Micro- and Nano-encapsulation

Technologies
Short Course on Micro- and Nano-encapsulation of Functional Ingredients in Food Products
World Congress on Oils & Fats and 31st Lectureship Series
31st Oct – 4th November 2015, Rosario, Argentina
CSIRO FOOD AND NUTRITION

Mary Ann Augustin & Luz Sanguansri

Outline

• Encapsulation Technology
• Applications in the Food Industry

• Nanotechnology & Nanoencapsulation

• Approaches for Control of Size and Assembly of Materials
• Applications in the Food Industry

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Encapsulation Technology

Applications in the Food Industry

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Role of Microencapsulation in the Food Industry

ENCAPSULATION HAS
Flavour & Health & AN IMPORTANT ROLE
Taste Wellness IN THE FOOD
INDUSTY

CONSUMERS ARE Convenience

Interactive
DEMANDING MORE
Foods & & Cost-
PRODUCT
Packaging effectiveness
ATTRIBUTES

Improved THE FOOD INDUSTRY

shelf life Food Safety IS LOOKING FOR
and product & Stability SUPERIOR
attributes INGREDIENT

Adapted from Perez & Gaonakar, Microencapsulation in the Food Industry,

2014, 543-549

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What are some of the things to think about?
Desired functionality of encapsulated ingredients in selected applications
Application Purpose Desired functionality of encapsulant matrix
Flavours Protection Provide protection against environment and undesirable ingredient
interactions
Controlled release Release flavour in the mouth in response to the desired trigger (e.g. shear
due to chewing for flavour burst, dissolution when in contact with saliva)
Bioactives Protection Provide protection against environment and undesirable ingredient
interactions
Decrease flavour Slow the release of undesirable flavours (e.g. bitterness of some nutrients,
release chalky taste of calcium salts)
Site-specific Protect against gastrointestinal tract conditions until targeted release site
delivery (e.g. protect probiotics and bioactive peptides against stomach conditions)
Controlled release Control rate of release (e.g. decrease size of microcapsules to improve bio-
accessibility or tailor the thickness of the wall material to increase resistance
to gastric/intestinal enzymes)
Leavening Controlled release Leavening control during baking

Perez & Gaonakar, Microencapsulation in the Food Industry, 2014, 543-549

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Microencapsulation for Food & Beverage Industry

Industry Segment Ingredient Function

Ready to Eat Meat Organic acids (et lactate) Improve shelf life
Increase resistance to bacteria (Listeria monocytogenes,
Clostridia, Salmonella)
Fat barrier stabilises flavours in ready-to-bake doughs
Bakery Flavours
(eg Flavourshure - Balchem)
Gums and candies Volatile anti-odour or anti- Minimize loss of volatile active components
microbial or taste-masking (eg TheraBreth (cinnamic aldehyde) – Wrigley;
formulations Trident (menthol) – Mondolez;
Instant coffee Thiols, unsaturated Flavour components
aldehydes, ketones
Dairy desserts Probioitics and vitamins Improve nutritional value

Range of dairy and food Omega-3 fatty acids / oil Improve nutritional value
products
Beverages Gas Gas-infusing or turbulence-inducing microparticles to
produce froth or foams (eg instant cappuccino)

Perez & Gaonakar, Microencapsulation in the Food Industry, 2014, 543-549

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Microencapsulation for Food & Beverage Industry
Ingredient Encapsulation Function Commercial
Mechanism Application
Flavour compounds (eg Heat resistant coating Flavour enhancement Tea, coffee, juice
thiols in coffee and esters Isoelectric precipitation
in fruit)
Heat and moisture- Flavor/odor masking and
Omega-3 fatty acids, Beverage, nutritional bar,
tolerant coating, protection from moisture
probiotics, prebiotics cereal
isoelectric precipitation and heat
Mint flavours Coacervates Flavour release and long Chewing gum
lasting
Cheese ripening enzymes Enzyme immobilisation Cheese ripening Cheese products

Probiotics Biopolymer matrix Stabilisation during Dairy products

storage and protection
through stomach
Carbonate, pressurised air Gas inclusion system / Foaming Beverages
biopolymers/ cyclodextrin
Spoilage by-product Nanocomposite / Colour change to indicate Interactive and intelligent
reacting agent Microencapsulation food safety packaging

Perez & Gaonakar, Microencapsulation in the Food Industry, 2014, 543-549

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Selected Examples from the
Literature

- Dairy encapsulants for hydrophobic,

hydrophilic and probiotic cores

- Plant protein-based micro- and nano-particles

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Dairy-based encapsulants used with hydrophobic
cores – Example 1
Encapsulated Dairy encapsulant Encapsulation Benefit(s) of Reference
component technique encapsulation
Orange oil WPI Spray drying Protection against Kim & Morr, 1996
oxidation
Soy oil Sodium caseinate Spray drying High encapsulation Hogan et al., 2001a
efficiency (89%)
CLA WPC Spray drying Protection against Jimenez et al., 2004;
oxidation 2006
Flaxseed oil WPI Spray drying Protection against Partanen, Raula,
oxidation Seppānen, Buchert,
Kauppinen, &
Forssell, 2008
AMF WPI Spray drying Protection against Moreau &
oxidation during Rosenberg, 1996
storage
AMF WPI, WPC-50, WPC- Spray drying High encapsulation Young et al., 1993a
75 efficiency (> 90%)
Retinol WPI Emulsification/ Gastroresistance and Beaulieu et al., 2002
Cold gelation /Air protection against
drying oxidation
Oregano, citronella SMP or WPC Spray drying Improved retention Baranauskienė et al.,
and marjoram of flavours during 2006
flavours spray drying
Augustin, M.A. and Oliver C.M..(2014) IN The Art and Science of Microencapsulation: An Application Handbook for the Food
Industry. (Eds. Anilkumar Gaonkar, Niraj Vasisht, Atul Khare, Robert Sobel), Academic Press, Chap 19, 211-226.

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 Dairy-based encapsulants used with hydrophilic
cores – Example 2
Encapsulated Dairy encapsulant Encapsulation Benefit(s) of encapsulation Reference
component technique
3-methylbutyr- WPC and sodium Double Improved retention of Brückner et al., 2007
aldehyde caseinate or SMP as emulsification/ aldehyde during storage
secondary Spray drying
emulsifier
Sumac Whey powder or Spray drying Improved retention of Bayram et al., 2008
concentrate SMP flavour during spray drying
Ascorbic acid Lactose Co-crystallisation Improved retention of Kim et al., 2001
ascorbic acid during co-
crystallization
Citric acid Casein Co-crystallisation Development of a novel, Abbasi & Rahimi,
efficient and cost-effective 2008
microwave encapsulation
technique that provided high
encapsulation efficiency
(100%)
IgY WPC as secondary Double emulsion Protected IgY from highly Cho et al., 2005
emulsifier /Gelation/Air acidic conditions and heat
drying treatment processes
Protease High melting milkfat Gel beads Increased rate of proteolysis Kailasapathy & Lam,
enzymes fraction during cheese ripening 2005
Caffeine WPC Hydrogels/Air Controlled release of Gunasekaran et al.,
drying caffeine 2006
Augustin, M.A. and Oliver C.M..(2014) IN The Art and Science of Microencapsulation: An Application Handbook for the Food
Industry. (Eds. Anilkumar Gaonkar, Niraj Vasisht, Atul Khare, Robert Sobel), Academic Press, Chap 19, 211-226.

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Dairy-based encapsulants used for probiotics –
Example 3
Encapsulated Dairy Encapsulation Benefit(s) of Reference
component encapsulant technique encapsulation
Lactobacillus sp Milkfat and/or Emulsification/ Improved cell viability in Picot & Lacroix,
denatured WPI Spray drying yogurt and after exposure to 2003; 2004
simulated gastrointestinal
fluids
Lactobacillus sp WPI Freeze drying Improved cell viability Kailasapathy &
during storage and in yogurt Sureeta, 2004

Bifidobacterium sp WPI Freeze drying Improved cell viability in Reid et al., 2005
simulated gastrointestinal
fluids
WPI Freeze drying Improved cell viability Reid et al., 2007
during the production and
storage of biscuits, and
improved pH stability
Milkfat Spray coating Improved cell viability Champagne et al.,
during storage 1995
Augustin, M.A. and Oliver C.M..(2014) IN The Art and Science of Microencapsulation: An Application Handbook for the Food
Industry. (Eds. Anilkumar Gaonkar, Niraj Vasisht, Atul Khare, Robert Sobel), Academic Press, Chap 19, 211-226.

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Plant protein-based micro- and nanoparticles for
food ingredient Delivery - 1
Type of particle Method Core
Zein microparticles Spray drying or supercritical Food grade antimicrobials:
anti-solvent method lysozyme, thymol, nisin
Spray or freeze drying Flax oil
Zein nanoparticles Liquid–liquid dispersion Polyphenols: curcumin,
method quercetin, tangeretin, cranberry
procyanidins
Phase separation or liquid– Essential oils: oregano, red
liquid thyme, cassia and carvacrol

Liquid–liquid dispersion Bioactive lipids: fish oil, DHA,

method or electrospraying Food coloring agents: curcumin,
indigocarmine
Zein-chitosan complex Low-energy phase Vitamin D3
nanoparticles separation method
SPI-zein complex Ca2+-induced cold gelation Riboflavin / Vitamin B12
microparticles / SPI method
nanoparticles
Wan et al. (2015) Food & Function 6, 2876 – 2889
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Plant protein-based micro- and nanoparticles for
food ingredient Delivery - 2
Type of particle Method Core
SPI/FA-conjugated SPI Ethanol solvation method Curcumin
SPI-CMCS complex Ca2+ induced co-gelation Vitamin D3
nanoparticles method
Soy protein-soy High-pressure Folic acid
polysaccharide complex homogenization and heating
nanogels
Soy lipophilic protein Ultrasonic treatment Conjugated linoleic acid
Gliadin nanoparticles Antisolvent precipitation All-trans-retinoic acid, vitamin E
method
Barley protein Pre-emulsifying process Fish oil, β-carotene
microparticles followed by microfluidizing
Barley protein nanoparticles High pressure β-Carotene
homogenization
Soy protein nanocomplex Ligand binding properties Vitamin B12, cranberry
polyphenols, curcumin, RES and
grape polyphenol

Wan et al. (2015) Food & Function 6, 2876 – 2889

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Nanotechnology and
Nanoencapsulation

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Nanotechnology
Nanotechnology is the ability to work at the atomic, molecular
and supramolecular level (in the order of 1-100nm) in order to
understand, create and use material structures, devices and
systems with fundamentally new properties and functions
resulting from their small structures Roco, Current Opinion in Biotechnology 2003, 14:337

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 Relevance of the concept of scale to food
materials – Link to Nanotechnology Concepts

Leser et al., IN Food colloids, biopolymers and materials (Eds Dickinson and
van Vliet, 2003), pp3-13

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Nanotechnology – Applications across Agrifood

http://www.bing.com/images/search?q=nanotechnology+in+food&view=detailv2&&id=B6E9F703FECEF1068BC82C8DFE20233396618D39
&selectedIndex=0&ccid=8LqON4nK&simid=608000695069049234&thid=OIP.Mf0ba8e3789ca59ba3e7f3eeadf1d949bH0&ajaxhist=0

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Concept of Size and Its Implications for Food
Materials, Processes and Products

Size relates to functionality in terms of the physical

properties of food materials
• Smaller size means bigger surface area for the purposes of water
absorption (solubility), chemical reaction (e.g. oxidation, digestion),
catalyst/enzyme activity, flavour release, bioavailability etc

Controlling the size and assembly of food components

provides opportunities for designing new food products
• Link b/w nanoscale and food microstructure
• Effects on nutritional and physiological functionality

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Nanoencapsulated particles
Nanoemulsions and Nanoparticles
- Developed using a range of materials
- Co-block polymer micelles, polyelectrolyte capsules, colloidosomes,
polymersomes, gelled macromolecules

Target release
- In response to environment (eg pH, salt concentration, ultrasound)

Target distribution
- Control of surface properties of polymers
- Control interaction between particle and cells in body

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New materials based on Nanotechnology

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Nanotechnology and
Nanoencapsulation

Approaches for Control of Size and

Assembly of Materials

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Top down and bottom up approaches

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Scientific Approaches for Modification of
Materials in Nanotechnology
• Top-down approach BioSilicon™

• Nanostructures are produced by breaking up bulk materials

with large structures into smaller ones
• Physical machining of materials to nanometre range by
grinding, milling, precision engineering, homogenisation
and lithography

• Bottom-up approach
• Nanostructures are built-up from individual atoms or
molecules that are capable of self-assembling

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 Top-Down Approach for
Size reduction of food
• Ball Milling and Jet Milling

• High Pressure Homogenisation

• Microfluidisation

• Ultrasound Emulsification

• Membrane Emulsification

Materials_ Microencapsulation | Augustin & Sanguansri

Solid Lipid Nanoparticles (SLNs)

SLNs are particles consisting of a matrix made of solid lipid shell

Weiss et al. (2008) Food Biophysics, 3, 146-154

25 | Materials_ Microencapsulation | Augustin & Sanguansri

 Emulsions – How the components assemble
will affect its functional properties

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 Bottom-up Approach in Nanotechnology
Building up products by assembly of molecules [Molecule-by-
molecule formation of hierarchical structures]
• Biomimetic Approach (Mimics strategy used by biological systems for
structuring of molecules)
– Nanometre scale self-assembly by autonomous organisation of components into
structures and patterns without human intervention
– Organisation of nanometre scale molecular assemblies into larger structures from
10 nm to sub-micrometre range)

A) Self-assembled polymer structures block co-polymer micelles

B) Polyelectrolyte capsules
C) Colloidosomes
D) Block co-polymer vesicles (polymersomes)

Forster & Konrad, J. Material Chemistry, 13, 2671, 2003

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Self-Assembled Nanoparticle of Common Food
Constituents That Carries a Sparingly Soluble
Small Molecule

Bhopatkar et al., JAFC 2015, 63 (17), pp 4312–4319

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Nanotechnology and
Nanoencapsulation

Applications in the Food Industry

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Nanotechnology in the Food Industry

Moraru et al., 2003. Food Technology, Vol 57(12), 24

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Potential benefits of nanotechnology in
Food
• Food safety and shelf life extension

• Enhancement of taste, flavour and texture

• Improvements in processing

• Improvement in absorption ratio of nutrients

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Nanotechnology in Food Safety
Sensing for safety
Creation of new materials and novel methods and devices for
sensing, diagnosis and analysis of pathogens and single
molecules for ensuring safety, quality and security of the food
supply in real time

• Interactions between biomolecules and molecular assemblies

with electronic structures or materials for nano- and
microfabrication of devices for improved methods and sensors
for detecting pathogens and improved diagnostics for food
allergens

• Formation of nanoparticles and quantum dots for biotagging or

barcoding within biological systems to design products with
electronic functionality in materials for use in intelligent
packaging of food materials and tracking food quality in supply
chains

• Silver Nanoparticles embedded in plastic that line storage bins

– Ag nanoparticles kill bacteria

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Bacterial detection in drinking water based
on gold nanoparticle–enzyme complexes
• Gold nanoparticles functionalized with
positively charged quaternary amine
headgroups bind to enzymes ⇒ inhibition
of enzymatic activity

• In the presence of bacteria, the

nanoparticles were released from the
enzymes and preferentially bound to the
bacteria
⇓
Increase in enzyme activity, releasing a
redox-active phenol from the substrate

Sensing of Escherichia coli and Staphylococcus aureus, resulting in a

rapid detection (<1 h) with high sensitivity (102 CFU mL−1)

Chen et al. Analyst, 140, 4991-4996

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Nanoencapsulation of essential oils to
enhance their antimicrobial activity in foods
• Under a fluorescent light, the
nanoemulsion droplets cannot be
distinguished when they are
dispersed in an aqueous system
due to their nanometric size

• When the nanoemulsion droplets

accumulate in the cell membrane
as well as the intracellular space,
the yeast cells became fluorescent
and can be observed

Brightfield (a and c) and fluorescence micrographs (b and

d) of S. cerevisiae cells exposed to nanoemulsion Terpene-
Soy lecithin captured by fluorescence microscopy after 5
min (a and b) and 24 h (c and d).
Donsi et al (2011) LWT - Food Sci Tech, 44,1908–1914

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Inactivation curve of L. delbrueckii suspended in
juice with terpenes nanoemulsion
Control
(a) orange juice treated with
1g/L terpene mixture
terpenes nanoemulsion

10g/L terpene mixture

Control
(b) pear juice treated with
1g/L terpene mixture
terpenes nanoemulsion

10g/L terpene mixture

Donsi et al (2011) LWT - Food Sci Tech, 44,1908–1914

35 | Micro and Nanoencapsulation Technologies | Augustin & Sanguansri

Nanocomposites used as antimicrobial films for
food packaging based on metallic silver

Nanoparticle release from nano-silver antimicrobial food

containers Echegoyen & Nerin (2103)
FOOD AND CHEMICAL TOXICOLOGY, 62, 16-22

• In all cases the total Ag migration is far below the

maximum migration limits stated by the European
legislation

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Detection and Quantification of Chloramphenicol in Milk and Honey
Using Molecularly Imprinted Polymers: Canadian Penny-Based SERS
(Surface-enhanced Raman Spectroscopy) Nano-Biosensor
Template molecule (CAP), functional
monomer (acrylamide), cross-linking
agent (ethylene glycol dimethacrylate),
initiator (2,2’-azobis(isobutyronitrile)),
and porogen (methanol) were employed
to form MIPs via “dummy” precipitation
polymerization

Canadian penny-based silver nano-structure was synthesized as SERS-active

substrate for determination of CAP in food matrices
⇓
Detects trace level of chemical hazards in food systems within 15 min
Gao et al (2014) JFS, 79(12) N2542-N2549

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 Nanotechnology for healthy foods
Development of healthier foods
Healthy foods and diets may be devised to promote health of consumers
and the understanding between genetic pre-dispositions, nutrition and diet
may be used to design diets for target populations

• New nanoscale technologies for the fabrication of materials, manufacture

and control of microencapsulated products have potential for improving
the quality of functional foods and target delivery of bioactives and desired
molecules

• Nanotechnology may be directed towards the manipulation of food surface

structure on a molecular scale to improve the metabolic consequences of
consuming processed foods

• Biotransformation for production of high value nutritional components and

food ingredients

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Nanoparticles for delivery of bioactives

McClements (2015) Journal of Food Science, 80(7), N1602-N1611,

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Stability of Bioactive in SLN & crystal structure of fat

• Crystallization behavior of SLN

depended on the bioactive and
surfactant type

• Oxidative stability of bioactives

depended on the crystal
structure

• Delivery systems need to be

designed specifically for each
bioactive compound

Salminen et al. Food Chem (2016), 928-937

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Nanotechnology encapsulation platforms used in
food applications

Examples:
• Novasol® range: ready to use liquid formulations
by Aquanova AG
• NutraLease®: nanosized self assembled liquid
structures (NSSL)

Chitosan hydrogels Whey protein nanostructures

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 Market drivers & trends - Encapsulation
GLOBAL FOOD ENCAPSULATION MARKET BY TECHNOLOGY Mmarket: $23 billion by the year 2014
2007 – 2014 ($MILLIONS)
Europe - CAGR of 9.1%
$25,000 North America - largest market share - 41% in
Macroencapsulation CAGR 2009-14
Hybrid Technologies 2014
Nano-encapsulation 2493.2 CAGR 6.5%
Microencapsulation
$20,000

4872.7 Target Groups/Markets:

CAGR 8.3%
Infant, functional and health food segments
$ Millions

$15,000 1823.9 Ageing population

1717.3
1619.1
6307.8 CAGR 7.7% Foods with disease prevention benefits.
3267.7
3025.8
2807.2
$10,000
4347.1
4049.1 Concerns – Nanoencapsulation:
3776.1 CAGR 7.8%
“Whilst this is in part due to potential and
$5,000
9070.5 unknown toxicity relating to nanoparticles, it is
5395.2 5788.7 6222.6 also the case that nanoencapsulation is rarely
the best solution”**
$0
2007 2008 2009 2014

Year
Source: Global food encapsulation market, MarketsandMarkets, 2009
** Global Business Insights – Innovations in delivery methods for nutraceutical food and drinks, 2011

Micro and Nanoencapsulation Technologies | Augustin & Sanguansri

Thank you
CSIRO Food & Nutrition
Mary Ann Augustin
Research Group Leader
t +61 3 9731 3486
e maryann.augustin@csiro.au