Current Drug Delivery, 2006, 3, 429-443 429

Multiple Emulsions: An Overview

Azhar Yaqoob Khan*, Sushama Talegaonkar, Zeenat Iqbal, Farhan Jalees Ahmed and
Roop Krishan Khar

*Department of Pharmaceutical Medicine, Faculty of Pharmacy, Jamia Hamdard, New Delhi 110062, India; Depart-
ment of Pharmaceutics, Faculty of Pharmacy, Jamia Hamdard, New Delhi 110062, India

Abstract: Multiple emulsions are complex polydispersed systems where both oil in water and water in oil emulsion exists
simultaneously which are stabilized by lipophillic and hydrophilic surfactants respectively. The ratio of these surfactants
is important in achieving stable multiple emulsions. Among water-in-oil-in-water (w/o/w) and oil-in-water-in-oil (o/w/o)
type multiple emulsions, the former has wider areas of application and hence are studied in great detail. Formulation,
preparation techniques and in vitro characterization methods for multiple emulsions are reviewed. Various factors affect-
ing the stability of multiple emulsions and the stabilization approaches with specific reference to w/o/w type multiple
emulsions are discussed in detail. Favorable drug release mechanisms and/or rate along with in vivo fate of multiple emul-
sions make them a versatile carrier. It finds wide range of applications in controlled or sustained drug delivery, targeted
delivery, taste masking, bioavailability enhancement, enzyme immobilization, etc. Multiple emulsions have also been em-
ployed as intermediate step in the microencapsulation process and are the systems of increasing interest for the oral deliv-
ery of hydrophilic drugs, which are unstable in gastrointestinal tract like proteins and peptides. With the advancement in
techniques for preparation, stabilization and rheological characterization of multiple emulsions, it will be able to provide a
novel carrier system for drugs, cosmetics and pharmaceutical agents. In this review, emphasis is laid down on formula-
tion, stabilization techniques and potential applications of multiple emulsion system.
Keywords: Multiple emulsion, carriers, complexation, stabilization, sustained, targeted, drug delivery.

1. INTRODUCTION drophobic) separates the internal and external aqueous

Drug delivery systems such as nanospheres, micro-
spheres, microemulsions, emulsion particles, self-
emulsifying drug delivery system (SEDDS), liposomes and
mixed micelles have been investigated as potential carrier
systems for the delivery and/or targeting of the drugs to spe-
cific sites in the body. Among these carrier systems, emul-
sions have always invited particular interest as a carrier for
lipophillic drugs due to their biocompatibility, satisfactory
stability and ease of manufacture on an industrial scale [1,2].
There are several types of emulsions depending on how
the oil and water phases are located in the dispersed system,
where the term oil and water represent the less polar and
more polar of the two immiscible phases respectively [3].
According to IUPAC 1972, “An emulsion is a dispersion of
droplets of one liquid in another one with which it is incom-
pletely miscible. In emulsions the droplets often exceed the
usual limits for colloids in size [4]”. Simple emulsions are
labeled as oil-in-water (o/w) when the oil phase is dispersed
in continuous aqueous phase, or water-in-oil (w/o) when the
water phase is dispersed in continuous oil phase. Biemulsions
are emulsions containing two internal phase droplets, either
of the same nature (but different size) or of different nature
(whatever the size). Multiple emulsions are symbolized as
w1/o/w2 or o1/w/o2 depending on whether an oil phase (hy
*Address correspondence to this author at the Department of Pharmaceuti-
cal Medicine, Faculty of Pharmacy, Jamia Hamdard, New Delhi, 110062,
India; Mobile No. +91 921322416; E-mails: azharykhan@yahoo.com,
s.talegaonkar@gmail.com
phases, or an aqueous phase (hydrophilic) separates the in-
ternal and external oil phases, respectively. Internal and ex-
ternal phases may or may not be identical (Fig. 1).
Multiple emulsions are complex polydispersed systems
where both oil-in-water and water-in-oil emulsion exists
simultaneously which are stabilized by lipophillic and hy-
drophilic surfactants respectively. The ratio of these surfac-
tants is important in achieving stable multiple emulsions.
Multiple emulsions are also called as emulsions of emulsions
or double or triple emulsions, since the internal phase itself
contains disperse globules, which are miscible with the con-
tinuous phase. These are also called as liquid membrane
system because the two miscible phases are separated by an
immiscible phase (liquid membrane) which acts as a thin
semi-permeable film through which solute must diffuse in
order to traverse from one phase to another [5,6].
Among water-in-oil-in-water (w/o/w) and oil-in-water-in-
oil (o/w/o) type multiple emulsions, the former has wider
areas of application and hence have been studied in great
detail. Since water-soluble drugs are enclosed in an inner
water phase of w/o/w emulsion, these emulsions can be used
as carriers of water soluble drug for sustained release deliv-
ery. Additionally, w/o/w emulsions possess many advantages
over w/o emulsions and have low viscosity due to the exter-
nal aqueous phase, which makes them convenient to handle
and use, especially in parenteral formulation as targeting
drug delivery system [5].

1567-2018/06 $50.00+.00 © 2006 Bentham Science Publishers Ltd.

430 Current Drug Delivery, 2006, Vol. 3, No. 4
Fig. (1). Two types of multiple emulsion systems.
1.1 Advantages
Although, multiple emulsions are relatively complex to
formulate, bulky and prone to various routes of physical deg-
radation, still there are certain advantages of multiple emul-
sions which make them a challenging drug delivery system.
Multiple emulsions have excellent and exciting opportunity
for slow or controlled release of active entrapped compound.
Other advantages include;
• A remarkable degree of biocompatibility.
• Complete biodegradability and the lack of toxic prod-
ucts resulting from carrier degradation.
• Avoidances of any undesired immune responses against
the encapsulated drug.
• Hydrophilic as well as hydrophobic drugs can be en-
trapped within the multiple emulsions.
• Protection of the loaded compound from inactivation by
the endogenous factors.
• Remarkable decrease in fluctuation of steady state con-
centration as compared to the conventional method of
drug administration which is a common advantage for
most of the new drug delivery systems.
• Considerable increase in drug dosing intervals with drug
concentration in therapeutic window for a relatively
longer time.
• Possibility of drug targeting to reticuloendothelial sys-
tem (RES) and other organs.
• Can be used in taste masking of bitter drug.
2. FORMULATION DEVELOPMENT
Emulsions are thermodynamically unstable systems and
hence emulsifiers, such as surfactants, copolymers and
polymers etc. are used to provide kinetic stability to maintain
their structure for relatively long period of time. The inten-
tion in preparing multiple emulsion system is to introduce
two different surfactants of opposite nature to the system.
Khan et al.
One surfactant stabilizes the w/o (lipophillic) emulsion while
the other stabilizes the o/w (hydrophilic) emulsion [5,7].
Emulsifiers get adsorbed at the surface of droplets during
emulsion formation and prevent them from drawing close
enough to aggregate. Phase diagrams can help in producing
multiple emulsions by using only one surfactant to stabilize
both the primary and secondary emulsion. The presence of a
liquid crystalline phase in the ternary system consisting of
water-emulsifier-oil, where the aqueous micellar solution of
the emulsifier and the oil rich solution of the emulsifier are
in equilibrium with each other, has been shown to greatly
improve the stability of emulsions [8].
2.1. Formulation Techniques
(A) Double Emulsification Technique / Two Steps Emulsi-
fication
It is the most common method for preparing multiple
emulsions because it is very easy and gives high yield with
reproducibility. This method involves re-emulsification of
primary w/o or o/w emulsion in the outermost continuous
phase using a suitable emulsifier. The first step involves,
obtaining an ordinary w/o or o/w primary emulsion which is
then re-emulsified with an excess of aqueous phase or oil
phase in presence of second emulsifier to get the multiple
emulsions of w/o/w or o/w/o type respectively (Fig. 2). The
effect of the ratio of hydrophobic surfactant (span 80) to that
of the hydrophilic emulsifiers influences the physicochemi-
cal properties of formulations [7,9].
Second emulsification step is very important as it can
lead to fracture of internal globules forming simple emulsion
of either o/w or w/o type depending upon various other fac-
tors [10]. In recent studies, a modified two-step emulsifica-
tion technique for the preparation of multiple emulsions is
being used whereby each step is being further divided in two
sub-steps (pre-emulsification by sonication and than stirring)
leading to a 2 x 2, two-step method of emulsification. This
modified method gives high yield and stable emulsion
[11,12]. Second step involving temperature and pressure as
production parameters influences the size of oil droplets in
Multiple Emulsions
A)
Fig (2). Two step preparation method of w/o/w multiple emulsions (A) Primary emulsification (B) Secondary emulsification.
w/o/w multiple emulsions. Fine w/o emulsions used for sec-
ond step emulsification produced high encapsulation effi-
ciency [13].
(B) Phase Inversion (One Step Technique)
The development of w/o/w system was first reported
during the phase inversion of the concentrated w/o emulsion
to o/w emulsion [14,15]. A w/o/w emulsion was prepared by
extruding through a polycarbonate membrane possessing
pores of 3 or 8 µm in diameter. This extruded multiple emul-
sion became semisolid and changed to w/o emulsion. This
resultant emulsion was easily re-dispersed into aqueous so-
lution of hydrophilic emulsifier to form w/o/w type emulsion
having a droplet size smaller and more homogenous than
original w/o/w emulsion. Mechanism of phase inversion can
be depicted as: w/o/w emulsion → formation of lamellar
structure → reversed micelle → w/o emulsion → re-
dispersed to formulate homogenized w/o/w emulsion [16].
Phase inversion of w/o/w emulsions takes place if total
emulsifiers HLB in system approaches the required HLB of
the oil [17-19].
Phase inversion technique has a wide globule size distri-
bution and indefinite number of encapsulated compartments.
This is why this technique is not being used for preparing
multiple emulsion of uniform size [20]. A single step emulsi-
fication process for the preparation of multiple emulsions by
introduction of two surfactants and two polymers has been
recently reported. Method consists of addition of aqueous
phase containing polyethylene glycol (PEG) and tween 20 to
an oil phase comprising 1-octanol, hydroxyl propyl cellulose
(HPC), and span 80 by stirring with a magnetic stirrer. This
method was more efficient in comparison to two-step emul-
sification, by using two polymers, which improves stability
of emulsion [21].
(C) Membrane Emulsification Technique
This is a new, convenient and more useful technique
where a microporous glass membrane with a defined pore
size can be used as emulsifying tool. It is based on the prin-
ciple of dispersing one immiscible phase (dispersing phase)
into other phase (continuous phase) by applying pressure.
Other methods of preparation give high variability in the
globule size but technique gives emulsion of desired size by
Current Drug Delivery, 2006, Vol. 3, No. 4 431
B)
passing through required membrane size [22,23]. The parti-
cle size of the w/o/w emulsion can be controlled with the
proper selection of porous glass membrane and the relation
between membrane pore size and particle size of emulsion
exhibits good correlation as described by the formula:
Y = 5.03 X + 0.19 (1)
Where X is the pore size and Y is the mean particle size of
the multiple emulsions prepared by using membrane emulsi-
fication technique [24,25].
A microporous glass membrane with narrow pore size
range was used successfully for preparing stable simple
(o/w) and water-oil-water (w/o/w) type emulsions. Narrow
range of pore size was used for controlling size of emulsion
and egg yolk phospholipids and soybean oil was employed
as oil phase. Both emulsions were stable for 6 weeks when
stored at 5°C [26]. A homogenous w/o/w emulsions encap-
sulating cytarabine, doxorubicin and vancomycin were pre-
pared by membrane emulsification. These emulsions were
having effective encapsulation and controlled release deliv-
ery [23].
(D) Microchannel Emulsification
Microchannel emulsification (MC) is a novel technique
devised for preparing monodispersed emulsions. A two-step
emulsification process employing MC emulsification as the
second step was used for preparing w/o/w emulsions. The
behavior of internal water droplets penetrating the MC was
investigated using ethyl oleate, and medium-chain triglyc-
eride (MCT) as oil phases. They observed successful MC
emulsification and prepared monodispersed oil droplets that
contained small water droplets. The w/o/w emulsion entrap-
ment yield was measured fluorometrically and was found to
be 91% [27].
2.2. Formulation Variables
There are various factors that affect the stability of w/o/w
emulsions which include method of preparation, the nature
of entrapped materials, the phase volume ratio, concentration
and the type of emulsifiers, nature of oil phase used, elec-
trolytes, and other additives in inner and outer aqueous phase
[5,7,28,29].
432 Current Drug Delivery, 2006, Vol. 3, No. 4
2.2.1. Emulsifiers
On the basis of the hydrophilic-lipophillic value (HLB)
of oil phase, optimized concentration of surfactant is used for
emulsification. For w/o/w emulsion, primary emulsifier be-
ing lipophillic in nature usually have HLB of 2-8 while sec-
ondary emulsifier being hydrophilic in nature have HLB of
6-16. Secondary emulsifier is generally less than 1/5th of
primary emulsifier concentration. At higher concentrations
the primary surfactant gets incorporated in secondary sur-
factant micelles and leads to instability of multiple emulsion.
Optimized concentration of emulsifiers is important in
achieving stabilized formulation. Type of emulsifier effects
the preparation and stability of a multiple emulsion [30, 31].
Non-ionic emulsifier alone or in combination are normally
preferred over other surfactants for pharmaceutical use be-
cause of their low toxicity and also because they are less
likely to interact with other compounds. Also, non-ionic sur-
factant gives better yield in comparison to ionic surfactants
[5,7,11].
2.2.2. Oils Used
Oils used in the preparation of pharmaceutical emulsions
are of various chemical types, including simple esters, fixed
and volatile oils, hydrocarbons, and turpenoid derivatives.
The oil itself may be the medicament or it may function as a
carrier for a drug, or even form part of a mixed emulsifier
system as in the case of some fixed oils that contain suffi-
cient free fatty acids. The most widely used oils in oral
preparations are nonbiodegradable mineral and castor oils
that provide a local laxative effect, and fish liver oils or vari-
ous fixed oils of vegetable origin (e.g., arachis, cottonseed,
and maize oils) as nutritional supplements. The mineral oils
give much higher yield than the vegetable oils [5,6,32].
Oil nature is another factor which influences stability
because it not only control the rheology of the multiple
emulsions but also the permeability of the membrane, hence
the diffusion of solute across oil membrane. Oils of varying
density and viscosity can be used for changing drug release
characteristics [33]. Recently, enzymatic hydrolysis of the oil
phase [octanoic acid triacylglycerol (C8TG)] and release of a
marker substance from the inner-aqueous phase to the outer-
aqueous phase was investigated using an artificial digestive
fluid and size of oil droplets in emulsion significantly af-
fected release from emulsion [34]. Semicrystalline oil phase
was used for modifying release characteristic of multiple
emulsions. Increase in the fat content of the oil phase or fats
of high melting point decreased the release of marker (L-
tryptophan) from emulsions [35].
2.2.3. Phase Volume
Primary phase volume (Φw/o) has no significant effect on
the yield of w/o/w emulsion, therefore wide range of the
primary phase volume can be used for the multiple emulsion
formulation. An optimal (25-50%) internal phase volume can
be utilized for the emulsion formulation. The secondary
phase volume (Φw/o/w) influences the yield of multiple drops
over a range of low volume fractions only [7,36]. Very high
phase volume ratio (70-90%) had also been reported to pro-
duce a stable multiple emulsion [37,38]. It is very important
to have proper order of phase addition while formulation and
Khan et al.
dispersed phase should be added slowly into continuous
phase for the formulation of a stable multiple emulsion.
2.2.4. Shear/Agitation
High shear disrupts the large percentage of multiple oil
drops and hence results in the instability of system due to
tremendous increase in effective surface area. Therefore,
with increased homogenization time, the yield of the system
falls rapidly. Generally, high agitation speed is used for pri-
mary and low speed is used for secondary emulsification for
the preparation of multiple emulsions. Very high and low
shear rate drastically affect stability of emulsion system
hence shearing/agitation time should be optimized. Higher
shear stress causes incorporation of air and excessive froth-
ing resulting in loss of surfactant at water-air interface while
low shear does not reduce size of globules. Thus high shear
combined with air lead to instability of multiple emulsions
[5,7,11,39].
2.2.5. Temperature
Temperature is a critical parameter during emulsion for-
mation and needs to be precisely controlled. Rise in tem-
perature augments the lipophillic character of the hydrophilic
emulsifier, as it tends to precipitate. Generally, for the pri-
mary emulsion formulation temperature is kept at 700C,
whereas for multiple emulsion preparation it is kept at 100C
[40,41]. Large temperature variations during manufacturing,
storage, transport and use leads to drastic modifications
within emulsions [42].
2.2.6. Rheology
The rheological properties of emulsions are influenced by
a number of factors, including the nature of the continuous
phase, the phase volume ratio, and to a lesser extent by parti-
cle size distribution. A variety of products ranging from mo-
bile liquids to thick semisolids can be formulated by altering
the dispersed phase volume and/or the nature and concentra-
tion of the emulsifiers. For low internal phase volume emul-
sions, the consistency of the emulsion is generally similar to
that of the continuous phase; thus, o/w/o emulsions are gen-
erally thicker than w/o/w emulsions, and the consistency of
an w/o/w system can be increased by the addition of gums,
clays, and other thickening agents that impart plastic or
pseudoplastic flow properties [43]. Micropipette aspiration
method has been used to evaluate elastic modulus of the oil
membrane [40].
3. IN VITRO CHARACTERIZATION
In-vitro evaluation of multiple emulsions is an important
step in developing a stable and effective drug carrier system,
which leads to predictive in vitro-in vivo correlation
(IVIVC). Significant changes in the following parameters
during preparation, storage or transport are indicators of poor
stability.
(i) Macroscopic Examination: Primary observations
like color, consistency and homogeneity are frequently used
to ensure formation of an emulsion. Type of multiple emul-
sions formed (w/o/w or o/w/o) can be validated by dilution
with the external phase [5].
Multiple Emulsions
(ii) Microscopic Examination: The multiplicity of the
multiple emulsions can be verified by light microscopic
and/or electron microscopic techniques [44,45]. Multiple
emulsion formation has also been studied inside a microcap-
illary using video microscopy [20]. Different filming tech-
niques (time-lapse, continuous and high speed cinemicrogra-
phy, photomicrography) had been used for characterization
[46]. In optical microscopy method, calibrated ocular and
stage micrometer can be used for globule size determinations
of both multiple emulsion droplets as well as droplets of in-
ternal dispersed phase. This method is simple and provides
useful information on the character of multiple emulsions
and allows determination of droplet size and size distribu-
tion. A suitable magnification may be used for the purpose.
However, this method has two main drawbacks; (1) simple
small drops passes to form large simple drop which gives a
false impression of multiple nature (2) if the internal droplets
are very small in size these can not be viewed due to reflec-
tion of light from the oil droplets surface [47].
Coulter-counter method is also used when multiple emul-
sion diameters are in narrow range. It can be used to measure
size of multiple globules entrapping the aqueous droplets.
This method has been used to investigate the effect of alter-
ing the primary emulsion concentration on water permeation
across multiple emulsions [48-50]. Freeze etching-electron
microscope technique has also been used for determining
external and internal droplets size distribution [51]. Recently,
Ursica and coworkers used this technique for determination
of multiple emulsion size and size distribution density [52].
(iii) Number of Globules: Number of globules is also an
important parameter for characterization of multiple emul-
sions. High multiple globule number suggests for a good and
stable emulsion after storage [53]. Number of globules per
cubic millimeter (mm3) can be measured using a hemocy-
tometer cell after appropriate dilution of the multiple emul-
sions. The globules in five groups of 16 small squares (total
80 squares) can be counted and the total number of globules
per cubic mm can be calculated using the formula [54];
No. of Globules / mm3 = No. of Globules X Dilution X 4000 …. (2)
No. of small squares counted
(iv) Entrapment Efficiency / Percentage Drug En-
trapment/ Emulsion Yield: Most common parameters for
evaluating multiple emulsion quality is the emulsion forma-
tion or yield of emulsion. Ratio of the lipophillic surfactant
in oil phase to hydrophilic surfactant in continuous phase,
affect w/o/w emulsion yield. This ratio should be more than
10 for achieving 90% or more yields of emulsions [7].
Two methods, size analysis and internal tracer technique
have been used for evaluating yield of multiple emulsion
droplets [32]. Particle size distribution analysis of simple and
multiple droplets of total system can be estimated for emul-
sion yield [49,55]. Several techniques (photomicrography,
coulter counter, electron microscopy) can be used for evalu-
ating particle size analysis. Second method is the use of the
internal tracer to establish entrapment efficiency of an im-
permeable marker molecule in the w1 phase of emulsion. The
method basically involves determination of unentrapped
marker or drug. Emulsion yield can be measured by dis-
solving a marker compound in the inner aqueous phase of
Current Drug Delivery, 2006, Vol. 3, No. 4 433
emulsion and measuring concentration of marker in the outer
aqueous phase. Yield can be determined by subtracting the
remaining amount of marker from the original amount
[18,56]. Dialysis [10,11], centrifugation, filtration and con-
ductivity measurement [57] are the four methods generally
used for determination of entrapment efficiency by this tech-
nique. Several markers have been used like glucose [7], hy-
drogen ions [58,59], electrolytes [60, 61], new coccine
[28,62], ionic drugs (Ephedrine HCl), dyes (sulphane blue,
polytartarazine), radioactive tracer (tritiated water), etc [63].
A new on line technique based on electrical conductivity
measurement has been used for continuously monitoring the
isotonic swelling behavior of w/o/w multiple emulsions liq-
uid membranes (ELMs) [64].
Formation percentage or yield of w/o/w multiple emul-
sions was improved by addition of various additives like
sodium alkylsulfonate, sodium alkylcarbonate, tweens, so-
dium carbonate, sorbitol in the internal aqueous phase. This
increase has been attributed due to decrease in interfacial
tension between the internal aqueous phase and oily phase of
emulsion [62]. Similar results had been reported by the addi-
tion of sodium chloride in inner aqueous phase on drug en-
trapment in w/o/w emulsions containing tryptophan as model
drug. Drug entrapment was decreased when sodium chloride
was added in the outer aqueous phase [65].
(v) Nuclear Magnetic Resonance (NMR): NMR is a
new technique to ensure the multiple character of the emul-
sion as the NMR signals of water protons are narrow and
singlet in simple emulsions while the signals are widened or
doubled in multiple emulsions. Therefore, NMR can be util-
ized for the identification of type of emulsion [47].
(vi) Rheological Analysis: Viscosity measurement is an
important parameter for the characterization and stability
studies of multiple emulsions. In case of an unstable emul-
sion system, oil layer ruptures and inner aqueous globules
merge with the continuous aqueous phase and disappear in-
stantly. Volume fraction of the dispersed phase decreases
gradually in emulsion with increasing oil globules rupture
lead to decrease in viscosity of emulsion system [66,67].
Solutes and water permeate through oil layer of vesicles
causing change in viscosity [68]. Newtonian viscosity of the
diluted w/o/w emulsion could be influenced by osmotic pres-
sure gradient between the inner and external aqueous phases.
This osmotic drift can be on either side of the emulsion de-
pending on viscosity of the aqueous phases and can be used
to estimate water permeation coefficient as well [9,69].
Rheological analysis of multiple emulsions is very important
as it determines the in vitro emulsion stability and in vivo
clinical performance. Brookfield, rotational or cone and plate
viscometers are widely used for viscosity determination of
multiple emulsions [11,57]. Three different types of tests (a)
oscillatory test (b) steady-state flow tests (c) turbulence
shock test were proposed to simulate in-vivo conditions. In-
terfacial film strength i.e. elasticity of (w/o) and (o/w) com-
ponents of (w/o/w) multiple emulsions was evaluated by
interfacial rheology measurement at the mineral oil/water
interface using an oscillatory surface rheometer. The effect
of adding hydrophilic surfactant on interfacial elasticity was
investigated which further provides an insight into stability
of the multiple emulsion system [70]. Negative thixotropic
434 Current Drug Delivery, 2006, Vol. 3, No. 4
flow of w/o/w emulsions was observed at low shear rate
which was more pronounced under increasing shear rate or
time or repeated shear [16]
(vii) Zeta Potential: The electrophoretic mobility of dis-
persed globules under the influence of applied voltage in
zeta-potentiometer can be used to measure the zeta potential
and surface charge of multiple emulsions. The apparatus
consists of cylindrically bored microelectrophoresis cell
equipped with platinum-iridium electrodes to measure the
electrophoretic mobility of the diluted w/o/w emulsion [71].
It can be calculated by using following equation
ζ=4Πηµ (3)
εΕ
Where, ζ = Zeta potential (mv)
η = Viscosity of the dispersion medium (poise)
µ = Migration velocity (cm/sec)
ε = Dielectric constant of the dispersion medium
Ε = Potential gradient (Voltage applied / distance
between electrodes)
(viii) Percentage Phase Separation: Phase separation is
a phenomenon by which one phase of emulsion gets sepa-
rated due to coalescence. Percentage phase separation is the
volume of phase in percentage separated from the total vol-
ume of emulsion after storage. 20 ml of freshly prepared
w/o/w emulsion is kept in 25 ml graduated glass cylinder and
allowed to stand for defined period at 40oC. The volume of
separated aqueous phase (Vsep) is observed periodically at
regular intervals and percentage phase separation (B) is cal-
culated as follows [72];
B (%) = 100 (Vsep/20) / [(V1+V2) / (V1+V2+V0)] ……… (4)
Where V1, V2, V0 represents volumes of inner aqueous
phase, dispersion phase and middle oil phase.
(ix) In-vitro Drug Release: The drug release from w/o/w
emulsion is generally determined by dialysis method using
cellophane tubing. Typically, 5 ml of (w/o/w) multiple emul-
sion is placed in the dialysis tube which is then tied at both
ends by thread and placed in basket (usually 100 rpm) and
dialyzed against specified dissolution media (usually 200 ml)
at 37 ± 1°C. A sink condition is maintained and samples are
withdrawn at different time intervals and replaced by fresh
dissolution media [73-75].
In vitro release study w/o/w emulsions containing theo-
phylline in inner aqueous phase of w/o/w emulsion was
studied by two methods. In method ‘A’, cellulose tube con-
taining the emulsion was placed in dissolution medium in
rotary basket and in method ‘B’ by dispersing the emulsion
in the dissolution medium and the system was then stirred by
a paddle, allowing the drug to permeate into a cellulose tube
placed in the dispersing medium [76].
4. MECHANISMS OF INSTABILITY
A stable emulsion is considered to be one in which the
dispersed droplets retain their initial character and remain
uniformly distributed throughout the continuous phase dur-
ing the desired shelf life. There should be no phase change or
Khan et al.
microbial contamination on storage, and the emulsion should
maintain elegance with respect to odour, colour, and consis-
tency. Instabilities of both chemical and physical origins can
occur in emulsion formulations. Emulsion stability is a phe-
nomenon, which depends upon the equilibrium between wa-
ter, oil and surfactants.
Main mechanisms by which instability occurs in multiple
emulsions of w/o/w type have been studied thoroughly and
can be summarized to following possible mechanisms [77-
79].
I. Aggregation of the internal and multiple emulsion drop-
lets,
II. Breakage or disruption of oil layer on the surface of in-
ternal droplets
III. Seepage of the contents from inner droplet in external
phase.
IV. Shrinkage and swelling of the internal drops due to os-
motic gradient across the oil membrane.
V. Phase separation
5. STABILIZATION OF MULTIPLE EMULSION
Inherent instability in multiple emulsions is a major
challenge for this delivery system. A number of approaches
had been tried out to improve the stability of multiple emul-
sion and many have been suggested for further investiga-
tions. Several reviews had been reported on the stabilization
and mechanisms of drug release that concentrates on differ-
ent mechanism and methods of stabilization [77-79].
5.1. Stabilization by Forming Polymeric Gel
Gelation of either internal or external phase or the oily
phase (membrane) has been employed in various studies to
stabilize the system. Stable w/o/w emulsion was formulated
by polymeric gel in the internal or external aqueous phase
either by cross linking of surfactant molecules through high
energy γ-radiation or by polymerization of monomers dis-
solved in aqueous phase. These methods are based on vis-
cosity enhancement of aqueous phase thereby improving
stability of emulsion droplets [80,81].
Stable multiple emulsion were achieved by incorporating
poly(acrylamide) [5], gelatin or aminoacid [82]. An emulsion
stabilized with anionic hydrocolloids in outer aqueous phase
was developed for antiperspirants, sustained release drug
formulations or personal hygiene products [83]. Synthetic
polymer (Carbopol 974P) and chemically modified cellulose
(Hydroxypropylcellulose) were used as gelling agent for
multiple emulsion preparation [84]. A stable multiple emul-
sion was developed by incorporating a co-emulsifier into the
external water phase. Incorporation of the co-emulsifier in-
creases the compaticity, elasticity and temperature stability
of multiple emulsions [85]. Multiple emulsions (w/o/w) of
arachis and olive oil were stabilized by cherry gum, in com-
bination with acacia and gelatin. Cherry gum was having
outstanding film-forming properties (liquid crystal-bearing
interfacial films) and the effect of its addition to aqueous
phase led to decreased coalescence of emulsion globules
[86]. Many thickening agents (acacia, tragacanth, sodium
Multiple Emulsions
alginate, methyl cellulose, carboxymethylcellulose, hy-
droxypropylmethylcellulose) were optimized for giving most
stable emulsion [87]. Several viscosifying agents were in-
corporated into outer aqueous phase were used and its effect
on the stability and multiplicity of multiple emulsions were
investigated. Klucel, viscosified formulation was most stable
while the one prepared with carbomer in outer phase showed
much better skin feel at lower concentrations [57].
5.2. Additives in Internal Aqueous Phase
Electrolytes have been used for stabilizing w/o/w emul-
sion. These electrolytes include ascorbic acid, acetic acid,
sodium chloride and sodium citrate. The addition of
lysozyme to the aqueous phase or stearylamine and oleic
acid to the oil phase effectively improves the stability of the
oil layer. The influence of various additives, e.g. glucose,
sucrose, acetic acid, citric acid, ascorbic acid, sodium chlo-
ride and sodium acetate on the stability of water-in-olive oil-
in-water multiple emulsion had been studied and it was ob-
served that glucose and sucrose could increase the viscosity
and as a result improves stability of emulsion system. Addi-
tives in the internal aqueous phase make it hypertonic and
stabilizes the system. Microscopic study conducted on such
emulsion indicated that as the dextrose concentration (0-
2.5%), in internal aqueous phase increased the stability re-
corded in terms of degree of coalescence of internal droplets
and rupture rate of interfacial oily layer, was also increased
[11, 88].
Addition of sodium chloride and hydrophilic surfactant
into the internal aqueous compartment of the multiple emul-
sions also influences the break down of the suspending ve-
sicular globules [28]. The stabilization of w/o/w emulsion by
making inner aqueous phase hypertonic, addition of chitosan
in the inner aqueous phase and phase inversion with porous
membrane and its application to transcatheter arterial emo-
bilzation (TAE) therapy was reviewed [79]. For longterm
stability of the double emulsion, it should have a balance
between the Laplace and osmotic pressures within emulsion
[89]. Recently, a substituted cyclodextrin (Hydroxypropyl-_-
cyclodextrin) in internal phase effectively stabilizes antioxi-
dant (Kaojic acid) in w/o/w multiple emulsions [90, 91].
5.3. Modulating Surfactant Concentration
Several attempts have been made for stabilizing multiple
emulsions by modulating emulsifiers systems [20,37,63,92].
A suitable surfactant with required HLB is used for prepar-
ing stable emulsion. Emulsifiers stabilize emulsions through
mechanical or electrical interfacial barrier formation or by
reduction of the interfacial tension or both [93]. For long
term emulsion stability, the strength of the mechanical bar-
rier is more important than interfacial tension [94]. There-
fore, an optimal HLB for a particular multiple emulsion sys-
tem depends on the concentration of the primary and secon-
dary emulsifiers [18]. Type and concentration of surfactant
are the primary factors which influence the stability, while
pH and ionic strength insignificantly influences stability of
emulsions [20].
Mixture of emulsifiers (spans and tween) was used to
improve stability of the multiple emulsions. Mixed emulsi-
fier gives higher film strength in comparison to single emul-
Current Drug Delivery, 2006, Vol. 3, No. 4 435
sions. This hypothesis was confirmed because addition of
bovine serum albumin (BSA) in internal aqueous phase did
not improve stability of emulsion [95].
The ratio of hydrophilic and hydrophobic surfactants is
very important in optimizing multiple emulsions. Long-term
stable water-in-oil-in-water multiple emulsions was devel-
oped by modulating concentrations of span 83 and tween 80
[96].
5.4. Formation of Interfacial Complex Films
Polymerized interfacial membrane has been found to be
more effective than previous methods in stabilizing multiple
emulsions. It is because resultant interfacial membrane re-
stricts surfactant migration from one interface to other,
which is one of the causes of instability in w/o/w emulsion
[92]. This mode of stabilization can be executed either
through in situ polymerization at the interface or by interfa-
cial interactions between a polymer and a surfactant. During
the search for stable w/o/w emulsion as vehicle for sustained
release preparation, numerous researchers have employed
interfacial complexation between non-ionic surfactants and
macromolecules [5]. Another approach to obtain stable
w/o/w multiple emulsions is based on the interfacial interac-
tions between a macromolecule, e.g. albumin [97] or poly-
acrylic acid or bovine serum albumin [98] in internal aque-
ous phase and lipophillic nonionic surfactant poloxamer 331
in middle oily phase, resulting into formation of stable w/o
emulsion. Hydrophilic poloxamer surfactant of high mo-
lecular weight, e.g. poloxamer 403 enhanced stability of
w/o/w emulsion of the above-mentioned primary emulsion.
Interfacial membrane was so strong that it withstood the
thinning of oil phase caused by swelling of internal aqueous
droplets because of osmotic pressure of influxed water [92,
99].
Interfacial complex films between polyvinyl alcohol,
acacia, polyvinylpyrrolidone and sorbitan monooleate were
used for preparing long-term stability of the emulsions
bearing rifampicin [44, 45]. Stable w/o/w emulsions were
prepared by interfacial films between various pluronic
F127:polyacrylacid complexes in the internal aqueous phase
and the lipophillic surfactant in oil phase. These emulsions
were capable for rapid or sustained drug delivery depending
on the pluronic F127:polyacrylacid complexes and type of
lipophilic surfactant used [100].
The significance of the inner and outer phase pressure, as
well as interfacial film strength on w/o/w multiple emulsions
stability was studied by using microscopy and long term
stability tests. Stability was correlated with the interfacial
film strength (measured by interfacial elasticity) of the hy-
drophobic surfactants at the mineral oil/external continuous
aqueous phase interphase. Authors concluded that metastable
dimpled structure and long term stability of multiple emul-
sion was dependent on the osmotic pressure of the inner
droplets. [101].
5.5. Pro-Multiple Emulsion Formation
In this approach, emulsion is stabilized by changing the
physical nature of emulsion from liquid to solid by using
adsorbent or solids. Whenever multiple emulsions is needed
436 Current Drug Delivery, 2006, Vol. 3, No. 4
this proemulsion is dispersed in specified quantity of water
to form multiple emulsion. This is a new approach and is still
under extensive investigations [102,103].
5.6. Steric Stabilization
Steric stabilization had been found to stabilize emulsions.
It is due to the ability of steric cloud to prevent their insta-
bility in emulsions by surrounding environmental challenges.
Steric stabilization can be achieved by using either adsorp-
tion or grafting of natural polymers or by using polymeric
surfactant or lipid grafted polymers. Multiple emulsions can
be stabilized by coating them with hydrophobized polysac-
charides [104,105].
6. IN VIVO STUDIES
Multiple emulsions besides solution, suspension, gel,
emulsion, micro-emulsion, self-emulsifying drug delivery
system (SEDDS) offer an excellent approach for administra-
tion of lipids [106]. Multiple emulsions had been used as
carrier for both lipophillic and hydrophilic drugs depending
on the intent of application, route of administration and fea-
sibility of formulation. Multiple emulsions can be adminis-
tered through oral, parenteral (i.v, i.p, s.c, i.m) and topical
routes for therapeutic purposes. In-vivo fate of multiple
emulsions differs from other delivery system in stability,
pharmacokinetic and pharmacodynamic perspectives [47].
6.1. Oral Administration
After oral administration, emulsion (lipid) gets digested
and absorbed by lymphatics and then transported to blood.
Both lipid absorption and digestion have taken great atten-
tion and many extensive reviews are available for the
mechanism of the lipid digestion and transport [107-109].
In brief, lipids firstly get hydrolyzed in the stomach and
small intestine resulting in fatty acid (FA) and monoglyc-
eride (MG), which get absorbed into the enterocyte, re-
esterified in triglyceride and packaged into intestinal lipo-
proteins (chylomicron and very low density lipoprotein).
These can also be absorbed directly through the intestinal
macrophage system and payer’s patches. These are finally
secreted into mesenteric lymph from where they reach the
systemic circulation via the thoracic lymph duct. Therefore,
drug transport through the intestinal lymphatics may be util-
ized (i) in modulating rate of drug absorption (Table 1) (ii) to
increase bioavailability by decreasing first pass metabolism
(Table 2) (iii) for targeting drugs to lymphatics (Table 3)
[110].
The enzymatic hydrolysis of the oil phase of w/o/w
emulsions and release of a marker compound from the inner-
aqueous phase was assessed by using an artificial digestive
fluid containing lipase and gall. Size of the oil droplets in the
w/o/w emulsion significantly affected the release profile and
enzymatic hydrolysis from emulsions [34].
Bioavailability of griseofulvin (500mg) from oil in water
emulsion (o/w), water-in-oil-in-water (w/o/w) and tablet
dosage form was evaluated and compared following oral
administration to eight healthy volunteers in a cross over
fashion. Urinary excretion data of major metabolite of grise-
ofulvin (6-desmethygriseofulvin), demonstrated significant
Khan et al.
increase in the rate and extent of absorption (p<0.05) in fol-
lowing order w/o/w>o/w> tablets [111]. Similarly, bioavail-
ability of nitrofurantoin (100mg) from tablet and w/o/w
emulsion was assessed by urinary excretion data of un-
changed drug in eight healthy volunteers in a crossover
study. Nitrofurantoin bioavailability was upto 70% and 50%
less in tablets comparison to arachis oil and soyabean oil
multiple emulsions. Multiple emulsions also reduced adverse
experiences of drug in comparison to tablets because of the
protection of the gastrointestinal wall from direct contact of
drug. Bioavailability of nitrofurantoin was further enhanced
by increasing the viscosity of formulation [112, 113]. Kho-
pade et al. reported an augmentation of 1.51 times in
bioavailability of isoniazid (100mg) when given in w/o/w
emulsions. This data was reported for six healthy male vol-
unteers and plain capsules of rifampicin were used for com-
parison [74].
The potential of multiple emulsions to circumvent the ill
effects of first pass metabolism, harsh GI enviornment and
also to get absorbed through lymphatic system has placed it
in an advantageous position for delivery of bioactive agents
like insulin and vancomycin which are normally not ab-
sorbed through oral route. The use of multiple emulsions as a
drug delivery device for insulin was reported by Engel in
1968. The study conducted in rats and gerbils (intraduodenal
administration) demonstrated a marked reduction in blood
glucose levels of 25% (rats) and 57% (gerbils) in comparison
to simple emulsion and drug solution after one hour sample
[114]. A similar study for increasing absorption of insulin
from colon and rectum by multiple emulsions containing
eicosapentaenoic acid and docosahexaenoic acid has also
been reported by Suzuki et al. [115].
In a study carried out by Matsuzawa et al., hypoglycemic
effects of insulin from the gelatin stabilized w/o/w emulsion
were observed after insulin administration to stomach, duo-
denum, jejunum, ileum and colon by using an in-situ loop
method in rats. In comparison to i.v. route, relative hypogly-
cemic efficacy from various regions was 0.65, 0.25, 0.49,
1.03 and 0.75 respectively. Clearly insulin was absorbed
predominantly from ileum region because of the presence of
lacteal and peyer’s patch [116]. Toorisaka et al. also reported
novel solid-in-oil-in-water (s/o/w) emulsion carrying insulin
which elicited a marked hypoglycemic effect for long time
when given orally to rats. This was probably due to an in-
creased absorption of insulin from small intestine through
micelles which were formed from degraded fatty acid of the
soyabean oil along with abundant lipases present in the in-
testine [117].
Shively et al. conducted a study with a w/o/w emulsion
of vancomycin, a glycopeptide water soluble drug, which
was prepared from solid state emulsion and then was admin-
istered to sprague-dawley male rats. When compared in
terms of absorption with an i.v route, it amounted to signifi-
cant 30% while the other dosage form given as suspension
did not show any absorption. The multiple emulsions also
depicted sustained and reproducible plasma concentration
[118]. Recently, an increased transport of a hydrophilic
marker through intestinal epithelial cell (Caco-2) monolayer
was observed by a multiple emulsions containing octanoic
acid triacyl glycerol (C8TG) as oil phase [119].
Multiple Emulsions
Oral salmon calcitonin (sCT), another poorly absorbed
polypeptide hormone was formulated as a water/oil/water
multiple emulsion, for administration to rat model. The mul-
tiple emulsion protected the peptide from enzymatic degra-
dation and increased bioavailability of sCT which was fur-
ther enhanced by incorporating the protease inhibitor,
aprotinin, in the outer aqueous phase [120].
In-vivo studies of rifampicin in w/o/w emulsions in brain
from the oral and nasal routes were carried out in rats in
comparison to drug solution. Prolonged plasma drug levels
for multiple emulsions from both the routes were observed
and significantly greater brain drug levels were seen at 4
hours after administration because of increased clearance of
drug from solution than multiple emulsions [121]. In-vitro-
in-vivo correlation of indomethacin from w/o/w multiple
emulsions was also established following an oral administra-
tion in male rabbits (10 mg/kg). Pharmacokinetic parameters
like C max, T max, AUC o-t concluded for prolonged drug action
over 12 hours. t 50% (time taken to release 50% of indometha-
cin in vitro) versus Cmax showed a good linear correlation
[75]. An oral sustained and prolonged release microsphere-
in-oil-in-water (s/o/w) formulation of Tegafur was evaluated
in rats. Mean residence time of tegafur from w/o/w emulsion
was two times greater than from a w/o emulsion or micro-
sphere system suggesting a sustained delivery [122].
6.2 Parenteral Administration
Parenteral administration of multiple emulsions could be
approached through a multitude of pathways like i.v, i.m, i.p
or s.c. Drug administration by i.p. route is important at pre-
clinical study to assess the relationship between in vitro and
in vivo performances of formulations. While drug admini-
stration by i.m route is convenient and fast absorption of
drugs is an added advantage [110]. Intravenous route offers
convenience and complete bioavailability for most of drug
solutions but particle sizes is constraint (<7 µm diameter)
[123]. The most important mechanism for multiple emulsion
from all the parenteral systems is the rapid clearance from
the systemic circulation by macrophages of reticuloendothe-
lial system (RES) that lines the sinusoids of liver, spleen and
bone marrow (non-specific defense mechanism) [105].
After i.v administration, emulsions get engulfed by cir-
culatory macrophages and transported to lymphatics and
liver for fat metabolism pathway while for other parenteral
routes, the emulsion droplets gain access to nearby lymphatic
node through interstitial spaces of lymphatic vessels (in con-
trast to low molecular weight water soluble compounds
which gets absorbed into blood) which are relatively porous
as compared to blood capillaries which have tight intracel-
lular junctions. Multiple emulsions has intrinsic lym-
photropic characteristic therefore it is being investigated for
the treatment of bacterial infections and cancer metastases
(lymphotropic drug targeting) [47].
Emulsions have attracted special interest in cancer ther-
apy [12, 25, 124]. Iodinated oil fatty acid has been widely
used as oily continuous medium and an important chemoem-
bolizing system for transcatheter arterial embolization (TAE)
therapy. Multiple emulsions is a special drug carrier system
for TAE therapy because of low viscosity and high encapsu-
Current Drug Delivery, 2006, Vol. 3, No. 4 437
lation. Multiple emulsions bearing Epirubicin (EPI) have
been used for TAE therapy. This emulsion showed low
hemolysis in comparison to simple emulsion because of the
entrapped drug inside the oily layer. The acute toxicity stud-
ies of emulsions after i.v administration were lower than
simple emulsion. After hepatic arterial administration, liver
toxicity of drug was also reduced [79].
Prolonged or sustained release potential of the multiple
emulsion has been explored for all the parenteral routes. In
vivo release of i.v. administration of water-soluble drugs,
cefadroxil and cephradine from water-in-oil-in-water
(w/o/w) multiple emulsion was explored in a study in rats
and was found to show a prolonged release characteristics in
comparison to aqueous solutions [125]. An in-vivo study of
5-fluorouracil from multiple emulsion in rats exhibited sus-
tained release profile from the injection site [126]. A multi-
ple emulsion system (w/o/w) having Iodine-131-labeled io-
dohippuric acid after intramuscular administration in rabbits
exhibited a sustained release potential [127].
Pharmacokinetics of vancomycin after parenteral admini-
strations such as intramuscular, intraperitoneal and intrave-
nous administration of vancomycin solutions and vancomy-
cin in w/o/w emulsions was studied. Pharmacokinetics of
vancomycin in w/o/w emulsion after i.m. administration
showed a prolonged release profile in comparison to vanco-
mycin solution which was rapidly absorbed. Firstly, w/o/w
emulsions were accumulated at the injection site and then the
release was initiated. After i.m. administration, vancomycin
was transferred into the blood circulation through lymph
nodes. Lymphatic accumulation of w/o/w emulsions was
larger than solution and accumulation of drug in lymph
nodes resulted in a prolonged drug release. After i.p admini-
stration, the release of vancomycin was sustained from the
w/o/w emulsions when compared to the solution form. This
was explained on the basis that emulsions remained at peri-
toneal cavity and were not absorbed into peripheral blood or
lymph nodes. The drug was then released in peritoneal cavity
and was absorbed by lymph resulting in continuous plasma
concentration. The i.v. administration of the w/o/w emulsion
was resulted in an initial accumulation in lung but then de-
creased gradually. While in liver the drug concentration in-
creased rapidly after one hour of administration. Little
amount drug also got accumulated in the spleen and kidney.
The administration of the w/o/w emulsion through i.v route
elicited significant differences in pharmacokinetic of vanco-
mycin and showed promising enhancement in plasma con-
centration for a prolonged period of time [12].
7. APPLICATIONS
In last 20 years, there had been a revolution in the emul-
sions technology and these systems had been investigated for
several industrial processes and as drug delivery systems.
Water-in-oil-in-water (w/o/w) multiple emulsions have sev-
eral potential applications in pharmaceuticals, food technol-
ogy [128], separation sciences [129] and in cosmetics [37].
The various pharmaceutical applications include immobili-
zation of enzymes, red blood cell substitute, transdermal
delivery, bioavailability enhancement, taste masking, drug
targeting, prolonged delivery of drugs etc.
438 Current Drug Delivery, 2006, Vol. 3, No. 4 Khan et al.

7.1. Prolonged/Controlled Drug Delivery
Multiple emulsions (w/o/w, ME) have been extensively
investigated for controlled/prolonged delivery of drugs. The
potential of using controlled release multiple emulsion has
already been reported by several workers and a number of
research papers are available, these have been summarized in
Table 1. Different categories of drugs have been tried out for
the sustained delivery having very short half-lives. ME de-
livers drugs in sustained fashion as the drugs are being in-
corporated in the inner aqueous phase of the w/o/w multiple
emulsions which is being separated by the middle oil phase
and drug has to partition through the oil membrane for re-
lease, leading to the sustained delivery of the drug. Both oral
and parenteral prolonged delivery had been investigated. In
parenteral delivery, multiple emulsions act as reservoirs of
the drug in blood and releases their content in controlled
fashion.
7.2. As Oxygen Substitute
A stable hemoglobin (Hb)-in-oil-in-water (Hb/o/w) mul-
tiple emulsion was prepared to simulate red blood cell (RBC)
properties. Multiple emulsions serve as the liquid membrane
across which gases (O2 and CO 2) are exchanged with hemo-
globin incorporated in the inner aqueous phase. Therefore,
this approach can further be utilized in future for the blood

Table 1. Multiple Emulsions Used for Prolonged/Controlled Drug Delivery

Category Drug investigated References

Analgesic and antipyretic agents Indomethacin 75, 130

Diclofenac sodium 131

Paracetamol 132

Anticancer agents Cytarabine 133

Doxorubicin 134

Etoposide 135

5-fluorouracil 10, 33, 97

Methotexate 136

Tegafur 122

Antimalarials Chloroquine 137

Antitubercular agents Rifampicin 44, 45, 71, 121

Isoniazid 73, 74

Antiasthmatic agents Salbutamol 138

Theophylline 100

Antibiotics Cefadroxil, Cephradine 125

Nitrofunrantoin 113

Antihistaminic Chlorpheniramine 87

Peptides Vancomycin 11, 12

Insulin 100, 117

Miscellaneous Salmon calcitonin 120

Prednisolone 31

Antipyrine, 4-Aminoantipyrine 125

Pilocarpine 139

Naltrexone 140

Phenylephrine 141

Pentazocine 142, 143

Multiple Emulsions Current Drug Delivery, 2006, Vol. 3, No. 4 439

substitute [144]. geting is an important concept, which minimizes the adverse

7.3. As Bioavailability Enhancer
Multiple emulsions have received great interest in facili-
tating the absorption of water soluble compounds through
gastrointestinal tract which do not normally gets absorbed
due to instability in the presence of certain physiological,
ionic/enzymatic environments in the gastrointestinal tract
like proteins, peptides etc. For highly polar drugs also, mul-
tiple emulsion system has found a great role in enhancing
absorption due to lipophillic characteristics. It is well-known
that lipid material like oils, fats, and lipophillic drugs are
being absorbed by lymphatic rather than portal system by-
passing the hepatic first pass metabolism and pouring the
drugs directly to the systemic circulation [145]. Multiple
emulsions have also been used to improve bioavailability of
those drugs, which have high first pass metabolism.
7.4. Targeted Drug Delivery System
Site specificity is very important prerequisite for any
parmacotherapy. Ideal approach for a drug delivery system is
to deliver drug only at the diseased tissue/organ and not af-
fecting other undiseased tissue (Drug targeting). Drug tar-
effects of drugs by specifically concentrating drug in dis-
eased tissue. Several micro particulate systems have been
used for drug targeting. It is of particular interest for the cy-
totoxic drugs (Anti-cancer agent) because of high toxicity for
non-diseased tissue. Multiple emulsions system had also
been prepared for several cytotoxic agents, targeting differ-
ent tumors. These can be used as lymphotropic carriers for
drug targeting. In vivo fate of multiple emulsion involve its
uptake by reticuloendothelial system (RES) thereby concen-
trating these oily vessels in lymphatic system and lymph
node. These are of immense potential for the lymphatic tar-
geting; other organs (Liver, brain etc.) have also been tar-
geted [149].
7.5. Taste Masking
Multiple emulsions had been investigated for the taste
masking potential of the bitter drugs. Water-soluble bitter
drugs had been incorporated in the inner aqueous phase of
w/o/w multiple emulsion, which is being surrounded by oil
layer and leading to masking the taste of the bitter drugs.
Several biocompatible/edible oils had been used for masking
taste and improving the masking potential. Taste masking of

Table 2. Multiple Emulsions Used in Bioavailability Improvement/Enhancement

Category Drug investigated References

Antidiabetic agents Insulin 114-116,146-148

Anti-fungal agent Griseofulvin 111

Antitubercular agents Isoniazid 73,74

Antibiotics Vancomycin 118

Nitrofurantoin 112, 113

Miscellaneous Pyrenetetrasulonic acid tetrasodium salt (PTSA) 119

Table 3. Multiple Emulsions Used for Drug Targeting

Target (Tissue /organ) Drug investigated References

Lymphatic system 5-fluorouracil 150

Iodohippuric acid 151

Bleomycin 152

Isoniazid 73

Tumor Bleomycin 153

Brain Rifampicin 121

Liver 5-fluorouracil 137

Epirubicin 79

Lungs Rifampicin 154

Inflammatory tissue Diclofenac sodium 155

440 Current Drug Delivery, 2006, Vol. 3, No. 4 Khan et al.

the chlorpromazine was carried out by multiple emulsions
[156]. Chloroquine, antimalarial agent is a known bitter drug
and can be used as model drug for taste masking. Multiple
emulsions have masked bitter taste of chloroquine signifi-
cantly [157-159].
7.6. Enzyme Immobilization
In recent years, multiple emulsions have become a new
tool in the field of biotechnology for the immobilization of
various enzymes, proteins, amino acids etc. Enzymatic con-
version of highly lipophillic, water insoluble substrates like
steroids have also been carried out (Table 4).
[29]. Ferreira and coworkers developed w/o/w multiple
emulsions carrying metronidazole and glucose and compared
their percutaneous release with w/o, o/w emulsions. Absorp-
tion of metronidazole was similar from w/o/w and o/w emul-
sions and was lower from w/o emulsion whereas in case of
glucose absorption was in following order o/w>w/o/w>w/o
[171-173]. Stability of ascorbic acid was enhanced by incor-
porating in multiple emulsions [174].
Table 5. Multiple Emulsions for Topical Drug Delivery
Drug envisaged References

Table 4. Multiple Emulsions Utilized for Enzyme Immobili- Metronidazole 171, 173
zation
Glucose 172, 173

Dihydralazine 175
Enzyme envisaged References
Hydrocortisone 176
Alcohol dehydrogenase 160 Ascorbic acid 174
Urease 161 Benzalkonium chloride 177
L-Phenyl alanine 162
7.9. As a Technique for Microsphere/Microcapsule
L-Leucine dehydrogenase 163 Preparation
Lipase 164 Now-a-days multiple emulsion system has been recog-
nized an intermediate step in the formulation of micro-
7.7. Drug Over Dosage Treatment/Detoxification spheres. Basic technique for microencapsulation involves
two-step emulsification method for microsphere preparation.
Multiple emulsions has not been much investigated for Finally, microspheres can be prepared by gelation of external
the overdosage treatment. For the first time, Liquid mem- phase, utilizing one of the following method (1) crosslinking
brane was proposed for the drug overdosage treatment in monomer in external aqueous phase by γ-radiations (2) heat
1978 [165]. This system could be utilized for the treatment denaturation of proteins (3) non-solvent addition following
of acidic drug overdosage (Barbiturates). When emulsion is cross linking (4) salt or alcohol addition. A novel technique
administered orally, acidic pH of stomach becomes external of w/o/o double emulsion solvent diffusion method was used
aqueous phase where barbiturate exists mainly in unionized for encapsulating hydrophilic drugs [178]. Recently, uniform
form which would easily transfer through the oil phase into size microcapsules were successfully developed by combin-
the inner aqueous phase across the concentration gradient. ing glass membrane and multiple emulsion-solvent evapora-
Basic buffer in inner phase ionizes the barbiturate and pre- tion method [179]. Microspheres of several drugs has been
vents its backward transport. Thus, entrapping excess drug in prepared and different polymer (ethylcellulose, eudragit,
multiple emulsions treats over dosage. Overdosage treatment polylactic acid, polyglycolic acid etc.) has been utilized
of quinine sulfate has been reported [166]. Multiple emul- based on the nature of the drug to be encapsulated (Hydro-
sions have been used for detoxification of blood [167, 168]. philic or lipophilic) (Table 6).
7.8. Topical Applications
Only few studies have been reported on topical admini- Table 6. Multiple Emulsions Used for Microencapsulation
stration of multiple emulsions (Table 5). First published
study shows that multiple emulsions release their contents Drug envisaged References
more slowly than solutions [169]. A typical three-phase
emulsion containing model compounds testosterone, caffeine Sulfadiazine 180
and tritiated water was formulated and the effect of perfluro- Leuprolide acetate 181
polymethylisopropylether (Fomblin HCl) on percutaneous
absorption was investigated [170]. Fomblin decreased the Pseudoephedrine 182
percutaneous absorption of testosterone but increased water Salbutamol sulphate 183
permeation while in case of caffeine the flux through skin
was not affected. A topical w/o/w emulsion containing active Theophylline, Propranolol, Acetaminophen, Tacrine 178
substance in each phase: sodium lactate (moisturizing agent) Retinol 184
in inner phase; spironolactone (anti acne agent) in intermedi-
ate oily phase; and chlorhexidine digluconate (antibacterial) Diclofenac sodium 185
in outer phase was prepared to understand formation mecha- Lysozyme 186
nisms. The compatibility of these ingredients in the system
was evaluated and release of sodium lactate was observed PTSA 187
Multiple Emulsions Current Drug Delivery, 2006, Vol. 3, No. 4 441

7.10. In Cosmetics [3] Salager J.L. In Pharmaceutical Emulsions and Suspensions, Drugs
and Pharmaceutical Sciences Series, Marcell & Dekker Inc., New
Emulsions are most useful agents in cosmetics and toilet- York, 2004, Vol. 105, pp. 19-71.
ries, where most formulations are based on either o/w or w/o [4] Israelachvili, J. Colloids Surf. A, 1994, 91, 1.
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Received: February 28, 2006 Revised: June 19, 2006 Accepted: June 29, 2006