Critical Reviews in Food Science and Nutrition

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Food-Grade Nanoemulsions: Formulation,

Fabrication, Properties, Performance, Biological
Fate, and Potential Toxicity

David Julian McClements & Jiajia Rao

To cite this article: David Julian McClements & Jiajia Rao (2011) Food-Grade Nanoemulsions:
Formulation, Fabrication, Properties, Performance, Biological Fate, and Potential Toxicity, Critical
Reviews in Food Science and Nutrition, 51:4, 285-330, DOI: 10.1080/10408398.2011.559558

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Critical Reviews in Food Science and Nutrition, 51:285–330 (2011)
Copyright
C Taylor and Francis Group, LLC
ISSN: 1040-8398 print / 1549-7852 online
DOI: 10.1080/10408398.2011.559558

Food-Grade Nanoemulsions:
Formulation, Fabrication, Properties,
Performance, Biological Fate, and
Potential Toxicity

DAVID JULIAN McCLEMENTS and JIAJIA RAO

Department of Food Science, University of Massachusetts, Amherst, MA

Nanoemulsions fabricated from food-grade ingredients are being increasingly utilized in the food industry to encapsulate,
protect, and deliver lipophilic functional components, such as biologically-active lipids (e.g., ω-3 fatty acids, conjugated
linoleic acid) and oil-soluble flavors, vitamins, preservatives, and nutraceuticals. The small size of the particles in nanoemul-
sions (r < 100 nm) means that they have a number of potential advantages over conventional emulsions—higher stability
to droplet aggregation and gravitational separation, high optical clarity, ability to modulate product texture, and, increased
bioavailability of lipophilic components. On the other hand, there may also be some risks associated with the oral ingestion
of nanoemulsions, such as their ability to change the biological fate of bioactive components within the gastrointestinal tract
and the potential toxicity of some of the components used in their fabrication. This review article provides an overview of
the current status of nanoemulsion formulation, fabrication, properties, applications, biological fate, and potential toxicity
with emphasis on systems suitable for utilization within the food and beverage industry.

Keywords nanoemulsions, emulsions, delivery systems, encapsulation, toxicology, stability

INTRODUCTION be useful for increasing the bioactivity of some nutraceuticals.

Nanoemulsions have a number of potential advantages over
conventional emulsions for particular applications within food
and beverage products. Nanoemulsions usually have better
stability to particle aggregation and gravitational separation
(Tadros et al., 2004). Nanoemulsions contain particles that only
scatter light waves weakly, and so they are suitable for incorpo-
ration into products that need to be optically clear or only slightly
turbid, such as fortified soft drinks and waters, and some kinds of
soups, sauces, and dips (Mason et al., 2006; Velikov et al., 2008;
Wooster et al., 2008). Nanoemulsions can be designed to form
highly viscous or gel-like systems at much lower droplet con-
centrations than conventional emulsions (Sonneville-Aubrun
et al., 2004; Tadros et al., 2004; Mason et al., 2006), which
may have interesting applications in some food products. Na-
noemulsions and other colloidal systems have been shown to
increase the bioavailability of certain types of lipophilic sub-
stances encapsulated within them (Acosta, 2009), which may
Address correspondence to D.J. McClements, Department of Food Science,
University of Massachusetts, Amherst, MA 01003. Tel.: 413 545 1019. E-mail:
mcclements@foodsci.umass.edu
285
This article provides an overview of the current status of na-
noemulsion formulation, fabrication, properties, applications,
biological fate, and potential toxicity, with specific emphasis on
edible systems that are applicable for application within the food
industry. Only oil-in-water nanoemulsions, which consist of oil
droplets dispersed within an aqueous phase, are considered in
this review since they currently have the greatest potential for
application within commercial products. Nevertheless, water-in-
oil nanoemulsions may also have some interesting applications
within certain types of food product.
Nanoemulsions: Terminology
Initially, we begin by describing and clarifying some of the
terminology commonly used to describe the characteristics of
emulsions, nanoemulsions, and closely-related colloidal disper-
sions (Table 1) (Mason et al., 2006). A conventional emul-
sion, also sometimes known as an emulsion or macro-emulsion,
typically has particles with mean radii between 100 nm and
100 µm. This type of colloidal dispersion is thermodynamically
286 D. J. McCLEMENTS AND J. RAO

Table 1 Comparison of thermodynamic stability and physicochemical properties of colloidal dispersions prepared from oil, water and emulsifier. The optical
properties are expressed for typical oil and water systems where there is a significant refractive index contrast, and the particle concentration is appreciable (e.g.,
> 0.1%)

System Droplet Radius Thermodynamic Stability Surface-to-Mass Ratio (m2 /g particles) Optical Properties

Emulsion 100 nm–100 µm Unstable 0.07−70 Turbid/Opaque

Nanoemulsion 10–100 nm Unstable 70−330 Clear/Turbid
Microemulsion 2–100 nm Stable 330−1300 Clear/Turbid

unstable, i.e., the free energy of the separated oil and water nanoemulsions, but much of the material covered will also be
phases is lower than that of the emulsion itself. Consequently, relevant to other types of colloidal dispersions.
conventional emulsions always have a tendency to breakdown Nanoemulsions, like conventional emulsions, can be clas-
over time. Conventional emulsions tend to be optically turbid or sified according to the relative spatial organization of the oil
opaque because the droplets they contain have similar dimen- and water phases. A system consisting of oil droplets dispersed
sions to the wavelength of light (r ≈ λ) and so they scatter light within a water phase is referred to as an oil-in-water (O/W)
strongly (provided the refractive index contrast between the oil nanoemulsion, whereas a system that consists of water droplets
and water phases is not very close to zero). A nanoemulsion, dispersed in an oil phase is referred to as a water-in-oil (W/O)
also referred to as a mini-emulsion, can be considered to be a nanoemulsion. The liquid that makes up the droplets is usually
conventional emulsion that contains very small particles, i.e., referred to as the dispersed phase, whereas the liquid that makes
mean radii between about 10 to 100 nm (Tadros et al., 2004; up the surrounding liquid is referred to as the continuous phase.
Mason et al., 2006). The relatively small particle size compared The particles in an O/W nanoemulsion can be considered to
to the wavelength of light (r << λ) means that nanoemulsions have a core-shell type structure, as represented schematically in
tend to be transparent or only slightly turbid (similar to mi- Fig. 1, with a core made of lipophilic material and a shell made
croemulsions). The very small particle size also means that of surface-active material. As mentioned previously, nanoemul-
they have much better stability to gravitational separation and sions are thermodynamically unfavorable systems because of
aggregation than conventional emulsions (Tadros et al., 2004; the positive free energy associated with creating an interface
Wooster et al., 2008). However, these systems are still thermo- between the oil and water phases due to the hydrophobic effect.
dynamically unstable, and so, will tend to breakdown over time. Consequently, they are metastable systems that tend to break-
In contrast, a micro-emulsion, also sometimes referred to as a down over time due to various physicochemical phenomenon
swollen micelle, is a thermodynamically stable system that typ-
ically contains particles with radii somewhere in the range of
2 to 100 nm. In this case, the free energy of the microemulsion is
lower than that of the phase-separated components from which it
is prepared, so it forms spontaneously. Nevertheless, there may
be kinetic energy barriers between the phase-separated com-
ponents and the microemulsion, which mean that some energy
must be inputted into the system before the microemulsion will
form, e.g., stirring. It is important to note that a microemul-
sion is only thermodynamically stable under a particular set of
environmental conditions (e.g., composition and temperature).
If these conditions are altered, then the microemulsion may no
longer be thermodynamically stable and will be converted to
another kind of system e.g., phase separated, liquid crystalline,
bicontinuous, nanoemulsion, or conventional emulsion. Never-
theless, if the system is brought back to the original conditions
(e.g., composition and temperature), then it should revert back
into a microemulsion, at a rate that depends on any kinetic en-
ergy barriers. Microemulsions are usually transparent because
the particle size is much smaller than the wavelength of light
(r << λ) so that light scattering is weak. From both a scientific Figure 1 Schematic representation of the core-shell structure of particles
and technical point of view, it is important to clearly distin- in nanoemulsions: a shell of amphiphilic material normally covers a core of
lipophilic material. The lipophilic material may contain a variety of different
guish between different kinds of colloidal dispersions since this
non-polar molecules, such as triacylglycerols, diacylglycerols, monoacylglyc-
determines the best approach to optimize their formation, sta- erols, and bioactive components. The amphiphilic shell may contain various
bility, physicochemical properties, and functional performance. surface-active molecules, such as surfactants, phospholipids, proteins, polysac-
In the remainder of this article, the focus will be primarily on charides or minerals.
 FOOD-GRADE NANOEMULSIONS
Table 2 Impact of various bulk physicochemical properties of oil and aqueous phases on the formation stability and performance of food-grade oil-in-water
nanoemulsions
Property Formation
Viscosity (η) – Efficiency of droplet disruption in high-energy
– Dispersed (ηD ) methods depends on viscosity ratio (ηD / ηC )
– Continuous (ηC )
Interfacial tension (γ ) – Low γ facilitates droplet disruption in high-energy
methods;
– Low γ facilitates droplet formation in
phase-inversion and spontaneous- emulsification
methods
Solubility – Water-solubility of surfactants and solvents affects
droplet formation in solvent displacement and
spontaneous-emulsification methods
Polarity – – Determines partitioning of
Density (ρ) – – Creaming rate increases with
Refractive index (n) – –
such as gravitational separation, flocculation, coalescence,
and/or Ostwald ripening (Dickinson, 1992; Friberg et al., 2004;
McClements, 2005). The rates at which these degradation
processes occur are often considerably different in nanoemul-
sions than in conventional emulsions because of differences in
their particle sizes. Nanoemulsions often have better stability
to gravitational separation, flocculation, and coalescence than
conventional emulsions, but are less stable to Ostwald ripening
(see later). A major concern of food scientists working with
this kind of system is therefore to create nanoemulsions with
sufficiently small particles and then to ensure that they have a
sufficiently long kinetic stability for commercial applications.
The kinetic stability of nanoemulsions can be improved by
controlling their microstructure (e.g., particle-size distribution),
or by incorporating substances known as stabilizers, such as
emulsifiers, texture modifiers, weighting agents, or ripening
retarders. (See the following section.)
Nanoemulsion Formulation
In this section, we provide an overview of the major compo-
nents that can be used to formulate food-grade nanoemulsions.
Oil Phase
The oil phase used to prepare nanoemulsions can be formu-
lated from various non-polar components, including triacylglyc-
erols, diacylgycerols, monoacylglycerols, free fatty acids, flavor
oils, essential oils, mineral oils, fat substitutes, waxes, weighting
agents, oil-soluble vitamins, and various lipophilic nutraceuti-
cals (such as carotenoids, curcumin, phytosterols, phytostanols,
and Co-enzyme Q). The formation, stability, and properties of
nanoemulsions often depend on the bulk physicochemical char-
acteristics of the oil phase, e.g., polarity, water-solubility, inter-
287
Stability Performance
– Creaming rate decreases with – Determines product formation,
increasing aqueous phase viscosity texture, shelf-life, and release
(ηC ) characteristics
– Low γ promotes droplet – Affects product formation and
coalescence; shelf-life
– Low γ may lead to poor emulsifier
affinity for droplet surfaces
– High water-solubility of an oil – Affects product shelf-life
phase promotes Ostwald ripening
– Affects release characteristics &
components between oil and flavor profiles
aqueous phases
– Determines product shelf-life
increasing density contrast
– Turbidity increases with increasing
refractive index contrast
facial tension, refractive index, viscosity, density, phase behav-
ior, and chemical stability (Tadros et al., 2004; McClements,
2005; Anton et al., 2007; Wooster et al., 2008; Anton et al.,
2009). These bulk physicochemical characteristics often limit
the type of homogenization method(s) that can be used to pre-
pare a nanoemulsion from a particular oil phase. A number of
ways in which the bulk physicochemical characteristics of the
oil phase may influence the formation, stability and performance
of nanoemulsions is summarized in Table 2.
In the food industry, it is often desirable to prepare na-
noemulsions using triacylglycerol oils due to their low cost,
abundance, and functional or nutritional attributes, e.g., corn,
soybean, sunflower, safflower, olive, flaxseed, algae, or fish oils.
Most of these oils primarily consist of long-chain triacylglyc-
erols (LCT), although medium-chain triacylglycerols (MCT)
and short-chain triacylglycerols (SCT) are being used in some
food applications. The formation of nanoemulsions using MCT
and LCT oils is often challenging due to their relatively low
polarity, high interfacial tension, and high viscosity. For ex-
ample, it is difficult to prepare nanoemulsions from these oils
using the phase-inversion temperature (PIT) method because
of their very high hydrophobicity (Witthayapanyanon et al.,
2006). Similarly, it is difficult to prepare nanoemulsions from
these oils using high-pressure homogenization methods because
their high viscosity limits droplet disruption within the homog-
enizer (Palamakula and Khan, 2004; Wooster et al., 2008; Do
et al., 2009). In these situations it is often necessary to modify
or optimize conventional homogenization methods to create the
very small droplets needed for nanoemulsion production (see
later). Once a nanoemulsion has been created using medium- or
long-chain triacylglycerol oils it is often highly physically sta-
ble. Conversely, some types of edible oils are highly effective at
forming nanoemulsions containing small droplets, but are much
less effective at stabilizing the droplets once they have been pro-
duced. For example, flavor and essential oils have a relatively
288 D. J. McCLEMENTS AND J. RAO
Figure 2 Nanoemulsions may breakdown through a variety of different physicochemical mechanisms, including gravitational separation, flocculation, coales-
cence, Ostwald ripening, and oiling-off.
high polarity, low interfacial tension, and low viscosity, which
facilitate the formation of very small droplets by both high pres-
sure homogenization and PIT methods. However, once this type
of nanoemulsion is formed it may rapidly breakdown because
of either Ostwald ripening (due to the high water-solubility of
the oil) or coalescence (due to the low interfacial tension of the
oil). Again, alternative strategies usually have to be developed
to improve the long-term stability of nanoemulsions created
using these oils, e.g., by adding ripening inhibitors to prevent
Ostwald ripening or by changing the emulsifier type to prevent
coalescence (see later).
Aqueous Phase
The aqueous phase used to prepare a nanoemulsion typically
consists primarily of water, but it may also contain a variety of
other polar components, including co-solvents (such as simple
alcohols and polyols), carbohydrates, proteins, minerals, acids,
and bases. The type and concentration of these components de-
termines the polarity, interfacial tension, refractive index, rheol-
ogy, density, phase behavior, pH, and ionic strength of the aque-
ous phase, which in turn will impact the formation, stability,
and physicochemical properties of the nanoemulsion produced
(Table 2). Careful control of the aqueous phase composition can
be used to optimize the formation or improve the stability of na-
noemulsions. The formation of small droplets in nanoemulsions
using high pressure-homogenization methods can often be facil-
itated by adding water-soluble cosolvents to the aqueous phase
to optimize the disperse-to-continuous phase viscosity ratio.
(See section titled “High Energy Approaches.”) The presence
of cosolvents may also improve the ability of small molecule
surfactants to form nanoemulsions using phase-inversion meth-
ods. The impact of specialized kinds of molecules, referred to
as “stabilizers,” which might be added to either aqueous or oil
phases, is discussed in the following section.
Stabilizers
If only an oil phase and an aqueous phase are homogenized
together, the system will normally rapidly breakdown through
a variety of different mechanisms, including droplet floccula-
tion, coalescence, Ostwald ripening, and gravitational separa-
tion (Fig. 2). For this reason, it is often necessary to add various
kinds of stabilizers to nanoemulsions to improve their long-term
stability.
Emulsifiers/Co-emulsifiers. The selection of an appropriate
emulsifier (or combination of emulsifiers) is one of the most
important factors to consider for the proper design of a
nanoemulsion. An emulsifier is a surface-active molecule
that is capable of adsorbing to droplet surfaces, facilitating
droplet disruption, and protecting droplets against aggregation
(McClements, 2005; Kralova et al., 2009). In high-energy ap-
proaches, the emulsifier facilitates droplet disruption within the
homogenizer by lowering the interfacial tension, which favors
the production of small droplets. In low-energy approaches,
the emulsifier facilitates the spontaneous formation of small
 FOOD-GRADE NANOEMULSIONS
droplets due to its ability to produce very low interfacial tensions
under certain environmental and solution conditions (see later).
The stability of a nanoemulsion to environmental stresses such
as pH, ionic strength, heating, cooling, or long-term storage is
often predominantly determined by the kind of emulsifier used.
In the food industry, the most important types of emulsifiers
are small molecule surfactants, phospholipids, proteins, and
polysaccharides. The type of emulsifier used has a major impact
on the type of homogenization approach that can be used to form
a nanoemulsion. Many small molecule surfactants are highly
effective at producing nanoemulsions using both high- and
low-energy methods. On the other hand, proteins and polysac-
charides are not typically suitable for producing nanoemulsions
using low-energy methods, and are not usually as effective as
surfactants at forming nanoemulsions with small droplet sizes
using high-energy methods. On the other hand, proteins and
polysaccharides have the advantage of being natural ingredients
that are often considered more “label-friendly,” and therefore
there is great interest in producing nanoemulsions with them.
It is often convenient to classify small molecule surfactants
according to their electrical characteristics—ionic, non-ionic,
and zwitterionic (McClements, 2005):
• Ionic surfactants: Most food-grade ionic surfactants are neg-
atively charged (such as CITREM, DATEM, and SLS), but
at least one positively charged surfactant is available for cer-
tain applications (i.e., lauric arginate). Ionic surfactants may
be used to form nanoemulsions by various low-energy and
high-energy approaches. Their utilization may be limited in
products where high-surfactant levels are required because
they tend to cause irritation (Sol et al., 2006; Solè et al.,
2006).
• Non-ionic surfactants: Non-ionic surfactants have been
widely used to form nanoemulsions due to their low toxicity,
lack of irritability, and the capacity to easily form nanoemul-
sions by both high-energy and low-energy approaches. Exam-
ples include sugar ester surfactants (e.g., sorbitan monooleate,
sucrose monopalmitate), polyoxyethylene ether (POE) sur-
factants (e.g., Brij 97), and ethoxylated sorbitan esters (e.g.,
Tweens and Spans) (Chiu, 2006; Liu et al., 2006; Jafari et al.,
2007; Henry et al., 2009).
• Zwitterionic surfactants: Zwitterionic surfactants have two
or more oppositely charged ionizable groups on the same
molecule. Consequently, they can have a net negative, neutral,
or positive charge depending on solution pH. Phospholipids
are common zwitterionic surfactants that have GRAS status,
which permits their use in food, e.g., lecithin. However, many
natural phospholipids are not particularly good at either form-
ing or stabilizing nanoemulsions when used in isolation, but
they may be effective when used in combination with co-
surfactants (Trotta et al., 1996; de Morais et al., 2006; Hoeller
et al., 2009).
The formation and stability of nanoemulsions can often be
improved by using combinations of emulsifiers, rather than us-
289
ing a single emulsifier. For example, employing a lipophilic and
hydrophilic surfactant in conjunction can facilitate the forma-
tion of small particles using both low-energy and high-energy
approaches. On the other hand, using mixed-emulsifier systems
can often improve the stability of nanoemulsions to particle ag-
gregation after formation. (See section titled “Finishing Tech-
niques.”)
Some methods of forming nanoemulsions using low-energy
methods require the presence of cosurfactants (such as short-
and medium-chain alcohols) or cosolvents (such as polyols like
propylene glycol, glycerol, and sorbitol) (Yaghmur et al., 2002;
Flanagan et al., 2006). Cosurfactants are amphiphilic molecules
that are surface active (often having a hydrocarbon chain and a
hydroxyl group), but that are not good at stabilizing emulsions
themselves due to the small size of the polar head group. A num-
ber of physicochemical mechanisms have been proposed to ac-
count for the ability of co-surfactants to facilitate nanoemulsion
formation— (1) Fluidization of the interface; (2) Optimization
of the disperse-to-continuous phase viscosity ratio; (3) Reduc-
tion of the electrical repulsion between the head groups of ionic
surfactants at an interface by acting as spacers; and, (4) Induc-
tion of the appropriate interfacial curvature (Gradzielski, 1998;
Garti et al., 2001; Shafiq-un-Nabi et al., 2007). Cosolvents are
highly polar molecules that are not particularly surface active
themselves, but that may alter the physicochemical properties of
the emulsifier molecules, such as their surface-activity, oil-water
partition coefficient, and the ability to form colloidal structures.
Cosolvents may do this by altering the structure and interac-
tions of the molecules in the aqueous phase, which alters the
magnitude of any hydrophobic interactions.
Texture Modifier. A texture modifier is a substance that thick-
ens or gels the aqueous phase (Imeson, 2010). Texture modi-
fiers are often incorporated into commercial products to improve
emulsion stability by retarding droplet movement, but they may
also be used to provide desirable textural characteristics, such as
“creaminess,” “richness,” “thickness,” or gel strength. Most tex-
ture modifiers used in the food industry are biopolymers, such
as proteins (e.g., egg, milk, vegetable proteins) or polysaccha-
rides (e.g., starch, pectin, alginate, carrageenan, xanthan, guar
gum).
Weighting Agent. A weighting agent is a substance that is
usually added to the oil droplets in O/W emulsions to match
their density to that of the surrounding aqueous phase (Mc-
Clements, 2005). Its purpose is to reduce the driving force for
gravitational separation (ρ) and therefore prevent or retard
creaming or sedimentation. Commonly used weighting agents
in the food and beverage industries include brominated veg-
etable oil (BVO), ester gum, damar gum, and sucrose-acetate
isobutyrate (SAIB) (McClements, 2005). The density of the
particles in nanoemulsions may also be matched to that of the
surrounding aqueous phase by coating them with thick dense
layers, or by partially crystallizing the lipid core. (See section
titled “Designing Functional Nanoemulsion Particles.”)
Ripening Retarder. The concept of a ripening retarder is
relatively new in the food industry, but they are particularly
290 D. J. McCLEMENTS AND J. RAO
important for stabilizing certain kinds of food-grade nanoemul-
sions, e.g., those containing oils susceptible to Ostwald ripening
(OR), such as flavor oils, essential oils, and short-chain triacyl-
glycerols. A ripening retarder is a highly hydrophobic material
that is incorporated into oil droplets to retard or inhibit Ostwald
ripening. OR is the process where large droplets grow at the
expense of smaller droplets due to diffusion of oil molecules
through the intervening aqueous phase. (See section titled
“Stability.”) Typically, a ripening retarder is a hydrophobic
component that is soluble in the oil phase but highly insoluble in
water, e.g., a long-chain triglyceride, mineral oil, or ester gum
(Kabalnov et al., 1987; Sonneville-Aubrun et al., 2004). A ripen-
ing retarder works by generating an entropy of mixing effect
that counteracts the OR effect. (See section titled “Stability.”)
NANOEMULSION FORMATION
There have been considerable advances in our understanding
of the methods that can be used to create nanoemulsions in the
past few years. Nanoemulsions can be fabricated using a num-
ber of different approaches, but these can usually be broadly
categorized as either high-energy or low-energy approaches de-
pending on the underlying principle (Tadros et al., 2004; Pouton
and Porter, 2006; Acosta, 2009; Anton et al., 2009; Leong et al.,
2009).
High-energy approaches utilize mechanical devices (“ho-
mogenizers”) capable of generating intense disruptive forces
that are capable of disrupting and intermingling the oil and
aqueous phases into tiny oil droplets, e.g., high pressure valve
homogenizers, microfluidizers, and sonication methods (Gutier-
rez et al., 2008; Velikov et al., 2008; Wooster et al., 2008; Leong
et al., 2009). At present, high-energy approaches are the most
common method used to prepare nanoemulsions in industrial
food operations because their utilization in the production of
conventional emulsions is already well-established, they are ca-
pable of large-scale production, and they can be used to prepare
nanoemulsions from a variety of different starting materials.
Low-energy approaches rely on the spontaneous formation of
tiny oil droplets within mixed oil-water-emulsifier systems when
the solution or environmental conditions are altered, e.g., phase-
inversion and spontaneous-emulsification methods (Bouchemal
et al., 2004; Tadros et al., 2004; Freitas et al., 2005; Chu et al.,
2007a; Anton et al., 2008; Yin et al., 2009). The minimum
particle size that can be produced using each approach depends
on many different factors, which are highlighted in the sections
below.
High-Energy Approaches
In general, the particle size produced by high-energy ap-
proaches is governed by a balance between two opposing pro-
cesses occurring within the homogenizer—droplet disruption
and droplet coalescence (Jafari et al., 2008). Only those mechan-
ical devices that are capable of generating extremely intense dis-
ruptive forces are capable of producing the tiny droplets required
to form nanoemulsions (Fig. 3), i.e., high-pressure valve homog-
enizers, microfluidizers, and ultrasonic devices (Tadros et al.,
2004; Leong et al., 2009). The reason that such intense energies
are needed is that the disruptive forces generated by the homog-
enizer must exceed the restoring forces holding the droplets into
spherical shapes (Walstra, 1993; Schubert and Behrend, 2003;
Schubert and Engel, 2004). These restorative forces are deter-
mined by the Laplace Pressure: P = γ /2r,which increases
with decreasing droplet radius (r) and increasing interfacial
tension (γ ). Thus, as the droplet radius becomes increasingly
smaller within a homogenizer, it becomes increasingly difficult
to break them up further. The smallest droplet size that can be
produced by a particular high-energy device depends on the ho-
mogenizer design (e.g., flow and force profiles), homogenizer
operating conditions (e.g., energy intensity, duration), environ-
mental conditions (e.g., temperature), sample composition (e.g.,
oil type, emulsifier type, concentrations), and the physicochemi-
cal properties of the component phases (e.g., interfacial tension,
viscosity) (Kentish et al., 2006; Wooster et al., 2008). Previous
studies have shown that the droplet size tends to decrease when
the energy intensity or duration increases, the interfacial ten-
sion decreases, the emulsifier adsorption rate increases, and the
disperse-to-continuous phase viscosity ratio falls within a cer-
tain range (0.05 < ηD /ηC < 5) (Walstra, 1993; Walstra, 2003;
Tadros et al., 2004). The extent of the ηD /ηC range where small
droplets can be produced depends on the nature of the disrup-
tive forces generated by the particular homogenizer used, i.e.,
simple shear versus extensional flow.
High-energy approaches are one of the most versatile means
of producing food-grade nanoemulsions because they can be
used with a wide variety of different oil and emulsifier types.
Provided that the homogenization conditions are suitably op-
timized, nanoemulsions can be produced using triacylglycerol
oils, flavor oils, and essentials oils as the oil phase, and proteins,
polysaccharides, phospholipids, and surfactants as emulsifiers.
Even so, the size of the particles produced depends strongly on
the characteristics of the oil and emulsifier used (see below).
For example, it is usually easier to produce very small droplets
when the oil phase has a low viscosity and/or interfacial tension
(e.g., flavor oils, essential oils, alkanes) than when it has a high
viscosity and/or interfacial tension (e.g., MCT or LCT).
High Pressure Valve Homogenizer
High pressure valve homogenizers are probably the most
common method of producing conventional emulsions with
small droplet sizes in the food industry (Schubert and Behrend,
2003; Schubert and Engel, 2004). They are more effective at re-
ducing the size of the droplets in pre-existing coarse emulsions,
than at creating emulsions directly from two separate liquids. A
coarse emulsion is usually produced using a high shear mixer
and is then fed directly into the inlet of the high pressure valve
homogenizer. The homogenizer has a pump that pulls the coarse
 FOOD-GRADE NANOEMULSIONS
Figure 3 Schematic representation of various mechanical devices that can be used to produce food-grade nanoemulsions using a high-energy approach: high
pressure valve homogenizer; microfluidizer: ultrasonic jet homogenizer; ultrasonic probe homogenizer.
emulsion into a chamber on its backstroke and then forces it
through a narrow valve at the end of the chamber on its forward
stroke (Fig. 3). As the coarse emulsion passes through the valve
it experiences a combination of intense disruptive forces that
cause the larger droplets to be broken down into smaller ones.
Different nozzle designs are available to increase the efficiency
of droplet disruption. The droplet size produced using a high
pressure valve homogenizer usually decreases as the number
of passes and/or the homogenization pressure increases. It also
depends on the viscosity ratio of the two phases (usually oil and
water) being homogenized. Small droplets can only usually be
produced when the disperse-to-continuous phase viscosity ratio
falls within a certain range (0.05 < ηD /ηC < 5) (Walstra, 1993;
Walstra, 2003; Tadros et al., 2004). Finally, it is important to have
sufficient emulsifier present to cover all the new droplet surfaces
formed during homogenization, and to use an emulsifier that can
rapidly adsorb to the droplet surfaces to prevent re-coalescence
(Jafari et al., 2008). Usually, there is a linear relationship be-
tween the logarithm of the homogenization pressure (P ) and the
logarithm of the droplet diameter (d): log d ∝ log P. The con-
stant of proportionality depends on the homogenizer—for large
homogenizers and low-viscosity fluids d ∝ P −0.6 ; for large ho-
mogenizers and high-viscosity fluids d ∝ P −0.75 ; and for small
(bench-top) homogenizers d ∝ P −1.0 (McClements, 2005). To
reduce the droplet size to the level required in nanoemulsions it
is usually necessary to operate at extremely high pressures and
to use multiple passes through the homogenizer. Even then, it
is only possible under certain circumstances to obtain droplets
291
less than 100 nm in radius (e.g., high emulsifier levels, low in-
terfacial tensions, and appropriate viscosity ratios). A number
of recent studies that have used high-pressure homogenization
to produce nanoemulsions that could potentially be used in the
food industry are summarized in Table 3.
Microfluidizers
A microfluidizer is somewhat similar in design to a high-
pressure homogenizer in that it involves using high pressures
to force an emulsion pre-mix through a narrow orifice to fa-
cilitate droplet disruption. However, the design of the channels
through which the emulsion is made to flow within the homog-
enizer is different (Fig. 3). Microfluidizers have traditionally
been used in the pharmaceutical industry to produce emulsion-
based products, but they have also been used in the food and
beverage industries to produce flavor emulsions, nutraceutical
emulsions, and homogenized milk. The microfluidizer works on
the principle of dividing an emulsion flowing through a chan-
nel into two streams, passing each stream through a separate
fine channel, and then directing the two streams at each other
in an interaction chamber. Intense disruptive forces are gener-
ated within the interaction chamber when the two fast-moving
streams of emulsion impinge upon each other, leading to highly
efficient droplet disruption.
A number of studies have examined the potential application
of microfluidizers for the production of model food nanoemul-
sions, such as beverage and dairy emulsions (Dalgleish et al.,
292 D. J. McCLEMENTS AND J. RAO

Table 3 An overview of recent research articles on O/W nanoemulsion formation using high pressure valve homogenizers

Oil Phase Bioactive component Surfactant/Co-surfactant Particle Diameter Reference

MCT β−carotene Tween 20,40,60,80 132–148 nm Yuan et al., 2008

MCT β−carotene Tween 20, decaglycerol monolaurate, whey 115–178 nm Mao et al., 2009
protein isolate, modified starch
Oleoresin Capsicum/ethyl Poly-caprolactone Pluronic F68 320–460 nm Choi et al., 2009
acetate
Olive oil Pluronic F68, Phospholipid 239–274 nm Wulff-Perez et al., 2010
MCT Curcumin Tween 20 160 nm Wang et al., 2008

1997; Henry et al., 2010; Klein et al., 2010). These studies have
shown that small droplets can be produced provided that con-
ditions are optimized to facilitate droplet disruption and inhibit
droplet coalescence. A number of recent studies carried out us-
ing a microfluidizer to produce nanoemulsions are highlighted
in Table 4. Like high pressure valve homogenizers, the droplet
size produced tends to decrease with increasing homogenization
pressure, number of passes, and emulsifier concentration. In ad-
dition, the disperse-to-continuous phase viscosity ratio should
be within a certain range to facilitate the formation of small
droplets (Wooster et al., 2008). As with the high pressure valve
homogenizer, there is usually a linear relationship between the
logarithm of homogenization pressure and the logarithm of the
droplet diameter: log d ∝ log P. Recently, our laboratory per-
formed experiments to elucidate the major factors that influence
the formation of nanoemulsions using a microfluidizer. The log-
arithm of the mean droplet diameter decreased linearly as the
logarithm of the homogenization pressure increased for both an
ionic surfactant (SDS) and a globular protein (β-lactoglobulin)
(Fig. 4). However, the slope of the log(d) vs. log(P ) relationship
was appreciably steeper for the surfactant (-0.57) than for the
protein (-0.29), which was attributed to the fact that the protein
may adsorb more slowly to the droplet surfaces, and that it may
form a viscoelastic coating that inhibits further droplet breakup.
The dependence of the mean droplet diameter on the viscosity
ratio (ηD /ηC ) was also examined by preparing emulsions us-
ing different oil-phase compositions (corn oil : octadecane) and
aqueous-phase compositions (water : glycerol) (Fig. 5). For the
ionic surfactant there was a distinct decrease in mean droplet
diameter with decreasing viscosity ratio, which suggested that
droplet disruption within the homogenizer became easier as the
viscosity of the two phases became more similar. On the other
hand, we found little change in mean droplet size with viscosity
ratio when a globular protein was used as an emulsifier, which
again may be due to the relatively slow adsorption of the pro-
tein and its ability to form a coating that inhibits further droplet
disruption.
Ultrasonic Homogenizers
Ultrasonic homogenizers utilize high-intensity ultrasonic
waves to create the intense disruptive forces necessary to
breakup up oil and water phases into very small droplets (Ken-
tish et al., 2006; Lin and Chen, 2008; Leong et al., 2009). The
energy input is provided using sonicator probes that contain
piezoelectric quartz crystals that expand and contract in response
to an alternating electrical voltage (Fig. 3). The tip of the son-
icator probe is placed within the liquids being homogenized,
where it generates intense mechanical vibrations that lead to
cavitation effects, i.e., the formation, growth, and collapse of
small bubbles in the liquid. The collapse of the micro-bubbles
formed by cavitation generates intense disruptive forces in the
immediate vicinity of the sonicator probe that leads to droplet
disruption. For efficient and uniform homogenization it is im-
portant to ensure that the emulsion spends sufficient time within
the region where droplet disruption occurs. Recently, batch and
flow-through ultrasonic homogenizers have been developed that
have special cells designed to optimize the efficiency of droplet
disruption (Leong et al., 2009). A number of recent studies car-
ried out using sonication to produce nanoemulsions are high-
lighted in Table 5. Experiments have shown that the size of the
droplets produced using an ultrasonic device tends to decrease
as the intensity of the ultrasonic waves is increased, or the resi-
dence time in the disruption zone is increased (Abismail et al.,

Table 4 An overview of recent research articles on O/W nanoemulsion formation using a microfluidizer

Oil Phase Bioactive component Surfactant/Co-surfactant∗ Particle Diameter Reference

WPC, WPH, SC, modified starch 150–600 nm Jafari et al., 2007

Peanut oil Tween 80, SDS 120 nm Wooster et al., 2008
Hexadecane Tween 80, SDS 80 nm Wooster et al., 2008
Corn oil/Ethyl acetate WPI 174 nm Lee and McClements, 2010
MCT CO Q10 Nonionic surfactant, soybean lecithin, glycerol 60 nm Hatanaka et al., 2008
Corn oil/Octadecane SDS, Tween20, beta-lactoglobulin 60–150 nm Qian and McClements, 2010
MCT α−Tocopherol Glycerol, Decaglyceryl monooleate Soybean 80–400 nm Hatanaka et al., 2010
lecithin
∗ Here the acronyms WPC, WPH, and SC mean whey protein concentrate, whey protein hydrolysate, and sodium caseinate, respectively.
 FOOD-GRADE NANOEMULSIONS 293

Figure 5 Influence of the droplet to continuous phase viscosity ratio on the

Figure 4 Influence of homogenization pressure on the mean particle diameter mean particle diameter of droplets produced using a microfluidizer using either a
of droplets produced using a microfluidizer using either a protein (BLG) or protein (BLG) or surfactant (SDS) as emulsifier (Qian and McClements, 2010).
surfactant (SDS) as emulsifier (Qian and McClements, 2010).

noemulsions are based on the low-energy approach, including

1999; Maa and Tsu, 1999). The homogenization efficiency also
depends on the type and amount of emulsifier present, and the
viscosity of the oil and aqueous phases (Maa and Tsu, 1999;
Jafari et al., 2006; Kentish et al., 2006; Leong et al., 2009).
All of these parameters must therefore be optimized to produce
nanoemulsions containing small droplets. There is also some
concern that the high local intensities involved in sonication
methods may lead to protein denaturation, polysaccharide de-
polymerization, or lipid oxidation during homogenization.
Low-Energy Approaches
spontaneous emulsification- and phase-inversion methods (Fer-
nandez et al., 2004; Pouton and Porter, 2006; Anton et al., 2008;
2009; Maestro et al., 2008).
Low-energy approaches are often more effective at producing
small droplet sizes than high- energy approaches, but they are
often more limited in the types of oils and emulsifiers that can
be used. For example, it is currently not possible to use proteins
or polysaccharides as emulsifiers in most of the low-energy
approaches used to form nanoemulsions. Instead, it is often
necessary to utilize relatively high concentrations of synthetic
surfactants to form nanoemulsions by these approaches, which
may limit their use for many food applications.

The formation of emulsions using low-energy approaches

relies on the spontaneous formation of oil droplets in oil-water-
emulsifier mixtures when either their composition or their en-
vironment is altered (Bouchemal et al., 2004; Tadros et al.,
2004; Freitas et al., 2005; Chu et al., 2007a; Anton et al., 2008;
2009; Yin et al., 2009). A number of methods for preparing na-
Spontaneous Emulsification
In this method an emulsion or nanoemulsion is spontaneously
formed when two liquids are mixed together (Miller, 1988; Pou-
ton and Porter, 2006; Anton et al., 2009). Systems prepared us-
ing this method are usually referred to as either self-emulsifying

Table 5 An overview of research articles on O/W nanoemulsions using Ultrasonic Homogenizer

Oil phase Surfactant/Co-surfactant Particle Diameter Reference

Medium-chain triglyceride Tween 80/ethanol 20–60 nm Amani et al., 2010

Sunflower oil, Canola oil Tween 80, Span 80, SDS 40 nm Leong et al., 2009
Flax seed oil Tween 40 150 nm Kentish et al., 2008
Olive oil, Sesame oil, Soybean oil Pluronic F68 < 500 nm Wulff-Perez et al., 2009
Soybean oil Phosphatidylcholine / Sodium palmitate, sucrose palmitate 58 nm Takegami, et al., 2008
294 D. J. McCLEMENTS AND J. RAO

Table 6 Some recent research articles on O/W nanoemulsions formed using the Spontaneous Emulsification method that may be suitable for food applications

Oil Phase Bioactive component Surfactant/Co-surfactant Particle Diameter Reference

MCT α-Tocopherol Span 80, Span85, Tween 20, Tween 80/Acetone, Ethanol, Ethyl acetate 100–600 nm Bouchemal et al., 2004
MCT Castor oil Soybean lecithin, Polyoxyl 35 castor oil 150–200 nm Kelmann et al., 2007
MCT Oligothymidylate Egg lecithin/glycerol 225 nm Martini et al., 2008
Palm oil Vitamin E Tween 80, Plurionic F-68 94 nm Teo et al., 2010

drug-delivery systems (SEDDS) or self-nano-emulsifying drug-
delivery systems (SNEDDS) in the pharmaceutical industry. A
number of recent studies using the spontaneous emulsification
approach are highlighted in Table 6. Practically, this method can
be carried out in a number of different ways—the compositions
of the two phases can be varied; the environmental conditions
can be varied (e.g., temperature, pH, and ionic strength); and/or,
the mixing conditions can be varied (e.g., stirring speed, rate of
addition, and order of addition). For example, an organic phase
consisting of a hydrophobic oil, a hydrophilic surfactant, and a
water-miscible organic solvent may be slowly added to water
(Anton et al., 2009). Alternatively, water may be added to an or-
ganic phase containing hydrophobic oil, water-miscible organic
solvent, and surfactant (Sonneville-Aubrun et al., 2009). The
water-miscible organic solvent is typically ethanol or acetone,
but in some cases it is possible to replace it with a water-miscible
surfactant instead (Anton et al., 2008; 2009).
A number of physicochemical mechanisms have been pro-
posed to account for spontaneous emulsification (Horn and
Rieger, 2001). Consider a situation where two phases are
brought into contact with each other. Each phase contains some
components that are immiscible with the other phase (e.g., one
phase contains oil and the other phase contains water), and one
of the phases contains another component that is partially misci-
ble with both phases (e.g., an amphiphilic alcohol or surfactant).
When the two phases are brought into contact, some of the com-
ponent that is partially miscible with both phases will move from
its original location in one phase into the other phase. As this
component moves it will cause an increase in oil-water inter-
facial area, interfacial turbulence, and spontaneous formation
of droplets (Fig. 6). The size of the droplets produced can be
controlled by varying the compositions of the two initial phases,
as well as the mixing conditions.
Previous studies using the spontaneous emulsification
method have used one or more synthetic surfactants. To the
knowledge of the authors, there are currently no studies where
proteins or polysaccharides have been used as an emulsifier in
the spontaneous emulsification approach. This may be because
of physicochemical limitations associated with the droplet-
formation mechanism in this method, i.e., the need for small oil
droplets to spontaneously form and bud-off from the oil-water
interface (Fig. 6). Interfaces covered with biopolymer emulsi-
fiers tend to have higher interfacial tensions and more elastic
properties than those formed with small molecule surfactants.
We recently compared the spontaneous emulsification
method of producing nanoemulsions with a high energy method.
The surfactant-oil-water system used consisted of 15.4 wt%
non-ionic surfactant, 23.1 wt% MCT, and 61.5% water, with the
surfactant containing a 50:50 mixture of a hydrophilic (Tween
80) and lipophilic (Tween 85) surfactant. Nanoemulsions were
prepared using two different methods: (i) High-energy method:
the surfactant, oil and water were blended together, and then
passed through a microfluidizer; (ii) Low-energy method: The
oil and surfactant were mixed together, and then simply added
to the water using slow stirring.
The microfluidization method produced droplets with
a diameter of about 110 nm, whereas the spontaneous-
emulsification method could produce droplets with diameters
around 140 nm. This simple experiment demonstrated that
nanoemulsions could be produced using the spontaneous

Figure 6 Schematic representation of proposed mechanism for spontaneous emulsification: oil droplets are spontaneously formed when an oil phase containing
a water-dispersible substance is mixed with an aqueous phase. The underlying mechanism is the movement of the water-dispersible substance from the oil phase
to the water phase (red arrows), leading to interfacial turbulence and spontaneous oil droplet formation.
 FOOD-GRADE NANOEMULSIONS
Figure 7 Schematic diagram of the temperature-dependence of the spontaneous curvature of surfactant monolayers and their influence on emulsion properties.
emulsification method, provided that the system composition
was optimized, i.e., surfactant, oil, and water contents.
Phase-Inversion Methods
A number of methods have been developed to formulate
nanoemulsions that depend on inducing a phase inversion in
emulsions from a W/O to O/W form (or vice versa), e.g.,
phase-inversion temperature (PIT), phase-inversion composi-
tion (PIC), and emulsion-inversion point (EIP) methods. We be-
gin by describing the principle underlying each of these meth-
ods, and then we conclude by highlighting a recent general
conceptual approach that has been developed to rationalize the
formation of emulsions based on the phase-inversion methods.
Phase-Inversion Temperature Methods. The phase-
inversion temperature method (PIT) relies on changes in the
optimum curvature (molecular geometry) or relative solubility
of non-ionic surfactants with changing temperature (Anton et
al., 2007; 2009; Gutierrez et al., 2008). Nanoemulsions can
be spontaneously formed using the PIT method by varying
the temperature-time profile of certain mixtures of oil, water,
and non-ionic surfactant. This type of phase inversion usually
involves the controlled transformation of an emulsion from one
type to another (e.g., W/O to O/W or vice versa) through an
intermediate liquid crystalline or biocontinuous microemulsion
phase. The driving force for this type of phase inversion is
changes in the physicochemical properties of the surfactant with
295
temperature (Fig. 7). The molecular geometry of a surfactant
can be described by a packing parameter, p (Israelachvili,
1992):
aT
p=
aH
where, aT and aH are the cross-sectional areas of the lipophilic
tail-group and hydrophilic head-group respectively. Due to the
hydrophobic effect, the surfactant molecules tend to sponta-
neously associate with each other in water and form monolayers
that have an optimum curvature that allows the most efficient
packing of the molecules (Israelachvili, 1992). The optimum
curvature of a surfactant monolayer depends on the packing pa-
rameter of the surfactant—for p < 1, the optimum curvature
is convex, for p = 1, monolayers with zero curvature are pre-
ferred, and, for p > 1 the optimum curvature is concave (Fig.
7). In an emulsion or nanoemulsion, the surfactant monolayer
is present at an oil-water interface. The type of emulsion that
a particular surfactant tends to favor depends on its molecular
geometry. Surfactants with p < 1 tend to favor the formation
of O/W emulsions, those with p > 1 favor W/O emulsions,
and those with p = 1 do not favor either O/W or W/O sys-
tems and instead lead to the formation of liquid crystalline or
bicontinuous systems (Fig. 7). Changes in the relative solubil-
ity of surfactants in oil and water phases with temperature are
also needed to understand the physicochemical properties of
non-ionic surfactants (Anton et al., 2007; Anton et al., 2009).
296 D. J. McCLEMENTS AND J. RAO

At low temperatures, the head group of a non-ionic surfactant

is highly hydrated and so it tends to be more soluble in water.
As the temperature is raised, the head group becomes progres-
sively dehydrated and the solubility of the surfactant in water
decreases. At a particular temperature (≈ PIT), the solubility of
the surfactant in the oil and water phases is approximately equal.
At higher temperatures, the surfactant becomes more soluble in
the oil phase than in the water phase.
The temperature dependence of the packing parameter or sol-
ubility of non-ionic surfactants accounts for the ability to form
nanoemulsions using the PIT method (Fig. 7). The temperature
at which an oil-water-surfactant system changes from an O/W
emulsion to a W/O emulsion is known as the phase inversion
temperature or PIT. At temperatures well below the PIT (≈ T <
PIT-30◦ C), the surfactant is soluble in water, its packing param-
eter is appreciably less than unity (p < 1), and it favors the
formation of O/W emulsions. As the temperature is raised, the
hydrophilic head-groups of the non-ionic surfactant molecules
become progressively dehydrated, which causes its water sol-
ubility to decrease, and its packing parameter to tend towards
unity. At the PIT the packing parameter equals unity (p =
1), and the emulsion breaks down because the droplets have
an ultralow interfacial tension and therefore readily coalesce
with each other. The resulting system may consist of excess oil,
excess water, and a third phase containing a mixture of surfac-
tant molecules, oil, and water organized into liquid crystalline
or bicontinuous microemulsion structures. To form nanoemul-
sions it is important to control the overall system composition
so that only this third phase is formed at the PIT. At tempera- Figure 8 Temperature dependence of the turbidity of tetradecane oil-in-water
nanoemulsions. The turbidity increases above the PIT due to droplet coalescence
tures sufficiently greater than the PIT (≈ T > PIT + 20◦ C), the and phase inversion.
surfactant is more soluble in oil than water, its packing param-
eter is larger than unity, and the formation of a W/O emulsion
is favored. A nanoemulsion can be formed spontaneously by sion was then measured as it was heated at a controlled rate.
rapidly cooling an emulsion from a temperature at or slightly At T << PIT, the turbidity was low because the oil droplets in
above the PIT to a temperature well below the PIT with contin- the nanoemulsion were so small that they did not scatter light
uous stirring. Recently, it has been proposed that the formation strongly. As the temperature approached the PIT, there was an
of nanoemulsions using the PIT method has a similar physico- increase in the turbidity because the optimum curvature tended
chemical basis as their formation using the spontaneous emulsi- towards unity, which increased the droplet coalescence rate and
fication method (Anton et al., 2009). Above the PIT, a non-ionic led to an increase in particle size. Above the PIT, the system was
surfactant/oil/water mixture consists of a W/O emulsion with the highly turbid since a water-in-oil emulsion was formed with rela-
surfactant molecules being present predominantly within the oil tively large droplets that scattered light strongly. If this emulsion
droplets because they are more oil-soluble than water-soluble at was again cooled rapidly to a temperature well below the PIT,
this temperature. When this system is quench-cooled below the a nanoemulsion could be formed again. This figure highlights
PIT, the surfactant molecules rapidly move from the oil phase one of the major limitations of the PIT method of producing
into the aqueous phase (just like the movement of water-miscible nanoemulsions—the droplets tend to be highly prone to droplet
solvent in the spontaneous-emulsification method), which leads coalescence when the temperature of the system is raised. This
to the spontaneous formation of small oil droplets because of could be a problem in many food and beverage applications that
the increase in interfacial area and interfacial turbulent flow require some form of thermal treatment, such as pasteurization,
generated (Fig. 7). sterilization, or cooking. Recently, we developed an approach
An example of the phase behavior of a mixture consisting to overcome this problem by forming nanoemulsions using a
of oil (20 wt% tetradecane), water (74%), and non-ionic sur- non-ionic surfactant with a relatively low PIT (Brij 30), and
factant (6% Brij30) upon heating is shown in Fig. 8 (Rao and then diluting the resulting nanoemulsions in a solution contain-
McClements, 2010). Initially, a nanoemulsion (d = 60 nm) was ing another surfactant (SDS or Tween 80) with none or a high
formed by heating this mixture above the PIT (≈ 36◦ C) and PIT (Rao and McClements, 2010). This second surfactant par-
then rapidly cooling it to 13◦ C. The turbidity of this nanoemul- tially replaced some of the original surfactant from the droplet
 FOOD-GRADE NANOEMULSIONS
Figure 9 Schematic representation of proposed mechanism for low-intensity emulsification by the catastrophic phase inversion method. The amount of water
added to a W/O emulsion is progressively increased, until a phase inversion occurs and an O/W emulsion is formed.
surfaces and increased the repulsive interactions between the
droplets, thereby inhibiting coalescence.
Phase Inversion Composition Methods. The phase-inversion
composition or PIC method is somewhat similar to the PIT
method, but the optimum curvature of the surfactant is changed
by altering the formulation of the system, rather than the tem-
perature (Anton et al., 2009). For example, an O/W emulsion
stabilized by an ionic surfactant can be made to phase invert to
a W/O emulsion by adding salt. In this case, the packing param-
eter is adjusted from p < 1 to p > 1 due to the ability of the
salt ions to screen the electrical charge on the surfactant head
groups (Maestro et al., 2008). Alternatively, a W/O emulsion
containing a high salt concentration can be converted into an
O/W emulsion by diluting it in water so as to reduce the ionic
strength below some critical level. Another way to prepare na-
noemulsions using the PIC method is by changing the pH to
alter the electrical charge and stability of emulsions. Fatty acids
may stabilize W/O emulsions at low pH (pH < pKa ) because the
carboxyl groups are uncharged so they have a relatively high oil
solubility and p > 1. However, they may stabilize O/W emul-
sions at high pH values because the carboxyl group becomes
ionized so they become more water-soluble and p < 1. Conse-
quently, nanoemulsions can be formed by increasing the pH of a
fatty acid-oil-water mixture from below to above the pKa value
of the carboxyl groups (Solè et al., 2006; Maestro et al., 2008).
Emulsion Inversion Point (EIP) Methods. In the emulsion
inversion point (EIP) methods the change from one type of an
297
emulsion to another (e.g., W/O to O/W or vice versa) is through
a catastrophic-phase inversion, rather than a transitional-phase
inversion as with the PIC or PIT methods (Fernandez et al., 2004;
Thakur et al., 2008). A transitional-phase inversion occurs when
the surfactant properties are altered by adjusting a formulation
variable, such as the temperature, pH, or ionic strength. On the
other hand, a catastrophic-phase inversion occurs when the ratio
of the oil-to-water phases is altered while the surfactant proper-
ties remain constant. Practically, catastrophic-phase inversion is
usually induced by either increasing (or decreasing) the volume
fraction of the dispersed phase in an emulsion above (or below)
some critical level. For example, a W/O emulsion consisting of
water droplets dispersed in oil is initially formed using a partic-
ular surfactant, and then increasing amounts of water are added
to the system with continued stirring (Fig. 9). At low amounts of
added water, additional water droplets are formed within the oil
phase. However, once a critical water content is exceeded, the
coalescence rate of water droplets exceeds the coalescence rate
of oil droplets, and so phase inversion occurs from a W/O to an
O/W system. The value of the critical concentration where phase
inversion occurs, as well as the size of the oil droplets produced,
depends on process variables, such as the stirring speed, the rate
of water addition, and the emulsifier concentration (Thakur et
al., 2008). The emulsifiers used in catastrophic-phase inversion
are usually limited to small molecule surfactants that can stabi-
lize both W/O emulsions (at least over the short term) and O/W
emulsions (over the long term).
298 D. J. McCLEMENTS AND J. RAO
Figure 10 Formulation-composition map for a typical surfactant-oil-water system.
Rationalizing Phase-Inversion Methods using the HLD Con-
cept. Recently, a new conceptual approach has been developed
to rationalize and generalize the phase-inversion behavior ex-
hibited by surfactant-oil-water (SOW) mixtures (Salager et al.,
2004; Queste et al., 2007). This approach involves describing
the behavior of a SOW system in terms of a formulation-
composition map (Fig. 10). The x-axis represents changes in
the “composition” of the emulsion, which is usually expressed
as the water-to-oil ratio (WOR = mass of water/mass of oil).
The y-axis represents changes in the “formulation” of the SOW
system, which is usually expressed as the hydrophilic-lipophilic
deviation (HLD). The HLD is a measure of the relative affin-
ity of the surfactant for the hydrophilic (water) phase and the
lipophilic (oil) phase (Salager et al., 2005; Witthayapanyanon
et al., 2008). Sometimes the surfactant-affinity difference (SAD)
is used to represent the formulation variable rather than the
HLD. These two terms are closely related to each other: HLD =
SAD/RT, where R is the gas constant and T is the absolute
temperature. The formulation-composition map is a convenient
means of specifying the types of emulsions that can be formed
by a given SOW system when either the surfactant properties
are altered (e.g., by altering pH, ionic strength, or temperature)
or the overall system composition is changed (e.g., by changing
the relative amounts of oil and water present).
The HLD value is a dimensionless parameter that character-
izes the behavior of a surfactant within a particular surfactant-
oil-water system (Leal-Calderon et al., 2007). It takes into ac-
count the influence of oil-phase properties (such as oil type),
aqueous-phase properties (such as salt or alcohol content), and
environmental factors (such as temperature) into the relative
affinity of a surfactant for the oil and water phases. The HLD
system is therefore more comprehensive than the traditional
hydrophilic-lipophilic balance (HLB) system that is commonly
used to describe surfactant behavior, since the HLB system only
usually takes into account the properties of the surfactant itself.
The HLD number of a SOW system can be calculated us-
ing the following empirical equation for ethoxylated non-ionic
surfactants (Queste et al., 2007):
HLD = [α − k × EACN – EON]
+ [a × CA + b × Cs + c × (T − T0 )] (1)
The first three parameters in this equation are mainly deter-
mined by the molecular characteristics of the surfactant and oil
used, while the last three parameters are mainly determined by
the influence of system conditions (composition and tempera-
ture) on surfactant properties. Here, α and k are constants that
depend on the surfactant type; EACN is the equivalent alkane
carbon number of the oil phase, and EON is the ethylene ox-
ide number of the surfactant head group. In addition, CA is the
 FOOD-GRADE NANOEMULSIONS
concentration of any alcohol present in the system (wt%), a is
a constant that depends on the type of alcohol present, CS is
the concentration of any salt present in the system (wt%), b is a
constant that depends on the type of salt present, T is the temper-
ature, T0 is a reference temperature (◦ C), and c is a constant that
depends on how the properties of a particular surfactant change
with temperature. Typically, the reference temperature is 25◦ C.
Values for the other parameters have been determined for a
number of surfactant-oil-water systems. For example, EACN
and other parameters have recently been reported for a number
of terpenes (Bouton et al., 2009) and for a number of triacyl-
glycerol oils (Phan et al., 2010). In general, the HLD number
of a particular class of non-ionic surfactants tends to increase
with increasing temperature, increasing salt concentration, de-
creasing EACN, and increasing EON. For certain surfactant-oil
combinations the HLD number can be made to change from neg-
ative to positive as the temperature is increased, which favors
transition from a O/W to W/O emulsion (see below). The tem-
perature at which this transition occurs is the phase-inversion
temperature (PIT) mentioned earlier. Triacyglycerol oils (MCT
and LCT) tend to have much higher EACN numbers than alka-
nes or flavor oils which accounts for the fact that it is difficult to
use the PIT method to form nanoemulsions from these oils—the
temperature required to make the HLD number go from negative
to positive is too high to be reached under normal circumstances.
For ionic surfactants the HLD number can be calculated from
the following expression (Salager at al 2005):
HLD = [α  − k  × EACN] + [a  × CA + b × ln(Cs )
+ c × (T − T0 )] (2)
Once these relationships have been established for a particu-
lar surfactant-oil-water combination, they can be used to predict
how the behavior of the surfactant will change when the system
composition (e.g., salt, alcohol, and oil) and environmental con-
ditions (e.g., temperature) are altered. This information can be
used to determine the optimum conditions required to form sta-
ble emulsions or nanoemulsions using phase-inversion methods.
The relationship between the HLD number of a surfactant
and its ability to stabilize emulsions is highlighted below:
• HLD < 0: The surfactant has: (i) A higher affinity for the water
phase than the oil phase; (ii) The surfactant tends to form
normal micelles or microemulsions in the aqueous phase;
(iii) The surfactant tends to stabilize O/W (rather than W/O)
emulsions. The more negative the HLD number, the greater
the affinity for the water phase.
• HLD = 0: The surfactant has: (i) An equal affinity for the water
and oil phases; (ii) The surfactant tends to form bicontinuous
microemulsion or liquid crystalline phases; (iii) The surfactant
tends to stabilize neither O/W nor W/O emulsions.
• HLD > 0: The surfactant has: (i) A higher affinity for the
oil phase than the water phase; (ii) The surfactant tends to
299
form reverse micelles or microemulsions in the oil phase;
(iii) The surfactant tends to stabilize W/O (rather than O/W)
emulsions. The more positive the HLD number, the greater
the affinity for the oil phase.
Knowledge of the HLD number, the water-to-oil ratio
(WOR), and the formulation-composition map for a particu-
lar system can be used to rationalize its behavior under different
conditions (Fig. 10). The formulation-composition map can be
conveniently divided into a number of different regimes, which
are designated by a letter and a sign (Leal-Calderon et al., 2007).
The sign determines the influence of formulation (HLD num-
ber) on the type of emulsion formed. In negative regimes (A-,
B-, and C-), HLD < 0, and the surfactant favors the formation
of O/W emulsions, whereas in positive regimes (A+, B+, and
C+), HLD > 0, and the surfactant favors the formation of W/O
emulsions (Salager et al., 2005; Witthayapanyanon et al., 2008).
The letter determines the influence of composition (WOR) on
the type of emulsion formed: A refers to a system where the oil
and water phases have fairly similar amounts (WOR ≈ 1) and
so formation of both O/W and W/O emulsions is equally favor-
able; B refers to a system where the oil phase is in excess and
so the formation of W/O emulsions is favored; and C refers to a
system where the water phase is in excess and so the formation
of O/W emulsions is favored. If both the formulation variable
(HLD number, sign) and the composition variable (WOR, let-
ter) favor the formation of a particular emulsion type (e.g., O/W)
then this emulsion is said to be “normal” and will tend to be
stable (Mira et al., 2003; Rondón-Gonzaléz et al., 2006). On the
other hand, if the formulation variable favors one emulsion type
(e.g., W/O) while the composition variable favors the other type
(e.g., O/W), then this system is said to be “abnormal” and will
tend to be unstable. In the formulation-composition map shown
in Fig. 10 there are four regimes where the emulsions should be
stable (A-, A+, B+, and C-) and two regimes where they should
be unstable (B- and C+). The emulsions formed in the abnormal
regimes are usually highly unstable to droplet coalescence and
phase separation, but multiple emulsions may be formed near
certain phase boundaries. For example, O/W/O emulsions may
be formed near the B- to A- boundary, whereas W/O/W emul-
sions may be formed near the C+ to A+ boundary (Fig. 10).
The formulation-composition map can be used to describe
both transitional- and catastrophic-phase inversions (Leal-
Calderon et al., 2007). For example, if one moves in a horizontal
direction from Regime B- to Regime A- by increasing the WOR,
the system would undergo a catastrophic-phase inversion from
a W/O to an O/W emulsion, passing through an intermediate
regime where a multiple emulsion (O/W/O) was formed. Con-
versely, if one moves in a vertical direction from Regime A+ to
Regime A- by decreasing the HLD value from a positive to nega-
tive value (e.g., decreasing temperature from above to below the
PIT), the system would undergo a transitional-phase inversion
from a O/W to a W/O emulsion, passing through an intermediate
regime where a bicontinuous system or liquid-crystalline phase
was formed. This concept is therefore an extremely valuable one
300 D. J. McCLEMENTS AND J. RAO
Figure 11 Various “finishing techniques” can be used to modulate the properties of nanoemulsions after they have been prepared in order to change their
stability or functional properties.
for rationalizing the conditions where it is possible to form stable
nanoemulsions using either type of phase-transition mechanism.
However, it does not indicate the potential size of the droplets
formed in each region, i.e., whether a nanoemulsion or an emul-
sion is formed. This will depend on the total amount of the emul-
sifier present, as well as the preparation method used to form the
emulsion (e.g., the energy intensity for a high-energy method).
Finishing Techniques
After a nanoemulsion has been formed using either a low-
energy or high-energy approach, it is possible to change the
droplet characteristics using a variety of methods, which we
refer to as finishing techniques (Fig. 11).
Interfacial Engineering
For some practical applications of nanoemulsions, it may be
advantageous to alter the nature of the interfacial layer surround-
ing the oil droplets after formation. For example, one type of
emulsifier may be particularly effective at producing very small
droplets during nanoemulsion preparation, but may not be that
effective at stabilizing the droplets after they have been formed.
This is true for nanoemulsions produced using the PIT method,
where the non-ionic surfactants used to form the emulsions are
not effective at preventing subsequent droplet coalescence at
elevated temperatures (Rao and McClements, 2010). Alterna-
tively, changing the emulsifier type after the formation of a
nanoemulsion may be useful in changing interfacial properties
such as composition, electrical charge, rheology, or thickness,
which may enable one to tune the functional performance of a
nanoemulsion for different applications. Interfacial properties
may be altered after nanoemulsion formation using a number of
approaches:
• Interfacial Displacement: A nanoemulsion may be mixed
with a solution containing an emulsifier that competes at
the oil-water interface thereby displacing some or all of the
original emulsifier from the droplet surfaces (Rao and Mc-
Clements, 2010). The interfacial composition will depend on
the relative concentrations and surface activities of the various
emulsifiers present in the system (McClements, 2005).
• Interfacial Deposition: The interfacial characteristics of the
droplets in a nanoemulsion may also be changed using inter-
facial deposition methods (Guzey and McClements, 2006a;
McClements, 2010). In this case, the droplets in a nanoemul-
sion are mixed with a solution containing a substance that
adsorbs to the surface of the existing emulsifier layer. For
example, if the original droplets are coated by an emulsifier
with an electrical charge, then oppositely charged substances
(such as polyelectrolytes or mineral ion) can be deposited onto
their surfaces using electrostatic deposition (Guzey and Mc-
Clements, 2006a; McClements, 2010). This technique can be
used to create multilayers around lipid droplets that have dif-
ferent functional properties than the original emulsifier layer.
• Interfacial Cross-linking: For certain types of emulsifiers it is
possible to cross-link them after they have adsorbed to the oil-
water interface, e.g., physically, chemically, or enzymatically.
This approach has been used to covalently cross-link adsorbed
protein layers (Dickinson, 1997; Bos and van Vliet, 2001;
Sandra et al., 2008) and polysaccharide layers (Littoz and
McClements, 2008).
 FOOD-GRADE NANOEMULSIONS
Solvent Displacement/Evaporation
The size of the droplets in an emulsion or nanoemulsion can
be reduced after an initial homogenization step using solvent-
displacement and/or solvent-evaporation methods (Horn and
Rieger, 2001; Tan et al., 2005a,b; Chu et al., 2007b). In these
methods, an oil-in-water emulsion is initially formed by ho-
mogenizing an organic phase (lipid and organic solvent) with
an aqueous phase (water and hydrophilic emulsifier). The or-
ganic solvent is selected based on its water miscibility, boiling
point, safety, and legal status. The lipid may be a liquid oil
(such as fish, corn, sunflower, or olive oil), a solid fat (such
as hydrogenated palm oil), a lipophilic nutraceutical (such as
β-carotene, lycopene, or curcumin), or a mixed system, but the
lipid must be capable of being dissolved within the organic
solvent prior to homogenization. This is because it is difficult
to form emulsions when the oil phase contains any crystalline
material. The organic solvent may be hydrophilic, lipophilic,
or amphiphilic (Horn and Rieger, 2001). After homogenization,
the oil droplets within the emulsion contain a mixture of lipid
and organic solvent. In the solvent-displacement method, an am-
phiphilic organic solvent that is partially miscible with water is
used, and the aqueous phase of the emulsion is initially saturated
with the solvent. When this emulsion is diluted with water, then
the organic solvent moves from the oil droplets into the aqueous
phase, which causes the droplets to shrink in size. If necessary,
the remaining organic solvent can then be removed by evap-
oration. In the solvent-evaporation method, a volatile organic
solvent that is either immiscible (lipophilic solvent) or partially
miscible (amphiphilic solvent) with water can be used (Horn
and Rieger, 2001). In this case, the organic solvent is removed
from the oil droplets by evaporation, which causes the droplets
to shrink in size. The final droplet size produced using these
two methods depends on the initial droplet size in the original
emulsion, the amount of organic solvent present, and the water-
solubility of the organic solvent. Some studies using the solvent
evaporation or displacement method are reported in Table 8.
Our laboratory recently used this approach to form na-
noemulsions containing protein-coated oil droplets (Lee and
McClements, 2010). An oil-in-water emulsion was formed us-
ing a lipid phase that consisted of corn oil and ethyl acetate (an
amphiphilic solvent), and an aqueous phase that contained water
Table 7 Reported applications of nanoemulsion in the food industry found on the internet (2010)
Food product name Company
Canola Active Oil Shemen, Haifa, Israel
Fortified Fruit Juice High vive, USA
NanoResveratrolTM Life Enhancement, USA
Nutri-NanoTM CoQ-10 3.1x Softgels Solgar, USA


R

R
Spray for Life Vitamin Supplements Health Plus International , Inc., USA
“Daily boost” Jamba Juice Hawaii, USA
“Color emulsion” Wild Flavors, Inc, USA
301
and a protein emulsifier (whey protein isolate, WPI). The size
of the droplets was then reduced by diluting these emulsions
with pure water to promote movement of ethyl acetate into the
aqueous phase (solvent displacement) and/or by heating them
to remove the ethyl acetate (solvent evaporation). The droplet
size produced could be controlled using these solvent removal
approaches by varying the ratio of ethyl acetate to corn oil in the
initial organic phase: the higher the fraction of ethyl acetate ini-
tially present in the organic phase, the smaller the final droplet
size produced (Fig. 12).
One limitation of this approach is that the volume of a droplet
is proportional to its diameter cubed (V ∝ d 3 ), so that to half the
droplet diameter one must reduce the droplet volume to 1/8th of
the original value. This means that a large amount of organic sol-
vent must be used compared to the amount of the encapsulated
component. The solvent removal method has been successfully
used to prepare dispersions of β-carotene nanoparticles from
food-grade ingredients (Tan and Nakajima, 2005a; 2005b; Chu
et al., 2007a). Various food-grade emulsifiers can be used to
prepare nanoemulsions based on this approach, including whey
proteins, caseinate, and small molecule surfactants (Tan et al.,
2005a,b; Chu et al., 2007a).
Lipid Phase Exchange
The composition of the oil phase within the droplets in an
O/W nanoemulsion may be changed after it has been prepared
by mixing it with another emulsion or microemulsion system
(Fig. 11). When an oil molecule has a finite solubility in the
aqueous phase, then it will be transported between lipid droplets
through molecular diffusion due to an entropy of mixing effect
(Pena and Miller, 2002; Capek, 2004). Alternatively, if the oil
molecules can be solubilized within surfactant micelles then
they may also be transported between the lipid droplets (Weiss
et al., 2000). Consequently, it is possible to mix an emulsion
containing droplets with one composition, with another
emulsion or microemulsion containing droplets of a different
composition. These compositional ripening effects will then
lead to the transport of oil between droplets until a steady-state
condition is reached (McClements and Dungan, 1993). This
method could be used to change the oil composition of the
Product description
Nanoencapsulation of fortified phytosterols
Nanoencapsulation of fortified vitamin, lycopene, Theanine, and
SunActive Iron
plant-based lipid, such as a solid triglyceride (fat), encased by a shell
consisting of a natural phospholipid, such as phosphatidylcholine
delivery system
30 nm O/W system
Nanoencapsulation of fortified vitamin beverage
Nanoencapsulation of fortified vitamin or bioactive components beverage
Beta-carotenal, apo-carotenal, or paprika nanoemulsions
302 D. J. McCLEMENTS AND J. RAO

Table 8 An overview of research articles where the properties in O/W nanoemulsions have been altered using finishing techniques, i.e., solvent displacement
of evaporation

Bioactive Particle Diameter Particle Diameter

Oil phase component Surfactant/Co-surfactant Before Finishing After Finishing References

Corn oil, Ethyl acetate — WPI 174 nm < 70 nm Lee and McClements, 2010
Hexane β−carotene Sodium caseinate 1000 nm 18 nm Chu et al., 2007
Acetone β−carotene Sodium caseinate Tween 20, — 30–206 nm (It depends on Yin et al., 2009
decaglycerol monolaurate, sucrose surfactant type)
fatty acid ester
Acetone β−carotene Gelatin or Tween 20/polylactic acid — 74 nm (gelatin) Ribeiro et al., 2008
77 nm (Tween 20)
Acetone β−carotene Tween 20 — 9–280 nm Silva et al., 2010
Hexane Hexane Tween 20 — 60–140 nm Tan and Nakajima, 2005

droplets in a nanoemulsion, e.g., an oil-soluble antioxidant, than when it is in its bulk state because of supercooling and cur-
antimicrobial, or nutritional component could be introduced vature effects (Turnbull and Cormia, 1961; Westesen, 2000).
into a nanoemulsion after it has been formed.

Lipid Crystallization DESIGNING FUNCTIONAL NANOEMULSION

The oil phase used in the preparation of nanoemulsions by ei-
ther the high-energy or low-energy methods must usually be liq-
uid. Nevertheless, it is possible to change the physical state of the
oil phase after a nanoemulsion has been formed (Fig. 11), e.g.,
by crystallizing the lipid phase (Weiss et al., 2007). The oil phase
can be fully or partially crystallized after nanoemulsion forma-
tion by controlling its composition and the temperature-time his-
tory of the product. In this case, a lipid phase is used that is crys-
talline at the final product temperature. Before a nanoemulsion
can be produced the lipid phase has to be heated above its melt-
ing point and maintained at this temperature throughout the man-
ufacturing process to ensure that it remains liquid. The resulting
nanoemulsion is then cooled at a controlled rate to crystallize the
oil droplets, without promoting product instability. By careful
selection of oil type and thermal history, it is possible to control
the type, concentration, and location of the crystals within the
droplets, which can lead to novel functional properties (Bum-
mer, 2004; Kesisoglou et al., 2007; Weiss et al., 2007). It should
be noted that fat crystallization usually occurs at a much lower
temperature when an oil is divided into nanometer-sized droplets
PARTICLES
The characteristics of the particles in nanoemulsions
ultimately determine the bulk physicochemical and functional
properties of nanoemulsion-based products, e.g., their optical
properties, stability, rheology, digestibility, and release charac-
teristics. In this section, we highlight the most important particle
characteristics that can be controlled to obtain nanoemulsions
with specific functional properties.
Particle Composition
The composition of the particles in nanoemulsions can be
controlled by appropriate selection of ingredients and process-
ing operations. The particles in oil-in-water nanoemulsions can
conveniently be considered to consist of a core of lipophilic
material surrounded by a shell of surface-active material (Mc-
Clements, 2005). A number of different non-polar food ingredi-
ents can be used to create the lipophilic core, including triacyl-
glycerols, diacylgycerols, monoacylglycerols, free fatty acids,

Figure 12 Nanoemulsions can be prepared by the homogenization/solvent evaporation method. An emulsion is prepared with an oil phase that contains a
mixture of oil and solvent. The solvent is then removed by evaporation, which causes the droplets to shrink in size. The photographs show whey-protein stabilized
5% oil-in-water emulsions before and after solvent evaporation: oil phase = 5% corn oil: 95% ethyl acetate.
 FOOD-GRADE NANOEMULSIONS 303

Figure 13a Predicted influence of droplet size and shell layer thickness on the effective composition of core-shell particles in nanoemulsions. As the core size
decreases or the shell layer thickness increases, the fraction of core in the particles decreases.

flavor oils, essential oils, mineral oils, fat substitutes, waxes,
weighting agents, oil-soluble vitamins, and nutraceuticals (such
S on r for nanoemulsions with
as carotenoids, curcumin, phytosterols, and Co-enzyme Q). As
mentioned earlier, these ingredients have different densities,
refractive indices, polarities, viscosities, and melting behavior,
which will impact the functional performance of the overall sys-
tem (Table 1). A variety of surface-active food ingredients can be
used to create the shell surrounding the lipophilic core, including
surfactants, phospholipids, proteins, polysaccharides, minerals,
and solid particles. The choice of these components plays a
critical role in determining particle properties, such as physical
stability, interactions, release characteristics, and digestibility.
In conventional emulsions, the shell thickness (δ) is much
smaller than the core radius (r), i.e., δ << r, and so the core-
shell particles consist primarily of the oil phase. On the other
hand, in nanoemulsions, the shell thickness may be of the same
order of magnitude as the core radius (δ ≈ r), and hence the shell
layer makes an appreciable contribution to the overall particle
composition (Tadros et al., 2004; Mason et al., 2006). This effect
can be described by the following equation:
r3
S = 1 − (3) particle composition on particle size will impact those particle
(r + δ)3
Here
S (= Vshell /Vtotal ) is the volume fraction of the shell
compared to the total particle, Vshell is the volume of the shell,
Vtotal is the total volume of the entire particle ( = Vcore + Vshell ),
r is the radius of the core, and δ is the thickness of the shell
(Fig. 1). The dependence of
different core radii and shell-layer thicknesses is shown in Fig.
13a. This figure shows that the composition of the particles in
small nanoemulsions may be quite different from those in con-
ventional emulsions made from the same ingredients due to the
large contribution of the shell. For example, a particle with a
core radius of 25 nm and a shell thickness of 10 nm is about 37%
core and 63% shell. The core-shell model used to calculate these
curves certainly over-simplifies the actual structure of the parti-
cles found in real nanoemulsions. In reality, the shell is not ho-
mogeneous but consists of a complex mixture of different kinds
of molecules that accumulate at the oil-water interface, such as
proteins, phospholipids, small molecule surfactants, bioactive
components, mineral ions, water, lipids, and polymers (Fig. 1).
In addition, its structure may be highly dynamic as molecules
move in and out of the interface, and undergo positional and
conformational changes at the interface. The precise structure
and orientation of these molecules at the particle surface may
be important in many practical applications.
The size-dependence of particle composition has a number
of important practical implications. First, the dependence of
properties (such as refractive index, density, and permeability)
that alter the bulk physicochemical properties and stabilities
of nanoemulsions (such as optical properties, creaming rate,
and release characteristics). Second, the loading capacity of a
lipophilic component within the particles in a nanoemulsion may
304 D. J. McCLEMENTS AND J. RAO
decrease as the core size decreases because of the reduction in
the fraction of lipophilic core present (Fig. 13a). Third, the de-
pendence of particle composition on particle size may influence
the accuracy of data obtained from instrumental techniques de-
signed to measure particle size, such as laser diffraction, spectro-
turbidity, and dynamic light scattering (McClements, 2005). The
mathematical models used by these instruments to calculate the
particle size distribution from some measured physical prop-
erty (such as a light-scattering pattern) usually assume that the
particles are homogeneous spheres with well-defined properties
(such as refractive index and density). Consequently, there may
be some errors in the reported particle size distributions in na-
noemulsions where the particles are core-shell structures rather
than simple homogeneous spheres.
Particle Concentration
In general, the particle concentration in a colloidal dispersion
is usually expressed as the number, mass, or volume of parti-
cles per unit volume or mass of the total system (McClements,
2005). In conventional oil-in-water emulsions, the particle con-
centration is usually characterized in terms of the disperse phase
volume fraction (φ), which is the volume of oil per unit volume
of emulsion. In oil-in-water nanoemulsions, the effective par-
ticle concentration may be quite different from the fraction of
pure disperse phase present, which can have a major impact on
their physicochemical and functional attributes (Tadros et al.,
2004; Mason et al., 2006).
As discussed in the previous section, the particles in
emulsions can be considered to have a core-shell structure (Fig.
1). Hence, the overall effective volume fraction (φ effective ) is the
sum of the volume fractions of the core (φ core ) and the shell
(φ shell ): φ effective = φ core + φ shell . An expression for the effective
volume fraction can be obtained from knowledge of the particle
dimensions:


δ 3
φeffective = φcore 1 +
r strength).
In conventional emulsions, the shell thickness is much smaller
than the oil core (δ << r), and so the effective droplet concen-
tration is similar to the oil-core concentration (φeffective ≈ φcore ).
However, in nanoemulsions the thickness of the emulsifier layer
may approach that of the droplet radius (δ ≈ r) and so the effec-
tive particle volume fraction may be appreciably higher than the
oil phase volume fraction (φeffective >> φcore ). The influence of
the shell layer on the effective particle volume fraction is shown
in Fig. 13b for nanoemulsions with different oil core sizes and
shell layer thicknesses. These calculations indicate that the
effective volume fraction of the particles in nanoemulsions may
be many times higher than the actual volume fraction of the
oil phase. This increase in effective particle concentration may
cause appreciable changes in the bulk physicochemical prop-
erties of nanoemulsions, such as rheology and stability. (See
section titled “Physicochemical Properties of Nanoemulsions.”)
For sterically-stabilized systems, the thickness of the shell
is determined primarily by the dimensions of the layer of ad-
sorbed surface-active molecules. For electrostatically-stabilized
systems, the effective shell thickness (δ) may be considerably
greater than the physical dimensions of the adsorbed layer
(Tadros et al., 2004; Mason et al., 2006). In these systems,
the thickness depends on the range of the electrostatic interac-
tions (Debye screening length, κ −1 ), which depends on the ionic
strength of the surrounding aqueous phase (McClements, 2005). √
For aqueous solutions at room temperatures, κ −1 ≈ 0.304/ I
nm, where I is the ionic strength expressed in moles per liter
(Israelachvili, 1992). For example, the ranges of the electro-
static interactions in monovalent salt solutions with different
ionic strengths are: κ −1 = 0.3, 0.96, 3, 9.6, and 30 nm for I =
1, 10−1 , 10−2 , 10−3 ,and 10−4 M, respectively. At low ionic
strengths, the Debye screening length may therefore be on the
same order as the particle radius in nanoemulsions, which can
have a major impact on the bulk physicochemical properties of
the system, such as the optical, rheological, and stability prop-
erties (Weiss et al., 2000; Weiss and McClements, 2001; Mason
et al., 2006; Kawada et al., 2010). The impact of this effect
on the rheology of nanoemulsions is discussed in section titled
“Rheological Properties”.
The particle concentration within a nanoemulsion is usually
controlled during the manufacturing process by controlling the
proportions of the two immiscible liquids (oil and water) used to
prepare it. However, the particle concentration can also be varied
after a nanoemulsion formation using dilution (e.g., by adding
more continuous phase) or concentration (e.g., by gravitational
separation, filtration, centrifugation, or evaporation) methods. It
should be noted that concentrating nanoemulsions by many of
these methods is much more difficult than concentrating con-
ventional emulsions because of their very small droplet size.
As discussed previously, the effective size of the particles in
a nanoemulsion may also be changed by using emulsifiers that
form different adsorbed layer thicknesses, or by altering solution
conditions that impact the range and magnitude of electrostatic
(4) interactions between charged particles (such as the pH or ionic
Particle Dimensions
The dimensions of the particles within a nanoemulsion are
one of its most important characteristics since they influence
its optical, rheological, stability, and release characteristics, as
well as potentially altering its biological fate. The dimensions of
the particles within a colloidal dispersion are usually character-
ized by the particle-size distribution (PSD), which represents the
fraction of particles within a range of discrete size classes (Wal-
stra, 2003; McClements, 2005). A PSD is typically presented as
a table or plot of particle concentration (e.g., volume or number
percent) versus particle size (e.g., diameter or radius). For many
purposes, it is more convenient to represent the full PSD using
a measure of the central tendency of the distribution (e.g., mean
or median) and a measure of the width of the distribution (e.g.,
standard deviation or polydispersity index).
 FOOD-GRADE NANOEMULSIONS
Figure 13b Predicted influence of droplet size and shell layer thickness on the effective concentration of core-shell particles in nanoemulsions. As the core size
decreases or the shell layer thickness increases, the effective particle concentration increases.
The effective radius of the particles in an oil-in-water na-
noemulsion is the sum of the radius of the lipophilic core and
the thickness of the shell: reffective = r + δ. There are appre-
ciable differences in the thicknesses of the layers formed by
food-grade emulsifiers—typically, small molecule surfactants
(such as Tweens and Spans) < globular proteins (such as egg,
whey, or soy proteins) < flexible proteins (such as caseinate or
gelatin) < polysaccharides (gum Arabic or modified starch). In
addition, the thickness of the shell layer may be increased by
adsorbing additional material to the droplet surfaces after they
have been formed, e.g., by electrostatic deposition of one or
more layers of charged biopolymers (or other charged species)
onto the surfaces of oppositely charged emulsifier-coated oil
droplets (Guzey and McClements 2006a; 2006b). In reality, it
may be difficult to accurately define the physical dimensions
of the particles in a nanoemulsion, since the shell may con-
sist of a polymeric or electrically-charged layer with no distinct
edge. In addition, the most appropriate shell thickness to use
in any mathematical calculations may depend on the partic-
ular situation encountered. For example, for a nanoemulsion
containing charged particles, δ may be taken to be the Debye
screening length for rheological purposes, but it may be taken to
be the physical thickness of the surfactant head-groups for light
scattering purposes. As discussed above, the fact that the shell
layer makes a significant contribution to the overall particle di-
mensions in nanoemulsions has important consequences for the
measurement and interpretation of particle-size distributions.
305
The PSD of an emulsion can usually be controlled by vary-
ing nanoemulsion preparation conditions and/or system com-
position. For example, in high-energy methods the droplet size
depends on the intensity and duration of the energy input, the
type and concentration of the emulsifier used, the interfacial
tension, and the relative viscosities of the disperse and contin-
uous phases (McClements, 2005; Jafari et al., 2006). Smaller
droplets can usually be produced by increasing the intensity or
duration of homogenization, by increasing the concentration of
the emulsifier used, or by controlling the viscosity ratio (Wal-
stra, 1993; Schubert and Engel, 2004; Wooster et al., 2008). In
low-energy methods, the droplet size depends on factors such as
system composition (such as surfactant-oil-water ratio, surfac-
tant type, and ionic strength) and environmental conditions (such
as temperature-time history, and stirring speeds) (Anton et al.,
2008; 2009). The most effective approach to control the size of
the particles within a specific nanoemulsion therefore depends
on the particular homogenization method used to prepare it. Fi-
nally, it should be stated that it is often important to stabilize the
particles within a nanoemulsion against aggregation or growth
after they have been formed, in order to prevent undesirable
changes in particle dimensions during storage or utilization.
Particle Charge
The droplets in nanoemulsions often have an electrical charge
because of adsorption of ionized emulsifiers, mineral ions, or
306 D. J. McCLEMENTS AND J. RAO

biopolymers to their surfaces (McClements, 2005). The sign

and magnitude of the electrical charge on the droplets plays
an important role in the functional performance and stability
of nanoemulsions, e.g., aggregation stability, interaction with
other food components, and ability to adhere to non-biological
(such as packaging materials) and biological surfaces (such as
the tongue, mucous layer, or other locations within the gastroin-
testinal tract). The electrical properties of a droplet are usually
characterized in terms of its surface-charge density (σ ), sur-
face potential (0 ), and/or ζ −potential (ζ ) (Hunter, 1986). The
surface-charge density is the number of unit electrical charges
per unit surface area. The surface-electrical potential is the free
energy required to increase the surface-charge density from zero
to σ by bringing charges from an infinite distance to the sur-
face through the surrounding medium. The surface-electrical
potential depends on the ionic composition of the surrounding
medium due to electrostatic screening effects and usually de-
Figure 14a The electrical charge on oil droplets can be varied by mixing
creases as the ionic strength of the aqueous phase increases. different kinds of ionic and non-ionic surfactants together. In this system we
The zeta-potential (ζ ) is the electrical potential at the “shear made a nanoemulsion containing lipid droplets stabilized by a non-ionic surfac-
plane,” which is defined as the distance away from the droplet tant (Tween 80), and then added different amounts of either an anionic (SDS)
surface below which the counter-ions remain strongly attached or cationic (LAE) surfactant after homogenization to vary droplet charge (data
to the droplet when it moves in an electrical field. Practically, provided by Khalid Ziani). The SDS concentration is expressed as a negative
number for convenience.
the ζ -potential is often a better representation of the electri-
cal characteristics of a droplet than the surface potential be-
acids) (Kralova et al., 2009), whereas those stabilized by cationic
cause it inherently accounts for the adsorption of any charged
surfactants have a positive charge (e.g., lauric arginate) (Asker
counter-ions. In addition, the ζ -potential is much easier to mea-
et al., 2009). It is possible to vary the magnitude of the droplet
sure than the surface potential or charge density, and therefore
charge in surfactant-stabilized systems by using different ratios
droplet charges are usually characterized in terms of ζ - poten-
of ionic and non-ionic surfactants to prepare them (Fig. 14a).
tial (Hunter, 1986). The range of the electrostatic interactions
The droplet charge can also be varied by using biopolymers
in a system is described by the Debye screening length (κ −1 ),
which decreases as the ionic strength of the aqueous solution
surrounding the droplets increases. (See section titled “Particle
Concentration.”)
Analytical instruments based on particle electrophoresis and
electro-acoustic are widely used to measure the ζ -potential of
the droplets in emulsions (McClements, 2005). These instru-
ments utilize mathematical models to convert the measured
electro-dynamic properties of a particle into a ζ - potential. These
models typically make assumptions about the size of the parti-
cles (r) relative to the thickness of the Debye screening length
(κ −1 ), e.g., it is often assumed that κ −1 << r. This assumption
may be valid for the relatively large droplets in conventional
emulsions, but may not be valid for the very small droplets in
nanoemulsions, particularly at low ionic strengths (high κ −1 ). It
is therefore important to take this effect into consideration when
determining the ζ -potential of droplets in nanoemulsions.
The electrical characteristics of nanoemulsion droplets can
be controlled by careful selection of particular emulsifier types.
In principle, droplets stabilized by non-ionic surfactants (e.g.,
Tweens and Spans) should have no droplet charge, but in prac- Figure 14b The electrical charge on oil droplets in nanoemulsions can be
tice they often have a significant negative charge which may be varied by using different kinds of biopolymer emulsifiers or biopolymer coat-
due to the presence of free fatty acids or other ionic impurities ings. This figure shows the electrical charge versus pH profiles for various kinds
remaining from their production or generated during storage of biopolymers in aqueous solutions. Proteins are amphoteric molecules whose
(Mun et al., 2005). Droplets stabilized by anionic surfactants charge goes from positive at low pH to negative at high pH, with a point of zero
charge depending on protein type. Polysaccharides may be anionic or cationic
have a negative charge (e.g., lecithin, DATEM, CITREM, fatty depending on their charge groups.
 FOOD-GRADE NANOEMULSIONS
with different electrical characteristics (Fig. 14b). Droplets sta-
bilized by commercial polysaccharide emulsifiers tend to have
a net negative charge (e.g., gum Arabic and modified starch)
due to the presence of anionic groups (e.g., sulfate or carboxyl)
on the polymer chains (Chanamai and McClements, 2002). Oil
droplets stabilized by proteins (e.g., whey protein, casein, soy
proteins, egg proteins) have a charge that depends on the solu-
tion pH relative to the isoelectric point (pI) of the protein (Gu
et al., 2005). Protein-coated droplets have a net positive charge
for pH < pI, no net charge at pH = pI, and a negative charge at
pH > pI. However, it should be stressed that protein molecules
have regions of both positive charge and negative charge on their
surfaces, and this heterogeneous charge distribution can greatly
impact their behavior (Cooper et al., 2005). Proteins with differ-
ent isoelectric points can be selected for particular applications,
e.g., β-lactoglobulin has a pI near 5, but lactoferrin has a pI near
8. The charge on emulsifier-coated droplets in nanoemulsions
can also be altered by adsorption of other charged substances
onto their surfaces, such as proteins, polysaccharides, phos-
pholipids, or multivalent ions (McClements, 2005; Guzey and
McClements, 2006a; Johnston et al., 2006; Quinn et al., 2008;
Ochs et al., 2010).
Interfacial Characteristics
Controlling the interfacial characteristics of particles is one of
the most powerful methods of designing nanoemulsion systems
with specific functional attributes (McClements, 2005; Mason
et al., 2006). We have already highlighted how some interfa-
cial characteristics of the particles in nanoemulsions can be
controlled, e.g., their composition (See section titled “Particled
Composition”), thickness (see section titled “Particle Dimen-
sions”), and electrical charge (see the previous section). There
are also a number of other interfacial properties that may also
be important for determining the overall functional attributes of
nanoemulsions, such as interfacial permeability, rheology, inter-
actions, and environmental responsiveness. The selective per-
meability of an interfacial layer to different kinds of molecules
depends on its porosity and specific interactions. Some surface-
active molecules may form interfacial layers that are so close-
packed and/or rigid that they are able to inhibit the diffusion of
other molecules through them, e.g., enzymes, lipids, and min-
eral ions. On the other hand, other surface-active molecules
may be able to form interfacial layers that have open and/or
dynamic structures that enable molecules to easily pass through
them. In principle, it is possible to design interfacial layers
that selectively allow only certain kinds of molecule through,
e.g., based on size, morphology, or charge. Consideration of
interfacial permeability may be important when designing sys-
tems that control the release of functional agents from inside
the particles or where it is important to keep two or more
chemically-reactive substances isolated from each other. The
rheology of interfacial layers can often be manipulated by con-
trolling the type and interactions of surface-active molecules
307
present. Some types of surface-active agents form relatively
low-viscosity interfacial layers (e.g., small molecule surfac-
tants, flexible biopolymers), whereas others form more viscous
or gel-like layers (e.g., surface- or thermally-denatured globular
proteins, or cross-linked biopolymers). Interfacial rheology may
be important in determining the particle stability to aggregation,
or in determining the bulk rheology in systems where there is
extensive interfacial layer overlap.
The interactions of the particles in a nanoemulsion with other
particles or surfaces can be controlled in a number of ways. Non-
specific electrostatic interactions can be controlled by control-
ling the electrical charge on the particle surfaces as described
in section titled “Particle Charge”. For example, a positively-
charged particle will tend to stick to a negatively-charged sur-
face, and vice versa. The surfaces of nanoparticles can also be
modified to alter their “stealth” character within the human body
by attaching hydrophilic polymers to them, e.g., polyethylene
glycol (PEG) (Howard et al., 2008). Particles with highly hy-
drophilic surfaces tend to be less prone to removal by the natural
defenses of the bodies, which increases the residence time of the
particles in the systemic circulation. Particle surfaces may also
be made more hydrophobic by using surface-active biopoly-
mers that have some non-polar character, such as surface- or
thermally-denatured globular proteins. Nanoparticles can also
be engineered so their surfaces have specific ligands attached
that are capable of binding to specific biological entities or or-
gans within the human body (Hashida et al., 2005; des Rieux
et al., 2006). These surface-modified nanoparticles can be used
for targeted- or sustained-release applications.
The environmental responsiveness of the particles in na-
noemulsions is also largely determined by their interfacial char-
acteristics. Particles may be designed so that they respond in
specific ways to changes in environmental conditions, such as
temperature, pH, ionic strength, and enzyme activity (Guzey
and McClements, 2006a). For example, a nanoemulsion may be
designed so that the entire particle remains intact under a certain
set of environmental conditions, but that either the shell and/or
core break down under another set of conditions. This feature is
particularly important in the development of food-grade deliv-
ery systems based on nanoemulsions where the delivery system
must remain stable in the food product, but becomes unstable in
a specific region of the GI tract (McClements et al., 2009).
The interfacial characteristics of the particles in a nanoemul-
sion can be controlled by selection of a particular emulsifier
type, such as a particular surfactant, phospholipid, protein, or
polysaccharide. For example, small molecule surfactants tend to
form fairly thin fluid-like layers; flexible biopolymers (such as
caseinate) tend to form relatively thick fluid-like layers; whereas
globular proteins (such as whey or soy protein) tend to form
thin elastic-like layers due to interfacial cross-linking (Dick-
inson, 1992; Dickinson, 2003). Interfacial characteristics may
also be controlled by using blends of different kinds of emulsi-
fiers in combination (Fig. 11), with the blending being carried
out either before or after homogenization. Finally, the interfacial
characteristics may be altered by depositing successive layers of
308 D. J. McCLEMENTS AND J. RAO
charged biopolymers (or other charged species) onto oppositely-
charged lipid droplets to form nanolaminated coatings around
them with different thicknesses, charges, or environmental re-
sponsiveness (Guzey and McClements, 2006a; Johnston et al.,
2006; Quinn et al., 2008; Ochs et al., 2010).
Particle Physical State
Nanoemulsions are normally prepared from oils that are liq-
uid, but it is possible to form them from oil phases that are
able to fully or partially crystallize at the final temperature of
application (Müller et al., 2002; Weiss et al., 2008). In these
cases, the oil phase is kept in liquid state during the forma-
tion of the nanoemulsion, i.e., by ensuring that the temperature
is above the melting point of the lipid. The oil droplets can
then be crystallized after their formation by cooling the na-
noemulsion sufficiently below the melting temperature of the
lipid (Walstra, 2003; Muller and Keck, 2004; Wissing et al.,
2004; McClements, 2005). The above approach has been used
to form solid lipid nanoparticles (SLN) or nanostructured lipid
carriers (NLC) (Fig. 11), which are nanoemulsions where the oil
phase has been fully or partly solidified (Müller et al., 2002). An
SLN contains lipid droplets that are fully crystallized and have
a highly-ordered crystalline structure. An NLC contains lipid
droplets with a less-ordered crystalline structure or an amor-
phous solid structure, which can aid in the encapsulation and
retention of highly lipophilic active components. The crystal-
lization temperature of an emulsified fat in a nanoemulsion may
be appreciably below that of the same fat in a bulk phase because
of supercooling effects (Gibout et al., 2007). In addition, the na-
ture of the crystals formed in a nanoemulsion may be different
from those formed by a bulk fat because of curvature affects,
the limited volume present for crystal growth in droplets, and
the lack of secondary nucleation sites (Muller and Keck, 2004;
Wissing et al., 2004). The concentration, nature, and location of
the fat crystals within the lipid droplets in an emulsion can be
controlled by careful selection of oil type (e.g., solid fat content
versus temperature profile, polymorphic forms), thermal history
(e.g., temperature versus time), emulsifier type (e.g., tail group
characteristics), and droplet size (Muller et al., 2000; Walstra,
2003; Muller and Keck, 2004). A major potential advantage
of SLN and NLC systems is that the rate of molecular diffu-
sion through the lipid phase can be reduced, which can slow
down chemical-degradation reactions and improve the stability
of encapsulated lipophilic components (Weiss et al., 2007).
PHYSICOCHEMICAL PROPERTIES
OF NANOEMULSIONS
Optical Properties
The impact of droplet characteristics on overall appearance
is often an important factor to consider when designing food
products (McClements, 2002; Mason et al., 2006). Some food
products should be clear or only slightly turbid (e.g., clear forti-
fied waters, soft drinks, juices, jellies, jams, deserts, dressings,
and sauces) and so the addition of any delivery system should
not make them look cloudy or opaque. Other food products
are optically opaque (e.g., turbid or opaque dressings, mayon-
naise, sauces, dips, creams, and yogurts) and light scattering by
the oil droplets in a delivery system may make an important
contribution to their overall appearance. In general, the optical
properties of a colloidal dispersion are characterized in terms
of its opacity and its color, which can be quantified in terms
of tristimulus color coordinates, such as the L ∗ a ∗ b∗ system
(McClements, 2002; McClements, 2005). In this color system,
L∗ represents lightness, and a∗ and b∗ are color coordinates:
where +a∗ is the red direction, −a∗ is the green direction; +b∗
is the yellow direction, −b∗ is the blue direction; low L∗ is
dark and high L∗ is light. The opacity of an emulsion can then
by characterized by its lightness (L*), and the color intensity
of an emulsion can then be characterized by its chroma: C =
(a ∗2 +b∗2 )1/2 . The higher the value of the chroma, the more
intense is the color. The optical properties of conventional emul-
sions (which are usually highly turbid or opaque) are typically
characterized using reflectance measurements, while the opacity
is expressed in terms of the lightness (L*). On the other hand, the
optical properties of nanoemulsions (which are usually clear or
slightly turbid) are typically characterized using transmission
measurements, and their opacity is expressed in terms of the
turbidity (τ ).
In general, the overall optical properties of nanoemulsions
are mainly determined by the relative refractive index, parti-
cle concentration, and particle-size distribution (McClements,
2002; McClements, 2002; McClements, 2005). The turbidity
or lightness of nanoemulsions tends to increase with increasing
refractive index contrast, with increasing particle concentration,
and have a maximum value at a particle size where light scat-
tering is strongest (i.e., r ≈ λ). The color intensity (chroma) is
inversely related to the degree of light scattering, i.e., when the
lightness or turbidity increases, the color intensity decreases. It
is for this reason that the color intensity of an emulsion may
decrease when the droplet concentration is increased or droplet
size is changed (Chantrapornchai et al., 1998; Chantrapornchai
et al., 1999; Chantrapornchai et al., 2001).
The dependence of the specific turbidity on particle diam-
eter for emulsions containing monodisperse (spread = 1) or
polydisperse (spread = 1.5, 2.0, or 2.5) droplets was calculated
using Mie theory at 600 nm (Fig. 15). The oil phase was as-
sumed to have a refractive index of 1.45 and the water phase
of 1.33. The specific turbidity was defined as the predicted tur-
bidity divided by the oil droplet concentration (expressed as a
volume percentage). From visual observation, emulsions can
be considered to be optically transparent when the turbidity at
600 nm is less than about 0.05 cm−1 for a 0.1 wt% oil-in-
water nanoemulsion, which corresponds to a specific turbidity
of about 0.5 cm−1 %vol−1 . For monodisperse emulsions, the pre-
dicted specific turbidity was relatively low (< 0.5 cm−1 %vol−1 )
 FOOD-GRADE NANOEMULSIONS 309

for droplets with diameters less than about 80 nm, but increased
appreciably when the droplets were larger than this size. As the
particle-size distribution became more polydisperse (increasing
spread from 1.0 to 2.5), the turbidity versus the particle diam-
eter relationship broadened. For polydisperse systems, a mean
particle diameter of < 30 nm (for a spread of 2.5), < 40 nm (for
a spread of 2.0) or < 60 nm (for a spread of 1.5) was required
for the systems to appear transparent, i.e., for specific turbidity
< 0.5 cm−1 %vol−1 . This effect can be attributed to the presence
of a population of larger particles in broad particle distribu-
tions that scatter light much more intensely than the smaller
particles. The precise particle-size dependence of the specific
turbidity depends on the particle size distribution of the emul-
sion in question. This highlights the importance of controlling
both the mean particle size and the width of the particle-size dis-
tribution in creating products that appear optically transparent.
These theoretical predictions are in agreement with the experi-
mental data of Wooster and co-workers, who reported that the
majority of droplets in a nanoemulsion should have a diameter
< 80 nm to obtain optically transparent systems (Wooster et al.,
2008). In summary, in order to prepare nanoemulsions that are
suitable for application in transparent food or beverage products
it is necessary to ensure that the majority of droplets fall below
some critical value (d ≈ 80 nm), which can be achieved by
having a small mean diameter and a narrow spread of the distri-
bution. These calculations also demonstrate that it is important
to prevent the droplets from growing during storage otherwise
the emulsions will become cloudy, e.g., by flocculation, coales-
cence, or Ostwald ripening.
Experimental measurements of the influence of droplet size
on the turbidity of protein-stabilized nanoemulsions are shown
in Fig. 16. The turbidity increment (i.e., the increase in turbid-
ity per cm per 1% increase in droplets) of the nanoemulsions
increases as the droplet diameter increases, particularly above
about 130 nm. For conventional emulsions, the lightness in-
creases steeply as the oil droplet concentration is increased from
0 to 5 wt%, but then increases more gradually at higher droplet
concentrations (Chantrapornchai et al., 1999). For nanoemul-
sions, the turbidity increases approximately linearly with in-
creasing droplet concentration for dilute systems, with the slope
depending on the droplet size (Lee and McClements, 2010).
The theoretical predictions shown in Fig. 15 were based on
the assumption that the particles in the nanoemulsions were ho-
mogeneous spherical oil droplets with a well-defined refractive
index. In practice, the particles in nanoemulsions consist of an
oil core surrounded by a shell of adsorbed material (e.g., emulsi-
fiers) that usually has different optical properties (refractive in-
dex) than the oil phase. For the small particles in nanoemulsions
this emulsifier layer makes up a relatively large volume fraction
of the coated particles (Fig. 1). To a first approximation, the

Figure 15 Calculated dependence of the specific turbidity of oil-in-water nanoemulsions and emulsions on mean particle size for systems with various particle
size distributions. It is assumed that the systems have a log-normal particle size distribution, which is characterized by its geometric mean and spread.
310 D. J. McCLEMENTS AND J. RAO

Figure 16 Turbidity increment versus mean droplet diameter for corn oil-in-water nanoemulsions stabilized by whey protein isolate produced using the
homogenization/solvent evaporation method. Adapted from Lee and McClements (2010).

refractive index of a coated droplet can be given by the weighted
average of the refractive indices of the different components:
nparticle =
S nS + (1 −
S )nC (5) of the droplets. A number of reviews of nanoemulsion rheology
Where n is the refractive index,
is the volume fraction, and additional information (Tadros et al., 2004; Mason et al., 2006).
the subscripts S and C stand for the shell (emulsifier layer) and
core (oil phase), respectively. The emulsifier layer is usually
comprised of surfactant and/or polymer molecules that have a
larger refractive index than either oil or water. Hence, the refrac-
tive index of the particles will tend to increase as the emulsifier
concentration in the interfacial layer increases, as the thickness
of the interfacial layer increases, or as the oil droplets become
smaller, which may appreciably alter the overall scattering prop-
erties of the particles. In practice, a more sophisticated mathe-
matical model of light scattering should be used that takes into
account the core-shell structure of the particles rather assuming
that they are homogeneous spheres (Quirantes et al., 1997).
Rheological Properties
Foods that contain lipid droplets may exhibit a wide variety
of different rheological characteristics (e.g., viscous liquids, vis-
coelastic liquids, viscoelastic solids, plastics, or elastic solids)
depending on their composition and microstructure (Quemada
and Berli, 2002; Walstra, 2003; Berli et al., 2005; McClements,
2005; Genovese et al., 2007). Nanoemulsions may have consid-
erably different rheological properties than conventional emul-
sions with the same oil content due to the relatively small size
have been published recently, which the reader is referred to for
The rheology of dilute colloidal dispersions is typically char-
acterized in terms of their shear viscosity (McClements, 2005;
Genovese et al., 2007). For dilute colloidal dispersions (φ <
0.05), the viscosity can be described by Einstein’s equation:
η
= 1 + 2.5φ (6)
η0
Here, η is the viscosity of the overall system, η0 is the viscosity
of the continuous phase, and φ is the disperse phase volume
fraction. This equation is not applicable at higher droplet con-
centrations due to droplet-droplet interactions. For concentrated
conventional emulsions, the viscosity can be described by the
following semi-empirical equation, which takes into account the
increase in viscosity that occurs in concentrated systems due to
droplet-droplet interactions:


η φ −2
= 1− (7)
η0 φc
Here, φ is the disperse phase volume fraction, and φc is a critical
disperse-phase volume fraction above which the droplets are so
 FOOD-GRADE NANOEMULSIONS
closely packed together that they cannot easily flow past each
other. Typically, φc has a value of around 0.4 to 0.6 depending on
the nature of the system. For conventional O/W emulsions φ is
usually taken to be the volume fraction of the oil phase present,
but for nanoemulsions φ should be replaced by the effective
disperse phase volume fraction (φeffective ) given in Eq. (2). As
discussed earlier, the effective disperse-phase volume fraction
of an O/W nanoemulsion may be much greater than the volume
fraction of the oil phase because of the presence of the adsorbed
layer (Fig. 13b).
The viscosity of a conventional emulsion increases with
increasing droplet concentration, gradually initially and then
steeply as the droplets become more closely packed (Fig. 17).
Around and above the droplet concentration where close pack-
ing occurs (φc typically around 0.4 to 0.6 for a non-flocculated
conventional emulsion), the emulsion exhibits solid-like char-
acteristics, such as visco-elasticity and plasticity (Quemada and
Berli, 2002; Berli et al., 2005; McClements, 2005). For na-
noemulsions, the oil concentration where the steep increase in
viscosity occurs can be considerably less than that for conven-
tional emulsions because the interfacial shell contributes ap-
preciably to the particles effective volume fraction (Fig. 17).
Experimental evidence of this effect is given in Fig. 18 (Weiss
et al., 2000), which shows that hydrocarbon oil-in-water emul-
sions undergo a transition from a liquid-like state when the
droplets are relatively large (d > 200 nm) to a gel-like state
when the droplets are relatively small (d < 80 nm). Similar
results have also been reported for silicone oil-in-water emul-
sions stabilized by an anionic surfactant (SDS), which can be
attributed to the “shell layer” around each oil droplet corre-
sponding to the Debye screening length (Wilking and Mason,
2007).
The impact of the droplet characteristics on the overall rhe-
ology of an emulsion may be an important consideration when
designing food products. Some products should have a rela-
tively low viscosity (such as beverages) and so the droplets
present should not appreciably increase the overall viscosity.
Other products should be highly viscous or gel-like (e.g., dress-
ings, dips, deserts) and the droplets may play an important role
by thickening the system or forming a gel network. The above
discussion suggests that it may be possible to prepare viscous
or gel-like nanoemulsions with rheological properties similar to
conventional emulsions of much higher fat content.
Stability
Like conventional emulsions, nanoemulsions are metastable
systems that tend to breakdown over time through a variety
of different physicochemical mechanisms, including gravita-
tional separation, flocculation, coalescence, Ostwald ripening,
and chemical degradation (Dickinson, 1992; Friberg et al., 2004;
McClements, 2005). In this section, a brief overview of the im-
pact of droplet size on the physical and chemical stability of
nanoemulsions is given.
311
Gravitational Separation
Gravitational separation is one of the most common forms of
instability in conventional emulsions, and may take the form of
either creaming or sedimentation depending on the relative den-
sities of the dispersed and continuous phases.Creaming is the
upward movement of droplets when they have a lower density
than the surrounding liquid, whereas sedimentation is the down-
wards movement of droplets when they have a higher density
than the surrounding liquid. Liquid edible oils (such as triacyl-
glycerol or flavor oils) normally have lower densities than liquid
water and so creaming is more prevalent in conventional oil-in-
water emulsions, whereas sedimentation is more prevalent in
water-in-oil emulsions. However, if an oil-in-water emulsion
contains crystalline lipids then they may be prone to sedimen-
tation because the density of lipids usually increases when they
crystallize. Oil-in-water nanoemulsions may also be prone to
sedimentation if they contain small oil droplets covered by a
relatively thick and dense shell (see below).
The velocity that a particle moves upwards in an emulsion or
nanoemulsion due to gravitational separation is given by Stokes’
law:
2
2grparticle (ρparticle − ρ0 )
v=− (8)
9η0
where, v is the creaming velocity, rparticle is the particle radius,
ρparticle is particle density, ρ0 is the continuous-phase density,
η0 is the continuous-phase viscosity, and g is the acceleration
due to gravity. For core-shell particles the radius is given by
rparticle = rcore + δ, and the particle density depends on particle
dimensions (rcore , δ) and the densities of the core and shell (see
below). Gravitational forces cause particles to move either up-
wards or downwards depending on their density relative to the
surrounding liquid. Hence, if only gravitational forces operated,
then the particles would accumulate at either the top or the bot-
tom of a sample. In practice, particles may also move due to
Brownian motion forces associated with the thermal energy of
the system. The root mean square distance moved by a√ particle
in a fluid due to Brownian motion is given by:  = (2Dt),
where D is the translational diffusion coefficient of the particle
and t is the time (Walstra, 2003). The dependence of the cream-
ing velocity on the particle size for oil droplets (ρOil = 920 kg
m−3 ) suspended in water (ηWater = 1 mPa s, ρWater = 1000 kg
m−3 ) is shown in Fig. 19 (assuming no shell). The distance the
particles are predicted to move in one day due to gravitational
forces and Brownian motion are plotted. The upward move-
ment of particles due to gravity increases with increasing par-
ticle size for gravitational forces, but decreases with increasing
particle size for Brownian motion. Consequently, particle move-
ment is dominated by gravity for the relatively large particles in
emulsions, but dominated by Brownian motion for relatively
small particles like those in some nanoemulsions. These calcu-
lations indicate that we would not expect to see any creaming in
nanoemulsions when the particle diameter fell below about 70
nm due to the domination of Brownian motion effects.
312 D. J. McCLEMENTS AND J. RAO

Figure 17 Calculated dependence of the relative viscosity of oil-in-water nanoemulsions on mean particle diameter for systems with a shell thickness of 10 nm.

As mentioned earlier, the shell layer may form an appreciable shell makes little contribution to particle density). This indicates
fraction of the total volume of the droplets in nanoemulsions that it should be possible to produce density-matched particles
(Fig. 1). The overall density of a particle consisting of a core by controlling the oil-core size and the thickness of the adsorbed
(oil) surrounded by a shell layer (emulsifier) is given by: emulsifier layer.

ρparticle =
S ρS + (1 −
S )ρC (9) Droplet Aggregation
Nanoemulsions typically have much better stability to par-
The shell layer usually has a higher density than the oil or
ticle aggregation (flocculation and coalescence) than conven-
aqueous phases, so that an increase in the volume fraction of
tional emulsions because of the influence of their small particle
the shell layer will tend to increase the overall particle density.
size on colloidal interactions (Tadros et al., 2004). The nature of
This may have important implications in preventing gravita-
the colloidal interactions operating in a particular nanoemulsion
tional separation in emulsions since it will reduce the density
system depends on the bulk physicochemical properties of the
contrast between the particles and surrounding fluid (Fig. 20).
various phases (e.g., dielectric constant and refractive index),
In addition, very small particles may actually sediment rather
particle-core characteristics (such as radius), shell charac-
than cream if they contain sufficiently thick and dense emulsi-
teristics (such as thickness, charge, packing, rheology, and
fier layers. A practical example of this effect is shown in Fig.
hydrophobicity), and the properties of the intervening fluid (such
21, which compares the pH-stability of protein-stabilized con-
as pH, ionic strength, osmotic pressure, and temperature). To a
ventional emulsions (d43 = 325 nm) and nanoemulsions (d43 =
first approximation the overall colloidal interactions between a
66 nm) to gravitational separation. The small particles in the
pair of droplets in an emulsion or nanoemulsion can be described
nanoemulsions tend to sediment because the overall particle
by the sum of the van der Waals (wVDV ), electrostatic (wE ), steric
density is greater than water (δ/r is relatively large, so the shell
(wS ), and hydrophobic (wS ) interactions (McClements, 2005):
makes a large contribution to particle density), whereas the large
particles in conventional emulsions tend to cream because their
overall density is less than water (δ/r relatively small, so the w(h) = wVDV (h) + wE (h) + ws (h) + wH (h) (10)
 FOOD-GRADE NANOEMULSIONS
Figure 18 Influence of particle radius on the rheology of SDS-stabilized oil-in-water nanoemulsions. The nanoemulsions gain solid like characteristics when
the droplet size falls below a critical level (Weiss and McClements, 2000).
The van der Waals and hydrophobic interactions are attrac-
tive, whereas the steric and electrostatic interactions are usually
repulsive. The steric interaction is a strong short-range repulsive
interaction, whereas the magnitude and range of the electrostatic
repulsion depends on the electrical charge on the droplets (ζ -
potential) and the ionic composition of the aqueous phase (I). In
general, the magnitude of both the attractive and repulsive col-
loidal interactions tends to increase with increasing droplet size
(McClements, 2005). Theoretical predictions of the influence
of ionic strength on the overall interaction potential between
protein-coated oil droplets are shown in Fig. 22 for nanoemul-
sions (r = 25 nm) and conventional emulsions (r = 250 nm). It
was assumed that the droplets had a relatively high ζ -potential
(+25 mV) so that there was a strong electrostatic repulsion be-
tween them in the absence of added salt. In the conventional
emulsions at low ionic strengths (I = 20 and 50 mM), there
was a high potential energy barrier (> 20 kT) due to electro-
static repulsion that should prevent droplets from coming close
enough to aggregate (Fig. 22a). On the other hand, at high ionic
strengths (200 and 500 mM), this energy barrier was greatly
diminished due to electrostatic screening effects and there was
a deep primary minimum. The droplets would therefore be ex-
pected to flocculate at high ionic strengths in conventional emul-
313
sions, which has been observed experimentally (Demetriades et
al., 1997a; 1997b). A similar kind of behavior was observed in
the nanoemulsions, but the attractive and repulsive interactions
were much weaker, and hence the height of the energy barrier
and the depth of the primary minimum were much smaller (Fig.
22b). In this case, one would expect there would be either little or
no droplet flocculation. This effect has recently been observed
in protein-stabilized corn oil-in-water nanoemulsions (d43 =
60 nm) in our laboratory, where they were found to be stable to
droplet aggregation and creaming from 0 to 1000 mM NaCl (data
not shown). Thus both theoretical predictions and experimental
measurements suggest that nanoemulsion delivery systems can
be designed that have much better stability to droplet aggrega-
tion and gravitational separation than conventional emulsions.
Ostwald Ripening
Ostwald ripening is the process whereby the mean size of the
droplets in an emulsion or nanoemulsion increases over time
due to diffusion of oil molecules from small to large droplets
through the intervening fluid (Kabalnov et al., 1992; Kabal-
nov, 2001). The driving force for this effect is the fact that the
314 D. J. McCLEMENTS AND J. RAO

Figure 19 Calculated influence of gravity and Brownian motion on the creaming velocity of differently sized oil droplets in oil–in–water emulsions.

water-solubility of an oil contained within a spherical droplet
increases as the size of the droplet decreases, which means that
there is a higher concentration of solubilized oil molecules in
the aqueous phase surrounding a small droplet than surrounding
a larger one (Kabalnov et al., 1992; McClements, 2005). Sol-
ubilized oil molecules therefore tend to move from around the
smaller droplets to around the larger droplets because of this
concentration gradient. This leads to an increase in droplet size
over time, which can be described by the following equation
once steady state has been achieved (Kabalnov et al., 1992):
32
d(t)3 − d(0)3 = ωt = αS∞ Dt
9 tains droplets comprised of two different lipid components—a
Here, d(t) is the number mean droplet diameter at time t, d0
is the initial number mean droplet diameter, ω is the Ostwald
ripening rate, α = 2γ Vm /RT, S∞ is the water-solubility of the
oil phase in the aqueous phase, D is the translational diffusion
coefficient of the oil molecules through the aqueous phase, Vm is
the molar volume of the oil, γ is the oil-water interfacial tension,
R is the gas constant, and T is the absolute temperature.
In practice, the main factor determining the stability of a na-
noemulsion to OR is the water-solubility of the oil phase (S∞ ).
For this reason OR is not usually a problem for nanoemulsions
prepared using oils with a very low water-solubility, such as
long-chain triglycerides (e.g., corn, soy, sunflower, or fish oils).
On the other hand, OR may occur rapidly for nanoemulsions
prepared using oils with an appreciable water-solubility, such as
short-chain triglycerides, flavor oils, and essential oils (Wooster
et al., 2008; Li et al., 2009). Ostwald ripening can be retarded
in these systems by adding a substance known as a ripening
inhibitor. A ripening inhibitor is a non-polar molecule that is
soluble in the oil phase but insoluble in the water phase, e.g., a
long-chain triacylglycerol. This type of molecule can inhibit OR
by generating entropy of mixing effect that counter-balances the
curvature effects. Consider an oil-in-water emulsion that con-
(11)
water-insoluble component and a water-soluble component. The
water-soluble molecules will diffuse from the small to the large
droplets due to OR. Consequently, there will be a greater con-
centration of water-insoluble molecules in the smaller droplets
than in the larger droplets after OR occurs. Differences in the
composition of emulsion droplets are thermodynamically un-
favorable because of entropy of mixing—it is more favorable
to have the two lipids distributed evenly throughout all of the
droplets, rather than be located in particular droplets. Conse-
quently, there is a thermodynamic driving force that operates in
the opposite direction to the OR effect. The change in droplet
 FOOD-GRADE NANOEMULSIONS 315

Figure 20 Predicted influence of droplet size and shell layer thickness on the effective density of the core-shell particles in nanoemulsions. It was assumed that
the density of the shell was 1200 kg m−3 and that of the oil phase was 920 kg m−3 .

size distribution with time then depends on the concentration
and solubility of the two components within the oil droplets.
This approach has previously been used to improve the stability
of food-grade nanoemulsions, such as those containing short-
chain triglycerides or essential oils (Wooster et al., 2008; Li et
al., 2009). An example of this effect is shown in Fig. 23, which
shows the change in droplet size during storage of orange oil-
in-water emulsions containing different levels of corn oil (the
ripening inhibitor). Orange oil (4-fold) has a relatively high sol-
ubility in water, and therefore is highly prone to OR, which leads
to an appreciable increase in mean droplet size during storage.
On the other hand, corn oil has very low solubility in water, and
therefore it can retard OR if it is incorporated into the oil phase
prior to homogenization. Our results show that incorporating ≥
10% corn oil into the lipid phase was sufficient for inhibiting
OR in these systems.
Chemical Stability
The small size of the lipid droplets in oil-in-water nanoemul-
sions may also impact the chemical degradation rate of any
encapsulated components, e.g., due to oxidation or hydrolysis.
First, the oil-water surface area increases as the droplet size de-
creases (A ∝ 1/r), which means that more of the lipid phase
will be exposed to the surrounding aqueous phase. If chemi-
cal degradation occurs primarily at the oil-water interface, then
reducing the lipid droplet size may promote the reaction rate.
Second, if chemical degradation is accelerated by the presence
of UV or visible light, then a transparent nanoemulsion contain-
ing small droplets may be more susceptible to degradation than
an opaque conventional emulsion because light can penetrate
more easily. Nevertheless, one must also take into account any
differences in the properties of the interfacial layer surrounding
the lipid droplets in nanoemulsions and emulsions.
Molecular Distribution and Release Characteristics
The ability of nanoemulsions to incorporate and release func-
tional components, such as flavors, antimicrobials, vitamins,
nutraceuticals, and drugs, depends on the molecular character-
istics of the functional components as well as the composition,
properties, and microstructure of the nanoemulsions themselves
(McClements, 2005). Some encapsulated components are solu-
ble in both oil and water phases and will move out of oil droplets
when the emulsions are diluted with water. In principle, the re-
lease of this kind of encapsulated component can be delayed by
316 D. J. McCLEMENTS AND J. RAO
Figure 21 Influence of pH on the stability of protein-stabilized 0.5 wt%
corn oil-in-water conventional emulsions (r43 = 163 nm) and nanoemulsions
(d43 = 33 nm) to gravitational separation. The particles in the nanoemulsions
moved downwards because their overall density was greater than water (due
to the thick adsorbed protein layer), whereas the particles in the conventional
emulsions moved upwards because their density was less than water (Lee and
McClements, 2010).
Figure 22a Predicted influence of changes in ionic strength on the total inter-
action potential between oil droplets coated by a globular protein: Nanoemulsion
(r = 25 nm, δ = 3 nm, ζ = 25 mV).
Figure 22b Predicted influence of changes in ionic strength on the total inter-
action potential between oil droplets coated by a globular protein: Nanoemulsion
(r = 25 nm, δ = 3 nm, ζ = 25 mV).
increasing the droplet size. However, the droplets in nanoemul-
sions are so small that they have little effect on the release rate
of encapsulated components through this mechanism. For en-
capsulated components that have a very low water-solubility,
one is typically interested in their release into the surrounding
aqueous phase in the human body, e.g., the mouth, stomach, or
small intestine. In this case, it is usually important to establish
any potential trigger mechanisms for release (e.g., dilution, pH,
Figure 23 Dependence of mean droplet diameter on storage time for 5% oil-
in-water emulsions stabilized by modified starch on ratio of orange oil to corn
oil in the oil phase (pH 3.5, 20◦ C).
 FOOD-GRADE NANOEMULSIONS
ionic strength, temperature, enzyme activity, etc.), as well as the
rate and extent of release. In an emulsion-based delivery system,
the release is usually characterized in terms of the increase in
concentration of the encapsulated compound in the continuous
phase or in some target organ (such as the mouth, nose, stom-
ach, or gastrointestinal tract) as a function of time (Aguilera,
2006). The ability of nanoemulsions to encapsulate and release
non-volatile components may be quite different from conven-
tional emulsions made from similar components. The solubility
of a component within an emulsion droplet usually increases
as its droplet size decreases. For a typical food oil contained
within a droplet surrounded by water (v = 10−3 m3 mol−1 ,
γ = 10 mJ m−2 ), the solubility increases 2.24-, 1.08-, 1.01-,
and 1.00-fold for oil droplets with radii of 10, 100, 1000, and
10,000 nm, respectively. This effect may alter the oil-water par-
tition coefficient (KOW ) of any substances encapsulated within
a nanoemulsion, and therefore their bioavailability.
PRACTICAL APPLICATIONS OF NANOEMULSIONS
Research into the development and application of food-grade
nanoemulsions has increased recently because of the growing
need for effective delivery systems to encapsulate, protect and
release functional food components. Indeed, a recent report
(“Food Encapsulation: A Global Strategic Business Report,”
Global Industry Analysts, Inc., San Jose, CA) estimated that the
world market for food encapsulation will be nearly $40 billion
by 2015. We have tabulated some commercial applications of
nanoemulsions found on the Internet (Table 9). In the remainder
of this section, we provide an overview of potential applications
of nanoemulsions in the food and beverage industries.
Encapsulation of Lipophilic Components
An important application of nanoemulsions in foods and bev-
erages is their ability to encapsulate lipophilic functional com-
ponents, such as vitamins, flavors, colors, antioxidants, preser-
vatives, and nutraceuticals (Graves and Mason, 2008; Given,
2009). Encapsulation of a lipophilic component may be carried
out for a number of reasons—to improve its ease of handling
and utilization; to facilitate its incorporation within a product;
to increase its bioavailability by improving the properties of
lipophilic compounds, such as solubility and efficient absorp-
tion through cells; to control the rate or location of its release;
Table 9 Some research articles on O/W nanoemulsions formed using the Emulsion Inversion Point method. Cremophor EL is a hydrogenated castor oil
surfactant
Oil Phase Surfactant/Co-surfactant
MCT Cremophor EL
Canola oil Span 20, Span 60, Span 80, Tween 20, Tween 60, Tween 80
Paraffin oil Tween 20/Span 20
Hexadecane/styrene Brij 98
317
or, to protect it from chemical degradation. One of the major ad-
vantages of nanoemulsions over conventional emulsions for this
kind of application is that they can be incorporated into clear or
slightly turbid products without altering their visual appearance.
(See section titled “Optical Properties.”)
Lipophilic components are usually blended with the oil phase
prior to nanoemulsion formation, so that they are trapped within
the oil core of the droplets once the nanoemulsion has been
produced. The physical location of lipophilic components within
nanoemulsions depends on their molecular and physicochemi-
cal properties, such as hydrophobicity, surface activity, oil-water
partition coefficient, solubility, and melting point. Highly non-
polar lipophilic components tend to be distributed within the
hydrophobic core of the particles, whereas more polar lipophilic
components may be incorporated within the amphiphilic shell.
The physical location of the lipophilic components may have
an important impact on the physical and chemical stability of
the system. For example, some lipophilic components are sus-
ceptible to chemical degradation when they come into contact
with particular water-soluble components in the aqueous phase.
In this case, it may be important to ensure that the lipophilic
components are trapped within the oil core, rather than within
the shell. An example of this effect is the chemical degradation
of citral molecules (a flavor oil component), which is promoted
when these lipophilic components come into contact with pro-
tons in the aqueous phase (Choi et al., 2009; 2010; Mei et al.,
2010). In practice, it is often difficult to determine the precise
location and orientation of lipophilic components within na-
noemulsion droplets. The analytical methods that can be used
are highly dependent on the nature of the lipophilic component
used. Some of the methods that have proved useful for the in situ
determination of the location of lipophilic components within
emulsion droplets include nuclear magnetic resonance (Hamil-
ton et al., 1996; Okamura et al., 2001; Nakaya et al., 2005;
Shen et al., 2005), fluorescence spectroscopy (Saito et al., 1996;
Tanaka et al., 2003) and Raman scattering microspectroscopy
(Day et al., 2010).
Nanoemulsions have been developed to encapsulate a num-
ber of different lipophilic components, such as β-carotene (Tan
and Nakajima, 2005a,b; Chu et al., 2007a; 2008; Yuan et al.,
2008a,b; Mao et al., 2009; Yin et al., 2009), citral (Choi et
al., 2009; 2010; Mei et al., 2010), capsaicin (Choi et al., 2009),
flaxseed oil (Kentish et al., 2006), tributyrin (Li et al., 2009), co-
Enzyme Q (Ozaki et al., 2010), and oil-soluble vitamins (Relkin
et al., 2009; Hatanaka et al., 2010).
Particle Diameter Reference
28–78 nm Sadurn et al., 2005
215–233 nm de Morais et al., 2006
80–180 nm Pey et al., 2006
36 nm Sadtler et al., 2010
318 D. J. McCLEMENTS AND J. RAO
Increased Bioavailability and Controlled Delivery
of Lipophilic Components
A number of studies have shown that the bioavailability of
highly lipophilic functional components encapsulated within
lipid droplets is increased when the droplet size is decreased
(Kesisoglou et al., 2007; Acosta, 2009). There are a number of
possible reasons for this increase in bioavailability. (See section
titled “Biological Fate, Bioavailability, and Potential Toxicity
of Nanoemulsions.”)
First, small droplets have a large surface area and may there-
fore be digested more quickly by digestive enzymes so that
their contents are released and absorbed more easily. Second,
the small droplets may penetrate into the mucous layer that
coats the epithelium cells within the small intestine, thereby
increasing their residence time and bringing them closer to the
site of absorption. Third, very small particles may be directly
transported across the epithelium cell layer by either paracellu-
lar or transcellular mechanisms. Fourth, the water-solubility of
highly lipophilic components increases as the droplet size de-
creases, which may enhance absorption. At present there is no
good understanding of the relative importance of these differ-
ent mechanisms for nanoemulsions with differing droplet sizes,
compositions, and surface characteristics.
Recently, it has been shown that the oral availability of cur-
cumin can be increased by incorporating it within nanoemul-
sions (Wang et al., 2008; Huang et al., 2010). Nanoemulsions
have also been shown to improve the bioavailability of various
lipophilic nutraceuticals and pharmaceuticals (Hatanaka et al.,
2008; Nielsen et al., 2008; Ozaki et al., 2010; Talegaonkar et al.,
2010). It has also been proposed that nanoemulsions can be used
to effectively deliver nutraceuticals to specific sites within the
human body, which may lead to improvements of their efficacy
(Vishwanathan et al., 2009; Huang et al., 2010). For example,
the Salvona Company developed a delivery system (MultiSalTM )
to deliver multiple active ingredients (water-soluble and fat-
soluble ingredients). It was proposed that this system enhances
the stability and bioavailability of a wide range of nutrients and
other ingredients, controls their release characteristics, and pro-
longs their residence time in the oral cavity, and thus prolongs
the sensation of flavors in the mouth. Recently, our laboratory
designed nanoemulsion-based delivery systems to control the di-
gestion and release of lipophilic food components (McClements
and Li, 2010).
Improved Emulsion Stability
As mentioned earlier, the stability of emulsions to grav-
itational separation, flocculation, and coalescence is greatly
improved when their size is reduced, which may help extend the
shelf-life of some emulsion-based products. (See section titled
“Stability.”) Nevertheless, it is important to design the system
so that Ostwald ripening does not occur since nanoemulsions
are highly susceptible to degradation through this mechanism
(Fig. 23). This can be achieved by preparing them from a carrier
oil that has very low water-solubility (Wooster et al., 2008;
Li et al., 2009), or by controlling the mechanical properties
of the interfacial layer to restrict changes in droplet size (Mun
and McClements, 2006). In addition, the very small size of
the droplets in nanoemulsions may promote the chemical
degradation of encapsulated components (Mao et al., 2009).
Nanoemulsions have very large specific surface areas so that
any chemical degradation reaction that occurs at the oil-water
interface may be promoted, such as lipid oxidation. In addition,
transparent nanoemulsions will allow UV and visible light to
penetrate them easily, which may promote any light-sensitive
chemical degradation reactions. Consequently, it may be nec-
essary to take additional steps to improve the chemical stability
of labile components encapsulated within nanoemulsions, e.g.,
by adding antioxidants or chelating agents.
Texture Modification
The fact that the droplets in nanoemulsions are very small
means that the interfacial coating surrounding them may make
an appreciable contribution to the particles overall volume
(Fig. 1). Consequently, it may be possible to promote the gela-
tion of a nanoemulsion at a much lower oil concentration than
for a conventional emulsion (Fig. 18). This may be desirable
for producing reduced fat products that need to be highly vis-
cous or gel like. At present, there is little work on utilizing
nanoemulsions to create novel textural effects in edible prod-
ucts. Nevertheless, recent studies with non-edible systems have
shown that transparent nanoemulsions with gel-like characteris-
tics can be prepared based on electrostatic repulsion effects, i.e.,
when the thickness of the electrical double layer is on the or-
der of the radius of the emulsion droplets (Wilking and Mason,
2007; Kawada et al., 2010). Potentially, this approach could be
used to produce highly viscous or visco-elastic nanoemulsions
using much lower oil contents than is required to produce sim-
ilar systems in conventional emulsions (Wilking and Mason,
2007).
Antimicrobial Activity
A number of studies suggest that nanoemulsions may be an
effective means of encapsulating and delivering antimicrobial
agents (Hamouda et al., 1999; 2001; Teixeira et al., 2007; Fer-
reira et al., 2010). Antimicrobial nanoemulsions have been de-
veloped for the decontamination of food packaging equipment
and for application to various food surfaces (Sekhon, 2010).
Nanoemulsions based on non-ionic surfactants, soybean oil and
tributyl phosphate, have been reported to be highly effective
against a variety of food pathogens, including bacteria, viruses,
fungi, and spores (Hamouda et al., 2001). However, they tend to
be much more effective against Gram-positive bacteria than
against Gram-negative bacteria, which was attributed to the
 FOOD-GRADE NANOEMULSIONS
influence of the cell-wall lipopolysaccharide that surrounds
Gram-negative bacteria. The mechanisms by which antimicro-
bial nanoemulsions inhibit microbial growth depend on the na-
ture of the encapsulated antimicrobial agents they contain (e.g.,
essential oils, proteins, surfactants), as well as on the nature of
the nanoemulsion droplets themselves (e.g., size, charge, com-
position). Nanoemulsions can be designed to contain a variety
of different antimicrobial agents that may act synergistically.
Nanoemulsion particles can be designed to deliver one or more
entrapped antimicrobial agents after they come into contact with
the surfaces of microorganisms. The interaction between the
droplets and the microorganism can be enhanced by controlling
the colloidal interactions in the system. For example, cationic
nanoemulsion particles can be made to bind strongly to anionic
bacteria surfaces through electrostatic attraction (Li et al., 2001),
which may improve their antimicrobial efficacy by bringing the
antimicrobial agents close to the site of action. Recently, it
has been shown that oil-in-water nanoemulsions prepared from
non-ionic surfactants and soybean oil are effective at inhibit-
ing bacterial growth in a variety of different biofilms (Teixeira
et al., 2007).
BIOLOGICAL FATE, BIOAVAILABILITY, AND
POTENTIAL TOXICITY OF NANOEMULSIONS
There has been growing concern about the increased utiliza-
tion of nanoparticles in foods and beverages because of poten-
tial toxic effects associated with their small dimensions (Hagens
et al., 2007; Chaudhry et al., 2008; Bouwmeester et al., 2009;
Souto et al., 2009). Reducing the dimensions of a material to
the nanometer region may alter its biological fate within the
human body, such as the absorption, distribution, metabolism,
and excretion (ADME) processes, thereby altering its potential
for promoting toxicity (Hagens et al., 2007; Bouwmeester et al.,
2009; Hu et al., 2009). The biological effects of nanoparticles de-
pend on their composition and physicochemical properties, such
as concentration, particle-size distribution, electrical charge, and
interfacial characteristics. There is likely to be a large difference
in the effects of non-digestible inorganic nanoparticles (such as
metals and minerals) compared to the effects of digestible or-
ganic nanoparticles (such as surfactants, lipids, proteins, and
carbohydrates). The particles in nanoemulsions usually consist
of a core of lipophilic material surrounded by a shell of ad-
sorbed material (Fig. 1). As mentioned previously, the lipophilic
core may be comprised of one or more non-polar components,
including triacylglycerols, diacylglycerols, monoacylglycerols,
flavor oils, essential oils, mineral oils, fat substitutes, weight-
ing agents, oil-soluble vitamins, and nutraceuticals (such as
carotenoids, phytosterols, and Co-enzyme Q). Some of these
lipophilic components are typically digested within the human
gastrointestinal (GI) tract (e.g., triacylglycerols and diacylglyc-
erols), some normally pass through the GI tract without being
absorbed (e.g., mineral oils and fat substitutes), while some
are absorbed without being digested (e.g., carotenoids and phy-
319
tosterols). The shell around nanoemulsion droplets may also
be comprised of one or more materials, including surfactants,
phospholipids, proteins, polysaccharides, and minerals, which
may be digested to different rates and extents within the GI
tract. For example, phospholipids and proteins may be digested
by phospholipases and proteases in the stomach and small in-
testine, whereas dietary fibers may not be digested until they
reach the large intestine. The biological fate of a nanoemulsion
particle may therefore depend on the nature of the core and shell
material. At present there is a knowledge gap in understanding
how specific particle characteristics influence the biological fate
of nanoemulsion particles within the GI tract, and it is clear that
more research is needed to establish their potential toxicity (Ha-
gens et al., 2007; Bouwmeester et al., 2009). In the remainder
of this section, some of the potential ways that nanoemulsion
droplets could promote toxic effects due to their small size are
discussed.
Biological Fate of Nanoemulsion Droplets
An improved understanding of the potential toxic effects of
ingested nanoemulsions depends on knowledge of the physico-
chemical and physiological processes that occur as nanoemul-
sion droplets pass through the human body (Singh et al., 2009;
McClements et al., 2009).
Digestion
In this section, a brief overview of the complex conditions
existing in different locations of the GI tract is given (Fig. 24),
and their potential influence on the properties of nanoemulsion
droplets prior to absorption is discussed (McClements et al.,
2009; Singh et al., 2009; van Aken, 2010b).
Mouth: After ingestion a nanoemulsion is mixed with saliva;
changes in pH, ionic strength, and temperature; is acted upon
by digestive enzymes; interacts with the surfaces of the tongue,
mouth and throat; and, experiences a complex flow/force profile
(van Aken, 2010a).
• As a result the nanoemulsion droplets may aggregate with
each other, or become coated with mucin molecules from the
saliva.
• Stomach: After passing through the esophagus and entering
the stomach, the “bolus” is exposed to highly acidic condi-
tions (pH 1 to 3); mixed with enzymes (e.g., proteases and
gastric lipases) and surface-active substances (e.g., phospho-
lipids and proteins); and, experiences a complex flow/force
profile (Kong and Singh, 2008; Ferrua and Singh, 2010).
Endogenous and exogenous surface-active substances may
displace the original surface-active materials from the lipid
droplet surfaces through competitive adsorption processes.
Gastric lipases initiate lipid digestion, while gastric proteases
initiate protein digestion.
320 D. J. McCLEMENTS AND J. RAO

Figure 24 Schematic diagram of the various physicochemical and physiological processes that may occur when nanoemulsions pass through the human GI
tract. The picture of the human body was obtained from http://en.wikipedia.org/wiki/Digestive tract (Copyright free).

• Small intestine: After entering the small intestine, the par-
tially hydrolyzed and emulsified lipids are mixed with al-
kaline digestive juices containing bile salts, phospholipids,
pancreatic lipase, colipase, and bicarbonate, which increase
the pH close to neutral. Surface-active substances present in
the small intestine compete with surface-active materials ini-
tially present at the lipid-droplet surfaces, and may displace
them. Various digestion enzymes may hydrolyze the com-
ponents present within a nanoemulsion droplet: pancreatic
lipase/co-lipase complex convert triglycerides and diglyc-
erides into monoglycerides and free fatty acids; phospho-
lipases convert phospholipids to free fatty acids and other
lipophilic substances; proteases convert proteins to peptides
and amino acids. Lipid digestion products are incorporated
within bile salt/phospholipid micelles and vesicles, and then
transported through the mucous layer to the surfaces of the
enterocytes where they are absorbed (Fig. 24). The mucous
layer has been reported to be about 30 to 100 µm thick (Roger
et al., 2010). Lipophilic components, such as nutraceuticals
or pharmaceuticals, which are incorporated into the mixed
micelles and vesicles, will also be transported to the surfaces
of the enterocytes.
As a result of exposure to these various physiological con-
ditions, the droplets in nanoemulsions may change appreciably
as they pass through the GI tract (Johnson, 2001; McClements
et al., 2007). For example, there may be changes in the com-
position, size distribution, aggregation state, electrical charge,
interfacial properties, and physical state of the nanoemulsion
droplets (Fig. 24). All of these changes need to be accounted for
when trying to establish the potential biological fate and poten-
tial toxicity of nanoemulsion delivery systems. At present, there
is a poor understanding of the impact of specific physiological
conditions on the fate of nanoemulsion droplets within the GI
tract.
Absorption
Once the nanoemulsion droplets and/or their digestion prod-
ucts reach the surfaces of the GI tract they may be absorbed (De
Jong et al., 2008; Acosta, 2009; Hu et al., 2009). There are two
main types of cells where particle absorption may occur: epithe-
lium cells and M-cells (des Rieux et al., 2006). Epithelium cells
are the most numerous type of cell lining the GI tract, but they
are not believed to be very efficient at absorbing particles. On
 FOOD-GRADE NANOEMULSIONS
Figure 25 The droplets in nanoemulsions, and any functional components encapsulated within them, may be absorbed to a different extent than the droplets in
conventional emulsions within the human gastrointestinal tract.
the other hand, M-cells are believed to be highly efficient at ab-
sorbing nanoparticles, but they are much less numerous (< 1%
of epithelium surface) being mainly located in specialized re-
gions called “Peyers patches.” Physiologically, the M-cells are
responsible for absorbing various kinds of antigens, including,
macromolecules, microorganisms, and certain types of particles,
which are then delivered to the underlying lymphoid system to
induce immune responses (des Rieux et al., 2006). Nanoparti-
cles may be absorbed by cells through a number of different
mechanisms (Sahay et al., 2010a; 2010b):
– Paracellular mechanism: Nanoparticles may be small enough
to pass between the narrow gaps (“tight junctions”) separating
neighboring epithelial cells (Fig. 25). This mechanism has
been suggested for the transport of solid lipid nanoparticles
across epithelium cells (Yuan et al., 2007).
– Transcellular mechanism: Nanoparticles may be small
enough to be absorbed directly through epithelial cell mem-
branes by either passive or active transport mechanisms
(Fig. 25). Typically, absorption occurs by an “endocytosis”
mechanism that involves the nanoparticle encountering the
cell membrane, the membrane wrapping itself around the
nanoparticle, and then part of the membrane budding-off to
form a vesicle with a nanoparticle trapped inside.
The pathway taken, and the rate and extent of nanoparticle
uptake, depends on the properties of the nanoparticles, e.g., size,
shape, charge, and interfacial chemistry (des Rieux et al., 2006;
Hu et al., 2009). Cationic nanoparticles have been shown to be
absorbed more readily than anionic nanoparticles (Hagens et al.,
2007; Bouwmeester et al., 2009). Nanoparticles (d < 1000 nm)
tend to be absorbed more readily by cells than microparticles
(d > 1000 nm), but that there is often an optimum intermediate
321
particle size for absorption in the nanometer range whose value
depends on particle type (Hu et al., 2009). Particle absorption
is also believed to depend on the surface hydrophobicity of
nanoparticles (des Rieux et al., 2006). The physicochemical
characteristics of a nanoparticles also determine their fate once
they have entered the epithelium cell: (i) They may be digested
by cellular enzymes into their constituent parts which may then
be absorbed; (ii) They may be transported directly out of the
cell and into the blood or lymph systems; or (iii) They may
accumulate within specific locations in the cell (Hu et al., 2009;
Sahay et al., 2010a,b).
Some types of nanoparticles have been shown to promote
cellular damage if they accumulate within biological cells (Un-
fried et al., 2007; Hu et al., 2009). The nanoparticles that
are transported out of the epithelial cells via the portal vein
or lymphatic system may circulate through the human body,
where they may then be metabolized, excreted, or accumulate
within certain tissues (Bouwmeester et al., 2009). As mentioned,
these processes depend on the precise physicochemical charac-
teristics of the nanoparticles involved. For example, smaller
(metallic) particles have been found to accumulate within a
wider range of tissues than larger particles (De Jong et al.,
2008).
One would expect that the direct absorption and accumu-
lation of the oil droplets in edible nanoemulsions would be
unlikely because they would be expected to be rapidly digested
within the stomach, small intestine, or epithelium cells. How-
ever, if the nanoemulsion droplets were formed from indigestible
oils (such as hydrocarbons or mineral oils) or if the droplets were
coated with indigestible shells (such as dietary fibers) then di-
rect absorption could occur. Direct particle absorption might
also occur if the nanoemulsion particles were able to diffuse
through the mucous layer so rapidly that only limited digestion
had time to occur. There is currently little empirical data on the
322 D. J. McCLEMENTS AND J. RAO
direct absorption of edible nanoemulsion particles by epithe-
lium cells, and further work is clearly needed. As mentioned
earlier, the physicochemical characteristics of the droplets in a
nanoemulsion may be altered considerably as they pass through
the GI tract (e.g., their size, aggregation state, charge, and com-
position), and these changes must be considered when studying
particle absorption mechanisms.
Metabolism, Distribution, and Excretion
If a nanoemulsion droplet is directly absorbed by the human
body (rather than being digested first), then its distribution, ac-
cumulation, metabolism, and excretion may be different from
that of normally digested components (Hagens et al., 2007;
Bouwmeester et al., 2009). Studies on metallic and inorganic
nanoparticles that are not digested in the GI tract have shown
that they may accumulate in different organs, with the extent
of retention depending on their size (De Jong et al., 2008). For
example, it was reported that smaller-sized gold nanoparticles
accumulated within a greater number of organs than larger-sized
ones. Further work is therefore needed to establish whether ed-
ible nanoparticles can be directly absorbed by the human body,
and if they can, what their biological fate is.
Biological Fate of Bioactive Components Encapsulated
within Nanoemulsion Droplets
Nanoemulsions are often used as delivery systems to en-
capsulate lipophilic components, such as ω-3 oils, carotenoids,
phytosterols, oil-soluble vitamins, and water-insoluble drugs
(McClements et al., 2007; Velikov et al., 2008; Acosta, 2009).
It is therefore important to understand how the properties of
nanoemulsions influence the release and absorption of these
encapsulated components.
The term bioavailability is defined as the fraction of an in-
gested component that eventually ends up in the systemic cir-
culation (Versantvoort et al., 2004). The overall bioavailability
(F ) of lipophilic components that have poor water-solubility
depends on a number of physicochemical and physiological
factors:
F = FB × FA × FM (12)
Here, FB is the fraction of lipophilic component released
from the delivery system into the lumen of the gastrointesti-
nal tract to become bioaccessible, FA is the fraction of the
released component that is absorbed across the intestinal ep-
ithelia, and FM is the fraction of the absorbed component that
reaches the systemic circulation without being metabolized.
The characteristics of the droplets present within nanoemul-
sions may affect these processes in a variety of ways (Acosta,
2009):
Transit Time: The time that a lipid particle spends within the
GI tract may affect the rate and extent of the enzymatic
hydrolysis of digestible lipids (such as triacylglycerols), as
well as the extent of absorption of any encapsulated lipophilic
substances. The particles in nanoemulsions may increase the
transit time in the GI tract compared to those in conventional
emulsions through a number of mechanisms (Bouwmeester
et al., 2009):
(i) The small size of the droplets in nanoemulsions means
they may be able to penetrate into the mucous layer
coating the enterocytes, whereas large droplets cannot
(Fig. 25);
(ii) If the droplets in a nanoemulsion have a positive charge
in the small intestine they may also become trapped
within the mucous layer (which consists of anionic
biopolymers).
An increase in transit time should enable more of the lipids to
be digested, and a greater fraction of the bioactive component
to be solubilized by micelles and absorbed by enterocytes
(Acosta, 2009).
Digestion: Lipid digestion occurs at the oil-water interface sur-
rounding the emulsified lipid droplets and so the rate of lipid
digestion tends to increase as the droplet size decreases, be-
cause this increases the surface area of lipid exposed to the
digestive enzymes (Armand et al., 1992; Lundin et al., 2008).
Thus nanoemulsions should be digested more rapidly than
conventional emulsions, which may lead to the more rapid
generation of lipid digestion products that can solubilize
bioactive components (see below), and to the faster release
of any encapsulated components. Nevertheless, the rate of
lipid digestion may not always be faster for nanoemulsions
than for conventional emulsions. Recent experiments in our
laboratory found that globular protein-stabilized nanoemul-
sions prepared using the homogenization/solvent evapora-
tion method were digested at a slower rate than conventional
emulsions, which was attributed to the fact that the nanoemul-
sions had a thicker protein coating than the conventional
emulsions.
The influence of the nature of the interfacial layer coating the oil
droplets on lipid digestion has also been shown in a number
of other studies. If the interface is able to prevent the lipase
molecules from adsorbing to the droplet surfaces, then the
rate and extent of lipid digestion may be retarded (Mun et al.,
2006; Chu et al., 2009). The composition of the carrier lipid
may also affect the rate and extent of digestion and absorp-
tion. Triacylglycerol oils (such as corn oil or fish oil) should
be fully digested within the stomach and small intestine,
whereas fat substitutes (such as Olestra) should not be. Thus,
more of a bioactive may be released from a triacylglycerol oil
than from a fat substitute. Lipid digestion also depends on the
molecular characteristics of digestible triacylglycerols oils,
e.g., medium-chain triglycerides (MCT) tend to be digested
more rapidly than long-chain triglycerides (corn oil) (Porter
et al., 2007).
Solubilization: The particle characteristics within a nanoemul-
sion may also influence the solubization of lipids and
lipophilic components within micelles and vesicles in the
 FOOD-GRADE NANOEMULSIONS
small intestine (Porter et al., 2006; Porter et al., 2007). First,
the presence of the lipid carrier itself stimulates the release
of bile salts and phospholipids from the bile duct, which
increases the number of micelles/vesicles present to solu-
bilize the lipophilic component. Second, the surface-active
digestion products from the lipid carrier material, such as
FFAs, MAGs, and DAGs, are incorporated into mixed mi-
celles thereby increasing their solubilization capacity.
Absorption: The relative bioavailability of highly lipophilic
compounds has been shown to increase appreciably when
the particle size is decreased below about 500 nm (Acosta,
2009). The precise physicochemical mechanism underlying
this process is currently unknown, but a number of mecha-
nisms have been proposed (Fig. 25):
(i) The water-solubility of highly lipophilic compounds in-
creases with decreasing particle size;
(ii) Small particles can diffuse into the mucous layer coating
the enterocytes, which increases their retention time and
their ability to interact with the enterocytes;
(iii) Very small particles may be able to cross the epithelia
by direct absorption into enterocytes through passive or
active transcellular routes or by passing between cells
by a paracellular route;
(iv) Some lipophilic components are crystalline at room
temperature and have to be dissolved in carrier oil be-
fore homogenization. When the carrier oil is digested
by lipase the lipophilic component may exceed its sol-
ubility limit in the oil phase and form crystals. The size
of the crystals formed will decrease as the size of the
particles within nanoemulsions decreases, which may
have important consequences for the absorption of the
lipophilic component.
Metabolism: If a nanoemulsion droplet is directly absorbed by
the human body (rather than being digested first) then the dis-
tribution, accumulation, metabolism, and excretion of any en-
capsulated components may be altered (Hagens et al., 2007;
Bouwmeester et al., 2009).
Potential Toxicity of Nanoemulsions
At present there is little experimental evidence on the poten-
tial toxicity of food-grade nanoemulsions. Nevertheless, there
are a number of physicochemical and physiological mechanisms
associated with the small particle size in nanoemulsions that
could potentially cause toxicity. Currently, there is no standard-
ized testing protocol specifically designed to assess the potential
toxicity of nanoemulsions intended for utilization within food
products (Maynard et al., 2006). An outline of a potential ap-
proach to evaluate the toxicity of nanoemulsions is highlighted
in Fig. 26.
Increased Bioavailability of Bioactive Components that are
Toxic at High Levels
As mentioned previously, the bioavailability of encapsulated
lipophilic components often increases when the size of the
323
droplets in emulsions falls below a critical level within the 100–
1000 nm range (Acosta, 2009). For many bioactive components,
an increase in bioavailability may be either desirable or may have
no adverse effects on human health. However, for certain bioac-
tive components there may be concerns about increasing their
bioavailability, particularly those that exhibit toxic effects when
consumed at high levels. If one of these bioactive components
normally has very low bioavailability but its absorption by the
human body is increased substantially by incorporation into a
nanoemulsion, then it could exhibit toxic effects that could not
be predicted from data obtained on the same material in macro-
scopic form. This is particularly true if the bioactive component
is incorporated into a product that is consumed regularly, such
as a soft drink or beverage emulsion.
Direct Absorption of Nanoemulsion Droplets
As mentioned above, there is evidence that non-digestible
nanoparticles, such as metals (silver and gold) and inorganic
materials (titanium dioxide and silicon oxide), can cross the
epithelial layer by various mechanisms, e.g., paracellular or
transcellular (passive or active) (De Jong et al., 2008). Once ab-
sorbed into an epithelium cell, a nanoparticle may be digested,
accumulate, or transported into the systemic circulation via the
blood or lymph systems (Hu et al., 2009). The nanoparticles
that are transported out of the epithelial cells may circulate
through the human body, where they may then be metabolized,
excreted, or accumulate within certain tissues (Bouwmeester
et al., 2009). These processes depend on the precise physic-
ochemical characteristics of the nanoparticles involved, e.g.,
composition, size, shape, charge, and interfacial chemistry. At
present, there is no evidence that edible nanoparticles fabri-
cated from components that are normally digested within the
GI tract are absorbed directly into the human body. Neverthe-
less, if the droplets in a nanoemulsion were prepared using an
indigestible oil (such as hydrocarbons or mineral oils) or if the
droplets were coated with an indigestible shell (such as dietary
fibers) then direct absorption could occur. Hence, further work
is needed to determine whether this mechanism is important in
humans.
Interference with Normal GI Function
Nanoparticles could alter the normal function of the GI tract
as they pass through the mouth, stomach, and small intestine
due to their small size, which could lead to some adverse effects
(Chaudhry et al., 2008). For example, it may be possible for very
small droplets to absorb directly through epithelium cells in the
mouth, esophagus, or stomach before being digested within the
small intestine. Very small droplets have a high specific surface
area and high curvature, and different surface reactivity than
bulk materials, which may alter the accumulation and activ-
ity of bile salts, lipase, and other digestive components at the
droplet surfaces, thereby altering the normal GI function. For
example, adsorption of proteins to particle surfaces, may lead
to denaturation and loss of normal function, which could have
324 D. J. McCLEMENTS AND J. RAO
Figure 26 Proposed protocol for testing the potential toxicity of food-grade nanoemulsions.
adverse effects on human health (Hoet et al., 2004). The particles
in nanoemulsions may be able to attach to cellular membrane
receptors, disrupting the cell’s normal metabolism and function.
In conclusion, the combination of small size, high-surface area,
and high-surface energy for the particles in nanoemulsions may
lead to effects in biological systems that are not predictable from
the bulk form of the same materials (Jiang et al., 2009).
Compositional Effects
Some of the components typically used to prepare nanoemul-
sions are toxic when consumed at sufficiently high levels, e.g.,
emulsifiers and solvents. A relatively large amount of emulsi-
fier is required to cover the large specific surface area associated
with the droplets in nanoemulsions. At present, the most widely
used emulsifiers for the preparation of nanoemulsions are small
molecule surfactants (and sometimes co-surfactants). This is
mainly because of the ability of surfactants to spontaneously
form nanoemulsions by low-energy methods (such as the PIT
method), and because of their ability to rapidly adsorb to droplet
surfaces and reduce the interfacial tension in high-energy meth-
ods (such as high-pressure homogenization). Natural emulsi-
fiers, such as proteins, polysaccharides, and phospholipids, tend
to be much less effective at forming nanoemulsions. Many small
molecule surfactants are known to exhibit toxic effects when
consumed at high levels (Kralova et al., 2009; He et al., 2010).
Consequently, the higher usage of surfactants in nanoemulsions
compared to conventional emulsions could lead to some adverse
health effects.
The preparation of nanoemulsions using some low-energy
methods (e.g., solvent displacement or evaporation methods)
involves the utilization of organic solvents, such as acetone,
hexane, or ethyl acetate (Horn and Rieger, 2001). These organic
solvents are usually removed by evaporation during the prepara-
tion of the nanoemulsion, but some residual solvent may remain
in the final product. It is therefore important to be aware of
the potential toxic effects associated with any residual organic
solvents if nanoemulsions are produced using this approach.
The potential toxicity of emulsifiers and solvents that are
suitable for utilization within foods, pharmaceuticals, and other
consumer products have been published by various organiza-
tions, including the World Health Organization (www.who.int),
the Food and Drug Administration (www.fda.gov) and the Eu-
ropean Food Safety Authority (www.efsa.europa.eu). These or-
ganizations carry out technical reviews of the toxicity data of
different additives and establish safe usage levels and analytical
protocols for testing toxicity.
CONCLUSIONS
Conventional oil-in-water emulsions are currently the
most widely used emulsion-based delivery systems in many
industrial applications. These emulsions contain droplets that
are relatively large (r > 100 nm), and so they are optically
opaque and susceptible to breakdown through gravitational
separation and droplet aggregation mechanisms. In contrast,
nanoemulsions contain relatively small droplets (r < 100 nm)
that do not scatter light strongly and that are highly stable to
gravitational separation and droplet aggregation. In addition,
the small size of the droplets in nanoemulsions means that they
are able to greatly increase the bioavailability of encapsulated
 FOOD-GRADE NANOEMULSIONS
lipophilic substances. Nanoemulsions may therefore have ap-
plications in the food, beverage, and pharmaceutical industries
as delivery systems, particularly in products that need to be
optically transparent, or when increased bioavailability of an
active component is important.
FUTURE TRENDS
Nanoemulsions are likely to become increasingly the focus of
research and development efforts because of their potential ad-
vantages over conventional emulsions for certain applications,
e.g., transparent foods and beverages, increased bioavailability,
and improved physical stability. Nevertheless, there are a num-
ber of challenges that need to be overcome before nanoemul-
sions are more widely used. First, suitable food-grade ingre-
dients must be identified for formulating food nanoemulsions.
Currently, many of the components used to fabricate nanoemul-
sions using both low-energy and high-energy approaches are
unsuitable for widespread utilization within the food industry,
e.g., synthetic surfactants, synthetic polymers, synthetic oils, or
organic solvents. Ideally, the food industry would like to prepare
nanoemulsions from commonly used and acceptable food grade
ingredients (such as flavor oils, triglyceride oils, proteins, and
polysaccharides). Second, suitable processing operations must
be identified to economically and robustly fabricate food-grade
nanoemulsions on an industrial scale. Many of the approaches
that have been developed within research laboratories may not
be suitable for scale-up to industrial production facilities. In ad-
dition, it would be advantageous to utilize existing equipment
and manufacturing lines, so as to reduce the costs of develop-
ing new manufacturing processes. Third, there may be some
safety concerns associated with the utilization of very small
lipid droplets in foods, e.g., they may change the extent or route
of absorption of lipophilic components. Thus, the bioavailabil-
ity or potential toxicity of a lipophilic component encapsulated
within nanometer-sized lipid droplets may be considerably dif-
ferent from when it is dispersed within a bulk lipid phase.
ACKNOWLEDGEMENTS
This material is based upon work supported by the Coop-
erative State Research, Extension, Education Service, United
States Department of Agriculture, Massachusetts Agricultural
Experiment Station, and United States Department of Agricul-
ture, NIFA Grant.
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