Nanoemulsion

THE IDEA
Proposed by Richard Feynman
in his book titled Theres Plenty
of Room at the Bottom
Feynman considered the
possibility of direct manipulation
of individual atoms as a more
powerful form of synthetic
chemistry than those used at the
time.
The idea of nanotechnology was
born.

contents:

contents
Introduction
Advantages & Disadvantages
Formulation aspect of nanoemulsion
Preparation of nanoemulsions
Characterization of nanoemulsion
Applications of nanoemulsion
Marketed formulations
Conclusion

INTRODUCTION
The term "Nanoemulsion" refers to a
thermodynamically
stable
isotropically
clear dispersion of two immiscible liquids,
such as oil and water, stabilized by an
interfacial film of surfactant molecules. A
Nanoemulsion is considered to be a
thermodynamically or kinetically stable
liquid dispersion of an oil phase and a
water phase, in combination with a
surfactant.
The dispersed phase typically comprises small
particles or droplets, with a size range of 5 nm200 nm, and has very low oil/water interfacial
tension.
5

NANOEMULSION
Nanoscale emulsion having size less than 100nm.
Due to their small droplet size, nano-emulsions may
appear transparent, and Brownian motion prevents
sedimentation or creaming, hence offering increased
stability.
In contrast to microemulsions, nanoemulsions are
metastable and can be diluted with water without changing
the droplet size distribution.
Nanoemulsion are thermodynamically stable system in
which the two immiscible liquid (water and oil) are mix to
form a single phase by means of appropriate surfactant .
6

Nanoemulsions
Emulsion
Thermodynamically unstable
Opaque
High energy required to form

Microemulsion
Thermodynamically stable
Clear
It forms spontaneously

Nanoemulsion
Thermodynamically or kinetically
stable
Clear
High shear application to form

Int. J. Nanomed. 2014,9,pp 1-8

The key difference between

emulsion,microemulsion and nanoemulsion
Emulsion
Microemulsion
Nanoemulsion
Thermodynamically
unstable

Thermodynamically stable Thermodynamically

stable Or Kinetically
stable

Appearance:
cloudy

Appearance: clear or
translucent

Form only after

application of the
large input energy

It forms spontaneously

Appearance:
clear or translucent
Form only after
application of the higher
shear

Nanoemulsions Classification

pending on the composition :

W : oil droplets dispersed in continuous aqueous phase

O : water droplets are dispersed in continuous oil phase

continuous : microdomains of oil and water are dispersed within the syste

J. Mat. Chem. 2012, 22, pp 9767-9773

Nanoemulsion:Lipi
d monolayer
enclosing a liquid
lipid core.

Liposome: Lipid
bilayer enclosing
an aqueous core.

10

Internal structures depend on relative

component amounts, concentrations and other
characteristics.

The relative oil and water domains that form in

nanoemulsion systems are usually so small (about
10-20 nm or less in diameter) that they do not
scatter light.

11

Nanoemulsions and
Subnanoemulsions
(1)nanoemulsion: d 10-100nm,
thermodynamic stability system
appearance
high Concentration of emulsifiers
(20-30% of dispersed phase)
mix with water and oil within
certain ranges
low viscosity
low surface tension
Formation mechanism has not been
clear completely.

(2)subnanoemulsion: d 100500nm, better stability when

compared with common
emulsions while poorer
stability when compared with
nanoemulsion
both can be used as drug
vehicles! sustained and
prolonged release!

 Macro emulsions are dispersions of at

least two non-miscible liquids.
They are thermodynamically unstable
systems that are stabilized kinetically.
Consequently, the stability of an emulsion
depends both on its composition and the
size the emulsion droplets.
Depending on the preparation method,
different droplet size distribution might be
achieved, explaining why the route of
preparation can have an influence on the
emulsion stability.

 Due to their small droplet size,

nanoemulsion may appear
transparent and Brownian motion
prevents sedimentation or creaming,
hence, offering increased stability.
The preparation of emulsions with
droplet sizes in the sub micrometer
range may be performed
mechanically, which involves highshear stirring, high-pressure
homogenizers or ultrasound
generators.

15

 DIFFERENCE BETWEEN MACROEMULSION

& NANOEMULSION
MACROEMULSIO
N
Thermodynamic Unstable but
ally
stabilized
kinetically.
Loading dose
Good
Preparation
Bottle method,
Method
wet gum method,
etc.,
Problems

Creaming,
flocculation and
sedimentation is
observed.

NANOEMULSIO
N
Stable

Poor
high-shear
stirring,
ultrasound
generators
No creaming,
flocculation and
sedimentation is
observed.
16

Method of preparation
1)High pressure homoginization:
By high pressure homoginizer or piston homoginizer which
produce NEs of exrtemly low particle size upto 1 nm.
2)Microfluidization:
This make use of microfluidizer.
This device use high pressure positive displacement
pump(500-20000 psi) which force the product through the
interaction chamber which consist of small micro channel.
Product flow throgh the micro channel on to the impigment
resulting in the formation of nano size droplet.

17

18

CHARACTERIZATION OF NANOPARTICALS
Nano-emulsions are not thermodynamically stable, and because of
that, their characteristics will depend on preparation method. Here
some parameters are discussed which should be analysed at the
time of preparation of nanoemulsion.
Phase Behavior Study

This study is necessary in characterization and optimization of

ingredients. This is used in case of NE formulation prepared by
phase inversion temperature method and self-emulsification
method.
Particle Size Analysis

Generally Dynamic Light Scattering(DLS) method are used.

Surface Charge Measurement

Surface zeta potential of NE droplets should be measured with.19

the

Contd
Transmission Electron Microscopy
TEM is used to observe the morphology in Nano-emulsion.
Viscosity
Viscosity should be measured to ensure the better delivery of
the formulation.
Morphology & structure
Morphology and structure of nanoemulsion can be studied
using TEM. The study of globule shape and surface can be
observed by TEM. To perform TEM observations, a drop of
the nanoemulsion is deposited on the holey film grid and
observed after drying.

20

Advantages of nanoemulsion
Nanoemulsions have a much higher surface area and free
energy than macroemulsions that make them an effective
transport system.
Are metastable and can be diluted with water without
changing droplet size distribution.
Do not show problems of inherent creaming, flocculation,
coalescence and sedimentation which are commonly with
marcoemulsions .
Can be formulated in variety of formulations such as foams,
creams, liquids and sprays.
Are non-toxic and non-irritant, hence, can be easily applied to
skin and mucous membranes.
Since nanoemulsions are formulated with surfactants, which
are approved for human consumption (generally regarded as
safe), they can be taken by enteric route.
Do not damage healthy human and animal cells, hence, are
suitable for human and veterinary therapeutic purposes. 21

Limitations Of Nanoemulsions
The manufacturing of nanoemulsion formulation is an expensive
process because size reduction of droplets is very difficult as it
required a special kind of instruments and process methods.
For example, homogenizer (instruments required for the
nanoemulsion formulation) arrangements is an expensive process.
Again microfluidization and ultrasonication (manufacturing process)
required high amount of financial support.
Stability of nanoemulsion is quite unacceptable and creates a big
problem during the storage of formulation for longer time of period.
Ostwald ripening is the main factor associated with unacceptability
of nanoemulsion formulations. This is due to high rate of curvature of
small droplets show greater solubility as compared to large drop with
22
a low radius of curvature.

Disadvantages
Nanoemulsions show poor drug
loading capacity.
Show drug expulsion after polymeric
transition during storage.
Relatively high water content of the
dispersions (70-99.9%) is observed in
nanoemulsion.

23

Nanoemulsions - Drug
Delivery

Advantages

Increase drug loading

Enhance drug solubility
Bioavailability
Controlled drug delivery
Protection of drug

Disadvantages

Expensive process
J. Phys. Chem. C2008,112(33),
Stability
12669-12676.
Solubility
Lack of understanding of interfacial chemistry
More proofs about more benefits compared to classical macroemulsions

Formulation additives

25

A Typical Formulation

26

Emulsifiers and coemulsifiers

(1)emulsifiers
natural ones: acacia, mucilage
tragacanth, gelatine, albumin,
casein, lecithin, SP, cholesterol
synthetic ones: Tweens,
poloxamers, Spans
(2)coemulsifiers: n-butanol,
ethylene glycol, ethanol,
propylene glycol, glycerin,
poly-glyceride

Preparation of nanoemulsion
(1)formation comditions
Emulsifiers :
20-30% of oil
adding coemulsifiers:
adjust the HLB value, insert the
interfacial film, form complex
aggregate film, enhance the degree
of rigidity and flexibility, further
reduce the interfacial tension,
increase the stability of
nanoemulsions

(2)steps of preparation
determine the formulation:
ternary phase diagram (p358)
note: keep constant
temperature
mixing according to the
determined ratios

preparation of subnanoemulsion
key instrument:
high pressure emulsifier
impact factors
stabilizer: oleic acid
enhance the strength of interfacial
film
increase the solubility of drug
increase the absolute value of
potential
complex emulsifiers:
lecithin+poloxamer

Method of preparation
High pressure homogeniser
Microfluidization
Phase inversion temperature
technique

31

Preparation of nanoemulsions
1. High pressure homogenization
. High pressure homogenization technique was initially
used for the production of solid lipid nanodispersions.
. This technique makes use of high pressure
homogenizer/piston homogenizer to produce
nanoemulsions of extremely low particle size (up to
1nm).
. There are two methods in high pressure
homogenization, i.e. hot homogenization and cold
homogenization.
. Higher stirring rates did not significantly change the
particle size, but slightly improved the polydispersity
index.
32

High-pressurehomogenization
This technique
makes use of
high-pressure
homogenizer/pis
ton homogenizer
to produce NEs
of extremely low
particle size (up
to 1nm)

33

2. Micro fluidization is a patented mixing technology, which

makes use of a device called micro fluidizer.
This device uses a high-pressure positive displacement
pump (500-20000 per square inch), which forces the product
through the interaction chamber, which consists of small
channels called micro-channels.
The product flows through the micro-channels which
resulting in very fine particles of sub-Micron range.
Micro fluidization The two solutions (aqueous phase and
oily phase) are combination together and processed in an
inline homogenizer to yield a coarse emulsion.
The coarse emulsion is into a micro fluidizer where it is
further processed to obtain a stable nanoemulsion.
The coarse emulsion is passed through the interaction
chamber of the micro fluidizer repeatedly until desired
particle size is obtained.
The bulk emulsion is then filtered through a filter under
nitrogen to remove large droplets resulting in a uniform
nanoemulsion .
34

MICROFLUIDIZATION:
It involves the use of device that is micro
fluidizer
It uses high-pressure positive displacement
pump of (500-20000)psi, which forces the
product through the interaction chamber, which
consists of small channels called micro
channels.
The product flows through the micro channels
on to an impingement area resulting in very fine
particles of submicron range. The two solutions
(aq. Phase and oily phase) are combined
together and processed to obtain a stable
nanoemulsion.
35

Microfluidization

www.ttlindia.com/images/microfluidics1.jpg

36

3. Phase inversion temperature technique: It involves

2 steps
STEP 1 result in the formation of a suitable system. This
procedure consists of magnetic stirring of all the
components-the ternary system- prepared in suitable
proportions of salt water(salt and water),non-ionic
surfactant and oil, with a rise from room temperature to
85 o c at a rate of 4 o c/minute.
A progressive cooling from 85 o c to 60 o c at a rate of 4 o
c/minute is performed.
Three temperature cycles (85-60-85-60-85 o c) are applied
to reach the inversion process.
STEP 2 is an irreversible shock induced by dilution with
cold water to the mixture maintained at the previously at
a defined temperature.
This fast cooling dilution process with cold distilled water
leads to nano objects. Afterwards, a slow magnetic stirring
37
is applied to the suspension for 5 minutes.

Quality evaluation
size and size distribution
drug concentration
stability

Nanoemulsions
Characterization/Evaluation

Surface characterization Morphology

Size, PDI

Refractive Index

Viscosity

Skin permeation

Characterization of
NEs:

11/26/15

Transmission eletron microscopy

Drolet size analysis
Viscosity
Refractive index
Skin irritation test
Thermodynamic stability study
Surface characteristics

Shakti Suthar

Characterization of nanoemulsion

Preliminary characterization such as

creaming, cracking or phase separation can be
observed visually for over 48 hours time period
in undisturbed standing conditions.
Morphology and structure Morphology and
structure of nanoemulsion can be studied
using TEM.
The study of globule shape and surface can be
observed by TEM.
To perform TEM observations, a drop of the
nanoemulsion is deposited on the holey film
grid and observed after drying.
41

Nanoemulsion droplet size

analysis
0.1ml of nanoemulsion is dispersed
in 50 ml (500 times dilution) of
double distilled water in a volumetric
flack and gently mixed by inverting
the flask.
Dynamic light scattering can be
performed at 25 o C, using heliumneon laser having particular
wavelength at an angle of 90 o on a
digital correlate.

42

 Refractive index The refractive index of nanoemulsion

formulation is determined using abbe type refractometer.
Viscosity determination The viscosity of the nanoemulsion
can be measured using Brookfield digital rheometer
temperature at 25 o C.

The surface charge can be determined by measuring the

zeta potential of the preparations . This is done by using an
aqueous phase containing NaCl and HCl and sodium
hydroxide to adjust the conductivity and the pH, respectively.
Zeta potential characterizes the surface charge of particle
and, thus, it gives information about repulsive forces
between particles and droplets. To obtain stable
nanoemulsions by preventing flocculation and coalescence of
the nano-droplets, zeta potential should usually reach a
value about 30mV.
43

Invitro skin permeation

studies

Franz diffusion cell

44

 Stability of nanoemulsion: A stability study of the

nanoemulsion can be performed as per ICH
guidelines. The samples were kept at fore different
conditions of temperature and relative humidity
(%RH) as 40 o C/75% RH, 30 o C/65% RH, 25 o
C/60% RH and refrigeration condition. The stability
is observed over a period of 3 months. The samples
are evaluated for particle size, viscosity, drug
content, surface charge, drug release, etc. Surface
charge .
Freeze-thaw cycling test can be carried out at
five alternative cycles each of 24 hour storage at
ambient and freezing conditions, high speed
centrifugation at various speeds starting from
2000 up to 5000 rpm each for 10 minutes.
45

Nanoemulsions Applications

Agriculture

Cleaning products

Cosmetic

Pharmaceutic

Biomedical

APPLICATIONS OF NANO-EMULSIONS
The compositional flexibility of nanoemulsions offers a wide range of
applications.
The incorporation of fluorescent dyes and other molecules into
nanoemulsions makes the interesting probes for exploring properties of living
cells and for drug delivery.
Nanoemulsion vaccine could inactivate and kill the virus and then
subsequently induce immunity to the virus that includes cellular immunity,
antibody immunity and mucosal immunity.
The deformable and liquid nature of the droplets may lead to discoveries of
new pathways for cellular uptake and dispersal. Both oil-soluble and watersoluble drug molecules can be incorporated into the nanodroplets of direct and
inverse nanoemulsions for potential pharmaceutical uses.
In the printing and data storage industries, one may imagine the resolution of
droplets.
47

Contd

In the personal care and food industries, nanoemulsions may provide

interesting alternatives as pleasantly transparent and soft solids that
possess plastic-like rheological properties. While being appealing from
an optical and rheological point of view, nanoemulsion also can deliver
moisturizers to the skin quite efficiently and also block ultraviolet
light without leaving a white residue.

The small size of the nano droplets will likely increase transport
efficiency of any active drugs or other molecules inside the droplets
across biological membranes, including the skin. Thus, nanoemulsions
may have significant applications in medical patches.

High-throughput production methodologies make nanoemulsions a

realistic commercial-scale alternative for diverse areas, including lotions
and pharmaceuticals.
48

 Applications Of Nanoemulsion
Use of nanoemulsions in cosmetics
Nanoemulsions as potential vehicles for
the controlled delivery of cosmetics and for
the optimized dispersion of active
ingredients, in particular, skin layers.
Due to their lipophilic interior, they are
more suitable for the transport of lipophilic
compounds than liposomes.
Similar to liposomes, they support the skin
penetration of active ingredients and thus,
increase their concentration in the skin.
Antimicrobial nanoemulsions
49

Prophylactic in bio-terrorism attack

Nanoemulsions as mucosal vaccine
Nanoemulsions as non-toxic disinfectant cleaner
Nanoemulsions in cell culture technology
Nanoemulsions in cancer therapy and in
targeted drug delivery
Nanoemulsions in the treatment of various
disease conditions
Nanoemulsions formulations for imported oral
delivery of poorly soluble drugs
Nanoemulsions as vehicle for transdermal
delivery SNEDDS(self nano-emulsifying drug
delivery systems)
50

Patented Nanoemulsions :
Some important patents related to nanoemulsions : Patent name:
Method of Preventing and Treating Microbial Infections.
Assignee: NanoBio Corporation (US).
US Patent number:6,506,803.
Patent name: nanoemulsions based on phosphoric acid fatty acid esters
and its uses in the cosmetics, dermatological, pharmaceutical, and/or
ophthalmological fields. Assignee: L'Oreal (Paris, FR).
US Patent number:6,274,150. Patent name: nanoemulsions based on
ethylene oxide and propylene oxide block copolymers and its uses in
the cosmetics, dermatological and/or ophthalmological fields.
Assignee: L'Oreal (Paris, FR). US Patent number: 6,464,990.
nanoemulsions of 5-aminolevulinic acid (6,559,183).
Assignee: ASAT AG Applied Science and Technology (Zug, CH). PCT
number: PCT/ EP99/08711. nanoemulsions of poorly soluble
pharmaceutical active ingredients and methods of making the same.
Patent no.: WO/2007/103294
51

Marketed formulations
Some commercially available
nanoemulsion formulations are:
Propofol (Dipivan);
Vitamins A,D,E,K (Vitalipid).

52

Marketed products:
Drug

Brand

Manufacturer Indication

Propofol

Diprivan

Astra zeneca

Dexamethazon
e

Limethasonn

Mitsubishi
Steroids
pharmaceutical
, Japan

Palmitate
alprostadil

Liple

Mitsubishi
Vasodilator
pharmaceutical
, Japan

Flubriprofen
axetil

Ropion

Kaken
NSAIDS
pharmaceutical
, Japan

Vitamines
A,D,E,K

Vitalipid

Fresenius
kabi,Europe

Anesthatic

Parenteral
nutrition
53

Commercially Available
Nanoemulsions

Conclusions

Nanoemulsions are receiving increasing attention as drug ca

Applicable for almost all routes of delivery

Wide range of molecules can be loaded

New technology could be developed to overcome the

oor absorption and miscibility of some phytopharmaceutica

Conclusion cont.
Nanoemulsion formulations offer several
advantages for not only the delivery of drugs,
but biological or diagnostic agents also.
Nanoemulsions can be formulated in a variety
of formulations such as foams, creams, liquids
and sprays.
Recently, nanoemulsions have been used in
several drug delivery applications such as
targeted drug delivery, mucosal vaccines, In
cell culture technology, anti-cancer, anti-HIV
therapy and diagnostic agents.
56