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Eect of Salts on Formation and Stability of Vitamin EEnriched Miniemulsions Produced by Spontaneous Emulsication
Amir Hossein Saberi, Yuan Fang, and David Julian McClements*,

Biopolymers and Colloids Laboratory, Department of Food Science, University of MassachusettsAmherst, Amherst,
Massachusetts 01003, United States

PepsiCo Global R&D, 100 Stevens Avenue, Valhalla, New York 10595, United States
ABSTRACT: Emulsion-based delivery systems are being utilized to incorporate lipophilic bioactive components into various
food, personal care, and pharmaceutical products. This study examined the inuence of inorganic salts (NaCl and CaCl2) on the
formation, stability, and properties of vitamin E-enriched emulsions prepared by spontaneous emulsication. These emulsions
were simply formed by titration of a mixture of vitamin E acetate (VE), carrier oil (MCT), and nonionic surfactant (Tween 80)
into an aqueous salt solution with continuous stirring. Salt type and concentration (01 N NaCl or 00.5 N CaCl2) did not have
a signicant inuence on the initial droplet size of the emulsions. On the other hand, the isothermal and thermal stabilities of the
emulsions depended strongly on salt levels. The cloud point of the emulsions decreased with increasing salt concentration, which
was attributed to accelerated droplet coalescence in the presence of salts. Dilution (26 times) of the emulsions with water
appreciably improved their thermal stability by increasing their cloud point, which was mainly attributed to the decrease in
aqueous phase salt levels. The isothermal storage stability of the emulsions also depended on salt concentration; however,
increasing the salt concentration decreased the rate of droplet growth, which was the opposite of its eect on thermal stability.
Potential physicochemical mechanisms for these eects are discussed in terms of the inuence of salt ions on van der Waals and
electrostatic interactions. This study provides important information about the eect of inorganic salts on the formation and
stability of vitamin E emulsions suitable for use in food, personal care, and pharmaceutical products.
KEYWORDS: emulsions, nanoemulsions, vitamin E, salts, stability, cloud point, spontaneous emulsication

INTRODUCTION
Emulsion-based delivery systems are nding increasing
utilization within food, personal care, and pharmaceutical
products to encapsulate, protect, and release lipophilic active
ingredients. This category of delivery system includes various
kinds of colloidal suspension that utilize lipid droplets as
building blocks, such as emulsions, solid lipid nanoparticles,
multilayer emulsions, multiple emulsions, and lled hydrogel
particles.1,2 Emulsions are the simplest type of emulsion-based
delivery system consisting of lipid droplets suspended in an
aqueous phase, with each lipid droplet being coated by a layer
of emulsier molecules. Nevertheless, the functional performance of emulsions can still be manipulated by altering oil phase
composition, emulsier type, and particle size distribution. For
example, reducing the particle size can lead to higher optical
clarity, greater stability to gravitational separation and droplet
aggregation, and higher oral bioavailability.35 Emulsions
containing very small droplets (d < 200 nm) are often referred
to as nanoemulsions, mini-emulsions, or ultrane emulsions. In
this paper, we will simply refer to them as emulsions for the
sake of convenience.
Emulsions can be prepared using several approaches, which
are usually categorized as either high-energy or low-energy
methods.6,7 High-energy methods use mechanical devices to
generate intense forces that intermingle and break up the oil
and water phases, such as high-pressure homogenizers,
microuidizers, and ultrasonic generators.3,8 Low-energy
methods rely on the spontaneous formation of ultrane
droplets within surfactantoilwater (SOW) mixtures when
2014 American Chemical Society

specic changes in composition and/or environmental


conditions occur.3,8 A number of low-energy methods have
been developed to form ultrane oil droplets including
spontaneous emulsication (SE), phase inversion temperature
(PIT), phase inversion composition (PIC), and emulsion
inversion point (EIP) methods.5,810 Among these methods,
spontaneous emulsication is one of the easiest and most costeective to implement because it simply involves metering an
oilsurfactant mixture into an aqueous solution with constant
agitation at a xed temperature.
Commercial food and beverage products typically contain a
variety of ingredients that may inuence the formation and
stability of emulsions, including sugars, cosolvents, and salts.
Salts are incorporated into foods for a number of purposes, for
example, to control avor, water activity, stability, and
nutritional prole. Salts have been reported to inuence various
physicochemical properties of nonionic surfactants, including
solubility and cloud point,1115 critical micelle concentration,16,17 emulsion phase inversion temperature,14,15,18,19 and
emulsion stability.11,19,20 Mei et al.19 suggested that salt may
facilitate the formation of emulsions using the PIT method.
Salts may alter the interlayer spacing of the liquid crystalline
phases formed during the intermediate steps of low-energy
emulsication, thereby leading to the formation of ultrane
Received:
Revised:
Accepted:
Published:
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August 10, 2014


October 22, 2014
October 24, 2014
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droplets.21 Furthermore, it has been reported that the storage


stability of emulsions stabilized by nonionic surfactants is
strongly dependent upon their PIT,2224 which depends on salt
concentration. Many previous studies on the impact of salt on
nanoemulsion formation have utilized simple organic solvents
or liquids as the oil phase. In this study, we utilized an oil phase
that is more representative of those that might be used in the
food industry, that is, vitamin E acetate dissolved in mediumchain triglycerides (MCT).
Recently, we demonstrated that stable vitamin E-enriched
emulsions containing very ne droplets (d < 60 nm) could be
produced by spontaneous emulsication.25 We also showed
that addition of cosolvents to the aqueous phase of these
emulsions had an appreciable inuence on their formation and
stability.26,27 The purpose of the present study was to
investigate the inuence of two salts commonly used in
industrial applications (NaCl and CaCl2) on the formation and
stability of vitamin E emulsions produced using spontaneous
emulsication. The knowledge gained from this study will be
useful for the successful incorporation of emulsion-based
delivery systems into commercial products.

Statistical Analysis. All experiments were carried out two or three


times using two freshly prepared samples, and the results are reported
as the calculated mean and standard deviation of these measurements.
Dierences between group means were determined by analysis of
variance (ANOVA) using statistical software (version 12.0; SPSS, Inc.,
Chicago, IL, USA). Means were subjected to Duncans test, and a P
value of <0.05 was considered statistically signicant.

RESULTS AND DISCUSSION


Eect of Salts on Initial Particle Size. Initially, the
inuence of salt type and concentration on the size of the oil
droplets formed in vitamin E emulsions produced by
spontaneous emulsication was determined. The oil phase
contained 80% vitamin E and 20% MCT. This composition
produced the smallest particle size as shown in a previous
study,25 and so we used this oil phase composition in the
present study. The aqueous phase contained either 01 N
NaCl or 00.5 N CaCl2. The inuence of dierent salt
concentrations on the initial mean droplet diameter of the
emulsions was measured. The mean droplet diameter remained
relatively constant at all salt levels used: 52 nm from 0 to 1 N
NaCl and 51 nm from 0 to 0.5 N CaCl2. Therefore, the
addition of salt did not have an appreciable eect on the mean
droplet diameter of the emulsions at any of the levels tested.
The particle size distributions of all these emulsions were also
similar, that is,, monomodal with similar modes and widths
(data not shown).
In principle, one might expect the presence of salts to aect
the size of the droplets produced by spontaneous emulsication
by altering the bulk physicochemical properties of the aqueous
phase (e.g., density, refractive index, viscosity, and interfacial
tension) or by altering its molecular properties (e.g., structural
organization). Changes in the viscosity of an aqueous solution
would be expected to aect the mass transport kinetics of
surfactant and oil molecules at the boundary between the
organic and aqueous phases, which could inuence the
spontaneous formation of small droplets at the phase
boundary.28 Changes in oil/water interfacial tension would
also be expected to inuence spontaneous droplet formation by
altering the dynamics of the phase boundary. Having said this,
the addition of NaCl and CaCl2 to aqueous solutions at the
highest levels used in this study (1 and 0.5 N, respectively)
should only slightly increase their interfacial tension, viscosity,
density, and refractive index (Table 1). Consequently, the
inuence of salts on the physicochemical properties of the

MATERIALS AND METHODS

Materials. Vitamin E acetate (VE) was kindly donated by BASF


(Ludwigshafen, Germany). MCT oil (MIGLYOL 812) was purchased
from Warner Graham Co. (Sasol Germany GmbH). Nonionic
surfactant (Tween 80), sodium chloride, calcium chloride anhydrous,
citric acid, and sodium benzoate were purchased from Sigma-Aldrich
Co. (St. Louis, MO, USA). Double-distilled water was used in the
preparation of all solutions and emulsions.
Emulsion Preparation. Emulsion formation was carried out using
the spontaneous emulsication procedure described in our previous
study.27 In brief, spontaneous emulsication was performed by the
addition of an organic phase to an aqueous phase with constant
magnetic stirring (600 rpm) at 25 C. The organic phase consisted of
VE, MCT, and surfactant, whereas the aqueous phase consisted of a
buer solution containing 0.8 wt % citric acid, 0.08 wt % sodium
benzoate, and dierent levels of NaCl or CaCl2 (pH 3.0). On the basis
of the results of our previous studies, we prepared emulsions with the
following composition: 8% vitamin E acetate; 2% MCT; 10% Tween
80; and 80% buer solution (pH 3). The oil (8 g VE + 2 g MCT) and
surfactant (10 g) were rst mixed together to form an organic phase,
which was then slowly poured into 80 g of aqueous phase over a 15
min period with continuous stirring.
Emulsions were diluted in three dierent ways to contain nal
systems with dierent droplet concentrations and aqueous phase
compositions: (i) with buer solution (pH 3); (ii) with salt solutions
(NaCl or CaCl2) with similar salt concentrations as the emulsions (pH
3); and (iii) with salt solutions containing various salt concentrations.
Particle Size Measurements. Particle size distributions were
measured using a dynamic light scattering instrument (Zetasizer Nano
ZS, Malvern Instruments, Malvern, UK). This instrument determines
the particle size from intensitytime uctuations of a laser beam (633
nm) scattered from a sample at an angle of 173. Each individual
measurement was an average of 13 runs. To avoid multiple scattering
eects, samples were diluted (500 times) before the particle size
measurements using buer solution (pH 3.0). The mean droplet
radius (Z-average) was calculated from the particle size distribution.
All measurements were conducted at ambient temperature.
Turbidity Measurements and Cloud Point Calculation. The
inuence of temperature on emulsion turbidity was determined using a
UVvisible spectrophotometer with temperature scanning capabilities
(Evolution Array, Thermo Scientic). The absorbance (A) at 600 nm
was measured as the temperature was increased from 20 to 95 C at 1
C/min. The cloud point was empirically dened as the temperature at
which the slope of the turbiditytemperature prole (d/dT) rst
reached a maximum value during heating.

Table 1. Eect of Salt Concentration (N Salts in Aqueous


Phase) on the Relative Viscosity, Refractive Index, Relative
Density, and Excess Interfacial Tension

water
NaCl
solution (1
N)
CaCl2
solution
(0.5 N)

relative
viscosity /
oa

refractive
index

relative
density
(kg/L)

excess interfacial
tensionb (mN/m)

1.000c
1.090c

1.333c
1.343c

1.000c
1.038c

0.0e
0.9e

1.070d

1.346c

1.044c

1.9e

a
Ratio of the absolute viscosity of solution at 20 C to the ratio of the
absolute viscosity of water at 20 C. bExcess interfacial tension is
dened as the dierence between interfacial tension of hydrocarbon/
salts solution and interfacial tension of hydrocarbon/water. cReference
50. dReference 51. eReference 52.

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aqueous phase might be expected to be relatively small.


Previous studies have shown that changes in the solubility and
optimum curvature of surfactants in surfactantoilwater
(SOW) mixtures play a crucial role in the spontaneous
formation of oil droplets using low-energy methods.8,29,30
The fact that we observed no inuence of salts on the initial
particle size in this study (Table 1 and Figure 1) suggests that
the salts did not have an appreciable inuence on surfactant
solubility or optimum curvature under the conditions used.
Nevertheless, this is likely to be highly dependent on the nature
of the surfactant used to form the emulsions.
Eect of Salts on Thermal Stability. Emulsion-based
delivery systems are usually exposed to elevated temperatures
during their manufacture, storage, or utilization, for example,
hot lling, pasteurization, or cooking. Therefore, information
about the thermal stability of emulsions is of particular
importance for many commercial applications. We therefore
investigated the impact of salt type and concentration on the
thermal stability of the vitamin E emulsions using a
spectrophotometric method. The turbidity of emulsions
containing dierent NaCl and CaCl2 concentrations was
measured as they were subjected to a temperature increase
from 20 to 95 C. The turbidity proles in the presence of
NaCl and CaCl2 were qualitatively similar, and therefore we
show only the results for the emulsion containing NaCl (Figure
1a).
The turbidity of the emulsions remained relatively low and
constant during heating from 0 to 75 C in the absence of
NaCl, then increased sharply around 7580 C, and then
remained high upon further heating from 80 to 95 C (Figure
1a). This steep rise in turbidity can be ascribed to an increase in
droplet size during heating. As the temperature increased, there
would be progressive dehydration of the polar headgroups of
the nonionic surfactant molecules (Tween 80), which would
reduce the hydration repulsion between the droplets, thereby
promoting droplet coalescence.31,32 Furthermore, dehydration
of the surfactant headgroups changes the optimum curvature of
the surfactant monolayer toward unity, which favors ultralow
interfacial tension, high interfacial exibility, and droplet
coalescence.3,33 The turbiditytemperature proles of emulsions containing salt were more complex than those containing
no salt (Figure 1a). For instance, for the emulsions containing
0.15 N NaCl, the turbidity remained low and constant from 20
to 74 C, increased steeply from 74 to 78 C, and remained
high from 78 to 88 C, which was qualitatively similar as the
turbiditytemperature prole measured in the absence of salt.
However, upon increasing the temperature further, there was a
sharp drop in the turbidity from 88 to 90 C, followed by
another steep increase from 90 to 93 C, after which the
turbidity remained high from 93 to 95 C (Figure 1a). The
turbiditytemperature proles observed for the emulsions
containing salt can be related to a transitional phase inversion
of the SOW system around the PIT. The phase inversion
temperature is governed by changes in the optimum curvature
and relative solubility of nonionic surfactants with temperature.29,34 The molecular geometry of a surfactant can be
dened by a packing parameter, p35
a
p= T
aH

Figure 1. (a) Inuence of temperature on the relative turbidity of


vitamin E-enriched mini-emulsion produced using 10% oil phase
(vitamin E/MCT, 8:2), 10% surfactant phase (Tween 80), and 80%
aqueous phase with dierent concentrations (N) of NaCl. To prevent
overlap of the graphs, the unit values of 4, 8, 12, 16, and 20 were added
to the turbidity data obtained for the emulsion with aqueous phase
containing 0.15, 0.25, 0.5, 0.75, and 1 N NaCl, respectively, and the
corresponding turbidities were reported as relative turbidity. (b)
Packing parameter (p) and optimum curvature of Tween 80 at
dierent temperatures. (c) Inuence of salt concentration on the cloud
point of vitamin E-enriched mini-emulsion produced using 10% oil
phase (vitamin E/MCT, 8:2), 10% surfactant phase (Tween 80), and
80% aqueous phase.

where aT and aH are the cross-sectional areas of the lipophilic


tail-group and hydrophilic headgroup, respectively. Initially, the
emulsions had relatively low turbidities at room temperature
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hydrophilic headgroups. In this study, we found that both NaCl


and CaCl2 reduced the PIT of the emulsions, which suggests
that they increased the packing parameter (p) by promoting
headgroup dehydration. Previously, this aect has been
attributed to the fact that these ions promote structure
formation of bulk water around the ions, thereby reducing
the amount of water molecules available to hydrate the
surfactant headgroups.18 In an alternative interpretation of
the Hofmeister series, Kabalnov et al.44 suggested that the
salting-in and salting-out phenomena have an interfacial origin
driven by preferential adsorption/desorption of salt ions at the
hydrophilic headgroup of the nonionic surfactant monolayers.
Adding NaCl as a nonadsorbing salt dehydrates the hydrophilic
headgroups through an osmotic eect associated with
preferential ion exclusion, 44 thereby altering the optimum
curvature and decreasing the PIT.
The fact that salt addition did not strongly inuence the size
of droplets during emulsion formation, but did inuence their
thermal stability (Figure 2), may have been due to their

because they contained very small droplets that did not scatter
light strongly.36 At these low temperatures the surfactant is
relatively hydrophilic, and the corresponding optimum
curvature (p < 1) favors the formation of oil-in-water
emulsions. As temperature increases, the headgroup of the
nonionic surfactant (Tween 80) undergoes progressive
dehydration, causing the packing parameter to increase (Figure
1b). As a result, the rate of droplet coalescence increases
because of the reduced hydration repulsion between droplets
and the increase in interfacial exibility, and so there is a
corresponding increase in turbidity. However, when the
temperature reaches the PIT, the surfactant monolayer has
zero curvature, the packing parameter is close to unity (p = 1),
and the interfacial tension becomes ultralow (Figure 1b). These
changes favor the formation of a bicontinuous microemulsion
phase containing very small structures that do not scatter light
strongly, thereby leading to a reduction in turbidity. Increasing
the temperature above the PIT further dehydrates the
headgroups of the surfactant molecules, which increases their
lipophilicity, increases the packing parameter (p > 1), and
changes the optimum curvature so that it favors the formation
of water-in-oil emulsions (Figure 1b). The water droplets in the
water-in-oil emulsions formed above the PIT are relatively large
and therefore scatter light strongly, leading to increased
turbidity.3,29
The impact of salt on the turbiditytemperature proles of
the emulsions can be attributed to the fact that salt decreases
the PIT of the emulsions. In the absence of salt, the PIT is
relatively high, and therefore we observed only the initial
droplet coalescence regimen. In the presence of sucient salt,
the PIT moved into the experimental temperature range, and
we could observe droplet coalescence, microemulsion formation, and phase inversion. A similar behavior was observed in
the turbiditytemperature proles of vitamin E-enriched
emulsions in the absence and presence of glycerol; that is,
glycerol reduced the PIT.26
The turbidity versus temperature proles were used to
determine the cloud points of the emulsions so that we could
establish the inuence of salt type and concentration on their
thermal behavior. The cloud point of the emulsions decreased
from around 78 to 65 C when the NaCl concentration was
increased from 0 to 1 M and decreased from around 78 to 70
C when the CaCl2 concentration was increased from 0 to 0.5
N (Figure 1c). The ability of NaCl14,15,20,37 and CaCl214,15,37 to
decrease the cloud points of nonionic surfactant has been
reported previously. Our results are also in agreement with
previous studies that have reported that the cloud point and/or
PIT of nonionic surfactants is a linear function of their salt
concentration.15,38,39 The decrease in thermal stability (cloud
point) of emulsions in the presence of NaCl and CaCl2 can be
attributed to the eect of these salts on the solubility and
packing characteristics of nonionic surfactants.
Specic anionic19,40,41 and cationic12,16,19,40 salt ions may
either increase (salting in) or decrease (salting out) the
solubility of nonionic surfactants due to their eects on the
structural organization of water molecules. Ions can be
classied as either structure-makers (salting out) or
structure-breakers (salting in) according to their position in
the Hofmeister series.42,43 The presence of mineral ions in an
aqueous solution may alter the PIT of a nonionic surfactant by
altering the solubility and packing characteristics of the
surfactant molecules. In particular, salts may aect the optimum
curvature of the surfactants by altering the hydration of their

Figure 2. Inuence of dilution on the cloud point of vitamin Eenriched mini-emulsion produced using 10% oil phase (vitamin E/
MCT, 8:2), 10% surfactant phase (Tween 80), and 80% aqueous
phase (0.5 M salt). Oil phase concentrations were 10, 5, 3.33, 2.5, and
1.67, and salt concentrations were 0.4, 0.2, 0.13, 0.1, 0.08, and 0.067 N
for the emulsion diluted with dilution factors of 1, 2, 3, 4, 5, and 6,
respectively.

inuence on the dehydration of the hydrophilic headgroups at


dierent temperatures. Emulsion formation was carried out at
low (room) temperature, and therefore the surfactant
molecules would have been highly hydrated. Presumably,
headgroup hydration was still sucient in the presence of salt
to generate a strong steric repulsion between the droplets and
to have a high surfactant curvature. However, once the
emulsions were heated, the eects of salts became more
pronounced because the surfactants were already partially
dehydrated, and any further change due to salt addition was
sucient to promote instability.
Inuence of Dilution on Thermal Stability. Emulsions
are diluted when they are incorporated into functional food and
beverage products. Our previous studies have shown that
emulsion dilution may have an appreciable inuence on
emulsion thermal stability.26,27,45 We therefore examined the
inuence of dilution with buer solution on the thermal
stability of vitamin E emulsions initially containing 0.5 N NaCl
or CaCl2 in the aqueous phase. Qualitatively, all of the diluted
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emulsions had similar turbiditytemperature proles as the


undiluted emulsions (Figure 1a): upon increasing the temperature from 20 to 95 C, the turbidity initially remained low,
then steeply increased, then sharply decreased to a minimum,
then steeply increased again, and then remained high (data not
shown). The cloud points determined from these proles
increased from 73 to 81 C for NaCl and from 70 to 81 C
CaCl2 as the emulsion dilution factor (DF) increased from 1 to
6 (Figure 2). As these samples were diluted with buer
solution, the observed results may have been due to a decrease
in salt concentration in the aqueous phase or due to a decrease
in oil droplet concentration.
We therefore carried out additional experiments in which the
emulsions were diluted with various salt solutions so as to
obtain samples where (i) the droplet concentration was kept
constant, but the salt concentration varied (Figure 3) or (ii) the

Figure 4. (a) Inuence of oil phase concentration on the cloud point


of diluted vitamin E-enriched mini-emulsions containing 0.4 N salts.
The initial emulsion was produced using 10% oil phase (vitamin E/
MCT, 8:2), 10% surfactant phase (Tween 80), and 0.4 N NaCl. (b)
Inuence of temperature on the turbidity of vitamin E-enriched
miniemulsions containing 0.4 N CaCl2 and dierent oil phase
concentrations. The initial emulsion was produced using 10% oil
phase (vitamin E/MCT, 8/2), 10% surfactant phase (Tween 80), and
0.4 N CaCl2.

Figure 3. Inuence of salt concentration on the cloud point of vitamin


E-enriched mini-emulsions containing 5% oil phase. The initial
emulsion was produced using 10% oil phase (vitamin E/MCT, 8:2),
10% surfactant phase (Tween 80), and 0.4 N salt.

droplet concentration varied, but the salt concentration was


kept constant (Figure 4). In the diluted emulsions with xed
droplet concentration, the cloud point increased from around
73 to 77 C for NaCl and from around 72 to 77 C for CaCl2
as the salt content decreased from 0.4 to 0.2 N, respectively
(Figure 3). These results suggest that at least part of the
increase in cloud point observed upon emulsion dilution
(Figure 2) was related to a reduction in salt concentration in
the aqueous phase. In the diluted emulsions with xed salt
concentration, the cloud point initially increased and then
decreased as the droplet content was reduced from 10 to 1%
(Figure 4a). This eect may have been an artifact of the cloud
point measurement at high droplet concentration. As the
droplet concentration increased, there was an increase in the
overall turbidity of the samples due to an increase in light
scattering by the droplets (Figure 4b). In the emulsions with
the highest droplet concentrations, the turbidity was so great
(>3 cm1) that it was not possible to observe the entire region
where rapid droplet coalescence occurred. Consequently, it was
not possible to accurately measure the cloud points for these
samples. In the more dilute emulsions (3% oil), the cloud
point decreased with increasing dilution (decreasing droplet
concentration), which may have been due to changes in the
nature of the microemulsion phase formed around the PIT
when the SOW ratio was changed. Taken together, these results

suggest that the increase in cloud point observed when saltcontaining emulsions were diluted with buer solution was
primarily due to the reduced salt concentration in the aqueous
phase, rather than the reduced droplet concentration.
Inuence of Salts and Dilution on Isothermal
Stability. Physical stability of emulsion-based delivery systems
during storage, transport, and utilization is required for most
commercial applications. Emulsions may become unstable
through a number of dierent physicochemical processes,
including occulation, coalescence, Ostwald ripening, and
gravitational separation.10,33 The mechanism and rate of
breakdown depend on emulsion composition and microstructure, as well as the environmental conditions experienced
during its lifetime. We therefore examined the inuence of salt
addition and dilution on the storage stability of vitamin E
emulsions. A series of emulsions was fabricated with dierent
droplet concentrations (2.5 or 10 wt %) and salt concentrations
(0, 0.2, and 0.4 N NaCl or CaCl2) by diluting an initial
emulsion with dierent types and amounts of salt solution.
After preparation, the resulting emulsions were stored for 1
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decreased their thermal stability by reducing the cloud point


(Figure 2). This independence of thermal and isothermal
stability of vitamin E emulsions has also been reported upon
posthomogenization addition of cosurfactants.46
There are a number of possible mechanisms that might be
responsible for the observed instability of the emulsions during
storage: occulation, coalescence, and Ostwald ripening.4,33 We
do not believe that occulation played an important role in
emulsion instability as we did not observe any ocs using an
optical microscope. We therefore postulated that the observed
instability was due to either coalescence or Oswald ripening. It
is often dicult to distinguish between these two mechanisms
because they both lead to a net increase in mean droplet
diameter over time.33,47 An increase in droplet growth rate with
increasing temperature could be attributed to the eects of
temperature on coalescence and/or Ostwald ripening. Increasing temperature can dehydrate the surfactant headgroups and
thereby increase droplet coalescence by (i) decreasing the
hydration repulsion between droplets and (ii) by changing the
optimum curvature of the surfactant monolayer toward
unity.3,31,48 A temperature rise may also increase the rate of
Ostwald ripening by increasing the diusion rate or solubility of
the oil molecules in the aqueous phase.49
Inorganic salts might inuence the isothermal storage
stability of oil-in-water emulsions stabilized by nonionic
surfactant through a number of mechanisms:
Eect of Salts on Surfactant Headgroup Dehydration.
Previous studies have reported that salts that decrease the cloud
point of emulsions stabilized by nonionic surfactant also
increase droplet coalescence.11,20 This eect was attributed to
the ability of the salts to promote dehydration of the surfactant
headgroup, thereby reducing the hydration repulsion between
droplets and altering the surfactant monolayer curvature.
However, the fact that we actually observed a decrease in
droplet growth during storage in the presence of NaCl and
CaCl2 suggested that this mechanism was not prominent under
the conditions used.
Eect of Salts on van der Waals Interactions. van der
Waals interactions are attractive forces that almost always act
between oil droplets suspended in water and, therefore,
promote droplet aggregation. The addition of salts to an
aqueous solution appreciably decreases the magnitude of the
van der Waals interaction due to an electrostatic screening
eect.33 This mechanism may therefore account for the
increased storage stability of the emulsions in the presence of
salt, because there would have been a reduction in the attractive
forces acting between the oil droplets. It should be noted that
salt can also decrease the electrostatic interaction between the
droplets through electrostatic screening, thereby increasing
droplet coalescence.
Eect of Salts on Oil Solubility. One of the major factors
inuencing the rate of Ostwald ripening in oil-in-water
emulsions is the solubility of the oil phase in the aqueous
phase.33 The addition of salts to the aqueous phase changes the
structural organization of the water molecules, which will alter
the equilibrium solubility of oil molecules. Salt addition causes
an increase in interfacial tension (Table 1), which would
increase the free energy associated with introducing a nonpolar
molecule into water, thereby decreasing its water solubility.
Consequently, addition of salt to the emulsions may have
reduced the rate of Ostwald ripening, which is consistent with
the reduction in droplet growth rate observed in the presence
of salt observed in this study.

month at three storage temperatures representing refrigeration


conditions (5 C), ambient storage in mild climates (20 C),
and ambient storage in hot climates (37 C).
The initial emulsion formed by spontaneous emulsication
contained 0% salt and 10% oil and was therefore designated 0/
10. The extent of droplet growth in this emulsion after 1
month of storage was appreciable and increased with increasing
storage temperature: 15% at 5 C, 19% at 20 C, and 54% at
37 C (Figure 5). The droplet size distribution of these

Figure 5. (a) Inuence of storage time and temperature on the particle


growth (%) of vitamin E-enriched mini-emulsions containing dierent
ratios of NaCl (N)/oil phase. The initial emulsions were produced
using 10% oil phase (vitamin E/MCT, 8:2), 10% surfactant phase
(Tween 80), and dierent NaCl concentrations (0, 0.4, and 0.8 N).
Means within dierent letters and the same number of stars are
signicantly (P < 0.05) dierent. (b) Inuence of storage time and
temperature on the particle growth (%) of vitamin E-enriched miniemulsions containing dierent ratios of CaCl2 (M)/oil phase. The
initial emulsions were produced using 10% oil phase (vitamin E/MCT,
8:2), 10% surfactant phase (Tween 80), and dierent CaCl2
concentrations (0, 0.2, and 0.4 N). Means within dierent letters
and the same number of stars are signicantly (P < 0.05) dierent.

emulsions remained monomodal throughout storage at all


temperatures (data not shown). The stability of the emulsions
to droplet growth improved signicantly (P < 0.05) as the level
of NaCl or CaCl2 added was increased, especially at the
elevated temperatures (Figure 5). In addition, reducing the
droplet concentration from 10 to 2.5% also led to a signicant
(P < 0.05) inhibition of droplet growth for emulsions
containing both types of salt (Figure 5). Overall, these results
suggest that the addition of NaCl or CaCl2 improved the
isothermal storage stability of the emulsions, even though it
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In summary, the addition of salt may have improved the


storage stability of the emulsions by decreasing the attractive
interactions between them (thereby decreasing coalescence) or
by reducing the water solubility of the oil phase (thereby
decreasing Ostwald ripening). Further studies are required to
establish the relative importance of these two dierent
physicochemical mechanisms. The improved stability of the
emulsions to droplet growth when the droplet concentration
was reduced can simply be attributed to the reduction in
droplet collision frequency in the more dilute system.
Conclusions. In this study, we examined the inuence of
salts on the formation, properties, and stability of vitamin Eenriched emulsions prepared by spontaneous emulsication.
We have shown that the addition of salts (NaCl and CaCl2) to
the aqueous phase of the emulsions did not change the initial
droplet size produced. However, the ability of salts to modulate
surfactant properties had an appreciable eect on their thermal
stability: the cloud point decreased with increasing salt
concentration. This eect was attributed to the ability of salts
to promote dehydration of the surfactant headgroup, which
alters the optimum curvature of the surfactant monolayer. The
isothermal stability of the emulsions was inuenced by storage
temperature, with the droplet growth rate increasing with
increasing temperature. This eect was attributed to the impact
of temperature on the rate of droplet coalescence and Ostwald
ripening. Addition of salts to the emulsions decreased the rate
of droplet growth during isothermal storage. This eect was
attributed to the ability of salts to reduce the attractive van der
Waals interactions between droplets (thereby reducing
coalescence) or to reduce the solubility of the oil molecules
in water (thereby reducing Ostwald ripening). These results
have important implications for the development of emulsionbased delivery systems for application in products containing
salts.

AUTHOR INFORMATION

Corresponding Author

*(D.J.M.) Phone: (413) 545-1019. Fax: (413) 545-1262. Email: mcclements@foodsci.umass.edu.


Funding

This material is based upon work supported by the Cooperative


State Research, Extension, Education Service, U.S. Department
of Agriculture, Massachusetts Agricultural Experiment Station
(Project 831), and by the U.S. Department of Agriculture, NRI
Grants (2011-03539 and 2013-03795).
Notes

The opinions expressed in this paper are those of the authors


and do not represent statements of position, intention, or
strategy of PepsiCo Inc.
The authors declare no competing nancial interest.

ACKNOWLEDGMENTS
We thank William Mutilangi of PepsiCo for useful advice and
discussions on this research.
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