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Fl00l0Ilt 0ätI0llä
Semlnar CnťŴ
MicroencupsuIution
Microencapsulation is defined as a technology of packaging
solids. liquids or gaseous materials in miniature. sealed
capsules that can release their contents at controlled rates
under the influences of specific conditions
{Anal & 8tevens. 2005·
!urpose for microencupsuIution
%o make liquids behave like solids
8eparate reactive materials
Reduce material toxicity
Provide environmental protection to compounds
Alter surface properties of the materials
Control release of materials
Reduce volatility or flammability of liquids
Mask the taste of bitter compounds
Þr|nc|p|e of Lncapsu|at|onť Membrane barrler lsolaLes cells from Lhe hosL lmmune svsLem whlle
allowlna LransporL of meLabollLes and exLracellular nuLrlenLsŦ Membrane wlLh slze selecLlve pores (30Ŵ
70 kua)Ŧ Sourceť lnC1LCP LncapsulaLlonŦ
Þr|nc|p|e of Lncapsu|at|onť Membrane barrler lsolaLes cells from Lhe hosL lmmune svsLem whlle
allowlna LransporL of meLabollLes and exLracellular nuLrlenLsŦ Membrane wlLh slze selecLlve pores (30Ŵ
70 kua)Ŧ Sourceť lnC1LCP LncapsulaLlonŦ
SecLlon of alalnaLe mlcrocapsules showlnať
A)Ŧ Lhe sLarch aralns ln cavlLlesţ 8)Ŧ lŦ oclJopbllosţ and C)Ŧ 8Ŧ lofootls locaLed ln Lhe
alalnaLe maLrlxŦ
Ŧ Jat funct|ons must te encapsu|ated |ngred|ents prov|de for te f|na| product?
2Ŧ Jat k|nd of coat|ng mater|a| sou|d be se|ected ?
3Ŧ Jat process|ng cond|t|ons must te encapsu|ated |ngred|ent surv|ve before
re|eas|ng |ts content ?
4Ŧ Jat |s te opt|mum concentrat|on of te act|ve mater|a| |n te m|crocapsu|e ?
5Ŧ |c mecan|sm ||| te |ngred|ent be re|eased from te m|crocapsu|e ?
6Ŧ Jat are te part|c|e s|zeţ dens|tţ and stab|||t requ|rements for te encapsu|ated
|ngred|ents ?
7Ŧ Jat are te cost constra|nts of te encapsu|ated |ngred|ent ?
żShahldl and Panţ 1993Ž
In des|gn|ng te encapsu|at|on processţ te fo||o|ng
quest|ons sou|d be askedť
In des|gn|ng te encapsu|at|on processţ te fo||o|ng
quest|ons sou|d be askedť
The terms immobilization and encapsulation were used interchangeably in
most reported literature
While encapsulation is the process of forming a continuous coating around an
inner matrix that is wholly contained within the capsule wall as a core of
encapsulated material. immobilization refers to the trapping of material
within or throughout a matrix
A small percentage of immobilised material may be exposed at the surface.
while this is not the case for encapsulated material
ifference between
ImmobiIizution und EncupsuIution
The microcapsule is composed of a semi permeable. spherical. thin and
strong membranous wall
Therefore the bacterial cells are retained within the microcapsules
Moreover. compared to an entrapment matrix. there is no solid or gelled
core in the microcapsule and its small diameter helps to reduce mass
transfer limitations
Advonfoqes of Micro-encopsuIofion
ImmobiIizofion/ Enfropmenf
The nutrients and metabolites can diffuse through the semi permeable
membrane easily
The membrane serves as a barrier to cell release and minimizes
contamination
The encapsulated core material is released by several mechanisms such as
mechanical rupture of the cell wall. dissolution of the wall. melting of the
wall and diffusion through the wall
EncopsuIofion of probiofics in
poIymer sysfems
EncopsuIofion of probiofics in
poIymer sysfems
Advonfoqes
nce entrapped/ encapsulated in matrix beads or in microcapsules. the
cells are easier to handle than in a suspension or in slurry
The number of cells in beads or microparticles can be quantified.
allowing the dosage to be readily controlled
Cryo and osmo-protective components can be incorporated into the
matrix. enhancing the survival of cells during processing and storage
inally. once the matrix beads/microcapsules have been dried. a further
surface coating can be applied
This outer layer can be used to alter the aesthetic and sensory properties
of the product and may also be functional. providing an extra level of
protection to the cells
In addition. the coating layer can have desirable dissolution properties.
which permit delayed release of the cells or release upon. for example. a
change in pH
u|ture 1ecn|que/Mecan|sm Þroduct keference
8Ŧ blflJomţ 8Ŧ lofootls Calclum alalnaLe Mavonnalse khalll and Mansourţ
1998
lŦ potocosel Mllk faL Cheddar cheese SLanLon eL alŦţ 1998
£otetococcos foeclom Mllk faL Cheddar cheese Cardlner eLalŦţ 1998
8Ŧ blflJomţ 8Ŧ
oJolesceotls
Cream WhlLe brlned
cheese
Choddusl and
8oblnsonţ 1998
8Ŧ blflJomţ 8Ŧ lofootlsţ
and 8Ŧ loooom
CalclumalalnaLe aels Crescenza cheese CobbeLl eL alŦţ 1997
lŦ loctls sobsppŦ loctls kŴCarraaeenan and locusL
bean aum
lresh cheese Sodlnl eL alŦţ 1997
lŦ cosel Llquld core alalnaLe
capsule
LacLlc acld ?oo eL alŦţ 1996
LacLobacllll AlalnaLe lrozen desserL Sheu and Marshallţ
1993
ImmobiIisution/ EncupsuIution of ceIIs for
food/biotechnoIoqicuI uppIicution
EncupsuIution of probiotics in k-
curruqeenun
Carrageenan is a natural polysaccharide that is extracted from marine
macroalgae and is commonly used as a food additive
Gelation of k-carrageenan is generally dependent on a change in
temperature.
The cell slurry is added to the heat-sterilized carrageenan solution at 40-45
0
C and gelation occurs by cooling to room temperature
The beads are formed after dropping the mixture of polymer and cells into
a potassium chloride (KCl) solution
The conventional encapsulation method. with sodium alginate in calcium
chloride (CaCl
2
). has been used to encapsulate L. acidophilus to protect
this organism from the harsh acidic conditions in gastric fluid
Studies have shown that calcium-alginate immobilized cell cultures are
better protected. shown by an increase in the survival of bacteria under
different conditions. than the non-encapsulated state
EncupsuIution of probiotics in
uIqinute systems
Alginic acid. a natural polymer. is a polyuronic acid that is extracted from
seaweeds and is composed of various proportions of 1-4 linked ß-D-
mannuronic (M) and o-L-guluronic (G) acids
These residues are present in various proportions depending on the source
of the alginic acid
Alginic acid and its salts are block copolymers. containing both MM and
GG homopolymer blocks and mixed blocks containing irregular sequences
of M and G units
The binding of divalent cations and the subsequent gel formation are
dependent on the composition and arrangement of the blocks of
residues
The GG blocks have preferential binding sites for divalent counter-
ions. such as Ca
2+
. and the bound ions interact with other GG blocks to
form linkages that lead to gel formation
n addition of sodium alginate solution to a calcium solution.
interfacial polymerization is instantaneous. with precipitation of
calcium alginate followed by a more gradual gelation of the interior as
calcium ions permeate through the alginate systems
EncupsuIution of probiotics in
ceIIuIose ucetute phthuIute (CA!
ecause of its ionizable phthalate groups. this cellulose derivative polymer
is insoluble in acid media at pH 5 and lower but is soluble at pH higher
than 6
In addition. CAP is physiologically inert when administered in vivo. and
is. therefore. widely used as an enteric coating material for the release of
core substances for intestinal targeted delivery systems
#ao. Shiwnarain. and Maharaj (1989) reported the encapsulation of .
pseudolongumin CAPusing an emulsion technique
Microencapsulated bacteria survived in larger numbers (10
9
cfu/mL) in an
acidic environment than non-encapsulated organisms. which did not retain
any viability when exposed to a simulated gastric environment for 1 h.
EncupsuIution of probiotics in proteins und
poIysucchuride mi×tures
Gelatin is useful as a thermally reversible gelling agent for encapsulation
ecause of its amphoteric nature. it is also an excellent candidate for
incorporating with anionic-gelforming polysaccharides. such as gellan gum
These hydrocolloids are miscible at pH >6. because they both carry net
negative charges and repel one another
However. the net charge of gelatin becomes positive when the pH is
adjusted below its isoelectric point and causes a strong interaction with
the negatively charged gellan gum
In a recent study. Guerin. Vuillemard. and Subirade (2003) encapsulated
ifidobacterium cells in a mixed gel composed of alginate. pectin and whey
proteins
They investigated the protective effects of gel beads without extra
membrane and gel beads coated with extra membranes. formed by the
conjugation of whey protein and pectin. in simulated gastric pH and bile
salt solutions on the survival of free and encapsulated . bifidum
EncupsuIution of probiotics in
chitosun
The biopolymer chitosan. the N-deacetylated product of the polysaccharide
chitin. is gaining importance in the food and pharmaceutical field because of
its unique polymeric cationic character. good biocompatibility. non-toxicity
and biodegradability
Chitosan can be isolated from crustacean shells. insect cuticles and the
membranes of fungi
The properties of chitosan vary with its source
The terms chitin and chitosan refer not to specific compounds but to two
types of copolymers. containing the two monomer residues anhydro-N-
acetyl-D-glucosamine and anhydro-D-glucosamine. respectively
Chitin is a polymer of b-(1-4)-2-acetamido-2- deoxy-D-glucopyranose
and is one of the most abundant organic materials on earth and second to
cellulose and murein. which is the main structural polymer of the
bacterial cell wall
In order to achieve sufficient stability. chitosan gel beads and
microspheres can be ionically cross-linked with Polyphosphates and
sodium alginate
EncupsuIution of probiotics in
sturch
Starch is a dietary component that has an important role in colonic physiology
and functions and a potential protective role against colorectal cancer
(Cassidy. ingham. & Cummings. 1994)
#esistant starch is the starch that is not digested by pancreatic amylases in
the small intestine and reaches the colon. where it can be fermented by human
and animal gut microflora
The fermentation of carbohydrates by anaerobic bacteria produces short
chain fatty acids and lowers the pH in the lumen
#esistant starch can be used to ensure the viability of probiotic populations
from the food to the large intestine
#esistant starch also offers an ideal surface for adherence of the probiotics to
the starch granule during processing. storage and transit through the upper
gastrointestinal tract. providing robustness and resilience to environmental
stresses.
acterial adhesion to starch may also provide advantages in new probiotic
technologies to enhance delivery of viable and metabolically active probiotics
to the intestinal tract
Talwalkar and Kailasapathy (2003) produced alginatee starch gel beads
by dropping a mixture of alginatee starch-bacteria into a CaCl
2
coagulation bath
The probiotic bacteria used for this study were L. acidophilus and .
lactis.
They found that encapsulation prevented cell death from oxygen toxicity
It is known that alginate gel beads restrict the diffusion of oxygen through
the gel. creating anoxic regions in the centre of the beads
8eneflL ÞroducL
laclllLaLes Lhe producLlon of oxvaenŴsenslLlve culLures urled probloLlc culLure
laclllLaLes Lhe recoverv of cenLrlfuaaLlonŴsenslLlve culLures urled probloLlc culLure
laclllLaLes Lhe recoverv of hlah LÞSŴproduclna culLures urled probloLlc culLure
Less conLamlnaLlon problems urled probloLlc culLure
CulLures can be alrŴdrled urled probloLlc culLure
lmproved survlval on exposure Lo aasLrlc soluLlons nuLraceuLlcal
lmproved survlval on exposure Lo blle soluLlons nuLraceuLlcal
lmproved sLablllLv durlna sLoraae ln drled form nuLraceuLlcal
lmproved acldlflcaLlon raLe urled sausaaes
lmproved survlval on heaLlna 8lsculLsţ powder
lmproved survlval on freezlna lce creamţ mllkŴbased medlum
lmproved reLenLlon ln Lhe flnlshed producL Cheese
ÞroLecLlon aaalnsL bacLerlophaaes lermenLed mllks
ÞroLecLlon aaalnsL veasL conLamlnanLs lermenLed mllks
lmproved survlval durlna sLoraae ?oahurLţ mllk
eneficioI effecfs of probiofic
microencopsuIofion
M|croencapsu|at|on
tecn|ques
1pes of mater|a|s for
coat|ng
Ma[or steps |n processes
SpravŴdrvlna WaLerŴsoluble polvmers (l) ÞreparaLlon of Lhe soluLlons lncludlna
mlcrooraanlsms
(ll) ALomlzaLlon of Lhe feed lnLo sprav
(lll) urvlna of sprav (molsLure evaporaLlon)
(lv) SeparaLlon of drled producL form
SpravŴconaeallna Waxesţ faLLv acldsţ waLerŴ
soluble and waLerŴlnsoluble
polvmersţ monomers
(l) ÞreparaLlon of Lhe soluLlons conLalnlna core
(eŦaŦ probloLlcs)
(ll) SolldlflcaLlon of coaL bv conaeallna Lhe molLen
coaLlna maLerlals lnLo nonŴsolvenL
(lll) 8emoval of nonŴsolvenL maLerlals bv sorpLlonţ
exLracLlon or evaporaLlon Lechnlques
lluldlzedŴbed
coaLlna/
alrŴsuspenslon
WaLerŴlnsoluble and waLerŴ
soluble polvmersţ llpldsţ
waxes
(l) ÞreparaLlon of coaLlna soluLlons
(ll) lluldlzaLlon of core parLlcles
(lll) CoaLlna of core parLlcles wlLh coaLlna
soluLlons
%echniques ond processes used for
encopsuIofinq probiofic microorqonisms
M|croencapsu|at|on
tecn|ques
1pes of mater|a|s for
coat|ng
Ma[or steps |n processes
LxLruslon WaLerŴsoluble and waLer
lnsoluble polvmers
(l) ÞreparaLlon of coaLlna soluLlon maLerlals
(ll) ulsperslon of core maLerlals
(lll) Coollna or passlna of coreŴcoaL mlxLures
Lhrouah dehvdraLlna llquld
CoacervaLlon/phase
separaLlon
Lechnlque
WaLerŴsoluble polvmers (l) Core maLerlal ls dlspersed ln a soluLlon of
coaLlna polvmerţ Lhe solvenL for Lhe polvmer
belna Lhe llquld manufacLurlna vehlcle phase
(ll) ueposlLlon of Lhe coaLlnaţ accompllshed bv
conLrolledţ phvslcal mlxlna of Lhe coaLlna and core
maLerlals ln Lhe vehlcle phase
(lll) 8laldlfvlna Lhe coaLlna bv Lhermalţ crossŴ
llnklna or desolvaLlon Lechnlquesţ Lo form selfŴ
susLalnlna mlcrocapsules
LlecLrosLaLlc meLhod CpposlLelv charaed
polvmers/ compounds
(l) Mlxlna of core and coaLlna maLerlals
(ll) LxLruslon of mlxLures of coreŴcoaLlna maLerlals
ln opposlLelv charaed soluLlons
(lll) lreezeŴdrv or ovenŴdrv of
mlcrocapsules/mlcrospheres/beads
praŴcoat|ng metods for te m|croencapsu|at|on of prob|ot|csŦ 1he Lhree Lechnoloales
lllusLraLed prlnclpallv dlffer ln Lhe Lvpe of alr fluldlzaLlon emploved and Lhe slLe ln Lhe
vessel where Lhe coaLlna maLerlal ls spravedŦ
praŴcoat|ng metods for te m|croencapsu|at|on of prob|ot|csŦ 1he Lhree Lechnoloales
lllusLraLed prlnclpallv dlffer ln Lhe Lvpe of alr fluldlzaLlon emploved and Lhe slLe ln Lhe
vessel where Lhe coaLlna maLerlal ls spravedŦ
CelŴparLlcle Lechnoloales for Lhe mlcroencapsulaLlon of probloLlcsŦ 1hree Lechnlques used for Lhe ML of
probloLlcs ln alalnaLe aelsŦ
ln Lhe exLruslon process (far lefL)ţ Lhe slze of Lhe parLlcles can be ad[usLed bv uslna vlbraLlna nozzles or
plezzo effecLsŦ WlLh Lhe emulslon processes (cenLre and rlahL)ţ aalLaLlon speed and concepLlon of Lhe
mlxers enable bead slze ad[usLmenLsŦ
1he emulslon processes are carrled ouL bv addlna an alalnaLe or carraaeenan cell suspenslon Lo an oll
phaseŦ SolldlflcaLlon Lhen occurs Lhrouah Lhe addlLlon of elLher a CaCl2 soluLlon or an acld soluLlonŦ
CoŴencapsulaLlon can be carrled ouL bv addlna Lhe second bloacLlve lnaredlenL Lo Lhe alalnaLe soluLlonţ
Lo Lhe polvmerlslna soluLlon or Lo Lhe coaLlna soluLlonŦ
ConcIusions und Future
Trends
Micro-encapsulation will assume importance in delivering viable strains of
probiotic bacteria in large numbers to consumers
It will be used as a tool to co-encapsulate both prebiotic ingredients and
probiotic bacteria within the same capsule to enhance growth and
multiplication of these bacteria through symbiotic effects when they are
released in the gastro-intestinal tract
In the future multiple-delivery may be developed. such as co-
encapsulating prebiotics and probiotics as well as nutraceuticals. thus a
new area of more complex nutritional matrices will need to be investigated
More in vivo studies should be conducted using human subjects to
confirm the efficacy of micro or nano encapsulation in delivering probiotic
bacteria and their controlled release in the gastro-intestinal system

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