Microemulsions Nanoemulsions Livestock Products

1. NANOEMULSIONS/MICROEMULSIONS
TECHNIQUES FOR DELIVERY OF
NUTRACEUTICALS THROUGH
LIVESTOCK PRODUCTS
Dr Anita Katekhaye
PhD Scholar, CVSc, Hyderabad

2. NUTRACEUTICALS
BACKGROUND AND HISTORY
LIQUID-LIQUID SYSTEMS
MICROEMULSIONS/NANOEMULSIONS
CHARACTERIZATION OF NANOEMULSIONS
TECHNOLOGY ADVANTAGES
FABRICATION OF NANOEMULSIONS
NUTRACEUTICALS VIA DAIRY PRODUCTS
NUTRACEUTICALS VIA MEAT PRODUCTS
REFERENCES
TRENDS IN NUTRACEUTICALS & FUNCTIONAL FOODS

3. TRENDS IN FUNCTIONAL FOOD &
NUTRACEUTICALS
• Global food encapsulation market is projected to reach about $39
billion by the year 2015 (GIA) Global Industry Analysis, Inc
• Currently, functional food market of India is estimated at $70 billion
Or
• 4% of processed food market and is growing at 3 times rate
• Japan is the single largest market followed by US and Europe
• In India, around 20 companies have record of producing
nutraceuticals & marketing them globally
Ref: A. A. Patel & A. K. Singh (2012)

4. • In India, Nutraceuticals are marketed as Indian
System of Medicine Drugs (ISMD) under the Over-the-Counter
(OTC) category
• No clinical validation of their safety or efficacy is required if
therapeutically usefulness is mentioned in the text
• Nutraceutical foods are not subject to the same testing and
regulations as pharmaceutical drugs
Ref: A. A. Patel & A. K. Singh (2012)

5. WHY THERE IS NEED OF
FORTIFICATION ?
Fortification of widely consumed staple foods offer one of the
simplest and most practical methods to alleviate “Hidden
Hunger”
1. In 1930s Vitamin D was added to milk in US to help prevent rickets in
children
2. In several countries liquid milk fortification with vit A and vit D is
mandatory
3. Dried milk often fortified with Vit A and D, Calcium and Iron
4. Milk based infant formula and weaning foods are fortified with a range
of Vitamins, Minerals and other nutrients such as PUFA
Investigation is carried out at NDRI suggest possibilities of fortification of
liquid milk with Ca and Iron

6. NUTRACEUTICALS
• Nutraceuticals is a hybrid or contraction of
Nutrition and Pharmaceutical
• Reportedly, it was coined in 1989 by Dr Stephen L De Felice,
founder and chairman of the “Foundation for Innovation in
Medicine” in Medicine, New Jersey, USA
• Term commonly used synonymously for Designer foods,
Health foods, Fortified foods, Medifoods, Vita foods, Pharma
foods, Functional foods and Dietary supplements

7. WHAT ARE NUTRACEUTICALS ?
A nutraceuticals is any substance considered as a food or its part
which in addition to its normal nutritional value provides
health benefits including prevention of disease or promotion of
health
Or
Any non-toxic food component that has scientifically proven
health benefits, including disease treatment or prevention
Functional component of the food must be standardized in
nutraceuticals product and produced under good
manufacturing conditions (GMPs)

8. CLASSIFICATION
Carotenoids, Saponins, Tocopherol, Terpenes
Tannin, Lignin, Flavones, Isoflavons, Courmarins
Amino acid, Indoles, Isothiocyanates, Choline
Ascorbic acid, Oligosaccharides, Non-starch PS
n-3PUFA, CLA, MUFA, lecithin, Sphingolipids
Ca, Se, K, Cu, Zn
Probiotics, Yeast

9. Summary of major Lipophilic Nutraceuticals Components that need to be
delivered into foods
NAME TYPES NUTRITIONAL BENEFITS
Fatty Acids n-3 fatty acids, CLA,
Butyric acids
Coronary Heart Disease, Bone
health, Immune response disorders,
Weight gain, Stroke prevention,
Mental health, Cancer, Visual acuity
Carotenoids β- Carotene, Lycopene,
Lutein and Zeaxanthin
Cancer, Coronary Heart Disease,
Macular degeneration and Cataract
Antioxidants Tocopherols, Flavonoids,
Polyphenols
Coronary Heart Disease, Cancer and
Urinary Tract Disease
Phytosterols Stigmasterol, β- sitosterol,
Campesterol
Coronary Heart Disease
Vitamins Vit D and Oil Soluble Vit Prevent Rickets and Osteomalacia

10. HISTORY
• Since beginning of last century Australian housewives used
Water/Eucalyptus oil/Soap flake/White spirit mixtures to wash wool
• Schulman et al.,(1959) :
First used microemulsion to describe multiphase system (water,
oil, surfactant and alcohol which form transparent solution)
• Rodawald, 1928:
Discovered liquid waxes the first commercial microemulsions
• In late 1970’s and early 1980’s interest in microemulsions really step up
when it was recognized that such systems could improve oil recovery and
when oil prices reached levels
• Last 20 years great deal of progress in understanding the properties of
Microemulsions

11. EMULSION
E.g. Mayonaise (W/O),
Butter
E.g. Milk, Meat Emulsion,
Cream

12. BACKGROUND
• Wide variety of delivery systems have been developed to encapsulate
lipophilic functional component including
e.g. Simple solution, Association colloids, Emulsions, Biopolymer
matrices, Powders etc
• Emulsion based delivery systems are getting more attention now days-
1. Due to their targeted delivery
2. More absorption in human gut
3. Solid matrix material can be added in liquid food system
Emulsion offers a viable option for drug delivery:- order of selective uptake
lymphatic regions Oil in Water> Water in Oil> Aqueous solution
(Nishioka & Yoshino 2001)

13. STRUCTURAL
DESIGN
PRINCIPLES

14. Structural components selection depends upon:-
TARGET APPLICATIONS
FINAL CONTINUOUS PHASE
CHARACTERISTCS OF BIO-ACTIVE
INGREDIENTS

15. LIPIDS
• Lipids are predominantly non-polar substances that are highly
hydrophobic
• In food Industry major source of lipids are Tryglycerols which
may come from animal, fish or plant origins
• Used to solubilize non-polar lipophilic components in foods
• In Microemulsions Diglycerides are used
• Nanoemulsions requires long chain triglycerides (LCT) with
some degree of unsaturation (Prevent Oswald Ripening and
Health effect also)

16. SURFACTANTS
• Are surface-active molecules that consists of a hydrophilic head group
and a lipophilic tail group
• Functional performance depends on the molecular characteristics of
head and tail groups
Head group characterize – Anionic, Cationic,
Zwitterionic, Non-ionic
Tail group characterize- number of tails (One or Two)
Synonyms - Emulsifier or Amphiphilic Compounds

17. SUITABLE NON IONIC
SURFACTANTS INCLUDE
1. Polysorbates –
Polyethoxyethylene sorbitan monoesters,
Tween 20, Tween 40, Tween 60, Tween 65,
Tween 80
2. Sugar Surfactant –
Sucrose nanopalmitate, sucrose monolaurate,
Sucrose monostearate Crodesta F-160, Sucrose
monopalmitate
3. Polyoxamers –
Polyoxyethylene-polyoxypropylene block
co-polymers
e.g. Monolan, Lutrol etc

18. Preferably surfactant that acts synergistically with the hydrophilic
non-ionic surfactant to alter the interfacial curvature
This lowers interfacial tension, permitting easier emulsion formation
Suitable food grade co-surfactant include:
Sorbitan fatty acid esters such as sorbitan monolaurate (Span
20), Span 40, Span 65, Span 60, Span 80, Span 85
Phospholipids egg/soy lecithin
Co-Surfactant

19. Purified or Ultrapure Water, Saline or Buffered Saline
More preferably balance of water 30 to 99.90 wt%
Co-solvent lowers the interfacial tension of the aqueous phase
which enables the formation of smaller emulsion droplet size
• Suitable Co-solvent include C1-C10 alcohols such as methanol,
ethanol ….decanol
• More preferably ethanol (C1-C4)
Aqueous Phase
Co-solvent

20. Additives are added to make the nanoemulsions last for
longer periods
Additives

21. DIFFERENT DELIVERY SYSTEMS
Fig: Different types of delivery systems that can be
created based on Emulsion Technology

22. Conventional oil-in-water (O/W) emulsions consist of emulsifier-
coated lipid droplets dispersed in an aqueous continuous phase
Formed by homogenization of oil and water phase together in the
presence of a hydrophilic emulsifier
Ref: Gaonkar et al., (2014)
CONVENTIONAL EMULSIONS
MULTIPLE EMULSIONS
Multiple water in oil in water (W/O/W) emulsions consist of small water
droplets contained within larger oil droplets that are dispersed in aqueous
continuous phase
Two steps of formation: Preparation of W/O emulsion
Homogenization of W/O emulsion and aqueous phase
and water soluble emulsifier

23. MULTILAYER
EMULSIONS
Fig. Schematic representation of formation of conventional and multilayer emulsions
Multilayer Emulsions are formed by adding polyelectrolytes to an emulsion containing
oppositely charged droplets so that they adsorb and form a nano-laminated coating

24. TYPICAL EMULSION-BASED LIQUID-
LIQUID SYSTEMS

25. ATTRIBUTES OF EDIBLE DELIVERY
SYSTEMS
EFFICIENTLY ENCAPSULATE AND
KEEPING IT ENTRAPPED
PROTECTION FROM CHEMICAL
DEGRADATION
CONTROL THE RELEASE OF THE
FUNCTIONALAGENT
COMPATIBLE WITH THE
SURROUNDING FOODS
Appropriate amount of functional agent
Functional Agent should be remain in its
active state
e,.g. Release rate or the specific
environmental stimuli that triggers
release
Beverages matrix, without causing adverse
effect affects on product Appearance,
Rheology, Mouth feel, Flavor and Shelf life

26. RESIST THE ENVIRONMENTAL
STRESSES
COMPLETELY PREPARED FROM
GRAS INGREDIENTS
NOT ADVERSELY IMPACT THE BIOAVAILABILITY OF THE
ENCAPSULATED MATERIAL
FOODS OR BEVERAGES EXPERIENCE DURING THEIR
PRODUCTION, STORAGE, TRANSPORT AND
UTILIZATION e.g HEATING, FREEZING ,CHILLING,
DEHYDRATION OR SHEARING
USING SIMPLE COST-EFFECTIVE PROCESSING
OPERATIONS
Ref: McClement, 2009

27. MICROEMULSIONS
Synonyms Swollen micelles, Micellar emulsion
Thermodynamically stable mixtures of water, oil and one or more
amphiphils which assemble spontaneously into nanometer-scale
droplets (Flanagan & Singh, 2006)
SPONTANEOUS FORMATION OF A MICROEMULSION ARISES FROM -
SYNERGISTIC INTERACTION OF &
TO EFFECTIVELY SOLUBILIZE OIL MOLECULES

28.  Water is continuous phase
 Shape of droplets or particles:
Spherical size or other structures possible
 Thermodynamically or 20 kinetically stable liquid:
Dispersion of an oil phase and a water
phase, in combination with a surfactant
 Transparent or Translucent in appearance:
Due to small particles or droplets dispersed
with size 5-100 nm

29.  Microemulsions form spontaneously:
Readily form without high energy input
Due to ultralow interfacial tension and favorable
energy of structure formation
 Microbiologically stable:
Due to water droplets are extremely small
 Long shelf life:
Particle sizes do not change with time (brownian
motion effect dominates gravity)
 Low calorific value:
Inclusion of water reduces calorific value of
emulsions and enables water-soluble ingredients
such as vitamins, flavors, salt, sugars, colorants
Regular ~ emulsions of larger water droplets can present a severe
microbiological problem

30. Theory of Formation
Under appropriate environmental conditions, certain
types of food components spontaneously assemble into well-
define structures since this minimizes the free energy of the
system
e.g. Micelles, vesicles, fibers, tubes, liquid crystals
SPONTANEOUS SELF ASSEMBLY
SYSTEM
(Bottom-up Approach)
Mcclements , 2009

31. Hydrophobic Effect - Primary driving force of microemulsions
formation, It minimizes contact between the non-polar tails of
surfactant and water

32. WINSOR CLASSIFICATION SYSTEM
Ref: Winsor, P. A. (1948)
Type I:
O/W Microemulsions system
which is in equilibrium with an upper
excess oil phase
Surfactant is preferentially soluble in
water (Winsor I) )
Type II:
W/O Microemulsions
system in equilibrium with a lower
excess aqueous phase
Surfactant is mainly in the oil phase
(Winsor II)

33. Flanagan & Singh (2006)
Type III:
Equal volumes of oil and water
are present
Bicontinuous structure is formed
Three-phase system where a
surfactant-rich middle phase
coexists with both excess water and oil
surfactant-poor phases
(Winsor III /Middle Phase
Microemulsion) Type IV:
Macroscopically single phase
Single-phase (isotropic) micellar
solution, that form upon addition of a
sufficient quantity of amphiphile
(Surfactant + Alcohol)
Thermodynamically stable
systems are primary interest to food
technologist

34. SINGLE PHASE MICROEMULSIONS
• Form at specific concentrations of oil, water and
surfactant (co-surfactant)
• Oil concentration (<30%): oil-in-water
• Aqueous concentration low : water-in-oil
• May comprise of:
1. Oil: 40-97wt~
2. Surfactant: 1-40wt~
3.Water phase: 0.1-25wt~
4. Co-surfactant: 0.1-10wt~
Preferred embodiment mole ratio of
Water: Surfactant is 5-70
Flanagan & Singh (2006)

35. STEPS IN MICROEMULSION PREPARATION
(Dissolving water soluble components)
(Solids dissolve possibly by heating)
(Undissolved material separated by centrifugation)
(Dissolving oil soluble component)
(Mixed in suitable vessel and given time to equilibrate)
(Using emulsification techniques - stirring, use of
membranes, applying shear, ultrasound etc)
Preparing Water Phase
Emulsifying Water & Oil Phases
Preparing Oil Phase

36. SHORTCOMINGS OF MICROEMULSIONS
Become physically unstable if its composition is changed e.g.
upon dilution, acidification or heating in food processing
1. Contains high amount of surfactants relative to amount of oil
2. Application of microemulsions in foods is limited by the
types of surfactants which are used to facilitate its formation
3. Many surfactants are not permissible in foods (bitter taste)
4. WHO and the FDA have placed restrictions on the daily intakes
of many of these surfactants
MAINTAINING OPTIMUM PHYSICAL
STABILITY
HIGH AMOUNT OF SURFACTANTS

37. NANOEMULSIONS
Synonyms -
(Sub-micron emulsions (SME), Mini-emulsions, Ultrafine emulsions)
Def:
A nanoemulsion is an emulsion which does not form
spontaneously, but instead formed by the application of shear to a
mixture of oil, water and surfactant
Oil droplet sizes >100nm (most preferably >60nm)
Droplet size is measured by Photon Correlation Spectroscopy
(Dynamic light scattering)

38. • It can be three types oil-in-water, water-in-oil, bicontinuous
• Kinetically stable
• Spontaneously do not form need high energy input to obtain
• Steric stabilization prevents any flocculation or coalescence
• Particle size may increase over time via coalescence,
flocculation and /or ostwald ripening
• Transparent / translucent appearance like microemulsions
• Metastable
SMALL SIZE AND HIGH KINETIC STABILITY MAKE NANOEMULSIONS SUITABLE FOR
DELIVERY OF ACTIVE COMPOUNDS IN BEVERAGES

39. FORMULATION TABLE OF
NANOEMULSIONS
Oil phase 0.01 to 40
Hydrophilic Non-ionic Surfactant 3 to 7
Co-surfactant 0.1 to 15
Aqueous phase 30 to 99.90
Co-solvent 15 to 45
Active Component 0.01 to 10
Additives 0 to 10
Active component may be 0.01 to 50 preferably 0.01 to 10 wt %
Components Most Preferably wt %

41. Also known as Bottom up Approach
Emulsion spontaneously form
Submicron emulsion can be obtained by employing the physico-
chemical properties of the systems generally referred to as low-energy
emulsification methods
LOW ENERGY APPROACHES
Phase Inversion Point
Membrane Emulsification
Spontaneous Emulsification
Solvent Displacement
Emulsion Inversion Point

42. (Leong et al., 2009)
 Disrupt oil & aqueous phases into tiny droplets using mechanical
devices
 Formation governed by the selected composition (Surfactant +
Functional compounds) & by the quantity of energy applied
HIGH ENERGY APPROACHES

43. • Smaller the droplet size more surfactant/energy is required
• Some chemical get easily degraded
• Homogenizers/ High-speed devices
• High Pressure Homogenizer/Microfluidizer
• Ultrasound
Devices used

44. Commonly used
Adaptable to different processing conditions
Variety of food ingredients can be used to
prepare Nanoemulsions
Effective in reducing size
More control of the size distribution and
composition of resultant Nanoemulsions
HIGH ENERGY APPROACH
Less Expensive
Energy Efficient alternative that takes the
advantage of the chemical energy stored
in the system
Suitable for some unstable molecules such
as Proteins and Peptides
LOW ENERGY APPROACH
Ref: Herrera M (2012)

45. PHASE TRANSITION
METHODS
Principle:-
Phase inversion occur when the structure of the emulsion is
inverts
Phase transition cause release of chemical energy that dispersed
the fine droplets through emulsification method
Ref: P. Fernandez et al. (2004)

46. Cross Ref: P. Fernandez et al. (2004)
Uson et al.,2004 –
Phase transitions produced during the
emulsification process which is carried out- At constant temperature
and changing the composition
OR
Izquierdo et al., 2001; Morales et al.,2003 –
At constant composition and changing the temperature
These methods make use of changing spontaneous curvature of the
surfactant

47. EMULSION PROCESSING:
HOMOGENIZER
Homogenization is a unit operation using a class of processing equipment referred
to as that are geared towards reducing the size of droplets in
liquid-liquid dispersions

48. HIGH PRESSURE HOMOGENIZERS
Principle -
The premixed liquid passed under a high
pressure (100-1500bar) through a minute gap of the
homogenizing valve
Such condition generates very high shear
stresses, acceleration, impact and intense turbulent
flows in the liquid resulting subdivision of the
dispersion phase particles or droplets and their
homogeneous distribution through the mixture
Poor Productivity, Component deterioration due to generation of heat, Oil-in-
water liquid nanoemulsions of less than 20% oil phase can be prepared

49. MICROFLUIDIZER PROCESSORS : FROM
LAB TO PRODUCTION

50. MICROFLUIDIZER

51. 1. Intensifying Pump -
Generate high pressure (500-20000 psi)
Forces the stream through micro-channels at high velocities
Into an impingement zone
Creating an intense turbulence mixing action
2. Interaction Chamber - (Heart of microfluidizer technique)
Reduces sizes of a pre-emulsion (coarse droplets)
Multiple Passes through the System is typically required to obtain the desired
fine emulsion with narrow droplet size distribution
Uniform emulsion compare to High Pressure Homogenization
Ref: Panagiotou T. and Fisher R. (2012). Review Article. Cahllenges 3(2), 84-113; doi:10.3390/challe3020084

52. ULTRASONIC HOMOGENIZERS
Principle-
Utilizes ultrasonic waves for homogeneous
distribution of the
dispersed phase by the actions:
1. Reducing sizes of the dispersed
particles/droplets (breaking)
2. Disintegrating the dispersed
particles agglomerates
3. Blending the dispersed phase
in the liquid
Use for small scale production

53. TECHNOLOGY
ADVANTAGES

54. Creation of transparent bioavailable beverages
containing ω-3 rich Oils, Phospholipids and
Minerals
Encapsulation of ω-3 fatty acids enhances the stability
and bioavailability of bioactive food ingredients
OXIDATIVELY STABLE BEVERAGES (Pinto et al., 2005)

55. PROTECTION OF NUTRACEUTICALS
Protect against flavor, taste and nutritional degradation
• Used as vehicles for targeted nutraceuticals
• Vehicles are expanded micelles in the size of<30 nm and used in
clear beverages without phase separation
• Potential application include Lycopene, beta-carotene, Omega-3
fatty acids, Phytosterols and Isoflavones
NANOSIZED SELF-ASSEMBLED LIQUID
STRUCTURES
(Garti and Aserin, 2007)

56. ENHANCE BIOAVAILABILITY
1. Greater ability to dissolve/disperse poorly soluble active
components at the interface
2. Faster rate of digestion by lipolysis & greater uptake
(more rapid release of emulsified active ingredients)
3. Below 100 nm have greater ability to penetrate epithelial layers (Skin and
Oral mucosa)
(OPTIMUM NUTRACEUTICALS DELIVERY )
TRANSPARENT TO TRANSLUCENT
1. Optically clear and applicable in beverages ( not alter visual appeal)
ENHANCE SOLUBILITY
1. Enhance solubility of bioactives at an interface

57. REDUCE VISCOSITY
REDUCE AMOUNT OF SURFACTANT
(NANOEMULSIONS)
HYDROPHOBIC BIOACTIVE
COMPONENT DELIVERY
1. Vehicle for Hydrophobic bioactive delivery
1. They do not show problem of creaming, flocculation,
coalescence and sedimentation
1. Mainly use for encapsulate a diverse range of Lipophilic
Compounds (Chen et al., 2006)
CONTROLLED DELIVERY OF NUTRACEUTICALS
To some extent amount of nutraceuticals release at target
organ can be controlled

58. CHARACTERIZATION
OF
NANOEMULSIONS

59. Determined by Photon Correlation Spectroscopy (PCS)
Light scattering was monitored at 25˚C at a scattering angle of 90˚
Determined by using Brookfield DV ultra V6.0 cone and Plate Rheometer at
25±0.3˚C
Determined by using an Abbes Type Refractometer
Morphology and structure of the nanoemulsions were studied using
Transmission Electron Microscopy (TEM)
Droplet Size Analysis
Viscosity Determination
Refractive Index
Transmission Electron Microscopy

60. Keeping the sample at refrigerator temperature (4˚C) and room temp (25˚C)
These studies performed for the specific period of time
Electrical characteristics of emulsion droplets can be measured by the
electrical charge (Z-potential) of oil droplets in emulsions
Measure in Shear-Plane
Z-potential predicts the stability of emulsion during storage
Color and Turbidity measurement
Optical Property
Stability Studies
Z-Potential

61. Scanning Electron Microscopy
Samples were fixed on stub using conductive double sided tape
and then made electrically conductive by coating in a vacuum with
thin layer of gold
Fig. SEM Encapsulated flavor oil in alginate solution and added in ice
cream
(Ashraf et al., 2015)

62. NUTRACEUTICALS
VIA
DAIRY PRODUCTS

63. FORTIFICATION IMPLICATIONS
IN DAIRY
1. It is widely consumed food cover large demographic
2. Various steps and process have measurable impact on some
specific nutrients
3. Fulfill deficiency of milk e.g. Fe, Cu, Fiber, LCF, Vit C, E
4. Easier quality control measure implementation
5. Cost affordable by target population
6. Higher stability and bioavailability of added micronutriens

64. NUTRACEUTICALS FORMS AND
NATURE OF EMULSION
Ref: Zuidam & Shimoni 2010
HYDROPHILIC NUTRACEUTICALS
LIPOPHILIC NUTRACEUTICALS
POLAR PARADOX
Water soluble Vit e.g. Ascorbic acid, Polyphenols
Water-in-Oil (W/O) type emulsions
Lycopene, β-carotene, Lutein, Phytosterols, Docosahexaenoic acid (DHA),
Carotenoids, Essential Fatty Acids can be encapsulated & delivered by Oil-in-
Water (O/W) type emulsions
Polar antioxidants are more active in bulk lipids than their nonpolar
counterparts
Nonpolar antioxidants are more effective in oil-in-water emulsion

65. UHT milk with DHA
Huimin et al (2014)
Studied formation and stability of Algae oil emulsion for application in
UHT milk and produce UHT milk enriched with DHA
- Starch hydrophobically modified with Octenyl Succinic Anhydride
(OSA starch) & Corn Syrup was used as emulsifier to stabilize algae
oil in UHT milk
- Oxidative stability measured after heat treatment and found Algae
oil-enriched milk was stable during 11 weeks of storage
- Even high temp storage 40˚C does not significantly increase the
oxidation process
Stable algae oil emulsion can be formed by OSA starches with Corn
syrup & can be applicable for enrichment of UHT milk

66. PROBIOTIC ENRICHMENT
Latha Sabhiki (2012)
Probiotics can be encapsulated in emulsion based delivery
system containing solution of alginate (discontinuous phase) and
dropped into vegetable oil (continuous phase) containing suitable
emulsifier (tween 80) and surfactant (Sodium lauryl sulfate) to form
beads
Later can be harvested by filtration.
Technique is Easy to scale up for large scale production and
provides both encapsulated and entrapped core materials

67. LIMITATIONS FOR
TECHNOLOGY
1. Residual oil, Emulsifier and Surfactant in the encapsulating
material can affect the growth of live probiotics
2. Interact with food components
3. Addition of oil may damage the organoleptic properties and texture of
the food products
4. Not suitable for low fat food products
Latha Sabhiki (2012)

68. Work conducted at NDRI, Karnal reports the effect of
microencapsulation using sodium alginate and starch on the tolerance
of probiotic by emulsion
Babu el al., (2009) Probiotic Lactobacillus paracasei S233
&
Sabikhi et al., (2010) Probiotic L. acidophilus LA1
Organisms survived well in the protected form at high
temperatures, high salt concentrations
&
At stimulated conditions of gastric pH and at high bile salt
concentrations

69. VITAMIN D FORTIFICATION
• Daily recommended intake of Vit D is
Adults- 5µg/Day
Children and Pregnant Woman- 10µg/Day
• Vit D is Fat soluble is hardly found in skim milk and low fat dairy products
consumed largely in modern societies being an important sources of Calcium and
Phosphate
• Vit D2 is largely used in pharmaceuticals industry
Ref: Patent: US 20090311329A1

70. • Novel approach for the nanoencapsulation and
stabilization of hydrophobic biologically active
compounds, particularly in non-fat or low fat edible
products
• Hydrophobic domains of casein micelles stabilize
the nutraceuticals in aqueous systems like milk
• Facilitate the enrichment of low fat and fat free
dairy and other food products with these bioactive
molecules ( vit D2, Vit K, Vit E, Vit A)
Patent: US 20090311329 A1 (2009)
Vit D2 Hydrophobic Nutraceuticals Adsorbed into Hydrophobic domains of
which tend to found in the core of the micelle

71. VIT D IN CHEESE
Tippetts et al., (2012)
Observed increased in retention of vit D 3 in
cheddar cheese by incorporating it as a part of an oil-in-
water emulsion using milk protein as emulsifier to obtain a
fortification level of 280 IU/serving
Significantly retention was greater in cheese curd
Vit D3 emulsified oil with NDM than control
Full fat cheese curd has little less retention compare to
whole milk cheese curd

72. Gaysinsky et al (2007)
Conducted research on the antimicrobial activity of Eugenol
microemulsions (Eugenol Encapsulated in Surfactant Micelles) in Ultra
high temp pasteurized milk containing different percentages of milk fat
(0, 2, 4%)
He studied antimicrobial effect on L monocytogens and E coli O157:H7
strains
Results suggest that food composition, especially fat level, may
affect the efficiency of targeting food borne pathogens with
surfactant-encapsulated antimicrobials

73. Cross Ref : Silva et al .,2012
Some of the giants of food industry such as Unilever and Nestle are
also applying this technology

74. MEAT PRODUCTS
FORTIFICATION

75. Salminen et al (2013)
Study provided systematic approach to evaluate the oxidative and
physical stability of both the encapsulated active ingredient and subsequent
inclusion into a meat product (Pork sausages)
Protein stabilized oil – (25wt% oil, 0.25 wt% WPI, 50mM Sodium citrate
buffer, pH 3.0) used, formed pre emulsion and incorporated in pork batter,
stuffed in casing and then steam cooked to internal temp 72˚C
Emulsion provide physical and oxidative stability to base emulsion
But addition in complex meat matrices may lead to break down of base
emulsion due to differing conditions present in the meat product

76. CLSM images of low fat pork sausages
with added with oil-in-water emulsion
(25 wt% oil, 0.25wt% WPI, 50mM sod
citrate buffer, pH3)
a) 0 day
b) 31 days of storage
CLSM fish-in-water type
emulsions (25 wt% oil, 0.25
wt% WPI, 50 mM sodium
citrate buffer, pH 3) during
storage
Salminen et al (2013)

77. SHORTCOMINGS IN
MEAT PRODUCTS
Consists of complex protein matrices, salts, minerals (ingredients used such
as ascorbic acid, phosphates, curing agents, spices)
1. Most food matrices differ in respect to their interactions to incorporated
emulsion delivery systems
2. Ingredient interactions affects emulsion stability
MEAT IS COMPLEX FOOD
Ref: McClements & Decker, 2000

78. Mixing, Heating, Freezing and Freeze-thawing of meat
products may likely influence the stability of emulsions based
delivery systems
Upon heating proteins stabilizing the emulsion may denature
cause release of droplets interfaces to become hydrophobic and thus
prone to aggregation
PROCESSING CONDITIONS
Ref: Salminen et al., (2013)

79. EFFECT OF SALT AND MINERALS
Overall electrostatic repulsion in emulsion droplets
dependent on the ionic strength
Presence of salt (150 mM NaCl) cause flocculation of
emulsion
In protein stabilized emulsions pH and ionic strength are
major factors in dictating their stability
Screening of protein charges in the emulsions due to high
salt and mineral concentration in the meat products may cause
flocculation or coalescence of oil droplets
Ref: Djordjevic et al., 2004

80. FUTURE PROSPECTS
1. Efficient Delivery of Herb Extract-
Technology can be used for Herb extract delivery in beverages
and meat products
2. Antimicrobial Nanoemulsions in Food Industry –
For decontamination of food, equipment, packaging of food
(Gruere et al., 2011)
3. Potential Delivery System for PUFA-
It is potential delivery system for PUFA in meat products
4. Improved Stability and Bioavailability of Nutraceuticals-
Are required for the delivery of sensitive ingredients
where direct contact with food matrix adversely affects
the performance of bioactive compounds
(Cournarie et al.,2004)

81. 5. Novel Products-
If these compounds encapsulated in multiple nanoemulsions
offer more scope for the control of encapsulation ingredient,
protection and release mechanism as compared to conventional
Emulsions (Sharma & Singh, 2012)
6. More Scope in Meat Products –
As it less studied topic in meat products in preparation of
functional meat products
7. Nanotherapy-
Targeted delivery of nutrients for people with deficiencies
8. Totally new Taste or Mouthfeel –
By encapsulating flavor

82. REFERENCES
1. Gosh et al.,2005. Book-Clinical Aspects of Functional Foods and Nutraceuticals , CRC Press, Pp. 63
2. Latha Sabhiki (2014), Dairy Technology Division, Special Training Course, NDRI, Karnal
3. Patent: US 20090311329A1, Yoav et al., (2007)-Casein micelles for nanoencapsulation of hydrophobic
compounds
4. Tippetts et al., (2012) J. Dairy Sci.Sep;95 (9):4768-74
5. Salminen et al., (2013). Meat Sci. Mar93 (3)659-67
6. P. Fernandez et al. (2004) / Colloids and Surfaces A: Physicochem. Eng. Aspects 251 :53–58
7. Flanagan & Singh (2006). Critical review in food science and nutrition,46:221-237
8. Cross ref: Winsor, P. A. Trans. Faraday Soc. 1948, 44, 376
9. R. Adjonu et al., (2014). Journal of food Engineering 122:15-27
10. Gaonkar et al., (2014). Book-Microencapsulation in the Food Industry: A Practical Implementation
Guide. Academic Press Elsevier Pp 151-152.
11. A. A. Patel & A. K. Singh (2012), Dairy Technology Division, National Training Program , NDRI, Karnal.
Pp 2-3.
12. Gavsinsky et al., (2007). Antimicrobial efficacy eugenol microemulsions in milk against Listeria
monocytogenes and E. coli O157:H7. J. Food Prot, Nov;70(11):2631-2638.
13. McCments, D V (2009). American Chemical Society. Washington DC, Pp 3-4.
14. Huimin et al., (2014). Food and Bioprocess Technology. Vol 7, Issue 2, Pp567-574.
15. P Sahrma & R R B Singh (2012). ), Dairy Technology Division, Special Training Course, NDRI, Karnal
Pp 90-92.