Nanoscience for Food

Value Metabolites

Dr. Pooja J Rao

Spice & Flavour Sciences
 Origin & Meaning - Nano
The prefix “nano” means “dwarf” in Greek.
A nanometer corresponds to one millionth part of
a millimeter.

atom nanoparticle solid

This scale corresponds to a size > molecule but <

bulk solid.
Therefore, a nanomaterial frequently exhibits
physical and chemical properties somewhere in
between or completely different and beyond our
imagination.
• High surface/volume ratio leads to unique properties
• The physical and chemical properties of nanoparticle lies in between that of a
single atom and that of bulk solid.

Sp. Surface Area = Surface Area / Volume

For sphere,
Surface Area = 4r2 and Volume = 4r3
3

If r = 9 nm If r = 3 nm If r = 1 nm
SSA = 0.33 x 109 m-1 SSA = 1 x 109 m-1 SSA = 3 x 109 m-1

Smaller the particle size greater is the sp. surface area

 Classification of nanomaterials
Classification NM classes Characteristics Examples
criterion
Origin (source) 1.Natural NMs Produced in nature through Proteins,
biochemical polysaccharides, DNA,
and biological processes in ribosomes,
organisms, enzymes, antibodies,
insects, plants, animals, etc.
and human bodies

2.Synthetic Synthesized by physical, Metal-based

(engineered) chemical, nanoparticles,
NMs biological, or hybrid polymer-based
methods nanoparticles, etc.

3. Incidental NMs naturally occurring Volcanic ash, ocean

nanomaterials or are spray, magnetotactic
produced incidentally as a bacteria, mineral
by-product of composites, etc.
industrial processes
Classification NM classes Characteristics Examples
criterion

Chemical Inorganic NMs Are mineral salts, carbon, Carbon-based

Composition nanomaterials: metal, and metal Nanoparticles of Ca21,
Oxide NPs Zn21, Mg21, Na1, etc.
Nanoparticles of Au, Ag,
Al, etc.
Nanoparticles of TiO2,
SiO2, etc.

Organic NMs Occur through physical Lipid-based

and chemical nanoparticles
interactions between Protein-based
organic component nanoparticles
Molecules Polysaccharide-based
nanoparticles
Polymer-based
nanoparticles
Surfactant-based
nanoparticles
Composite NMs obtained by combining
several organic Conjugated/complexed
and inorganic materials biopolymers-based
nanomaterials
Classification NM classes Characteristics Examples
criterion

Dimensions Zero-dimensional All the dimensions of Nanoparticles,

NMs nanomaterial are nanocrystals
measured within the
nanoscale

One dimension is Nanotubes, nanorods,

One-dimensional outside the nanoscale and nanowires
NMs
Two dimensions are Graphene, nanofilms,
outside the nanoscale nanolayers, and
Two-dimensional Nanocoatings
NMs
All dimensions are Bulk powders,
outside the nanoscale dispersions of
nanoparticles,
Three-dimensional bundles of nanowires,
NMs multinanolayers
Methods of Nanoparticles Preparation
• Encapsulation
 A process of enclosing a flavor or bioactive (core) in an inert encapsulating agent (carrier).
• Types
 Microencapsulation and Nanoencapsulation
• Advantage
 Protection against evaporation, temperature, chemical reactions (such as flavor-flavor
interaction, light-induced reactions, oxidation)
 Enhances retention of flavor property during processing and storage, allows controlled release
• Techniques
 Chemical technique – Coacervation, Molecular inclusion, Liposomes
 Mechanical technique – Spray drying, Spray chilling, spray cooling, Extrusion,
homogenization, sonication
 Type of nanomaterials

• Nanoemulsions - Lipophilic molecules

Enhancement in nutritional, therapeutic and flavor qualities
• Liposomes - Hydrophilic molecules
Enhancement in biological property and visual appeal
• Dendrimers - Drug delivery system and gene therapy
Enhanced stability of antibodies, peptides
• Quantum dots - Semiconductor fluorescent nanomaterials
Optical imaging, display devices, bioassay
• Metal nanoparticles – nanospehres, nanoshells, nanocages
Application in biosensors, generate radiation used to target microbes
Delivery systems Preparation Methods

Nanoparticles Emulsification and cross-linking and drying

Solid lipid nanoparticles Hot emulsification of high melting liquid

Nanostructured lipid carriers Both solid (fat) and liquid (oil) lipids

Nanoemulsion HPH, Ultrasound assisted method

Micelles Self-assembly after dissolving of surface

active compounds ternary mixture of
emulsifier, oil, water

Liposome Reverse phase evaporation

 Nanoemulsion
Nanoemulsions – These are isotropic and kinetically stable emulsion systems in which the oil
droplets containing hydrophobic bioactives are stabilized by a thin layer of emulsifier.
• The substance that make up droplet in emulsions – dispersed or internal phase
• The substance that make up surrounding liquid – continuous or external phase
• Concentration of droplets – dispersed phase volume fraction ( )
• Droplet diameter - <200 nm ----- Transparent
• Droplet diameter – upto 500 nm ---- Milky
• Types- Oil-in-Water (O/W); Water-in-Oil (W/O); Water-in-Oil-in-Water (W/O/W)
What is the requirement?
• Mechanical emulsion – Creaming, sedimentation, flocculation, coalescence, Ostwald ripening
Probable solution
• Use of surfactant – Being amphiphilic they adsorb at interface and reduce the colloid droplet
fusion
Properties
• Nanoemuslions are thermodynamically unstable but kinetically stable
• Stability can be enhanced by reducing the droplet size while increasing the density of oil and
viscosity of water phase (use of emulsifier or thickening agent)
 Food and Nanomaterials
• Foods contain both natural nanomaterials and engineered nanomaterials
• Natural -
Casein – 100 nm; α
Lactoalbumin (α-LA) (1.8
nm); β Lactoglobulin dimer
(β-LG) (3.5 nm); Bovine DNA
serum albumin (BSA) (4
nm) and immunoglobulin G
(IgG) (6 nm) (de Wit,
1998).

• Engineered nanomaterials - Deliberately added in order to increase food quality and safety,
such as nanoparticle-based delivery systems

• EFSA for agrofood – Nanostructured materials (metals – Ag, Au, Al, Fe; metal oxide – TiO 2,

SiO2, ZnO), nanoparticles and their aggregation at nanoscale, nanoencapsulates and

nanoproducts, Applications in delivery of pesticides, herbicides, plant growth hormones
• USFDA for food industry – Nanomaterials considered as GRAS
• Applications as food additives, nutraceutical protection, enhancement in bioavailability
• Packaging and sensors – enhancement in barrier property, detection of toxins, pathogen and
pesticides
 Emulsifier & Stabilizer
 Surface-active molecules adsorbing to the surface of freshly formed droplets
during homogenization, forming a protective membrane around the droplets.
 Prevents the aggregation of molecules by coming close together
 Two ends with different affinity
 Polar head and Non-polar tail
 Arrange according to the dispersion medium
 Concentration an important factor (critical micelle concentration)
 Low – exist separately (sub-colloidal form)
 High – Aggregates to form micelles
Stabilizer
• Enhances the
stability of
emulsions
• May be an
emulsifier or a
thickening agent
• Based on nature of interaction between dispersed phase and dispersion medium
 Lyophilic colloids ----- Colloids having affinity towards dispersion medium (intrinsic colloid)
 Stable and does not precipitate easily
 Presence of 2 layers (i) positive/negative charge (ii) layer of solvent
 Hydrophilic Colloids – insulin, albumin, acacia, gelatin, starch
 Lipophilic Colloids – rubber, polystyrene, resin, curcumin, lutein
 Lyophobic colloids --- Colloids having negligible or no affinity towards dispersion medium
(extrinsic colloid)
 Not stable and precipitate easily
 Presence of 1 layer - positive/negative charge
 Dispersion of inorganic particles in water – Au, Ag, AgI solution
 Needs special method for preparation (size reduction and aggregation)
 Amphiphilic colloid ----- Colloids having two ends with different affinity
 Polar head and Non-polar tail
 Arrange according to the dispersion medium
 Concentration an important factor (critical micelle concentration)
 Low – exist separately (subcolloidal form)
 High – Aggregates to form micelles
 Nanoemuslions are thermodynamically unstable but kinetically stable

• From a purely thermodynamic point of view, an emulsion is an unstable system because the
interfacial tension is always greater than zero so it always costs energy to increase the
interfacial area.
• Thus, a natural tendency for a liquid/liquid system to separate and reduce its interfacial area
and, hence, its interfacial energy. However, emulsions exist because of kinetic stability (i.e.,
they are stable over a period of time).
• A system may not be able to reach equilibrium during the time scale of an observation
because of the presence of an energy barrier between two states.
Elow = Lower free energy state
Ehigh = Higher free energy state
ΔE* = Energy barrier
Elow is thermodynamically favourable
thus greater fraction of molecules likely
to occupy

McClements, D. J. (2015). Food emulsions: Principles,

practices, and techniques, third edition (3rd ed.). Boca
Raton: CRC Press, Taylor and Francis Group.
• The oil/water emulsions are classified on the basis of their degree of kinetic stability.
1. Emulsions separate in a few minutes, and the separated water is free water
2. Nano emulsions separate in tens of days
3. Microemulsions separate (sometimes only partially) in days or even months

• Water-in-oil emulsions are considered to be special liquid-in-liquid colloidal dispersions.

Their kinetic stability is a consequence of small droplet size and the presence of an
interfacial film around water droplets and is caused by stabilizing agents (or emulsifiers).
These stabilizers suppress the mechanisms involved that would otherwise break down an
emulsion. Such mechanisms include:
1. Sedimentation
2. Aggregation or flocculation
3. Coalescence
4. Phase separation
Ref.: https://petrowiki.spe.org/Stability_of_oil_emulsions
 Surface films and stability to coalescence
The ability of surfactants to adsorb at the interfacial layer helps in forming a film around the
water droplets at the oil/water interface.
These films enhance the stability of an emulsion by increasing the interfacial viscosity. Highly
viscous interfacial films retard the rate of oil-film drainage during the coalescence of the water
droplets by providing a mechanical barrier to coalescence, which can lead to a reduction in the
rate of emulsion breakdown.

A good correlation exists between the presence of incompressible interfacial film and emulsion
stability. These films are classified into two categories on the basis of their mobilities.
•Rigid or solid films are like an insoluble, solid skin on water droplets characterized by very
high interfacial viscosity. There is considerable evidence that very fine solids stabilize these
films. They provide a structural barrier to droplet coalescence and increase emulsion stability.
These films also have viscoelastic properties.
•Mobile or liquid films are characterized by low interfacial viscosities. Liquid films are formed,
for example, when an emulsifier is added to an emulsion. They are inherently less stable than
 Techniques & Equipment for Nanoemulsion

• Top-down Approach • Bottom- up Approach

 Ultrasonication  Solvent demixing
 High pressure homogenization Self-emulsification
 Microfluidization (spontaneous emulsification)
 Nanospray drying  Phase inversion assays
 Membrane technology
 Ball milling
• High-energy method - Microfluidization
• Initially, a high shear mixer is used to form a coarse O/W emulsion by blending oil, emulsifier, and water
together. Coarse emulsion is forced through an interaction chamber in the microfluidizer using pneumatic
pressure.
• The microfluidizer works on the principle of dividing a pressure stream into two parts, passing each part
through a fine orifice, and directing the flows at each other in the heart of the microfluidizer, that is, the
interaction chamber.
• The microfluidizer uses high pressure to guide the flow stream through microchannels toward the
impingement area, which creates a very high shearing action that provides an exceptionally fine emulsion.
• Within the interaction chamber, cavitation, along with shear and impact, reduces emulsion droplet size
(Jafari et al., 2006).
• By using a microfluidizer the distributions of produced particle sizes appear to be narrower and smaller
than the products of traditional homogenization. A schematic illustration of a microfluidization system for
nanoemulsion preparation is given in Fig. 13.1.

Ref.: Nanoscaled
Dispersed Systems Used i
n Drug-Delivery Applicatio
ns
Catalina Natalia Cheaburu-
Yilmaz et 1l., 2019
High-energy method - High Pressure Homogenization

• HPH is typically performed by forcing a liquid through a narrow nozzle at a high pressure.
• High shear stress is created.
• Pressures are between 15 and 40 bars or 50-5000 psi.
• Droplet size can be adjusted by the levels of pressure, energy input and the number of pass.
• Weak bonds in macromolecules can be broken by applying sufficiently high pressures, that is,
both physical and chemical changes can be controlled.

Mechanism of HPH
• Cavitation theory
• Globule dispruption by turbulent eddies
(micro whirls)
• High-energy method - Ultrasonication
• When ultrasonic waves pass through a liquid medium, a large number of microbubbles
form, grow, and collapse in very short times, about a few microseconds.
• Ultrasonication generates alternating low-pressure and high-pressure waves in liquids,
leading to the formation and violent collapse of small vacuum bubbles.
• This phenomenon is termed cavitation and causes high-speed impinging liquid jets and
strong hydrodynamic shear-forces.
• These effects are used for the deagglomeration of nanometer-size materials.
• In this aspect, ultrasonication is an alternative to high-speed mixers and agitator bead mills.
Ultrasonication has been used as a technique to disperse nanomaterials in different matrices

Ref.: Nanoscaled
Dispersed Systems Used in Drug-Deli
very Applications
Catalina Natalia Cheaburu-Yilmaz et
1l., 2019
• Low-energy method – Spontaneous emulsification
• The lipophilic bioactive, oil polymer, and optionally phospholipids are dissolved in a water-
soluble solvent (e.g., acetone).
• The above solution is poured into an aqueous solution containing a nonionic surfactant (e.g.,
poloxamer 188).
• The fast diffusion of solvent into water provokes the spontaneous emulsification of the oily
solution in the form of nanodroplets.
• The dissolved polymer will form a film around the droplets that contain the drug.
• The principal drawback of this method is the polymer aggregation observed whilst working
with high polymer concentration or low organic solvent/water ratio.

Ref.: Nanoscaled
Dispersed Systems Used in Drug-Deli
very Applications
Catalina Natalia Cheaburu-Yilmaz et
1l., 2019
• When changes in the surfactant spontaneous curvature are produced during the emulsification
process from negative to positive (to obtain O/W emulsions) or vice versa (for W/O
emulsions), they are designated as “phase inversion” methods.

• These are classified as phase inversion temperature (PIT) and phase inversion composition
(PIC) methods if emulsification is triggered by a change in temperature or composition,
respectively.

• Nature and structure and phase behavior of surfactant is important when these emulsification
methods are used, since the phases involved in the emulsification process are determinant to
obtain nanoemulsions with minimum droplet size and low polydispersity.

• Emulsification in very small droplets is favored, the emulsions are kinetically unstable,
means agglomeration occurs soon.
• Nanoliposomes- The conventional methods for preparing nanoliposomes include solubilizing
the lipids in organic solvent, drying down the lipids from organic solution, dispersion of
lipids in aqueous media, purification of resultant liposomes and analysis of the final product
 Characterization of Nanoemulsion

• Particle size measurement

• Zeta potential
• Creaming index
• Turbidity
• Phase separation

Zeta Potential

Ref.:Zetasizer Nano series technical note

Additional information
https://www.sciencedirect.com/science/article/pii/S002364381530164X

https://www.sciencedirect.com/science/article/pii/S0308814621008827