MULTIPLE EMULSIONS:

Multiple emulsion are complex system which consist of

both w/o and o/w at the same time.
There are potential matrices for the encapsulation of
bioactive compounds and for the controlled release
compounds
w/o/w multiple emulsion are system where small water
droplets are entrapped within larger oil droplets and they
are disperse continuous water phase
Because of presence of reservoir phase that can be use to
prolong release of active ingredients.

 This is made possible by double emulsification hence

the
systems are also called as Double
emulsions.
These are also called as Liquid membrane systems
as the liquid film which separates the liquid phases
acts as a thin semi permeable film through which
solute must diffuse in order to transverse from one
phase to another.
Multiple emulsions are thermodynamically
unstable.
They are often stabilized by using a combination of
hydrophilic & hydrophobic surfactants.
The ratio of these surfactants is important in achieving
stable multiple emulsions.

 TYPES OF MULTIPLE EMULSIONS

1. Water/Oil/Water (w/o/w)
2. Oil/Water/Oil
(o/w/o)

W/O/W Type of emulsion

 WATER/OIL/WATER (W/O/W)
Water/Oil/Water (w/o/w) multiple emulsions consist of
dispersed oil globules containing smaller aqueous
droplets; each inner aqueous droplet is separated from
the outer aqueous phase by an oil phase layer.
The presence of atleast two surfactants is required.
One of them is predominantly Lipophilic for stabilizing
the primary w/o emulsion & the other is Hydrophilic for
the secondary o/w emulsion.

O/W/O type of emulsion

 OIL/WATER/OIL(O/W/O)
Oil-in-water-in-oil (o/w/o) multiple emulsions contain an
inner oil phase, a water phase, and outer oil phase.
The inner oil phase is first dispersed in water to form an oilin-water (o/w) emulsion.
Then the o/w emulsion is further dispersed in the outer oil
phase to form the o/w/o type multiple emulsion.

Formulation of Multiple
Emulsions

(1) emulsification equipment;

(2) nature of the oil phase;
(3) volumes of the two dispersed phases;
(4) nature and quantity of the
emulsifying agents;
(5) nature of entrapped materials,
including the drug substance; and
(6) added stabilizing components
(gelling agents, etc.).

Formulation of Double
Emulsion
o The formulate a double emulsoin , it is necessary to
choose ,at least , an oil and two surfactants ,one low in
HLB and one high in HLB. eg. span surfactant(HLB<5)
and Tween surfactant(HLB>10) and with a vegetable oil
(caprylic / capric triglyceride)
o The first stage involves making state diagram that
provide the means for pre-selecting formulas . Those
selected for in-depth study are the very while emulsions
that cream slowly and/or 0moderately.
o HLB (blend)=f * HLB (A)+(1-f)*HLB (B)

 Multiple emulsion w/o/w contain two

emulsifiers:
1 Low HLB surfactant (Hydrophobic in
nature)
2 High HLB surfactant (Hydrophilic in
nature)
Low HLB surfactant use in disperse phase
High HLB surfactant use in continuous
phase.

EMULSIFYING AGENTS
Emulsifier or surface active agent
[SAA] is molecule which has two parts,
one is hydrophilic and the other is
hydrophobic. Upon the addition of
SAA, it tends to form monolayer film at
the oil/ water interface.

surfactants

Anionic

Sodium oleate
Pottasium
oleate
Glyceryl
monostearate
cationic
SLS
Cetrimide

Non ionic

span 80
span20
Tween 20
Tween 60
Tween 80

 Different techniques used for the

preparation of Multiple Emulsions
Two step emulsification technique,
Phase inversion technique,
Membrane Emulsification Technique,
Latest
Design of Double Emulsion by Osmotic Pressure Tailoring,
Micro encapsulation of Multiple-layer Emulsion with high-voltage
electrostatic field.

TWO STEP EMULSIFICATION

TECHNIQUE

This method involve re-emulsification of primary W/O or

O/W emulsion using a suitable emulsifier agent.

Multiple emulsions, either W/O/W or O/W/O emulsions,

are generally prepared using a Two-step procedure, the

primary emulsion (W/O) is first prepared using water
solution in oil.

and a low-HLB surfactant

In the second step, the primary emulsion (W/O) is

reemulsified in an aqueous solution of a high-HLB

surfactant to produce a W/O/W multiple emulsion.

 The first step-that is, the preparation of the primary

emulsion-is usually carried out in a high-shear device
to produce a very fine droplets.
The second emulsification step is carried out in a lowshear device to avoid rupturing the multiple droplets.

TWO STEP
EMULSIFICATION/
DOUBLE
EMULSIFICATION

 EXAMPLE
The primary W/O emulsion was prepared by adding an aqueous
solution containing sodium salicylate to a Span 83 solution in light
mineral oil at an equal volume ratio and stirring with a magnetic
mixer (1000 rpm) for 15 minutes.
The concentration of Span 83 in light mineral oil varied from 0.1% to
40% wt/vol.
In second step, the W/O primary emulsion was reemulsified in a
Tween 80 solution containing concentrations of Twee80 from 0.1%
to 10% wt/vol at an equal volume ratio and stirred for 5 minutes at
600 rpm to produce the W/O/W multiple emulsion.

Modified two-step emulsification

Recently ,a Modified two-step emulsification technique for the preparation

of W/O/W emulsion was reported.

This method is different from the conventional two-step technique.

Firstly, sonicated and stirring are used to produce fine, homogenous and
stable W/O emulsion.

Secondly, a continuous phase is poured into a dispersed phase for

preparing W/O/W emulsion.

The composition of internal aqueous phase-oily phase-external phase

is fixed at 1:4:5,which produces most stable formulation of W/O/W
emulsions.

 PHASE INVERSION TECHNIQUE

The development of W/O/W system during the phase

inversion of the concentrated W/O emulsion.
In this technique, an increase in volume concentration of
dispersed phase may increase in phase volume ratio,
which subsequently leads to the formation of multiple
emulsion.
The method typically involves the addition of an aqueous
phase containing the hydrophilic emulsifier examples
-Tween 80 or
-Sodium dodecyl sulphate or
-CTAB (Cetyl trimethylammonium bromide)
to an oil phase consisting of liquid paraffin and
containing lipophilic emulsifier ex Span 80.

A well-defined volume of oil phase is placed in a

vessel of pin mixer.

An aqueous solution of emulsifier is then

introduced to the oil phase in the vessel at a rate
of 5ml/ min, while the pin mixer rotates steadily
at 88 rpm at room temperature.

When volume fraction of the aqueous solution of

hydrophilic emulsifier exceeds 0.7, the continuous
oil phase is substituted by the aqueous phase
containing a number of the vesicular globules
among the simple oil droplets, leading to phase
inversion and formation of W/O/W multiple
emulsion.

MEMBRANE EMULSIFICATION
TECHNIQUE

This method uses low shear forces to produce emulsions.

A porous glass membrane with controlled and homogenous
pores is used in this method.
Particle size of the emulsion can be controlled with the
proper selection of the porous glass membrane.

This is based on the use of microspores with a very narrow

pore size distribution on the membrane.

The phase which is to be dispersed is pressed through

membrane pores.

The droplets formed at the membrane

surface are detached by the continuous
external aqueous phase flowing across the
membrane surface.
To support the emulsification and prevent
coalescence of droplets, a surface active
compound must be added to the continuous
phase.
It can be successfully applied to make
multiple emulsions as drug delivery
systems.

Stability of Multiple Emulsions

Emulsion stability is a phenomenon, which depends upon the equilibrium
between water, oil and surfactant. Unfortunately multiple emulsions are
thermodynamically unstable. The possible indications of instability
includes:
Leakage of the contents from the inner aqueous phase.
Expulsion of internal droplets in external phase.
Constriction or distension of the internal droplets due to osmotic gradient
across the oil membrane.
Flocculation of internal aqueous phase and multiple emulsion droplets.
Disruption of oil layer on the surface of internal droplets.
Phase separation.

Stabilization of multiple
emulsions
1) coalescence of the internal aqueous
droplets;
2) coalescence of the oil droplets;
3) rupture of the oil film resulting in
the loss of the internal aqueous
droplets, and
4) passage of the water and watersoluble substances through the oil
layer between both water phases

Methods to Stabilize Multiple

Emulsions
The followings are some of the attempt or
studies made to
restore or strengthen the stability of multiple
emulsions :
Liquid crystal stabilized multiple emulsion.
Stabilization in presence of electrolytes.
Stabilization by forming polymeric gel.
Stabilization by interfacial complexation
between
non- ionic surfact andmacromolecules.
Stearic stabilization

EVALUATION OF MULTIPLE
EMULSION: In-vitro characterization
1) Average globule size and size
distribution,
2) Area of interfaces
3) Number of Globules
4) Creaming volume measurement
5) Conductivity test,
6) Rheological Evaluation,
7) Measurement of Zeta potential.

Average Globule Size and

Size Distribution
o The optical microscopy method using calibrated ocular and stage
micrometer can be utilized for globule size determinations of both multiple
emulsion droplets as well as droplets of internal dispersed phase.
o Bright field micrographs equipped with differential interference contrast
optics have been used to characterize the internal droplet of multiple
emulsions.
o Various other techniques used to characterize colloidal carriers like Coulter
counter, freeze-fracture electron microscopy and scanning electron
microscopy are also used to determine average globule size and size
distribution of multiple emulsions.
o Recently, NMR self-diffusion methods are adapted to multiple emulsion
characterization.

Area of Interfaces

o The average globule diameter determined can be

used in the calculation ofthe total area of
interface using the formula
S = 6/d
where,
S =Total area of interface (sa, cm)
d= Diameter of globules (cm)

Number of Globules
Number of globules per cubic meter can be
measured using the haemocyto meter cell. The
globules in five groups of 16 small squares(total 80
small squares) are counted and the total number of
globules in per cubic mm are calculated using the
formula (Chatterj ee, 1985) :

No.of globules/mm3= no.of globuledilution4000

no. of small squares
counted

Rheological Evaluation
The rheological of multiple emulsions is an important
parameter as it relates to emulsion stability and clinical
performance.
The viscosity and interfacial elasticity are two major
parameters, which relate to product rheology. The viscosity of
the multiple emulsions can be measured by Brookfield
rotational Viscometer.
Interfacial rheology (i.e., interfacial elasticity at the oil-aqueous
interface)can be investigated at the mineral oil/water interface
using an Oscillatory Surface rheometer.

Creaming volume measurement

The creaming volume was defined as
the relative difference in volume of the
multiple emulsion and the volume of
the creamed phase.

 Conductivity test
This test was found to be an important parameter to
study the stability and yield of W/O/W emulsions.
The conductivity was determined by means of a
systronics digital conductivity meter.
The entrapment percent / yield value (E%)
was calculated according to the following equation
E% = 100 . (Ci Ct )/ Ci
Ci = conductivity of the internal aqueous phase
Ct = conductivity value of multiple emulsion at a
given time t.

Zeta Potential
The zeta potential measurements are pivotal in the designing of
surface modified orligand anchored multiple emulsion systems.
The zeta potential and surface charge can be calculated using
Smoluchowskis equation from the mobility and electro phoretic
velocity of dispersed globules using the Zeta-potentiometer. Zeta
potential was calculated using following formula:

= 4 103
E

Where,
Zeta potential( mv )
Viscosity of the dispersion medium(poise)
Migration velosity (cm/s)
Di electric constant of the dispersion midium
Potential gradient(voltage applied/distance b/w electrodes)

Percent Drug Entrapment

Percent entrapment of drug or active moiety in
the multiple emulsion is generally determined
using dialysis, centrifugation, filtration and
conductivity measurements.
The% Entrapment can be calculated using the
following equation :

In Vitro Drug Release

The drug released from the aqueous inner
phase of a W/O/W emulsion can be
estimated using the conventional dialysis
technique.
Aliquots were withdrawn at different time
intervals and estimated using standard
procedure and the data were used to
calculate cumulative drug release profile.

In Vitro Stability Studies

Emulsion stability is determined by phase separation on
storage of W/O/W emulsions .
Freshly prepared multiple emulsion allowed to stand for
one week at room temperature and the volume of
aqueous phase separated ( Vsep ) is measured at
suitable time intervals and percent phase separation is
calculated using following formula:

Drug Release Mechanisms & Models

Some of the mechanisms includes :
Diffusion of unionized drug (hydrophobic species) through the
oil layer
Carrier medicated transport
Micellar transport
Rupture of oil membrane
Thinning of oil membrane

APLLICATONS OF MULTIPLE EMULSIONS

Multiple emulsions finds wide range of applications in
controlled or sustained drug delivery, targeted
delivery, taste masking, bioavailability enhancement,
enzyme immobilization, etc.
It will be able to provide a novel carrier system for
drugs, cosmetics and pharmaceutical agents.

Applications in Therapeutics
&Cosmetics
Multiple emulsion systems are finding unlimited
uses because of theirvesicular structure with
innermost phase closely similar to that of
liposomalvesicles and the selective
permeability characteristic of liquid membrane.

Biomedical & Pharmaceutical Applications of Multiple Emulsions

APPLICATIONS
ENTRAPPED
Enhanced oral bioavailability
Masking action
Drug over dosage treatment
Barbiturates,
Quinine Sulphate
Vaccine adjuvant
Separation and extraction technique
in the fabrication of micro capsulated
dosage form
In cancer therapy and drug targeting
Bleomycin
Other applications

DRUG
Insulin
Chloroquin
Salicylates ,

Influenza virus
Different hydrocarbons
Diclofenac sodium
5-Fluorouracil,
Food and cosmetics

1. Hemoglobin multiple emulsion having specified properties

is suitable for provision of oxygen as a blood substitute
and other oxygen transfer processes.
2. W/O/W multiple emulsions are systems of potential
interest in the oral administration of insulin. Although it
has been shown that a single oral administration of
insulin-loaded W/O/W multiple emulsion to diabetic rats
led to the significant decrease of blood glucose.
3. (w/o/w) multiple emulsions and polymeric nanoparticle
formulations containing influenza virus surface antigen
hemagglutinin (HA) are thought to be suitable carriers for
a vaccine delivery system.

4. Using a water-in-oil-in-water multiple emulsion system

developed for pulmonary drug targeting, the
effectiveness of tetrandrine as an antifibrotic agent.
5. To develop a prolonged and sustained release
preparation , an albumin micro-sphere-in-oil-in-water
emulsion was prepared. Tegafur was used as a
model drug.
6. Vitamin c has been widely used in formulations of
skin care products.
7. Multiple emulsions are also used in topical
application ex a w/o/w emulsion of
Metronidazole.