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Content Introduction Classification Defination and synonyms Structure of Nanoemulsion Physical properties of a Nanoemulsion Methods of preparation Components of Nanoemulsion Evaluation of Nanoemulsion Application Conclusion References 2

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Nanotechnology C omprises technological developments on the nanometer scale, usually 0.1 to 1000 nm . The pharmaceuticals  developed on the basis of nanotechnology are termed as ‘ NANOPHARMACEUTICALS ’. Introduction

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Emulsion: “They are thermodynamically unstable system consisting of atleast two immissible liquid phase, one of which is dispersed as a globules in the other liquid phase which is continuous phase.” 4


CLASSIFICATION : It is based upon the nature of Dispersion Phase, Oil-in-water Emulsion(o/w) (0.1-100 µm) Water-in-oil Emulsion (w/o) (0.1-100 µm) Micro Emulsion (0.01 µm) Nano Emulsion (0.1-0.5 µm) 5

The various nanopharmaceuticals currently being used : _:

The various nanopharmaceuticals currently being used : _ Nanoemulsion Nanosuspension Nanospheres Nanoshells Nanocapsules Lipid Nanoparticle Dendrimers 6


Nanoemulsion “Nanoemulsion can be defined as a oil in water(o/w) emulsion with mean droplet daimeters ranging from 50 to 1000nm.Usually the average droplet size is between 100-500nm.” Synonyms Sub micron size emulsion Mini emulsion Ultrafine Emulsion 7

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transparent or translucent O/W or W/O emulsion droplet diameters ranging from 50-1000 nm . [ avg droplet size is between 100-500 nm ] kinetically stable unlike microemulsions which are thermodynamically stable Nanoparticles can exist core of particle w/o form water o/w form oil Ostwald ripening is the primary instability process : Can be reduced by the addition of a second less soluble oil phase and/or addition of a strongly adsorbed and water insoluble polymeric surfactant.

Single nanoparticle:

The inner core consisting of any oil Drug or nutrient molecules within the core The emulsifier layer surrounding the nano-particle . Single nanoparticle Nanoemulsion : Lipid monolayer enclosing a liquid lipid core. Liposome : Lipid bilayer enclosing an aqueous core.

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Structure of Nanoemulsion 10 Fig-1 Fig-2 Fig-3

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11 Types of Nanoemulsion

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Physical Properties of Nanoemulsion The relative transparency of nanoemulsion . Their response to mechanical shear or ‘rheology’. The enhanced shelf stability of nanoemulsion against gravitationally driven creaming. 13

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A nanoemulsion (a) and A macroemulsion (b) with droplet diameters of less than 100 nm and more than 1000 nm, respectively. Nanoemulsion and Macroemulsion 14

Difference between Emulsion and Nanoemulsion:

Difference between Emulsion and Nanoemulsion Themodynamically unstable. Millky appearance. Droplet upto few micrometer. Thermodynamically stable. Transluscent,isotropic. Droplet 0.1-0.5 µm. Emulsion Nanoemulsion 15

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Advantages Of Nanoemulsions 1 . R eduction of globules as the potential to: Increase surface area Enhance solubility Increase oral bioavailability More rapid onset of therapeutic action Decrease the dose needed 2 . They do not show the problems of inherent creaming, flocculation, coalescence and sedimentation . 3. They are non-toxic, non-irritant hence can be easily applied to skin and  mucous membranes. 4 . T hey can be taken by enteric route becoz they are formulated with surfactants, which are approved for human consumption (GRAS),

Limitations of Nanoemulsion:

Limitations of Nanoemulsion The manufacturing of Nanoemulsion is an expensive process. Stability of Nanoemulsion is a unacceptable and creates a big problem during the storage of formulation for the longer period time. Less availability of surfactant and cosurfactant required for the manufacturing of nanoemulsion . 17

Preparation of Nanoemulsion:

Preparation of Nanoemulsion Drug poorly water soluble drug eg : CBZ,Diclofenac,Ramipril Oil phase eg : dimethicone oil , castor oil , Soyabean oil Aqueous phase Surfactant Cremphore , lecithin Cosurfactant : Propylene glycol, polysorbate 80, cetylphosphate,hydrogenated caster oil

Representative pseudoternary phase diagram of surfactant and cosurfactant (Smix) mixture showing oil/water nanoemulsion area (shaded area):

Representative pseudoternary phase diagram of surfactant and cosurfactant (Smix) mixture showing oil/water nanoemulsion area (shaded area)

Thermodynamic Studies:

Thermodynamic Studies Prepare number of formulations Formulations centrifuged for specific period. select stable formulation kept under heating and cooling cycle. select stable formulation subjected to a freeze-thaw cycle test select stable formulation for further mfg

Methods Of Preparation Of Nanoemulsions::

Methods Of Preparation Of Nanoemulsions : High-Energy 1. High pressure homogenization 2 Microfludization 3 Ultrasound energy Undesirable for labile drugs and macromolecules (proteins and nucleic acids) (for lab & industrial preparation) Low-Energy 1. Spontaneous 2 . Solvent-diffusion 3 . Phase inversion temperature (PIT) a. Rapid cooling of selected ME state b. Dilution with water (for lab preparation)

1.High-Pressure Homogenisation :

1.High-Pressure Homogenisation In a high-pressure homogenizer, the dispersion of two liquids (oily phase and aqueous phase) is achieved by forcing their mixture through a small inlet orifice at very high pressure (500 to 5000 psi), which subjects the product to intense turbulence and hydraulic shear resulting in extremely fine particles of emulsion.

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Dia : High-Pressure Homogenisation Advantage : produce nanoemulsions of extremely low particle size (up to 1nm). Disadvantage : high energy consumption and increase in temperature of emulsion during processing.

2.Microfluidization :

2.Microfluidization patented mixing technology, use of a device called microfluidizer . This device uses a high-pressure positive displacement pump (500 to 20000psi), which forces the product through the interaction chamber, which consists of small channels called ‘ microchannels ’. The product flows through the microchannels on to an impingement area resulting in very fine particles of sub-micron range.

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Coarse emulsion The two solutions (aqueous phase and oily phase) are combined together Interaction chamber Microfluidizer Filtrations desired particle size Stable & uniform nanoemulsion .

Lab scale preparation::

Lab scale preparation: High shearing Energy

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Solvent evaporation method 16

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Phase inversion method 29

Stability of Nanoemulsion:

Stability of Nanoemulsion Brownian movement Low rate of sedimentation – due to the small particle size low gravitational force As particle s become smaller, the attractive force of van der waals will be smaller. High Zeta potential- more electrostatic repulsive forces ;resulted in reducing the coalescence or coagulation of emulsion droplets Optimum Thermodynamically stability ;but high kinetic stability

Evaluation Of Nanoemulsion:

Evaluation Of Nanoemulsion

Evaluation parameters of Nanoemulsion:

Evaluation parameters of Nanoemulsion A) A verage Globule Size And Size Distribution:- Method used are-a)Transmission electon Mcroscopy. b)Droplet size analysis. c)Light scattering. 32 B) Rheological Evaluation :- 2 major parameters : -Viscosity. -Refractive Index.

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C ) Zeta Potential : - It is used to determine surface charge by the help of mobility & electrophoretic velocity of dispersed globules. 33 D) Area Of Interfaces : It can be determined by following formula. S = 6/d where, S = Total area of Interface ( d = Diameter of Globules (cm) E Analytical test (UV,HPLC) F Biological studies G In vitro drug release

Transmission Electron Microscopy :

Transmission Electron Microscopy Morphology and structure of the nanoemulsion were studied using transmission electron microscopy (TEM) To perform the TEM observations, a drop of the nanoemulsion was directly deposited on the holey film grid and observed after drying

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eg:TEM positive image of aceclofenac nanoemulsion showing the size of some oil droplets. . Scanning Electron Microscopy picture of nanoemulsion .

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Droplet size Analysis Determined by photon correlation spectroscopy that analyzes the fluctuations in light scattering due to Brownian motion of the particles, using a Zetasizer 1000 HS (Malvern Instruments) Viscosity determination Brookfield viscometer Refractive Index Abbe -type refractometer Zeta potential measurement The zeta potential was measured by electrophoretic mobility using Malvern Nanosizer / Zetasizer ® nano -ZS ZEN 3600 (Malvern Instruments, USA). Biological studies Skin Irritation Test Plasma-time profile

Applications of Nanoemulsion:

Applications of Nanoemulsion Use of Nanoemulsion in cosmetics. Antimicrobial Nanoemulsion. Nanoemulsion as Non-toxic disinfectant cleaner. Nanoemulsion in cell culture technology. NE as a vehicle for Transdermal drug delivery. NE in cancer therapy and targeted drug delivery. Nanoemulsion in the treatment of various other disease condition Nanoemulsion as a mucosal vaccines. Nanoemulsion as a vehicle for a occular delivery 37

1.Use Of Nanoemulsions In Cosmetics :

1.Use Of Nanoemulsions In Cosmetics Due to their lipohilic interior, nanoemulsions are more suitable for the transport of lipophilic compounds High skin penetration due to small size Body Moisturiser Nanoemulsion Face Lotion with Vitamin Nanoemulsion Face Cream for Night Use with Vitamin Nanoemulsion

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2.Antimicrobial Nanoemulsions Antimicrobial nanoemulsions are oil-in-water droplets that range from 200-600 nm. The nanoemulsion has a broad spectrum activity against -bacteria (e.g., E. coli, Salmonella, S. aureus ), -enveloped viruses (e.g., HIV, Herpes simplex), -fungi (e.g., Candida, Dermatophytes ), -spores (e.g., anthrax). nanoemulsion particles are fuse with lipid-containing organisms. Due to electrostatic attraction between the cationic charge of the emulsion and the anionic charge on the pathogen active ingredient and the energy releases destabilize the pathogen lipid membrane cell lysis and death

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In the case of spores

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A unique aspect of the nanoemulsions is their selective toxicity to microbes at concentrations that are non-irritating to skin or mucous membrane. The safety margin of the nanoemulsion is due to the low level of detergent & have sufficient energy in each droplet to destabilize the targeted microbes without damaging healthy cells. As a result, the nanoemulsion can achieve a level of topical antimicrobial activity that has only been previously achieved by systemic antibiotics.

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Nanoemulsions as a prophylactic medication A human protective treatment, to protect people exposed to bio-attack pathogens such as Anthrax ,Hepatitis and Ebola . The technology has been tested on gangrene and clostridium botulism spores and even used on contaminated wounds to salvage limbs

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3.Nanoemulsions As Mucosal Vaccines (Under Trial) Nanoemulsions are being used to deliver either recombinant proteins or inactivated organisms to a mucosal surface to produce an immune response. nanoemulsion causes proteins applied to the mucosal surface facilitates uptake by antigen presenting cells systemic and mucosal immune response that involves the production of specific IgG and IgA antibody as well as cellular immunity The first applications, an influenza vaccine and an HIV vaccine, can proceed to clinical trials.

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4.Nanoemulsions In Cell Culture Technology Cell cultures are used for in vitro assays or to produce biological compounds, such as antibodies or recombinant proteins it has been very difficult to supplement the media with oil-soluble substances , only small amounts of these oil-soluble substances ( lipophilic compounds ) could be absorbed by the cells. Nanoemulsions are a new method for the delivery of oil-soluble substances to mammalian cell cultures. These nanoemulsions are transparent and can be passed through 0.1 µm filters for sterilization. Nanoemulsion droplets are easily taken up by the cells. The advantages of using nanoemulsions in cell culture technology are · Better uptake of oil-soluble supplements in cell cultures. · Improve growth and vitality of cultured cells. · Allows toxicity studies of oil-soluble drugs in cell cultures

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Nanoemulsion in cancer Therapy

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“ Anti-oxidant Synergy Formulation”(ASF) suppresses malignancy: Nanoemulsions improve its effectiveness in culture ASF forces differentiation of neuroblastoma (most common solid tumor in children)...Like neurons, if they begin to differentiate, they no longer multiply

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The neuroblastoma in the nude mice Tumor No Tumor Control (no treatment) ASF Nano -emulsion

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Nanoemulsion delivery of Tamoxifen to breast cancer cell lines

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The Effect of Tamoxifen (T ) and Nanoemulsion Preparation of Tamoxifen (NT) on Cell Proliferation Control (no treatment) Tamoxifen (T) Nanoemulsion Preparation of Tamoxifen Days of Culture

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Nanoemulsion delivery of anti- inflammatory agent Aspirin in mice

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Parenteral nanoemulsions Carbamazepine IV injection (Under trial) widely used anticonvulsant drug, is a poorly soluble drug with no parenteral treatment available for patients. Solubility of drug increases by decreasing the particle size upto nanometer treatment available for patients i.e. CBZ nanoemulsion given by IV route

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O ral lipid nanoemulsion of primaquine Primaquine is one of the most widely used antimalarial and is the only available drug till date to combat relapsing form of malaria especially in case of Plasmodium vivax and Plasmodium ovale . application of PQ in higher doses is limited by severe tissue toxicity including hematological and GI related side effects which are needed to be minimized. when incorporated into oral lipid nanoemulsion having particle size in the range of 10–200 nm showed effective antimalarial activity against Plasmodium infection in swiss albino mice at a 25% lower dose level as compared to conventional oral dose.

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PATENTED NANOEMULSIONS Some important patents related to nanoemulsions : 1.Patent name: Method of Preventing and Treating Microbial Infections.Assignee : NanoBio Corporation (US) US Patent number: 6,506,803 2. Patent name: Non-toxic Antimicrobial Compositions and Methods of Use.Assignee : NanoBio Corporation (US) US Patent number: 6,559,189 and 6,635,676, 3. Patent name: Nanoemulsion 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 4. Patent name: Nanoemulsion based on oxyethylenated or non- oxyethylenated sorbitan fatty esters, and its uses in the cosmetics, dermatological and/or ophthalmological fields. Assignee: L'Oreal (Paris, FR) US Patent number: 6,335,022

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5. Patent name: Nanoemulsion 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 6. Patent name: Nanoemulsion based on glycerol fatty esters, and its uses in the cosmetics, dermatological and/or ophthalmological fields. Assignee: L'Oreal (Paris, FR) US Patent number: 6,541,018 7. Patent name: Nanoemulsion based on sugar fatty esters or on sugar fatty ethers and its uses in the cosmetics, dermatological and/or ophthalmological fields . Assignee: L'Oreal (Paris, FR) US Patent number: 6,689,371 8. Patent name: Transparent nanoemulsion less than 100 NM based on fluid non-ionic amphiphilic lipids and use in cosmetic or in dermopharmaceuticals.Assignee : L'Oreal (Paris, FR)US Patent number: 5,753,241


Conclusion NE formulation offer several advantages for the delivery of drugs, biologicals, or diagnostic agents. Less availability of surfactant and cosurfactant required for the manufacturing of nanoemulsion . Nano-emulsion has a various application as a drug delivery systems because of their capacity of solublizing non-polar active compound. There is no success for a delivery system if only academic research groups are developing it. Success can only be possible if also pharmaceutical industry takes up developments. 57


References Vyas S.,Khar R., 2002, Targeted And Controlled Drug Delivery System , 1 st Edition, CBS Publication, 303-329. Jain N.K, 2001, Controlled And Novel Drug Delivery , 1 st Edition, CBS Publication,381-399. R.S.R. Murthy, Vesicular and Particulate Drug Delivery Systems, 1 st Edition,Career Publication,105-140. Leon Lachman,Herbert A.Liberman,Joseph L.Kanig,The theory and practice of industrial pharmacy,Varghese publishing house,Third edition,502-545. Monzer Fanun,Colloids in the drug delivery,CRS press group,221-244. Website : 58

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Thank You 59

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