Songklanakarin J. Sci. Technol.

36 (6), 651-661, Nov. - Dec. 2014

http://www.sjst.psu.ac.th

Review Article

Factor affecting the properties of water-in-oil-in-water emulsions for

encapsulation of minerals and vitamins
Nattapong Prichapan and Utai Klinkesorn*

Department of Food Science and Technology, Faculty of Agro-Industry,

Kasetsart University, Chatuchak, Bangkok, 10900 Thailand.

Received: 15 November 2013; Accepted: 20 October 2014

Abstract

The direct fortification of minerals and vitamins into food may induce chemical degradation, change the level of
bioavailability or decrease the sensory quality of food products. The strategy to solve these problems is encapsulation
technology. Numerous investigations described the use of water-in-oil-in-water (W/O/W) emulsions as encapsulation system.
The properties and encapsulation efficiency of W/O/W emulsions are influenced by emulsion components, the emulsifica-
tion processes, and environmental conditions. The recently published results of research done on the factors affecting the
properties of W/O/W emulsions for encapsulation of minerals and vitamins including form and concentration of core
materials, concentration of inner water phase and lipophilic emulsifier, type and concentration of oil phase, type and concen-
tration of hydrophilic emulsifier and stabilizer and the pH of the outer water phase have been reviewed in this article.

Keywords: water-in-oil-in-water emulsions, encapsulation, minerals, vitamins

1. Introduction The direct fortification of minerals and vitamins into

food is not straightforward. Some micronutrients may cause
Minerals and vitamins are very important to sustain adverse effects to the sensory quality of the food such as
body functions and are essential for all parts of the body. color changes, undesirable flavor, and a sandy texture, while
Although minerals and vitamins are present in various foods some micronutrients may accelerate chemical reactions
from both animal and plant sources, many people still have leading to decreased nutritional quality of the food and may
mineral and vitamin deficiencies. This does not only occur also lead to toxicants. Furthermore, some micronutrients are
with poor people in developing countries who do not have very sensitive to high temperatures, exposure to light, changes
enough food to eat, but also with other people around the in pH, high levels of oxygen, and moisture content, which
world. These mineral and vitamin deficiencies may appear in can lead to degradation during preparation, processing, and
some groups of people such as the elderly, pregnant and storage (Ball, 2006; Mehansho et al., 2006; Jimenez-Alvarado
lactating women, some people with limited consumption et al., 2009; Liu, 2009; Lutz et al., 2009; Stevanovic and
choices, such as vegetarians or vegans and may also come Uskokovic, 2009; Guzun-Cojocaru et al., 2010; Lakkis et al.,
from changing food consumption behaviors to eating high 2011; Giroux et al., 2013).
processed food more than fresh whole foods (Bonnet et al., The strategy to solve these problems is encapsulation
2009; Guzun-Cojocaru et al., 2010; Kroner, 2011). technology, a technique that entraps sensitive material, e.g.
minerals and vitamins, in the carrier or coating material before
being added to food. Emulsion technology is one promising
technology for the encapsulation of such materials. The
* Corresponding author. advantage of an emulsion system is it allows nutrients to be
Email address: [email protected] quite stable and easily dispersed in the medium.
652 N. Prichapan & U. Klinkesorn / Songklanakarin J. Sci. Technol. 36 (6), 651-661, 2014

Usually, emulsion systems consist of two or more 2. Health Benefits, Deficiency and Fortification of Minerals
immiscible liquids, such as oil and water, where one of liquids and Vitamins
is dispersed as small spherical droplets in the other. Emulsion
systems are conventionally classified into two main groups; Minerals and vitamins have different health benefits
simple or single emulsions and multiple or double emulsions. and they are important for normal functions. Minerals and
Oil-in-water (O/W) emulsions consist of oil droplets dispersed vitamins are essential for all parts of the body. Deficiency of
in a continuous aqueous phase (Figure 1A), and water-in-oil minerals and vitamins may cause dysfunction and lead to
(W/O) emulsions that consists of water droplets dispersed in ill-health. Thus, we gathered the health benefit, deficiency
a continuous oil phase (Figure 1B) are examples of a single problem, and food source of some minerals and vitamins that
emulsion. Whereas, water-in-oil-in-water (W/O/W) emulsions have the important health effect on our body including iron,
containing W/O droplets dispersed in a continuous aqueous calcium, magnesium, ascorbic acid, thiamine, riboflavin, and
phase (Figure 1C), and oil-in-water-in-oil (O/W/O) emulsions cobalami as shown in Table 1.
where O/W droplets are dispersed in a continuous oil phase Fortification of minerals and vitamins in foods may
(Figure 1D) are classified as multiple emulsions. solve the problem of mineral and vitamin deficiency. However,
W/O emulsions and W/O/W emulsions are normally direct fortification of some minerals and vitamins into foods
used to encapsulate water-soluble substances. The W/O is not straightforward because it may not be stable or may
emulsions are suitable used for nutrient fortification in cause adverse effects on the food products (Table 2). In
oil-based foods, while W/O/W emulsions are properly used addition, bioavailability of the nutrients is also an important
in water-based food products. O/W emulsions and O/W/O point and should be considered. Bioavailability is the amount
emulsions are mostly used for encapsulation of oil-soluble of nutrients that can be absorbed and utilized by our body.
substances. The O/W emulsions encapsulate nutrients for There are three main factors that affect bioavailability,
fortification in water-based foods products, while O/W/O including personal factors, nutrient stability, and dissolution
emulsions are used for fortification in oil-based foods. or release of nutrients. The personal factors include age, sex
Furthermore, W/O/W emulsions and O/W/O emulsions can and physiologic state (e.g. pregnancy) which affect the
be used as a co-encapsulation system which is encapsula- gastric residence time and permeability through the gastro-
tion together of both water-soluble and oil-soluble intestinal tract. Formulation (form of nutrients, pH and
substances. For example, W/O/W can entrap water-soluble presence of other nutrients), food processing, and storage
nutrients in the inner water phase and also entrap oil-soluble conditions (temperature, light and oxygen) are factors that
substances in the oil phase of the same system (Benichou affect nutrient stability.
et al., 2002; Benichou et al., 2007; Bonnet et al., 2009; Lutz The factors that influence dissolution or release of
et al., 2009; Bonnet et al., 2010a; Bonnet et al., 2010b; nutrients are the encapsulation process and formulation such
O’Regan et al., 2010; Li et al., 2012; Giroux et al., 2013). as coatings or fillers excipient and surfactants. Thus, the
The aim of this work is to offer an overview of the design and utilizing the proper encapsulation technique may
health effects of some minerals and vitamins including iron, be a promising way to enhance the fortification and bioavail-
calcium, magnesium, vitamin C (ascorbic acid), vitamin B1 ability of nutrients in food products by protecting them until
(thiamine), vitamin B2 (riboflavin), and vitamin B12 (cobalamin). the time of consumption and delivery them to the target site
In addition, the use of W/O/W emulsions for encapsulation within the human body (Yetley, 2007; Fang and Bhandari,
and the factors affecting the properties of W/O/W emulsions 2010).
including form and concentration of core material, concen-
tration of inner water phase and lipophilic emulsifier, type 3. Encapsulation Technology
and concentration of oil phase, type and concentration of
hydrophilic emulsifier and stabilizer, and pH of outer water Generally, encapsulation technology is a technique
phase are also highlighted. that entraps sensitive materials, solid, liquid, or gas state, in
wall material in small sealed capsules to protect them from
adverse environmental conditions such as high temperatures,
high or low pH, exposure to UV light, and high levels of
dissolved oxygen. Moreover, the freeing of this substance
from the encapsulated capsule can be controlled to release
at certain rates and specific conditions. The size of the
encapsulated particle may cover a range from sub-micron to
millimeters. Encapsulation technology can be also used to
reduce the evaporation of aromatic compounds, mask un-
Figure 1. Schematic diagram of oil-in-water (O/W) emulsions (A), desirable flavors and tastes of some substances or improve
water-in-oil (W/O) emulsions (B), water-in-oil-in-water the dispersion ability of some substances by making them
(W/O/W) emulsions (C), and oil-in-water-in-oil (O/W/O) disperse more uniformly throughout the medium (Fang and
emulsions (D) (not to scale). Bhandari, 2010; Abbas et al., 2012).
Table 1. Summary of food sources, health benefit, and deficiency problem of some minerals and vitamins.

Nutrients Food sources Health benefit Deficiency problem References

Iron Meats, poultry, fish, shellfish, Essential for blood circulatory Anemia, high risk of heart attack, slow Kroner, 2011
and some plants system, transports oxygen development in children, and complications
throughout the body of pregnancy
Calcium Milk, cereals, and their products. Essential for bone and teeth, Calcium release from bones, high risk of Theobald et al., 2005;
muscle contraction, digestion, osteoporosis, and fracture of bones Saeidy et al., 2013
and neurotransmitter secretion
Magnesium Vegetables, fruits, and pulses Essential for protein synthesis, Cardiovascular diseases, hypertension, Bonnet et al., 2009; Kroner, 2011
enzymatic reactions, and muscular muscular weakness, and diarrhea
contraction
Ascorbic acid Fruits and vegetables such as Degenerative chronic diseases, Dry and splitting hair, rough, dry and Liu, 2009; Lutz et al., 2009;
(vitamin C) citrus fruits, guava, potatoes, involved in synthesis of collagen hemorrhage skin, decreased wound Stevanovic and Uskokovic, 2009;
and tomatoes and epinephrine, amino acid and healing rate, weakened tooth enamel, Kroner, 2011
cholesterol metabolism, against scurvy, gingivitis, anemia, and high risk of
oxidative damage and cancers infections
Thiamine Meat, whole grain cereals, nuts, Role in cellular energy production, Symptom to two systems including Bates, 2007; Osiezagha et al., 2013
(vitamin B1) and beans and helps neuron to be normal cardiovascular system and nervous system
actives
Riboflavin Dairy products, cereals, offal, Protecting against cancers, Night blindness, high risk of cancer & Powers, 2003
(vitamin B2) fish, and dark-green vegetables cardiovascular diseases, and plays cardiovascular disease, peripheral
a role in thyroxine metabolism neuropathy, neurodegeneration, and
mental illness
Cobalamin Found in animal tissues Essential for production of blood Anemia, irritability, memory loss, weakness, Kroner, 2011
(vitamin B12) cells and DNA, and necessary for trembling and unstable movements, and
N. Prichapan & U. Klinkesorn / Songklanakarin J. Sci. Technol. 36 (6), 651-661, 2014

normal neurologic function psychosis

653
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Table 2. Summary of fortification form, recommended daily dosages and direct fortification problem of some minerals and vitamins.

Nutrients Form for fortification Recommended daily dosages * Direct fortification problem
Iron Ferrous sulphate, ferrous gluconate, ferric 1-3 years 7 mg, 4-8 years 10 mg, 9-13 years 8 mg, Sensitive to reaction, color change, lipid
ammonium citrate, sodium iron EDTA, girls 14-18 years 15 mg, boys 14-18 years 11 mg, oxidation, precipitation (Guzun-Cojocaru
ferrous bisglycinate, ferrous fumarate, women 19-50 years 18 mg. Pregnancy 27 mg, et al., 2010; Zimmermann and Windhab,
ferric saccharate, ferrous citrate, ferric lactation 9 mg, after 51 years 8 mg 2010)
citrate, carbonyl iron, and ferric
pyrophosphate (Lynch, 2005; Kroner, 2011)
Calcium Calcium carbonate, calcium triphosphate,
0-6 months 210 mg, 6-12 months 270 mg, 1-3 years Flocculation, sandy texture, precipitation
and calcium malate (Theobald, 2005; 500 mg, 4-8 years 800 mg, 9-18 years 1,300 mg, (Lakkis et al., 2011)
Kressel et al., 2010; Kroner, 2011) 19-50 years 1,000 mg, 50 years and older 1,200 mg,
pregnancy and lactation: 18 years or younger
1,300 mg, 19-50 years 1,000 mg
Magnesium Magnesium phosphate, magnesium 0-6 months 30 mg, 7-12 months 75 mg, 1-3 years Protein aggregation, nutrients degrada-
pyrophosphate, and magnesium potassium 80 mg, 4-8 years 130 mg, 9-13 years 240 mg, men: tion, unfavorable taste (Bonnet et al.,
phosphate (Skelcey et al., 1974) 14-18 years 410 mg, 19-30 years 400 mg, more than 2009)
30 years 420 mg, women: 14-18 years 360 mg,
19-30 years 310 mg, over 30 years 320 mg,
pregnancy: 14-18 years 400 mg, 19 to 30 years
350 mg, over 31 years 360 mg, lactation:14-18 years
360 mg, 19 to 30 years 310 mg, over 31 years 320 mg
Ascorbic acid (vitamin C) Ascorbic acid, ascorbate salt, and Men 90 mg and women 75 mg Very unstable to high pH, high tempera-
esterified ascorbate tures, UV light, dissolved oxygen (Ball,
(Liu, 2009; Kroner, 2011) 2006)
Thiamine (vitamin B1) Thiamine hydrochloride 9-13 years 0.9 mg, boys 14-18 years 1.2 mg, girls Unstable to high pH, high temperatures,
(Benichou et al., 2002) 14-18 years 1.0 mg, men 1.2 mg, women 1.1 mg, high moisture content (Ball et al., 2006)
pregnancy & lactation 1.4 mg
Riboflavin (vitamin B2) Riboflavin, and riboflavine- 1-3 years 0.5 mg, 4-8 years 0.6 mg, 9-13 years 0.9 Unstable to light and high pH (Ball et al.,
5'-monophosphate (Owusu et al., 1992) mg, boys 14-18 years 1.3 mg, girls 14-18 years 1.0 2006)
mg, men 1.3 mg, women 1.1 mg, pregnancy 1.4 mg
Cobalamin (vitamin B12) Cobalamin and adenosylcobalamin 1-3 years 0.9 µg, 4-8 years 1.2 µg, 9-13 years 1.8 µg, Unstable to High pH (Ball et al., 2006)
N. Prichapan & U. Klinkesorn / Songklanakarin J. Sci. Technol. 36 (6), 651-661, 2014

(Collins, 2003) 14-18 years 2.4 µg, men & women 2.4 µg, pregnancy
2.6 µg, lactation 2.8 µg

*(Standing Committee on the Scientific Evaluation of Dietary Reference Intakes and its Panel on Folate, Other B Vitamins, and Choline and Subcommittee on Upper
Reference Levels of Nutrients Food and Nutrition Board Institute of Medicine, 1998; Kroner, 2011; Osiezagha et al., 2013)
 N. Prichapan & U. Klinkesorn / Songklanakarin J. Sci. Technol. 36 (6), 651-661, 2014 655

Many methods and techniques can be applied for example, Li et al. (2012) produced W/O/W emulsions for use
encapsulation of nutrients including spray drying, fluid bed as an encapsulation system consisting of riboflavin in the
coating, spray chilling or cooling, melt extrusion, coacerva- inner water phase and -tocopherol in the oil phase.
tion, liposome, inclusion encapsulation, cocrystallization,
nanoencapsulation, freeze drying, yeast encapsulation and 4.1 Preparation of W/O/W emulsions
emulsions (Stevanovic and Uskokovic, 2009; Fang and
Bhandari, 2010; Zuidam and Shimoni, 2010; Abbas et al., W/O/W emulsions are usually produced using a two-
2012). step emulsification method (Figure 2). The first step is the
An emulsion encapsulation technique can be used to production of W/O emulsions by homogenizing an oil phase
encapsulate both water-soluble and oil-soluble liquid and the inner water phase together in the presence of a water-
substances. The size of the liquid droplets in emulsion is soluble core material at high pressure or high shear rate. In
usually in the range from 0.1-5,000 µm. Preparation of multi- this step, a lipophilic emulsifier or oil-soluble emulsifier was
layer around the droplets in emulsions may obtain a more used to stabilize the water-oil interface. The second step is
stable and effective encapsulated core. The liquid droplets the production of W/O/W emulsions by homogenizing the
in emulsion, especially oil-soluble substances, can be former W/O emulsion and an outer water phase at a pressure
produced in a dry powder form by spraying or freeze drying or shear rate lower than that used in the first stage to avoid
their emulsion. The advantage of this technique is that it is rupture or expulsion of inner droplets. To stabilize the oil-
flexible because it can be used to encapsulate both hydro- water interface, a hydrophilic emulsifier or water-soluble
philic and lipophilic substances (Fang and Bhandari, 2010; emulsifier was used in this step (Van der Graaf et al. 2005;
Zuidam and Shimoni, 2010; Abbas et al., 2012). McClements, 2012).
Abbas et al. (2012) mentioned that there are many
methods that can be applied for encapsulation, but there are 4.2 Factors affecting the properties of W/O/W emulsions
no techniques that are effective for all food systems. Each for encapsulation
technique has different advantages and limitations. However,
W/O/W emulsions are the one encapsulation technique that There are many factors that can affect the properties
is suitable to fortify nutrients in beverages and water-based of W/O/W emulsions. In this review, we categorize them into
foods because this technique can keep the nutrients to be five groups: form and concentration of core material, con-
quite stable and easily dispersed in an aqueous phase. centration of inner water phase and lipophilic emulsifier, type
Moreover, it is easily to be scaled up to industrial size and concentration of oil phase, type and concentration of
(Krasaekoopt et al., 2003). hydrophilic emulsifier and stabilizer and pH of outer water
phase.
4. Encapsulation of Minerals and Vitamins in Water-in-oil-
In-water Emulsions 4.2.1 Form and concentration of core material

The water-in-oil-in-water or W/O/W emulsion system The form and concentration of the core material affects
(Figure 1C) consists of small internal water droplets the properties of W/O/W emulsions. For the form of the core
embedded in larger oil droplets that are also dispersed in material, different forms of core material lead to differences
another outer water continuous phase. This can be referred in properties such as size, binding capacity, and hydrophilic
to as emulsions of emulsions that is to say water-in-oil-in- property that all affect W/O/W emulsions characteristics.
water emulsions consisting of water-in-oil emulsions (Figure If the core material has the form of a bigger molecule, the
1B) dispersed in a continuous water phase. The W/O/W
emulsions have two types of interfaces; these are inner
water-oil interface and oil-outer water interface, so it has to
use oil-soluble emulsifiers to stabilize the inner water-oil
interface and water-soluble emulsifiers to stabilize the oil-
outer water interface (Benichou et al., 2002; Dickinson, 2011;
McClements, 2012).
For fortification of minerals and vitamins into food
products that have water as a continuous phase, W/O/W
emulsions are mostly used with different components as
summarized in Table 3. Moreover, W/O/W emulsions may
also be used for co-encapsulation, which is encapsulation
for both water-soluble substances and oil-soluble substances
together by an encapsulating water-soluble substance in the Figure 2. Schematic diagram of two-step procedure for preparation
inner water phase and encapsulating an oil-soluble substance of water-in-oil-in-water (W/O/W) emulsions (not to
in the oil phase of the same W/O/W emulsion system. For scale).
 656

Table 3. Examples of minerals and vitamins encapsulated in water-in-oil-in-water emulsions.

Nutrients Hydrophilic emulsifier and stabilizer Oil phase;lipophilic emulsifier Hydrophilic emulsifier and References
in inner water phase stabilizer in outer water phase
Iron - Whey protein isolate (WPI) - Corn oil; polyglycerol polyricinoleate - Polyoxyethylene Sorbitan Choi et al., 2009
- Panodan SDK (esters of ester (PGPR) monolaurate (Tween20)
monoglycerides and diglycerides - Mineral oil; PGPR - Whey protein concentrate
of diacetyl tartaric acid) (WPC) and gum arabic or
mesquite gum or low methoxyl
pectin (LMP) Jimenez-Alvarado et al., 2009
Calcium - - Sunflower oil; PGPR - Xanthan gum Marquez and Wagner, 2010
- - Canola oil; glycerol monostearate - Gelatin and agar Saeidy et al., 2013
(GMS)
Magnesium - - Olein, miglyol, olive oil or rapeseed oil; - Sodium caseinate Bonnet et al., 2009
PGPR
- - Olive oil; PGPR - Sodium caseinate Bonnet et al., 2010a
Ascorbic acid - - Medium chain triglyceride or limonene; - WPI and modified pectin Lutz et al., 2009
(vitamin C) - Panodan SDK andgellan gum PGPR - Mesquite gum, maltodextrin Carrillo-Navas et al., 2012
- Chia essential oil; PGPR and WPC
Thiamine - - Medium chain triglyceride; PGPR and - WPI and xanthan gum Benichou et al., 2002
(vitamin B1) monoglyceride oleate
Riboflavin - - Soy oil; PGPR - WPI and LMP or ê-carrageenan Li et al., 2012
(vitamin B2)
Cobalamin - Gelatin - Medium chain triglyceride; PGPR - Sodium caseinate or sodium O’Regan and Mulvihill, 2010
(vitamin B12) caseinate-maltodextrin conjugate
- - Butter oil; PGPR - Skim milk or sodium caseinate Giroux et al., 2013
N. Prichapan & U. Klinkesorn / Songklanakarin J. Sci. Technol. 36 (6), 651-661, 2014
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encapsulation efficiency will be higher than for smaller forms. but after 28 days there was no significant difference. The
If the core material attaches to the molecule that has a higher difference in release rate of fresh emulsions containing differ-
binding capacity, the encapsulation efficiency will be higher, ent core concentrations was not discussed by the authors.
because it will occur with a lower free form of the core However, in our opinion, this may due to the effect of the
material, so it is harder to release. Moreover, the higher hydro- osmotic gradient. The higher core material concentration led
philic property of the core material also leads to lower release, to the higher osmotic gradient, resulting in emulsion droplet
because higher hydrophilicity molecules are harder to diffuse swelling. This occurs from diffusion of the water phase from
through the oil phase. the outer water phase to the inner water phase to balance the
Marquez and Wagner (2010) encapsulated different osmotic gradient (Dickinson, 2011). The intensity of water
calcium salts in the inner water phase of W/O/W emulsions diffused from the outer phase to the inner phase was higher
using sunflower oil as the oil phase, PGPR as a lipophilic than the diffusion of water from the inner phase to the outer
emulsifier, soybean milk as the outer water phase, and phase resulting in a low release of sodium ascorbate in fresh
xanthan gum as the stabilizer. They found that emulsions emulsions. However, for a longer time storage, the diffusion of
that used calcium lactate as the core material had lower G water from the outer phase to the inner phase may cause some
and G and higher tan  than emulsions that used calcium droplets broken and release core material resulting in a high
chloride as the core material. The lower G and G and the release of sodium ascorbate from emulsion droplets after 28
higher tan  values mean that the W/O/W emulsions have a days.
weaker structure. This may due to the calcium lactate, which
is a bigger molecule, and has a lower release value than 4.2.2 Concentration of inner water phase and lipophilic
calcium chloride, which occurs with a lower crosslink to the emulsifier
soybean proteins and therefore has a weaker structure.
In the same way, Bonnet et al. (2010b) encapsulated The concentration of the inner water phase and lipo-
different magnesium salts in the inner aqueous phase of philic emulsifier affects the properties of W/O/W emulsions.
W/O/W emulsions using olive oil as the oil phase, PGPR as Higher concentrations of the inner water phase lead to lower
lipophilic emulsifier, and sodium caseinate as the hydrophilic release rates of the core material. In the same way, higher
emulsifier. They found that the encapsulated MgCl2 had the concentrations of lipophilic emulsifier lead to lower release
highest release rate and completed release in a 15 day- rates of the core material. An example for the effect of
storage, since small molecules of magnesium can easily dispersed aqueous phase content was shown by Marquez
transfer through the oil phase. The encapsulated gluconic and Wagner (2010). They found that higher dispersed
acid hemimagnesium salt was found to have a lower release aqueous phase content led to the formation of lower G and
value than MgCl2, but a higher release value than phosvitin G and higher tan  value emulsions. The lower G and G
with MgCl2. This may due to the fact that gluconic acid has a and the higher tan  meant that the W/O/W emulsions were
lower binding capacity than phosvitin and is quite a small thinner. This can explain why higher dispersed aqueous
molecule, so it can easily pass through the oil phase. Whereas, phase content reduced the calcium concentration leading to
encapsulated phosvitin with MgCl2 was found to have the reduced osmotic gradient and calcium release, so there was
lowest release value because the hydrophilicity and high less crosslink with soybean proteins producing thinner
molecular weight of phosvitin cause it to be unable to pass emulsions. For the effect of lipophilic emulsifier concentra-
though the oil phase. tion, Marquez and Wagner (2010) found that higher PGPR
For the effect of concentration of core material on concentrations produced lower G and G and higher tan 
encapsulation efficiency of W/O/W emulsions, there was value emulsions. This may be due to increases in the PGPR
report that higher concentrations of the core material can lead concentration which reduced the water droplet size of the
to higher encapsulation efficiency. For example, Benichou W/O droplets, so reduced calcium release led to less
et al. (2002) entrapped thiamine hydrochloride in the W/O/W crosslink and less elastic behavior of the emulsion.
emulsions that used medium chain fatty acid triglycerides as
the oil phase, PGPR as the surfactant, monoglyceride oleate 4.2.3 Type and concentration of oil phase
for droplet size reduction and better stability, and whey
protein isolate and xanthan gum as hydrophilic emulsifiers. The type and concentration of the oil phase affects
They found that increasing vitamin B1 concentrations caused the properties of W/O/W emulsions. Different types of oil
a decrease in vitamin B1 release. This may due to the fact that phase lead to variations in properties such as viscosity, and
thiamine hydrochloride has an amphiphilic structure, so it saturated fatty acid content affecting the encapsulation
has surface properties to reduce interfacial tension. efficiency of W/O/W emulsions. An oil phase with higher
In a similar way, Lutz et al. (2009) encapsulated viscosity values lead to higher encapsulation efficiency
sodium ascorbate in the W/O/W emulsions using PGPR as because the viscous oil phase retards diffusion and expulsion
emulsifier, and WPI and modified pectin as a hydrophilic of the inner water phase with the core material inside. More-
emulsifier. They found that fresh emulsions had a reduced over, oil types with higher saturated fatty acid levels yield
release rate with increasing sodium ascorbate concentration, higher hydrophobicity or lower compatibility with water
658 N. Prichapan & U. Klinkesorn / Songklanakarin J. Sci. Technol. 36 (6), 651-661, 2014

causing a faster expulsion of the inner water phase that has caseinate-maltodextrin conjugate was significantly higher
the core material inside. than that of sodium caseinate, especially in the case of malto-
For example, Bonnet et al. (2009) encapsulated MgCl2 dextrin with a dextrose equivalent (DE) of about 10. This was
in W/O/W emulsions using olein, miglyol, olive oil or because the protein-polysaccharide conjugate formed a more
rapeseed oil as the oil phase, PGPR as a lipophilic emulsifier, bulky polymeric layer at the interface and some parts of the
sodium caseinate as a hydrophilic emulsifier, and lactose to polysaccharide protruding toward the continuous phase
match the osmotic pressure between the inner and outer caused better steric stabilization against droplet flocculation
water phase to avoid water transfer. They found that double and coalescence.
emulsions produced with miglyol had significantly faster The greater molecular weight of polysaccharides lead
magnesium release than other oils. This can be explained to more effective protection of the core material than lower
since miglyol has the lowest viscosity and the highest satu- ones, for example, Jimenez-Alvarado et al. (2009) entrapped
rated fatty acid of these four oils, so the lower viscosity leads ferrous bisglycinate in the W/O/W emulsions using mineral
to easier material transfer and release, and higher saturated oil as the oil phase, and protein-polysaccharide complexes
fatty acid leads to higher hydrophobicity and lower com- (whey protein concentrate (WPC) with gum arabic (GA) or
patibility with the water causing faster magnesium release. mesquite gum (MG) or low methoxyl pectin (LMP)) as water-
In contrast, double emulsion produced with rapeseed oil, soluble surfactants in outer water phase. They found that the
which has a quite high viscosity and the lowest saturated encapsulation yield and ferrous ion content of W/O/W
fatty acid content, had the highest magnesium retention. emulsions stabilized with 5 wt% total concentration of WPC:
In the same way, Lutz et al. (2009) found that double MG and 0.7 wt% total concentration of WPC:LMP were
emulsions produced from medium chain triglycerides had a significantly highest at the initial time. This may be due to
lower sodium ascorbate release rate than produced from R(+)- formation of a thick layer at the interface of the droplet, but
limonene. This can be explained since R(+)-limonene has a only ferrous ion content of water-in-oil-in-water emulsion
lower viscosity which leads to easier release of the sodium stabilized with 5 wt% total concentration of WPC:MG were
ascorbate from the inner water phase to the outer water significantly higher at the end of storage time which may be
phase. due to the molecular weight of the polysaccharides. Mesquite
For the effect of lipid phase content on property of gum has a greater molecular weight than gum Arabic and low
W/O/W emulsions, there was a report that higher lipid phase methoxyl pectin, respectively. Since mesquite gum has a high
content lead to get more viscous W/O/W emulsions. Marquez molecular weight and can form a thick layer at the interface,
and Wagner (2010) showed that increased lipid phase content therefore it was the most effective polysaccharide to protect
led to the formation of higher G and G and lower tan  against ferrous bisglycinate oxidation in between these
value emulsions. The higher G and G and the lower tan  polysaccharides.
values mean that the W/O/W emulsions were more viscous. The higher charge of polysaccharides has both posi-
This can be explained in a way that the higher lipid phase tive and negative effects on the properties of W/O/W emul-
content produced in the more water-in-oil droplets led to sions. For the positive effect, Lutz et al. (2009) encapsulated
tighter packing and a viscous systems. sodium ascorbate in W/O/W emulsions that used different
modified pectin types U63 and C63, the modified pectin types
4.2.4 Type and concentration of hydrophilic emulsifier and U63 and C63 are 63% degree of esterification, but U63 has
stabilizer a more negative zeta potential and more intermolecular
interactions than C63. They found that double emulsion
The type and concentration of the hydrophilic stabilized with the WPI/U63 complex had lower sodium
emulsifiers and stabilizers affect the properties of W/O/W ascorbate release rates than found in a double emulsion stabi-
emulsions. Some hydrophilic emulsifiers and stabilizers have lized with the WPI/C63 complex. They explained that U63
a synergistic effect on the stabilization of emulsion when pectin have more charges which leads to more interaction
used together. Different types of hydrophilic emulsifiers and and causes more elastic and a greater stiffness complex with
stabilizers, which have different properties such as molecular WPI. Thus, the WPI/U63 complex was more effective to
weight and charge, result in different effects on the properties prevent the release of sodium ascorbate than the WPI/C63
of the W/O/W emulsions. Using two hydrophilic emulsifiers complex.
together or hydrophilic emulsifiers with stabilizers may In contrast, Li et al. (2012) encapsulated riboflavin in
increase the encapsulation efficiency more than using either the W/O/W emulsions with hydrophilic emulsifiers and stabi-
alone. For example, O’Regan and Mulvihill (2010) encapsu- lizers of whey protein isolate and low methoxyl pectin (LMP)
lated vitamin B12 in W/O/W emulsions with gelatin to solidify or -carrageenan (KCG), which has a higher minus charge
the inner water phase with medium chain triglyceride oil as than low methoxyl pectin. They found that the encapsulation
the oil phase, PGPR as the lipophilic emulsifier, sodium efficiency of the WPI-LMP complex stabilized emulsion was
caseinate or sodium caseinate-maltodextrin conjugate as the higher than that of the WPI-KCG complex when the WPI: PS
hydrophilic emulsifier. They found that the encapsulation ratio was 5:0.5. They explained that the low charge poly-
efficiency of double emulsion stabilized with sodium saccharide chain has a flexible worm-like form that easily to
 N. Prichapan & U. Klinkesorn / Songklanakarin J. Sci. Technol. 36 (6), 651-661, 2014 659

curvature and interact with protein molecule, while the high

charge polysaccharide chain has a rigid stick-like form, so
low methoxyl pectin, which has lower charge, has a more
flexible chain, and can more easier bind with proteins to create
a dense and seal interface leading to more encapsulation
efficiency than that of the WPI-KCG complex.
For the effect of hydrophilic emulsifier or stabilizer
concentration, the higher hydrophilic emulsifier or stabilizer Figure 3. Schematic diagram of emulsion droplet stabilized by
concentration should have higher encapsulation efficiency protein and anionic polysaccharide at pH << pI (A), and
because increased hydrophilic emulsifier levels lead to pH >> pI (B) (not to scale).
stronger interfacial films against core material release. For
example, Li et al. (2012) found that increasing the polysac- lower vitamin B1 release, but at a pH of 3.5 vitamin B1 was
charide concentration caused the encapsulation efficiency to released less than at pH 2 because at pH values more than
increase. In the same way, Benichou et al. (2002) found that about 4.5 (above the isoelectric point of the whey protein
increases in the xanthan gum ratio reduced the vitamin B1 isolate) this caused the whey protein isolate to have a minus
release, because more xanthan gum content led to the pro- zeta potential as xanthan gum, so the whey protein isolate
duction of a more synergistic and rigid film around the and xanthan gum did not interact (Figure 3B) which led to a
droplet. less sealed interface and higher vitamin B1 release. On the
However, increasing the concentration of some hydro- other hand, at pH 2 xanthan gum had less minus zeta potential
philic emulsifiers can lead to decreases in encapsulation and a reduced different of zeta potential between whey
efficiency. For example, Bonnet et al. (2010a) encapsulated protein isolate and xanthan gum, which led to an interaction
magnesium in the W/O/W emulsions by using olive oil as an between them being less than at pH 3.5. This had a greater
oil phase, PGPR as a lipophilic emulsifier, and sodium difference in zeta potential between the whey protein isolate
caseinate as a hydrophilic emulsifier. Surprisingly, the higher and xanthan gum, so the complex interface at pH 3.5 was
sodium caseinate concentrations led to higher magnesium more sealed and had lower vitamin B1 release.
release. This may due to increased levels of sodium caseinate
meaning more sodium ions leading to more osmotic pressure 5. Conclusions
in the external water phase. When the external osmotic
pressure is higher than the internal osmotic pressure, the W/O/W emulsions can be used as an entrapping and
water may migrate from the internal to the external water delivering system for minerals and vitamins. However, there
phase. are many factors that affect the stability and efficiency of
these emulsion systems that have to be considered such as
4.2.5 pH of outer water phase volume and properties of oil and inner or outer water phase,
concentration and form of core material, type and concentra-
When two ionic hydrophilic emulsifiers or stabilizers tion of emulsifiers and stabilizers, and also process condi-
are used together, the pH of outer water phase will affect the tions. For encapsulation of minerals and vitamins, W/O/W
encapsulation efficiency of W/O/W emulsions. The pH that emulsions should have both high encapsulation efficiency
should be used is one that allows two ionic hydrophilic and physicochemical stability, including being stable to
emulsifiers or stabilizers to have different charges in order to environmental stress and chemical reactions. Moreover, it
let them exhibit electrostatic attraction. For example, when should still be stable when it is applied to food products and
protein and anionic polysaccharide are used as a hydrophilic should not decrease the quality of those products.
emulsifier and stabilizer, respectively, the pH of the system
should be lower than pI (isoelectric point) of protein in order Acknowledgements
to obtain the cationic structure of protein that can exhibit
electrostatic attraction with an anionic polysaccharide The authors would like to thank the Thailand Research
structure (Figure 3A). The larger difference in charge inten- Fund under The Royal Golden Jubilee Ph.D. Program (PHD/
sity between the ionic hydrophilic emulsifiers or stabilizers 0001/2555: 2.F.KU/55/I.1.R.03) for their financial support.
lead to increased electrostatic attraction and provides a Special thanks to S. Watmough for his assistance in writing
thicker and denser interfacial layer. the manuscript.
Benichou et al. (2007) entrapped thiamine hydro-
chloride in W/O/W emulsions by using medium chain fatty References
acid triglyceride as the oil phase, PGPR as the lipophilic
emulsifier, monoglyceride oleate and glycerol for droplet size Abbas, S., Wei, C.D., Hayat, K. and Xiaoming, Z. 2012.
reduction and better stability. Whey protein isolate and Ascorbic acid: microencapsulation techniques and
xanthan gum was used as hydrophilic emulsifier and stabili- trends—a review. Food Reviews International. 28/4,
zer, respectively. They found that lower pH levels led to 343-374.
660 N. Prichapan & U. Klinkesorn / Songklanakarin J. Sci. Technol. 36 (6), 651-661, 2014

Ball, G.F.M. 2006. Vitamins in foods: analysis, bioavailability, Krasaekoopt, W., Bhandari, B. and Deeth, H. 2003. Evalua-
and stability, CRC Press, United States. tion of encapsulation techniques of probiotics for
Bates, C. J. 2007. Thiamine. In Handbook of vitamins, J. yoghurt. International Dairy Journal. 13, 3-13.
Zempleni, R. B. Rucker, D. B. McCormick and J. W. Kressel, G., Wolters, M. and Hahn, A. 2010. Bioavailability
Suttie, editors. CRC Press, United States, pp. 253-287. and Solubility of Different Calcium-Salts as a Basis
Benichou, A., Aserin, A. and Garti, N. 2002. Double emulsions for Calcium Enrichment of Beverages. Food and
stabilized by new molecular recognition hybrids of Nutrition Sciences. 1, 53-58.
natural polymers. Polymer for Advanced Technologies. Kroner, Z. 2011. Vitamins and Minerals, Greenwood, United
13, 1019-1031. States.
Benichou, A., Aserin, A. and Garti, N. 2007. W/O/W double Lakkis, J.M., Anton, G. and Maria, J. 2011. Microcapsules
emulsions stabilized with WPI-polysaccharide containing salts for food products, Spain, EP2292102
complexes. Colloids and Surfaces A: Physicochemical A1.
and Engineering Aspects. 294, 20-32. Li, B., Jiang, Y., Liu, F., Chai, Z., Li, Y., Li, Y. and Leng, X.
Berger, N., Sachse, A., Bender, J., Schubert, R. and Brandl, 2012. Synergistic effects of whey protein–polysaccha-
M. 2001. Filter extrusion of liposomes using different ride complexes on the controlled release of lipid-
devices: comparison of liposome size, encapsulation soluble and water-soluble vitamins in W1/O/W2 double
efficiency, and process characteristics. International emulsion systems. International Journal of Food
Journal of Pharmaceutics. 223, 55-68. Science and Technology. 47, 248-254.
Bonnet, M., Cansell, M., Berkaoui, A., Ropers, M.H., Anton, Liu, C. 2009. Vitamin C: Dietary requirements, dietary sources,
M. and Leal-Calderon, F. 2009. Release rate profiles and adverse effects. In Handbook of Vitamin C
of magnesium from multiple W/O/W emulsions. Food Research, H. Kucharski and J. Zajac, editors. Nova
Hydrocolloids. 23, 92-101. Science Publishers, United States, pp. 127-152.
Bonnet, M., Cansell, M., Placin, F., Anton, M. and Leal- Lutz, R., Aserin, A., Wicker, L. and Gerti, N. 2009. Release of
Calderon, F. 2010a. Impact of sodium caseinate con- electrolytes from W/O/W double emulsions stabilized
centration and location on magnesium release from by a soluble complex of modified pectin and whey
multiple W/O/W emulsions. Langmuir. 26, 9250-9260. protein isolate. Colloids and Surfaces B: Biointerfaces.
Bonnet, M., Cansell, M., Placin, F., David-Briand, E., Anton, 74, 178-185.
M. and Leal-Calderon, F. 2010b. Influence of ionic Lynch, S. R. 2005. The impact of iron fortification on nutri-
complexation on release rate profiles from multiple tional anaemia. Best Practice & Research Clinical
Water-in-Oil-in-Water (W/O/W) emulsions. Journal of Haematology. 18/2, 333-346.
Agricultural and Food Chemistry. 58, 7762-7769. Marquez, A.L. and Wagner, J.R. 2010. Rheology of double
Collins, D.A. 2003. Adenosyl-cobalamin fortified composi- (w/o/w) emulsion prepared with soybean milk and
tions. United States. US 2003/0018009 A1. fortified with calcium. Journal of Texture Studies. 41,
Dickinson, E. 2011. Double Emulsions Stabilized by Food 651-671.
Biopolymers. Food Biophysics. 6, 1-11. McClements, D.J. 2012. Advances in fabrication of emulsions
Fang, Z. and Bhandari, B. 2010. Encapsulation of polyphenol. with enhanced functionality using structural design
Trends in Food Science & Technology. 21, 510-523. principles. Current Opinion in Colloid & Interface
Giroux, H.J., Constantineau, S., Fustier, P., Champagne, C.P., Science. 17, 235-245.
St-Gelais, D., Lacroix, M. and Britten, M. 2013. Cheese Mehansho, H. 2006. Iron fortification technology develop-
fortification using water-in-oil-in-water double emul- ment: new approaches. The Journal of Nutrition. 1059-
sions as carrier for water soluble nutrients. Inter- 1063.
national Dairy Journal. 29, 107-114. O’Regan, J. and Mulvihill, D. M. 2010. Sodium caseinate–
Guzun-Cojocaru, T., Cayot, P., Loupiac, C. and Cases, E. maltodextrin conjugate stabilized double emulsions:
2010. Effect of iron chelates on oil–water interface, Encapsulation and stability. Food Research Inter-
stabilized by milk proteins: The role of phosphate national. 43, 224-231.
groups and pH. Prediction of iron transfer from Osiezagha, K., Ali, S., Freeman, C., Barker, N.C., Jabeen, S.,
aqueous phase toward fat globule surface by changes Maitra, S., Olagbemiro, Y., Richie, W. and Bailey, R.K.
of interfacial properties. Food Hydrocolloids. 24, 364- 2013. thiamine deficiency and delirium. Innovations
373. in Neuroscience. 10/4, 26-32.
Jimenez-Alvarado, R., Beristain, C.I., Medina-Torres, L., Owusu, R.K., Zhu, Q. and Dickinson, E. 1992. Controlled
Roma´n-Guerrero, A. and Vernon-Carter, E.J. 2009. release of L-tryptophan and vitamin B2 from model
Ferrous bisglycinate content and release in W1/O/W2 water/oil/water multiple emulsions. Food Hydro-
multiple emulsions stabilized by protein–polysaccha- colloids. 6/5, 443-453.
ride complexes. Food Hydrocolloids. 23, 2425-2433. Powers, H. J. 2003. Riboflavin (vitamin B-2) and health. The
American Journal of Clinical Nutrition. 77, 1352-1360.
 N. Prichapan & U. Klinkesorn / Songklanakarin J. Sci. Technol. 36 (6), 651-661, 2014 661

Saeidy, S., Keramat, J. and Nasirpour, A. 2013. Microencapsu- Van der Graaf, S., Schroen, C.G.P.H. and Boom, R.M. 2005.
lation of calcium using water in oil in water double Preparation of double emulsions by membrane emul-
emulsion method. Journal of Dispersion Science and sification—a review. Journal of Membrane Science.
Technology, DOI:10.1080/01932691.2013.788453. 251, 7-15.
Skelcey, J.S., Richards, D.W. and Hart, A.M. 1974. Cereal Xia, S., Xu, S. and Zhang, X. 2006. Optimization in the
products containing magnesium compounds as nutri- preparation of coenzyme Q10 nanoliposomes. Journal
tional supplements. United States. 3, 852,497. of Agricultural and Food Chemistry. 54, 6358-6366.
Standing Committee on the Scientific Evaluation of Dietary Yetley, E.A. 2007. Multivitamin and multimineral dietary
Reference Intakes and its Panel on Folate, Other B supplements: definitions, characterization, bioavail-
Vitamins, and Choline and Subcommittee on Upper ability, and drug interactions. The American Journal
Reference Levels of Nutrients Food and Nutrition of Clinical Nutrition. 85, 269S-276S.
Board Institute of Medicine. 1998. Dietary Reference Zimmermann, M.B. and Windhab, E.J. 2010. Encapsulation
Intakes for Thiamin, Riboflavin, Niacin, Vitamin B6, of iron and other micronutrients for food fortification.
Folate, Vitamin B12, Pantothenic Acid, Biotin, and In Encapsulation Technologies for Active Food Ingre-
Choline, National Academies Press, Washington, dients and Food Processing, N. J. Zuidam and V. A.
D.C., pp. 58-356. Nedovic, editors. Springer, London, pp. 187-209.
Stevanovic, M. and Uskokovic, D. 2009. Encapsulation Zuidam, N.J. and Shimoni, E. 2010. Overview of microencap-
devices for vitamin C. In Handbook of Vitamin C sulates for use in food products or processes and
Research, H. Kucharski and J. Zajac, editors. Nova methods to make them. In Encapsulation Technolo-
Science Publishers, United States, pp. 185-211. gies for Active Food Ingredients and Food Process-
Theobald, H.E. 2005. Dietary calcium and health. British ing, N.J. Zuidam and V.A. Nedovic, editors. Springer,
Nutrition Foundation Nutrition Bulletin. 30, 237-277. London, U.S.A., pp. 3-29.