, explaining the larger variability in
droplet size. Furthermore, the droplet rupture does not affect uni-
formly all the droplets, since the droplets in locations closer to the
probe are exposed to a much more intense cavitation than those
more distant from it [31]. So, part of the droplets will keep the orig-
inal size or have lower reduction, and part of them will be signifi-
cantly smaller. This also corroborates to the presence of the two
peaks in size distribution.
After 24 h of emulsions preparation, analyses were repeated for
comparison. In coarse emulsions, diameters were significantly
increased after this period, which may indicate droplet coalescence
with time. Although creaming was observed in the sonicated emul-
sions (further discussed in Section 3.2), the remaining droplets in
the continuous phase presented little or no variation in the mean
diameters and size distribution, as it can be seen in Table 1 and
in Fig. 2. Even with no statistical significance, there was a trend
towards reduction of emulsions Span after 24 h, which may be evi-
dence of larger droplets migrating to the cream phase.
Rotor-stator equipments have the ability to mix liquids from
separated phases or to reduce droplet size of a coarse emulsion.
When fed into the device, the liquid flows through a narrow gap
between a rotating disc (rotor) and a static disk (stator). Droplets
are formed or reduced when colliding against the stator wall by
the shear stress in the gap [32], but sizes smaller than 1 lm are
not generally reached. In the sonication process, the mechanism
for droplet rupture is the acoustic cavitation phenomenon, which
consists of growth and collapse of micro bubbles of gas dissolved
in the medium. The collapse of these bubbles produces powerful
shock waves that radiate near the probe and propagate into the
medium, leading to droplets breakdown. This energy can also be
converted into thermal energy, resulting in increased temperatures
[32,33]. The amount of energy employed in rotor-stator devices is
usually significantly lower than that required for sonication pro-
cesses and, therefore, the mean diameter of emulsions droplets
obtained by ultrasound are expected to be very smaller. The results
found in this work are in accordance with this observation and
with those of previous works [21,22,31,34].
The achievement of a saturation diameter with the increase in
sonication time, which was found in our work, follows the trend
described by other authors that also used ultrasound to produce
oil-in-water emulsions [31,34,35].
The main differences in the trends between D[4,3] and D[3,2]
diameters may be related to the polydispersity of samples. The
presence of droplets with larger diameters has higher impact on
the calculation of D[4,3] (Eq. (2)) than on D[3,2](Eq. (1)) [36]. There-
fore, the volume-weighed mean diameter is frequently referred to
as more sensitive to the detection of fat droplet aggregation when
compared to the volume-surface mean diameter [20,21,37]. This
same reason also explains the increase in D[4,3] when sonication
time was incremented from 7 to 11 min at 148 W of power: the
last treatment resulted in the broadest size distribution of all,
and the parameter calculation was then affected. The larger distri-
bution may have occurred due to droplet coalescence favored by
the excessive amount of energy employed in this treatment. This
phenomenon is also referred as over-processing [31].
The micrographs obtained by confocal laser scanning micro-
scopy (Fig. 3) confirmed that our process produced oil-in-water
emulsions, where the oil droplets are red-labeled by the Nile Red
dye. Variations in size distributions, already discussed in this sec-
tion, can also be clearly observed.
3.2. f-potential and kinetic stability
Immediately after preparation, the f-potential of emulsions var-
ied from 54.596 to 48.247 mV (Table 2), with no significant dif-
ference among assays.
The negative charge occurred mainly because of sodium casei-
nate, a dairy protein which acted as stabilizing agent in the system.
Sodium caseinate is negatively charged at pH 7.5 (
50 mV data
not shown), at which emulsions were prepared. The carbohydrates
used also have negative charge at this pH value, although con-
tributing little compared to the protein. Systems with net charge
between 30 and 60 mV, as were emulsions here obtained, are gen-
erally considered moderately stable due to electrostatic repulsion
[38].
The kinetic stability was observed visually by taking the height
measurements of the cream and the continuous layers of emul-
sions placed in glass tubes, in a period of 24 h after preparation.
776 L. Consoli et al. / Ultrasonics Sonochemistry 38 (2017) 772782

Fig. 1. Optical micrographs of emulsions prepared at different process conditions. Scale bar: 5 lm.

Results are expressed as the creaming index (CI) and are presented
in Fig. 4.
Creaming index of sonicated emulsions is more strongly
affected by power than by sonication time. Power of 72 W showed
low effectiveness against phase separation at all sonication times.
The same observation is valid for 3 min sonication, at any power
used. For emulsions sonicated for more than 3 min, the higher
the power employed, the lower the creaming index. The most
stable emulsions were those with smallest droplet sizes (148 W,
with 7 or 11 min of sonication).
At 148 W of power, the use of a sonication time higher than
7 min did not imply significant reduction of creaming index. This
is in accordance with the observations found in Section 3.1, where
the same variation on the process conditions resulted in similar
droplet size. This result is also important because shows that
equivalent stability properties may be achieved even when signif-
icantly lower energy is employed. Considering these findings, we
assume that the condition of 148 W of power and 7 min of sonica-
tion time had the best performance and will be set for the continu-
ity of the work.
It is interesting to notice that, although the CI obtained for the
coarse emulsion was close to those obtained for some fine emul-
sions, a more intense separation was observed in it. Fig. 5 illus-
trates the pictures of the coarse emulsion and that sonicated for
7 min at 148 W, observed after 24 h of preparation. As can be seen,
the sonicated emulsion remained wholly turbid, and the line
 L. Consoli et al. / Ultrasonics Sonochemistry 38 (2017) 772782 777

Fig. 2. Droplet size distribution diagrams of emulsions obtained using different process conditions.

between the cream layer and the continuous phase is difficult to
visualize. For the coarse emulsion, the separation is much more
clear, and the continuous phase shows a very low turbidity. The
upper phase is more intensively yellow colored, as is the palm
oil, which demonstrates that separation was more intense.
Emulsions stabilized by proteins are principally affected by four
main destabilization mechanisms: creaming, coalescence, floccula-
tion and Ostwald ripening, but the later occurs less frequently in
oil-in-water food emulsions, since oils commonly used are mainly
composed of long chain tryacylglycerols that show low solubility
in water, and thus do not diffuse through the water phase [39,40].
Creaming is the instability mechanism that occurs by gravita-
tional separation, due to the lower density of the disperse phase.
It is one of the most common mechanisms of emulsion destabiliza-
tion found in the food industry. Flocculation is the process by
which droplets associate to each other, without losing their indi-
vidual characteristics [41]. In some cases, the formation of droplet
aggregates by flocculation may accelerate the creaming process,
because those aggregates can move to the top of the emulsion
more rapidly than individual droplets [27].
The relative concentration of protein-to-oil is one of the main
parameters when considering flocculation in protein-stabilized
778 L. Consoli et al. / Ultrasonics Sonochemistry 38 (2017) 772782

Fig. 3. Micrographs obtained by confocal laser scanning microscopy for emulsions obtained by the rotor-stator treatment and sonication at 72 W, 105 W and 148 W for
7 min. Scale bar: 5 lm.

Table 2
f-potential measured for oil-in-water emulsions immediately and 24 h after
preparation.

Power Sonication time f-Potential (mV)

(W) (min)
Immediately after 24 h after
preparation preparation
0 54.60 5.19aA 41.53 8.27aB
A
72 3 49.66 4.82 41.64 4.18aB
7 49.14 6.04aA 44.31 9.17aA
11 51.82 0.76aA 45.83 2.89aB
105 3 48.25 3.23aA 43.73 4.94aA
7 48.38 1.51aA 44.39 0.65aB
11 52.76 0.98aA 42.41 2.75aB
148 3 51.21 4.28aA 43.47 4.74aB
7 51.04 1.11aA 46.07 4.63aB
11 50.28 1.32aA 46.46 2.32aB
Fig. 4. Creaming index obtained for coarse and sonicated emulsions after 24 h of
Values are averages of two experiments, with each individual sample analyzed in preparation. The bar of 0 W power and 0 min of sonication corresponds to coarse
triplicate. Different lowercase letters in each column and different capital letters in emulsion, i.e., the rotor-stator treatment. Same letters represent statistically similar
each row represent statistically significant difference (p 6 0.05). results with p P 0.05.
 L. Consoli et al. / Ultrasonics Sonochemistry 38 (2017) 772782
Fig. 5. Comparison between coarse emulsion (A) and emulsion sonicated for 7 min
at 148 W, after 24 h of preparation.
emulsions, because it directly affects the interdroplet interactions.
Bridging flocculation is known to occur when the amount of pro-
tein in the system is not sufficient to cover the surface of all dro-
plets, and the layer recovering droplets is then insufficient to
cause electrostatic repulsion among them, which leads to the irre-
versible formation of clusters [27]. On the other hand, depletion
flocculation represents the opposite situation, when excessive
amount of protein is present in the medium. In this case, the
unbound protein in the solvent self-associates, forming flocs. This
type of interaction is weaker compared to the bridging flocculation,
and thus considered as reversible [42].
Besides protein:oil ratio, other factors such as pH, calcium ions,
ionic strength, and the presence of alcohol, non-ionic surfactants,
sucrose or polysaccharides may also interfere in the stability of
emulsions [27].
In Figs. 1 and 3, the micrographs suggest that the formation of
the cream layer in the emulsions, after 24 h of their production,
has occurred not only for creaming, but also for droplets floccula-
tion, due to the aggregation that can be observed. Indeed, from
Table 2 we can see that, in the period of 24 h, f-potential of emul-
sions from most of the assays underwent a significant reduction.
Decrease in charge is associated with the flocculation destabiliza-
tion mechanism, since, once charge is reduced, the electrostatic
repulsion among droplets may not be sufficient to keep them sep-
arated, leading to the formation of droplet aggregates [39]. Our
results are consistent with previous reported works [21,43], in
which flocculation has been also mentioned as one of the main
instability mechanisms in sodium caseinate stabilized emulsions
produced by sonication.
The presence of ethanol in emulsified systems stabilized by pro-
teins can also contribute to the occurrence of flocculation. Ethanol
is supposed to flatten the protein layer over droplets surface in oil-
in-water emulsions, which impairs the steric repulsion. Since it is a
poor solvent to dairy proteins, aggregation is another mechanism
779
that can be related to emulsions destabilization induced by ethanol
[44]. Studies concerning the effect of ethanol on the properties of
sodium caseinate stabilized emulsions have shown that only con-
centrations higher than 25% [45] 30% (v/v) [46] or 40% [44] of this
component would cause significant destabilization in the systems.
Inversely, ethanol can also act as a cosurfactant in oil-in-water
emulsions, decreasing the interactions between surfactant mole-
cules and the interface energy [47]. Its combination with calcium
ions under a restrict concentration range in casein stabilized
emulsions also showed the ability to enhance stability to depletion
flocculation [45,48]. However, all the ethanol concentrations previ-
ously studied are much higher than that used in our work
(0.4 g/100 g of emulsion), and this amount can thus be considered
insufficient to affect caseinate structure [47].
Another factor that must be regarded is the use of polysaccha-
rides. The occurrence of flocculation is highly frequent in systems
containing hydrocolloids, such as food starch derivatives, which
were used in our work. Both mechanisms of bridging or depletion
flocculation are supposed to occur, depending on the interactions
happening between the hydrocolloids and the protein, but the lat-
ter is favored when the non-spherical shape of polysaccharides
molecules increase, as is our case. Furthermore, systems with
low oil concentrations (510%), like ours, are also more susceptible
to flocculation [49].
In spite of the formation of the cream layer on the top of the
emulsions within the period of 24 h, it is important to highlight
that the kinetic stability reached for some assays can be considered
satisfactory for spray drying, since this process is usually per-
formed in a short period after emulsion preparation. Furthermore,
it has been already demonstrated that the infeed solids content is
the main factor that contributes to the retention of volatiles and
encapsulation efficiency of food oils during spray drying. The
mechanism by which the concentration of solids influences the
retention of the active materials is by reducing the time required
to form a semi-permeable membrane at the surface of the drying
particle [6,7]. Also, previous works have indicated that the use of
low molecular weight carbohydrates, as is DGS in comparison to
maltodextrin, can provide better oil retention in spray dried
microparticles. So, the application of the formulation evaluated in
our work for the microencapsulation of oils and flavors can provide
good retention results [5052].
3.3. Rheological behavior and viscosity
Apart frequency, power and sonication time, which are process-
ing related parameters of ultrasound emulsification, viscosity of
the phases plays an important role on the characteristics of the
final product. Continuous phases with higher viscosities favor the
production of emulsions with smaller droplets. Similar results are
usually obtained when low viscosity oils are incorporated as dis-
perse phases in oil-in-water emulsions. Furthermore, the relation
between the viscosities of disperse and continuous phases is also
likely to be a parameter even more important than the individual
viscosity of each phase itself [17].
The rheology of caseinate emulsions is straightly related to the
colloidal interactions among the droplets, which depend on the
structure and composition of the adsorbed layer at the oil-in-
water interface [53]. Rheological measurements are expected to
bring valuable information about emulsion flocculation. Generally,
shearing can disrupt the flocs formed by excess of protein in the
medium, as is the case of depletion flocculation. In this situation,
a shear-thinning behavior is more likely to occur, since the floc
separation at higher shear rates decreases the apparent viscosity
[54].
In our system, emulsions prepared by rotor-stator and ultra-
sound devices presented Newtonian flow behavior (R2
0.9998).
780 L. Consoli et al. / Ultrasonics Sonochemistry 38 (2017) 772782
Table 3
Apparent viscosity determined for palm oil-in-water emulsions by fitting the Newtonian model to flow curves data (R2 > 0.9995 for all assays). Results are expressed in mPa.s.
Sonication time (min) Power (W)
0 72
0 13.84 0.12*
3 13.45 0.08aA
7 14.03 3.82aA
11
0 W power corresponds to the rotor-stator treatment. Values are averages of two experiments, with each individual sample analyzed in duplicate. Different lowercase letters
in each column and different capital letters in each row represent statistically significant difference (p 6 0.05).
*
Coarse emulsion viscosity was statistically similar (p P 0.05) to all other treatments.
Fig. 6. Correlation between D[3,2] and energy density (ED) required for each
emulsion.
The apparent viscosity ranged between 13.43 and 14.96 mPa.s
(Table 3), with no significant differences in flow behavior induced
by the sonication process. These values are a result of the high con-
centration of solids, especially the carbohydrates (maltodextrin
and DGS), which are 5-fold concentrated in relation to the protein.
Such carbohydrates are frequently added to emulsions as thicken-
ing agents, due to their ability to increase viscosity and thus to
retard creaming [49]. However, the carbohydrates concentration
and the increased viscosities were not sufficient to hinder flocs for-
mation and creaming. This result is in accordance with findings
previously reported by Liang et al. [55], who observed that the
use of maltodextrin in combination with sodium caseinate did
not imply improvements in stability of emulsions. Although the
addition of maltodextrin and DGS were not as effective as expected
when regarding the improvement of stability properties, their use
remains necessary because of the increase in solid concentration
provided by these components, because the use of higher solid
concentrations contributes for obtaining better process yield in
spray drying [5].
3.4. The effect of energy density (ED) on emulsions droplets mean
diameter
Fig. 6 relates ED obtained for each assay to the respective D[3,2],
measured immediately after emulsion preparation.
The ED delivered to the rotor-stator device was about 40 times
lower than that employed on the mildest condition on the ultra-
sound treatment (72 W of power, 3 min), and almost 300 times
lower than the most drastic one (148 W of power, 11 min), thus
corresponding to the point marked on the y axis. As one of the pro-
cessing conditions was increased, the mean diameter D[3,2]
105 148
13.43 0.25aA 13.83 0.25A
13.98 0.28abA 13.98 0.22A
14.73 2.23aA 14.96 0.21bA 14.30 0.21A
Table 4
Parameters found from fitting power and logarithmic functions (Eqs. (9) and (10),
respectively) to experimental data for energy density and volume-surface mean
diameter D[3,2], of coarse and sonicated emulsions. Parameters are dimensionless.
Function Parameter R2
a b
Power 4.56 0.29 0.9931
Logarithmic 4.34 0.66 0.9897
reduced, until apparent stabilization. This finding matches the
observations reported in Section 3.1, where a saturation diameter
was reached when sonication time was increased. That was
already expected since, as described in Section 2.4, sonication time
is accounted in the energy density calculations. Such results con-
firm that it is possible to achieve emulsions with sub-micron dro-
plets and low creaming index without using drastic process
conditions, i.e., saving energy and reducing operational costs.
Table 4 shows the parameters found by fitting power and loga-
rithmic functions to the experimental data presented in Fig. 6.
Although there is not yet a common sense among authors, param-
eter a is generally related to the properties of the fluid and param-
eter b is linked to droplet disruption mechanism [30], with this
correlation only postulated for the power fit. Indeed, the power
function better correlated to the data in comparison to the loga-
rithmic fit, but the latter also showed a good determination coeffi-
cient. It seems reasonable to associate parameter a in Eq. (10) with
the fluid properties, as it is done for Eq. (9), since both fits pre-
sented very similar values, and the formulation used was the same
for all assays.
Karbstein and Schubert [30] first stated that the reduction on
the volume-surface mean diameter D[3,2] as a power function was
applicable only to continuous emulsification systems, when the
retention time in the dispersion zone is in the order of millisec-
onds. However, such approximations have been done for ultra-
sound equipments in batch operation [31,56].
Reduction of droplet size as a function of energy input was also
observed by Kaltsa et al. [19], but the authors did not achieve a sat-
uration diameter in the process conditions that were evaluated.
Energy input influence on droplet diameter has also been
expressed as function of time, amplitude and power [20,57]. Since
sonication time is directly related to energy input (Eq. (7)), curves
generally assume the same shape as the experimental data pre-
sented in Fig. 6, when it is plotted against mean diameter
[35,56,57]. If data are plotted in logarithmic scale, a linear depen-
dence of D[3,2] with time or energy input is usually observed.
Besides mean diameter, the saturation diameter was expressed
as a monoexponential function of time [35].
The prediction of droplets mean diameter produced by mechan-
ical devices is only one of the factors that make energy density an
important concept when producing emulsions, so that Delmas
et al. [35] suggested that energy density is the major control
 L. Consoli et al. / Ultrasonics Sonochemistry 38 (2017) 772782
parameter in ultrasound emulsification, and not sonication time or
power isolated. Furthermore, it is also useful for the design,
scaling-up and process control of emulsifying devices. Another
application in which this concept may be very helpful is in the
comparison of products properties of different mechanical emulsi-
fication processes [58].
4. Conclusions
Sonication was found to be effective in producing sub-micron
emulsions with high solids concentration. Both sonication time
and power amplitude showed significant effects on emulsions
properties. However, we could see that the energy density
employed on the process, which results from a combination of
time and power, is a parameter of high relevance as well, and must
then be regarded when designing emulsions. After supplying a
given amount of energy to the system, a saturation diameter was
achieved, and thus emulsions presented also similar kinetic stabil-
ity characteristics. Then, operation at 148 W of power for 7 min
was chosen for the continuity of the work, since it produced emul-
sions with analogous properties of those produced with increased
energy inputs.
Acknowledgements
The authors are grateful to FAPESP (EMU 2009/54137-1 and
2015/11984-7) for the financial support, for CNPq (140273/2014-
0 and 140271/2014-7) for the PhD fellowships and for the National
Institute of Science and Technology on Photonics Applied to Cell
Biology (INFABIC) at the State University of Campinas, for the Con-
focal Laser Scanning Microscopy.
References
[1] D.J. McClements, Encapsulation, protection, and release of hydrophilic active
components: potential and limitations of colloidal delivery systems, Adv.
Colloid Interface Sci. 219 (2015) 2753.
[2] A. Sosnik, K.P. Seremeta, Advantages and challenges of the spray-drying
technology for the production of pure drug particles and drug-loaded
polymeric carriers, Adv. Colloid Interface Sci. 223 (2015) 4054.
[3] A. Singh, G. Van den Mooter, Spray drying formulation of amorphous solid
dispersions, Adv. Drug Deliv. Rev. 100 (2016) 2750.
[4] Z. Fang, B. Bhandari, Spray drying, freeze drying and related processes for food
ingredient and nutraceutical encapsulation, in: N. Garti, D.J. McClements
(Eds.), Encapsulation Technologies and Delivery Systems for Food Ingredients
and Nutraceuticals, Woodhead Publishing, Cambridge UK, 2012, pp. 73109.
[5] G.A. Reineccius, Multiple-core encapsulation: the spray drying of food
ingredients, in: P. Vilstrup (Ed.), Microencapsulation of Food Ingredients,
Leatherhead Publishing, Surrey, 2001.
[6] S.M. Jafari, E. Assadpoor, Y. He, B. Bhandari, Encapsulation efficiency of food
flavours and oils during spray drying, Drying Technol. 26 (2008) 816835.
[7] G.A. Reineccius, The spray drying of food flavors, Drying Technol. 22 (2004)
12891324.
[8] C.-H. Tang, X.-R. Li, Microencapsulation properties of soy protein isolate and
storage stability of the correspondingly spray-dried emulsions, Food Res. Int.
52 (2013) 419428.
[9] E.C. Frascareli, V.M. Silva, R.V. Tonon, M.D. Hubinger, Effect of process
conditions on the microencapsulation of coffee oil by spray drying, Food
Bioprod. Process. 90 (2012) 413424.
[10] H.C.F. Carneiro, R.V. Tonon, C.R.F. Grosso, M.D. Hubinger, Encapsulation
efficiency and oxidative stability of flaxseed oil microencapsulated by spray
drying using different combinations of wall materials, J. Food Eng. 115 (2013)
443451.
[11] V.M. Silva, G.S. Vieira, M.D. Hubinger, Influence of different combinations of
wall materials and homogenisation pressure on the microencapsulation of
green coffee oil by spray drying, Food Res. Int. 61 (2014) 132143.
[12] C. Turchiuli, M.T. Jimenez Munguia, M. Hernandez Sanchez, H. Cortes Ferre, E.
Dumoulin, Use of different supports for oil encapsulation in powder by spray
drying, Powder Technol. 255 (2014) 103108.
[13] A.G.S. Carvalho, V.M. Silva, M.D. Hubinger, Microencapsulation by spray drying
of emulsified green coffee oil with two-layered membranes, Food Res. Int. 61
(2014) 236245.
[14] J.H. Chew, N. Fu, T. Gengenbach, X.D. Chen, C. Selomulya, The compositional
effects of high solids model emulsions on drying behaviour and particle
formation processes, J. Food Eng. 157 (2015) 3340.
781
[15] M.D.R. Hernndez Snchez, M.-E. Cuvelier, C. Turchiuli, Effect of a-tocopherol
on oxidative stability of oil during spray drying and storage of dried emulsions,
Food Res. Int. 88 (2016) 3241, Part A.
[16] J. Chandrapala, C. Oliver, S. Kentish, M. Ashokkumar, Ultrasonics in food
processing, Ultrason. Sonochem. 19 (2012) 975983.
[17] S. Abbas, K. Hayat, E. Karangwa, M. Bashari, X.M. Zhang, An overview of
ultrasound-assisted food-grade nanoemulsions, Food Eng. Rev. 5 (2013) 139
157.
[18] M. Sivakumar, S.Y. Tang, K.W. Tan, Cavitation technology a greener
processing technique for the generation of pharmaceutical nanoemulsions,
Ultrason. Sonochem. 21 (2014) 20692083.
[19] O. Kaltsa, C. Michon, S. Yanniotis, I. Mandala, Ultrasonic energy input influence
on the production of sub-micron o/w emulsions containing whey protein and
common stabilizers, Ultrason. Sonochem. 20 (2013) 881891.
[20] O. Kaltsa, I. Gatsi, S. Yanniotis, I. Mandala, Influence of ultrasonication
parameters on physical characteristics of olive oil model emulsions
containing xanthan, Food Bioprocess Technol. 7 (2014) 20382049.
[21] C. Huck-Iriart, V.M. Pizones Ruiz-Henestrosa, R.J. Candal, M.L. Herrera, Effect of
aqueous phase composition on stability of sodium caseinate/sunflower oil
emulsions, Food Bioprocess Technol. 6 (2013) 24062418.
[22] C. Huck-Iriart, J.A. Rincon-Cardona, M.L. Herrera, Stability of whey protein
concentrate/sunflower oil emulsions as affected by sucrose and xanthan gum,
Food Bioprocess Technol. 7 (2014) 26462656.
[23] G. de Figueiredo Furtado, R.A. Mantovani, L. Consoli, M.D. Hubinger, R.L. da
Cunha, Structural and emulsifying properties of sodium caseinate and
lactoferrin influenced by ultrasound process, Food Hydrocolloids 63 (2017)
178188.
[24] I.A.M. Appelvist, M. Golding, R. Vreeker, N.J. Zuidam, Emulsions as delivery
systems in foods, in: J.M. Lakkis (Ed.), Encapsulation and Controlled Release
Technologies in Food Systems, Blackwell Publishing, Ames, 2007, pp. 4181.
[25] E. Dickinson, Caseins in emulsions: interfacial properties and interactions, Int.
Dairy J. 9 (1999) 305312.
[26] C. Huck-Iriart, M.S. lvarez-Cerimedo, R.J. Candal, M.L. Herrera, Structures and
stability of lipid emulsions formulated with sodium caseinate, Curr. Opin.
Colloid Interface Sci. 16 (2011) 412420.
[27] E. Dickinson, Flocculation of protein-stabilized oil-in-water emulsions,
Colloids Surf. B 81 (2010) 130140.
[28] O.M.o.A.o.A. International, Official Method 960.52, in: Gaithersburg, MD, USA,
1998.
[29] C. Noello, A.G.S. Carvalho, V.M. Silva, M.D. Hubinger, Spray dried
microparticles of chia oil using emulsion stabilized by whey protein
concentrate and pectin by electrostatic deposition, Food Res. Int. 89 (Part 1)
(2016) 549557.
[30] H. Karbstein, H. Schubert, Developments in the continuous mechanical
production of oil-in-water macro-emulsions, Chem. Eng. Process. 34 (1995)
205211.
[31] S.M. Jafari, Y.H. He, B. Bhandari, Production of sub-micron emulsions by
ultrasound and microfluidization techniques, J. Food Eng. 82 (2007) 478488.
[32] S.M. Jafari, E. Assadpoor, Y. He, B. Bhandari, Re-coalescence of emulsion
droplets during high-energy emulsification, Food Hydrocolloids 22 (2008)
11911202.
[33] S. Mahdi Jafari, Y. He, B. Bhandari, Nano-emulsion production by sonication
and microfluidizationa comparison, Int. J. Food Prop. 9 (2006) 475485.
[34] M. Tabibiazar, S. Davaran, M. Hashemi, A. Homayonirad, F. Rasoulzadeh, H.
Hamishehkar, M.A. Mohammadifar, Design and fabrication of a food-grade
albumin-stabilized nanoemulsion, Food Hydrocolloids 44 (2015) 220228.
[35] T. Delmas, H. Piraux, A.-C. Couffin, I. Texier, F. Vinet, P. Poulin, M.E. Cates, J.
Bibette, How to prepare and stabilize very small nanoemulsions, Langmuir 27
(2011) 16831692.
[36] A. Rawle, The importance of particle sizing to the coatings industry. Part 1:
Particle size measurement, Adv. Colour Sci. Technol. 5 (2002) 112.
[37] P. Relkin, S. Sourdet, Factors affecting fat droplet aggregation in whipped
frozen protein-stabilized emulsions, Food Hydrocolloids 19 (2005) 503511.
[38] A.M. Chuah, T. Kuroiwa, S. Ichikawa, I. Kobayashi, M. Nakajima, Formation of
biocompatible nanoparticles via the self-assembly of chitosan and modified
lecithin, J. Food Sci. 74 (2009) N1N8.
[39] R.J.B.M. Delahaije, H. Gruppen, M.L.F. Giuseppin, P.A. Wierenga, Towards
predicting the stability of protein-stabilized emulsions, Adv. Colloid Interface
Sci. 219 (2015) 19.
[40] T.J. Wooster, M. Golding, P. Sanguansri, Impact of oil type on nanoemulsion
formation and Ostwald ripening stability, Langmuir 24 (2008) 1275812765.
[41] D.J. McClements, Critical review of techniques and methodologies for
characterization of emulsion stability, Food Sci. Nutr. 47 (2007) 611649.
[42] E. Dickinson, M. Golding, Depletion flocculation of emulsions containing
unadsorbed sodium caseinate, Food Hydrocolloids 11 (1997) 1318.
[43] M.S. lvarez Cerimedo, C.H. Iriart, R.J. Candal, M.L. Herrera, Stability of
emulsions formulated with high concentrations of sodium caseinate and
trehalose, Food Res. Int. 43 (2010) 14821493.
[44] S.O. Agboola, D.G. Dalgleish, Effects of pH and ethanol on the kinetics of
destabilisation of oil-in-water emulsions containing milk proteins, J. Sci. Food
Agric. 72 (1996) 448454.
[45] E. Dickinson, M. Golding, Influence of alcohol on stability of oil-in-water
emulsions containing sodium caseinate, J. Colloid Interface Sci. 197 (1998)
133141.
[46] L. Medina-Torres, F. Calderas, J.A. Gallegos-Infante, R.F. Gonzlez-Laredo, N.
Rocha-Guzmn, Stability of alcoholic emulsions containing different
782 L. Consoli et al. / Ultrasonics Sonochemistry 38 (2017) 772782
caseinates as a function of temperature and storage time, Colloids Surf. A 352
(2009) 3846.
[47] J.-L. Feng, Z.-W. Wang, J. Zhang, Z.-N. Wang, F. Liu, Study on food-grade
vitamin E microemulsions based on nonionic emulsifiers, Colloids Surf. A 339
(2009) 16.
[48] S.J. Radford, E. Dickinson, M. Golding, Stability and rheology of emulsions
containing sodium caseinate: combined effects of ionic calcium and alcohol, J.
Colloid Interface Sci. 274 (2004) 673686.
[49] E. Dickinson, Hydrocolloids at interfaces and the influence on the properties of
dispersed systems, Food Hydrocolloids 17 (2003) 2539.
[50] S.A. Hogan, B.F. McNamee, E.D. ORiordan, M. OSullivan, Emulsification and
microencapsulation properties of sodium caseinate/carbohydrate blends, Int.
Dairy J. 11 (2001) 137144.
[51] S. Drusch, S. Berg, M. Scampicchio, Y. Serfert, V. Somoza, S. Mannino, K.
Schwarz, Role of glycated caseinate in stabilisation of microencapsulated
lipophilic functional ingredients, Food Hydrocolloids 23 (2009) 942948.
[52] L. Sanguansri, L. Day, Z. Shen, P. Fagan, R. Weerakkody, L.J. Cheng, J. Rusli, M.A.
Augustin, Encapsulation of mixtures of tuna oil, tributyrin and resveratrol in a
spray dried powder formulation, Food Funct. 4 (2013) 17941802.
[53] E. Dickinson, Structure formation in casein-based gels, foams, and emulsions,
Colloids Surf. A 288 (2006) 311.
[54] E. Dickinson, M. Golding, Rheology of sodium caseinate stabilized oil-in-water
emulsions, J. Colloid Interface Sci. 191 (1997) 166176.
[55] Y. Liang, G. Gillies, H. Patel, L. Matia-Merino, A. Ye, M. Golding, Physical
stability, microstructure and rheology of sodium-caseinate-stabilized
emulsions as influenced by protein concentration and non-adsorbing
polysaccharides, Food Hydrocolloids 36 (2014) 245255.
[56] T.S.H. Leong, T.J. Wooster, S.E. Kentish, M. Ashokkumar, Minimising oil droplet
size using ultrasonic emulsification, Ultrason. Sonochem. 16 (2009) 721727.
[57] B. Abismal, J.P. Canselier, A.M. Wilhelm, H. Delmas, C. Gourdon, Emulsification
by ultrasound: drop size distribution and stability, Ultrason. Sonochem. 6
(1999) 7583.
[58] H. Schubert, R. Engel, Product and formulation engineering of emulsions,
Chem. Eng. Res. Des. 82 (2004) 11371143.