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Micro- and Nano-encapsulation TechnologiesCSIRO FOOD AND NUTRITIONMary Ann Augustin & Luz SanguansriShort Course on Micro- and Nano-encapsulation of Functional Ingredients in Food ProductsWorld Congress on Oils & Fats and 31st Lectureship Series31st Oct 4th November 2015, Rosario, ArgentinaOutline Encapsulation Technology Applications in the Food Industry Nanotechnology & Nanoencapsulation Approaches for Control of Size and Assembly of Materials Applications in the Food IndustryMicro and Nanoencapsulation Technologies | Augustin & Sanguansri2 |Encapsulation TechnologyApplications in the Food Industry3 | Micro and Nanoencapsulation Technologies | Augustin & SanguansriRole of Microencapsulation in the Food Industry Micro and Nanoencapsulation Technologies | Augustin & Sanguansri4 |Adapted from Perez & Gaonakar, Microencapsulation in the Food Industry, 2014, 543-549Health & WellnessENCAPSULATION HAS AN IMPORTANT ROLE IN THE FOOD INDUSTYFlavour & TasteInteractive Foods & PackagingCONSUMERS ARE DEMANDING MORE PRODUCT ATTRIBUTESConvenience& Cost-effectivenessFood Safety & Stability THE FOOD INDUSTRY IS LOOKING FOR SUPERIOR INGREDIENTImproved shelf life and product attributesWhat are some of the things to think about?Desired functionality of encapsulated ingredients in selected applicationsApplication Purpose Desired functionality of encapsulant matrixFlavours Protection Provide protection against environment and undesirable ingredient interactionsControlled 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 interactionsDecrease flavour release Slow the release of undesirable flavours (e.g. bitterness of some nutrients, chalky taste of calcium salts)Site-specific deliveryProtect against gastrointestinal tract conditions until targeted release site (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 bakingMicro and Nanoencapsulation Technologies | Augustin & Sanguansri5 |Perez & Gaonakar, Microencapsulation in the Food Industry, 2014, 543-549Microencapsulation for Food & Beverage IndustryIndustry Segment Ingredient FunctionReady to Eat Meat Organic acids (et lactate) Improve shelf lifeIncrease resistance to bacteria (Listeria monocytogenes, Clostridia, Salmonella)Bakery Flavours Fat barrier stabilises flavours in ready-to-bake doughs (eg Flavourshure - Balchem)Gums and candies Volatile anti-odour or anti-microbial or taste-masking formulationsMinimize loss of volatile active components(eg TheraBreth (cinnamic aldehyde) Wrigley;Trident (menthol) Mondolez; Instant coffee Thiols, unsaturated aldehydes, ketonesFlavour componentsDairy desserts Probioitics and vitamins Improve nutritional valueRange of dairy and food productsOmega-3 fatty acids / oil Improve nutritional valueBeverages Gas Gas-infusing or turbulence-inducing microparticles to produce froth or foams (eg instant cappuccino)Micro and Nanoencapsulation Technologies | Augustin & Sanguansri6 |Perez & Gaonakar, Microencapsulation in the Food Industry, 2014, 543-549Microencapsulation for Food & Beverage IndustryIngredient EncapsulationMechanismFunction CommercialApplication Flavour compounds (eg thiols in coffee and esters in fruit)Heat resistant coatingIsoelectric precipitationFlavour enhancement Tea, coffee, juiceOmega-3 fatty acids, probiotics, prebioticsHeat and moisture-tolerant coating, isoelectric precipitationFlavor/odor masking and protection from moisture and heatBeverage, nutritional bar, cerealMint flavours Coacervates Flavour release and long lastingChewing gumCheese ripening enzymes Enzyme immobilisation Cheese ripening Cheese productsProbiotics Biopolymer matrix Stabilisation during storage and protection through stomachDairy productsCarbonate, pressurised air Gas inclusion system / biopolymers/ cyclodextrinFoaming BeveragesSpoilage by-productreacting agentNanocomposite / MicroencapsulationColour change to indicatefood safetyInteractive and intelligentpackagingMicro and Nanoencapsulation Technologies | Augustin & Sanguansri7 |Perez & Gaonakar, Microencapsulation in the Food Industry, 2014, 543-549Selected Examples from the Literature- Dairy encapsulants for hydrophobic, hydrophilic and probiotic cores- Plant protein-based micro- and nano-particlesMicro and Nanoencapsulation Technologies | Augustin & Sanguansri8 |Micro and Nanoencapsulation Technologies | Augustin & SanguansriDairy-based encapsulants used with hydrophobic cores Example 19 |Encapsulated componentDairy encapsulant Encapsulation techniqueBenefit(s) of encapsulationReferenceOrange oil WPI Spray drying Protection against oxidationKim & Morr, 1996Soy oil Sodium caseinate Spray drying High encapsulation efficiency (89%)Hogan et al., 2001aCLA WPC Spray drying Protection against oxidationJimenez et al., 2004; 2006Flaxseed oil WPI Spray drying Protection against oxidationPartanen, Raula, Seppnen, Buchert, Kauppinen, & Forssell, 2008AMF WPI Spray drying Protection against oxidation during storageMoreau & Rosenberg, 1996AMF WPI, WPC-50, WPC-75Spray drying High encapsulation efficiency (> 90%)Young et al., 1993aRetinol WPI Emulsification/Cold gelation /Air dryingGastroresistance and protection against oxidationBeaulieu et al., 2002Oregano, citronella and marjoram flavoursSMP or WPC Spray drying Improved retention of flavours during spray dryingBaranauskien et al., 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.Micro and Nanoencapsulation Technologies | Augustin & SanguansriDairy-based encapsulants used with hydrophilic cores Example 210 |Encapsulated componentDairy encapsulant Encapsulation techniqueBenefit(s) of encapsulation Reference3-methylbutyr-aldehyde WPC and sodium caseinate or SMP as secondary emulsifierDouble emulsification/Spray dryingImproved retention of aldehyde during storageBrckner et al., 2007 Sumac concentrateWhey powder or SMPSpray drying Improved retention of flavour during spray dryingBayram et al., 2008Ascorbic acid Lactose Co-crystallisation Improved retention of ascorbic acid during co-crystallizationKim et al., 2001Citric acid Casein Co-crystallisation Development of a novel, efficient and cost-effective microwave encapsulation technique that provided high encapsulation efficiency (100%)Abbasi & Rahimi, 2008IgY WPC as secondary emulsifierDouble emulsion /Gelation/Air dryingProtected IgY from highly acidic conditions and heat treatment processesCho et al., 2005Protease enzymesHigh melting milkfat fractionGel beads Increased rate of proteolysis during cheese ripeningKailasapathy & Lam, 2005Caffeine WPC Hydrogels/Air dryingControlled release of caffeineGunasekaran et al., 2006Augustin, 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.Micro and Nanoencapsulation Technologies | Augustin & SanguansriDairy-based encapsulants used for probiotics Example 311 |Encapsulated componentDairy encapsulantEncapsulation techniqueBenefit(s) of encapsulationReferenceLactobacillus sp Milkfat and/or denatured WPIEmulsification/Spray dryingImproved cell viability in yogurt and after exposure to simulated gastrointestinal fluidsPicot & Lacroix, 2003; 2004Lactobacillus sp WPI Freeze drying Improved cell viability during storage and in yogurtKailasapathy & Sureeta, 2004 Bifidobacterium sp WPI Freeze drying Improved cell viability in simulated gastrointestinal fluidsReid et al., 2005WPI Freeze drying Improved cell viability during the production and storage of biscuits, and improved pH stabilityReid et al., 2007Milkfat Spray coating Improved cell viability during storage Champagne et al., 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.Plant protein-based micro- and nanoparticles for food ingredient Delivery - 1Micro and Nanoencapsulation Technologies | Augustin & Sanguansri12 |Type of particle Method CoreZein microparticles Spray drying or supercritical anti-solvent methodFood grade antimicrobials: lysozyme, thymol, nisinSpray or freeze drying Flax oilZein nanoparticles Liquidliquid dispersion methodPolyphenols: curcumin, quercetin, tangeretin, cranberryprocyanidinsPhase separation or liquidliquidEssential oils: oregano, red thyme, cassia and carvacrolLiquidliquid dispersion method or electrosprayingBioactive lipids: fish oil, DHA, Food coloring agents: curcumin, indigocarmineZein-chitosan complex nanoparticlesLow-energy phase separation methodVitamin D3SPI-zein complex microparticles / SPI nanoparticlesCa2+-induced cold gelation methodRiboflavin / Vitamin B12Wan et al. (2015) Food & Function 6, 2876 2889 Plant protein-based micro- and nanoparticles for food ingredient Delivery - 2Micro and Nanoencapsulation Technologies | Augustin & Sanguansri13 |Type of particle Method CoreSPI/FA-conjugated SPI Ethanol solvation method CurcuminSPI-CMCS complex nanoparticlesCa2+ induced co-gelation methodVitamin D3Soy protein-soy polysaccharide complex nanogelsHigh-pressure homogenization and heatingFolic acidSoy lipophilic protein Ultrasonic treatment Conjugated linoleic acidGliadin nanoparticles Antisolvent precipitation methodAll-trans-retinoic acid, vitamin EBarley protein microparticlesPre-emulsifying process followed by microfluidizingFish oil, -caroteneBarley protein nanoparticles High pressure homogenization-CaroteneSoy protein nanocomplex Ligand binding properties Vitamin B12, cranberry polyphenols, curcumin, RES and grape polyphenolWan et al. (2015) Food & Function 6, 2876 2889 Nanotechnology and Nanoencapsulation14 | Micro and Nanoencapsulation Technologies | Augustin & SanguansriNanotechnologyNanotechnology 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 structures15 |Roco, Current Opinion in Biotechnology 2003, 14:337Micro and Nanoencapsulation Technologies | Augustin & SanguansriRelevance of the concept of scale to food materials Link to Nanotechnology ConceptsLeser et al., IN Food colloids, biopolymers and materials (Eds Dickinson and van Vliet, 2003), pp3-13Micro and Nanoencapsulation Technologies | Augustin & Sanguansri16 |Nanotechnology Applications across Agrifood Micro and Nanoencapsulation Technologies | Augustin & Sanguansri17 |http://www.bing.com/images/search?q=nanotechnology+in+food&view=detailv2&&id=B6E9F703FECEF1068BC82C8DFE20233396618D39&selectedIndex=0&ccid=8LqON4nK&simid=608000695069049234&thid=OIP.Mf0ba8e3789ca59ba3e7f3eeadf1d949bH0&ajaxhist=0Concept 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 etcControlling 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 functionalityMicro and Nanoencapsulation Technologies | Augustin & Sanguansri18 |Nanoencapsulated particlesNanoemulsions and Nanoparticles- Developed using a range of materials- Co-block polymer micelles, polyelectrolyte capsules, colloidosomes, polymersomes, gelled macromoleculesTarget 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 body19 | Micro and Nanoencapsulation Technologies | Augustin & SanguansriNew materials based on Nanotechnology20 | Micro and Nanoencapsulation Technologies | Augustin & SanguansriNanotechnology and NanoencapsulationApproaches for Control of Size and Assembly of Materials21 | Micro and Nanoencapsulation Technologies | Augustin & SanguansriTop down and bottom up approachesMicro and Nanoencapsulation Technologies | Augustin & Sanguansri22 |Scientific Approaches for Modification of Materials in Nanotechnology Top-down approach 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-assemblingBioSiliconMicro and Nanoencapsulation Technologies | Augustin & Sanguansri23 |http://pubs.acs.org/journals/mdd/about.htmlhttp://pubs.acs.org/journals/mdd/about.htmlTop-Down Approach for Size reduction of food Ball Milling and Jet Milling High Pressure Homogenisation Microfluidisation Ultrasound Emulsification Membrane EmulsificationMaterials_ Microencapsulation | Augustin & Sanguansrihttp://www.avestin.com/c5page.htmlhttp://www.avestin.com/c5page.htmlSolid Lipid Nanoparticles (SLNs)Materials_ Microencapsulation | Augustin & Sanguansri25 |SLNs are particles consisting of a matrix made of solid lipid shell Weiss et al. (2008) Food Biophysics, 3, 146-154Emulsions How the components assemble will affect its functional propertiesMicro and Nanoencapsulation Technologies | Augustin & Sanguansri26 |Bottom-up Approach in NanotechnologyBuilding 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 micellesB) Polyelectrolyte capsulesC) Colloidosomes D) Block co-polymer vesicles (polymersomes)Micro and Nanoencapsulation Technologies | Augustin & Sanguansri27 |Forster & Konrad, J. Material Chemistry, 13, 2671, 2003Self-Assembled Nanoparticle of Common Food Constituents That Carries a Sparingly Soluble Small MoleculeMicro and Nanoencapsulation Technologies | Augustin & Sanguansri28 |Bhopatkar et al., JAFC 2015, 63 (17), pp 43124319Nanotechnology and NanoencapsulationApplications in the Food Industry29 | Micro and Nanoencapsulation Technologies | Augustin & SanguansriNanotechnology in the Food IndustryMoraru et al., 2003. Food Technology, Vol 57(12), 24Micro and Nanoencapsulation Technologies | Augustin & Sanguansri30 |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 Micro and Nanoencapsulation Technologies | Augustin & Sanguansri31 |Nanotechnology in Food SafetySensing for safetyCreation 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 bacteriaMicro and Nanoencapsulation Technologies | Augustin & Sanguansri32 |Bacterial detection in drinking water based on gold nanoparticleenzyme complexesMicro and Nanoencapsulation Technologies | Augustin & Sanguansri33 | 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 bacteriaIncrease in enzyme activity, releasing a redox-active phenol from the substrate Sensing of Escherichia coli and Staphylococcus aureus, resulting in a rapid detection (Nanoencapsulation of essential oils to enhance their antimicrobial activity in foodsMicro and Nanoencapsulation Technologies | Augustin & Sanguansri34 |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, 44,19081914 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 observedInactivation curve of L. delbrueckii suspended in juice with terpenes nanoemulsionMicro and Nanoencapsulation Technologies | Augustin & Sanguansri35 |Donsi et al (2011) LWT - Food Sci Tech, 44,19081914(a) orange juice treated with terpenes nanoemulsion (b) pear juice treated with terpenes nanoemulsion ControlControl1g/L terpene mixture1g/L terpene mixture10g/L terpene mixture10g/L terpene mixtureNanocomposites used as antimicrobial films for food packaging based on metallic silverMicro and Nanoencapsulation Technologies | Augustin & Sanguansri36 |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 legislationhttp://apps.webofknowledge.com/full_record.do?product=UA&search_mode=GeneralSearch&qid=12&SID=W1xug7lYTivM8qS8SYs&page=9&doc=84&cacheurlFromRightClick=noCanadian penny-based silver nano-structure was synthesized as SERS-active substrate for determination of CAP in food matricesDetects trace level of chemical hazards in food systems within 15 minMicro and Nanoencapsulation Technologies | Augustin & SanguansriDetection and Quantification of Chloramphenicol in Milk and Honey Using Molecularly Imprinted Polymers: Canadian Penny-Based SERS (Surface-enhanced Raman Spectroscopy) Nano-Biosensor37 |Gao et al (2014) JFS, 79(12) N2542-N2549Template 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 polymerizationNanotechnology for healthy foodsDevelopment of healthier foodsHealthy 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 ingredientsMicro and Nanoencapsulation Technologies | Augustin & Sanguansri38 |Nanoparticles for delivery of bioactivesMicro and Nanoencapsulation Technologies | Augustin & Sanguansri39 |McClements (2015) Journal of Food Science, 80(7), N1602-N1611, Micro and Nanoencapsulation Technologies | Augustin & SanguansriStability of Bioactive in SLN & crystal structure of fat 40 | 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 compoundSalminen et al. Food Chem (2016), 928-937Nanotechnology encapsulation platforms used in food applicationsChitosan hydrogels Whey protein nanostructuresExamples: Novasol range: ready to use liquid formulations by Aquanova AG NutraLease: nanosized self assembled liquid structures (NSSL)Micro and Nanoencapsulation Technologies | Augustin & Sanguansri41 |Market drivers & trends - Encapsulation5395.2 5788.7 6222.69070.53776.1 4049.14347.16307.82807.23025.83267.74872.71619.11717.31823.92493.2$0$5,000$10,000$15,000$20,000$25,0002007 2008 2009 2014MacroencapsulationHybrid TechnologiesNano-encapsulationMicroencapsulationGLOBAL FOOD ENCAPSULATION MARKET BY TECHNOLOGY2007 2014 ($MILLIONS)Source: Global food encapsulation market, MarketsandMarkets, 2009** Global Business Insights Innovations in delivery methods for nutraceutical food and drinks, 2011CAGR 7.8%CAGR 7.7%CAGR 8.3%CAGR 6.5%CAGR 2009-14Mmarket: $23 billion by the year 2014Europe - CAGR of 9.1% North America - largest market share - 41% in 2014 Target Groups/Markets:Infant, functional and health food segments Ageing populationFoods with disease prevention benefits.Concerns Nanoencapsulation:Whilst this is in part due to potential and unknown toxicity relating to nanoparticles, it is also the case that nanoencapsulation is rarely the best solution** $ MillionsYearMicro and Nanoencapsulation Technologies | Augustin & SanguansriThank youCSIRO Food & NutritionMary Ann AugustinResearch Group Leadert +61 3 9731 3486e maryann.augustin@csiro.auMicro- and Nano-encapsulation TechnologiesOutlineSlide Number 3Role of Microencapsulation in the Food Industry What are some of the things to think about?Microencapsulation for Food & Beverage IndustryMicroencapsulation for Food & Beverage IndustrySlide Number 8Slide Number 9Slide Number 10Slide Number 11Plant protein-based micro- and nanoparticles for food ingredient Delivery - 1Plant protein-based micro- and nanoparticles for food ingredient Delivery - 2Slide Number 14NanotechnologyRelevance of the concept of scale to food materials Link to Nanotechnology ConceptsNanotechnology Applications across Agrifood Concept of Size and Its Implications for Food Materials, Processes and Products Nanoencapsulated particlesSlide Number 20Slide Number 21Top down and bottom up approachesScientific Approaches for Modification of Materials in NanotechnologyTop-Down Approach for Size reduction of food Solid Lipid Nanoparticles (SLNs)Emulsions How the components assemble will affect its functional propertiesBottom-up Approach in NanotechnologySelf-Assembled Nanoparticle of Common Food Constituents That Carries a Sparingly Soluble Small MoleculeSlide Number 29Nanotechnology in the Food IndustryPotential benefits of nanotechnology in FoodNanotechnology in Food SafetyBacterial detection in drinking water based on gold nanoparticleenzyme complexes Nanoencapsulation of essential oils to enhance their antimicrobial activity in foodsInactivation curve of L. delbrueckii suspended in juice with terpenes nanoemulsionNanocomposites used as antimicrobial films for food packaging based on metallic silverSlide Number 37Nanotechnology for healthy foodsNanoparticles for delivery of bioactivesSlide Number 40Nanotechnology encapsulation platforms used in food applicationsMarket drivers & trends - EncapsulationSlide Number 43

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