e emulsification: an innovative continuous process to create ......marianna petronela vamanu, serena...
TRANSCRIPT
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Membrane emulsification: an innovative continuous process to
create highly uniform capsules/particles
Dr Marijana Dragosavac Loughborough University
Chemical Engineering Department
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• How can we produce drops and turn them into particles?
• Conventional ways to produce drops
• Drop by Drop devices to produce drops • Microchannel & Membrane emulsification
• Specific encapsulations – from drops to particles
PRESENTATION LAYOUT
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• Highest value products - complex formulated systems
• To produce uniform particles - novel methods for producing - novel formulations
• Droplets with D >10 µm have become increasingly popular
• Additional treatment of the droplets is needed to produce particles
PARTICLE FORMULATION BACKGROUND
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IF WE COULD CREATE A DROP THEN WITH THE ADITIONAL
TREATMENT WE COULD GET THE SPHERICAL PARTICLE
HOW TO MAKE A DROP?
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They all apply more energy than needed for the production of droplets, and give droplets with wide size distribution if larger
droplets are wanted.
Karbstein H. and Schubert H. 1995
MAKING EMULSIONS - CONVENTIONAL WAY
High-pressure systems Stirring vessels
Colloidal mills
Toothed disc dispersing machines
Ultrasound systems
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They use low energy per unit volume and give monosized distribution.
Microchannel emulsification
Injection of dispersed phase through microgrooves.
Kawakatsu et. al. 1997
Membrane emulsification
Injection of dispersed phase through membrane.
MEMBRANE
Nakashima et. al. 1991
Patent - Asher and Tsien 1980
MAKING EMULSIONS - DROP-BY-DROP
MICROCHANNEL
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D50=200 µm
Hydrophilic nickel membrane
Microchannel emulsification Membrane emulsification NO shear
Apply shear smaller droplets obtained
Apply shear No change in droplet size
MAKING EMULSIONS - DROP-BY-DROP
Dragosavac 2012 Kosvintsev 2008
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Silicon Wafer Membranes Nanomi Holland
SPG membranes SPG Technology Co.,Ltd Japan
Ceramic membranes
MEMBRANES AVAILABLE AT THE MARKET
0.2 - 40 μm
TAMI industries (France) Various other producers
0.3 - 40 μm
0.5 - 250 μm
Drop size
Drop size
Drop size
Fragile
9
Marijana Dragosavac 9
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METAL MEMBRANES
10
Stainless steel membranes Nickel membranes
100 µm
used at Loughborough
Marijana Dragosavac 10
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MEMBRANE SURFACE
O/W and W/O/W
W/O and O/W/O
Hydrophilic surface
Hydrophobic surface
Marijana Dragosavac 11
Oil droplet
Water droplet
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MODELLING
Marijana Dragosavac 12
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caF - Capillary force
Fca MEMBRANE
dF - Drag force
),,( max drfF pd t=
),( pca rfF g=2
2
max 29 p
ddp rddd -÷
øö
çèæ= ptgp
dca FF =
2rp
Fb Fd
Dispersed phase
Continuous phase
FORCE BALANCE MODEL
dhwt 1825.0max transr=
Kosvintsev et al. 2008
Dragosavac et al. 2008 dd
13
Marijana Dragosavac 13
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SHEAR STRESS ON THE MEMBRANE SURFACE
Movements of continuous phase: § STIRRING § CROSS FLOW § PULSATIONS OF THE CONTINUOUS PHASE
Movements of the membrane: § VIBRATIONS § ROTATIONS § TORSIONAL MOVEMENTS
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Dispersed phase
Continuous phase
Sheer to detach droplets
MEMBRANE
EMULSION PRODUCTION
Biodegradable polymeric particles (PCL)
Solvent evaporation
100 µm
Porous inorganic silica particles
Drying
100 µm
Complex coacervation
Coating with the shell /
polymerisation
SOME OF THE PARTICLES PRODUCED UP TO DATE AT LOUGHBOROUGH
Cell encapsulation
Gelation
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COMPLEX COACERVATION O/W emulsion
Motivation for the work: Currently batch production High polydispersity of the product and usually too big droplet size Need for pig gelatine alternative
16
Piacentini, Emma, et al. "Microencapsulation of oil droplets using cold water fish gelatine/gum arabic complex coacervation by membrane emulsification." Food research international 53.1 (2013): 362-372.
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1. Drop production in hydrocolloids solution
2. Coacervation (phase separation) implying the formation of a coacervate phase – pH adjustment
3. Wall formation by aggregation of the hydrocolloid around
droplets of the emulsified hydrophobic material – time, room temperature
4. Wall-hardening, which is generally achieved by cross-linking the hydrocolloid forming the wall
COMPLEX COACERVATION
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COMPLEX COACERVATION
100 µm 100 µm
100 µm 100 µm
1. 2.
3. 3. & 4.
Drop production pH adjustment
Time
Wall-hardening
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• ROOM TEMPERATURE - less energy compared to alternative gelatine types and
• New possibilities for encapsulation of VOLATILE COMPOUNDS
• Increased CONSUMER consent for religious or diet reasons and health safety
WHY FISH GELATINE?
19
Marijana Dragosavac
FISH GELATINE CAPSULES
COMPLEX COACERVATION – Oil encapsulation
Piaccentini et al., 2013 ITM-CNR @ University of Calabria, Rende
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DIFFERENT RATIOS OF FG:GA – PARTICLE SIZE
Shear = 6 Pa Injection rate =1 L h-1
pH=3.5 Room temperature
20
Marijana Dragosavac
Piaccentini et al., 2013
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100 µm 100 µm
100 µm 100 µm
(a) (b)
(c) (d)
(a) 30:70; (b) 40:60; (c) 80:20; and (d) 50:50.
DIFFERENT RATIOS OF FG:GA FOR MICROCAPSULES
FG:GA
ROOM TEMPERATURE
21
Marijana Dragosavac
Piaccentini et al., 2013
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PARTICLE SIZE CONTROL WITH SURFACTANT ADDED
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Shear = 6 Pa Injection rate =1 L h-1
20 µm pore size
Marijana Dragosavac
Piaccentini et al., 2013
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3rd Latin-America Symposium on Microencapsulation
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(a)
100 µm 100 µm
(c)
(b)
(d)
100 µm 100 µm
Freeze dried particles (to enable future volatile compound encapsulation) Silica particles or diatomaceous earth added to produce free flowing particles
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FRAGRANCE OIL ENTRAPMENT
Marijana Dragosavac
Piaccentini et al., 2013
23
3rd Latin-America Symposium on Microencapsulation
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DISPERSION CELL, PULSATING & VIBRATING SYSTEM Dispersed phase: Sunflower oil
Continuous phase: Fish gelatine (FG) and Gum Arabic (GA)
Piaccentini et al., 2013 Flow through the pulsed membrane = 3L h-1
SPAN for all experiments below 0.5
24
Marijana Dragosavac
COMPLEX COACERVATION
24
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CONTINUOUS ENCAPSULATION/SHELL FORMATION
CURRENT WORK - COMPLEX COACERVATION
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1
3
9
4 8
6
Cross-linking
CONTINUOUS ENCAPSULATION/SHELL FORMATION
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ANTYCANCER DRUG ENCAPSULATION O/W & W/O/W
Motivation for the work: Currently batch production Low uniformity of the produced particles using conventional emulsification methods Need for higher encapsulation efficiency Anticancer drug - extremely expensive & temperature sensitive)
Marijana Dragosavac 3rd Latin-America Symposium on Microencapsulation
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ENCAPSULATION OF WATER SOLUBLE PEPTIDE USING BIODEGRADABLE POLYMER
1. Dispersion phase – polymer (PLGA) mixed with DCM (volatile oil phase)
2. Mixing the peptide with previously prepared dispersion phase
3. Injecting through the membrane into 1% PVA solution
4. DCM will evaporate from the particles leaving only peptide within the spherical PLGA particles
Marijana Dragosavac 28
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O/W EMULSIONS
ENCAPSULATION OF WATER SOLUBLE PEPTIDE USING BIODEGRADABLE POLYMER
W/O/W EMULSIONS
HPLC - ENCAPSULATION EFFICIENCY (EE) OF PEPTIDE POLYMER
CONCENTRATION (%) EE (%)
COSOLVENT METHOD (O/W)
10 40
20 50
W/O/W 10 70
20 85
Dragosavac 2012, Unpublished material
Commercially available 14 day kit
(~1g particles/ 70mg peptide)
~200£
üCancer treatment
29
Marijana Dragosavac 29 3rd Latin-America Symposium on Microencapsulation
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G. Gasparini et. al, 2010, Colloids and Surfaces B: Biointerfaces
O/W EMULSIONS
W/O/W EMULSIONS
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ENCAPSULATION OF WATER SOLUBLE PEPTIDE USING BIODEGRADABLE POLYMER
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HIGH THROUGHPUT - SINGLE PASS SYSTEM
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0.5 mm
0.5 mm OIL phase
EMULSION
HIGH THROUGHPUT - SINGLE PASS SYSTEM
OIL phase
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SINGLE PASS SYSTEM (9 mm insert) Dispersed phase Sunflower oil; continuous phase 2% Tween 20
Drop diameter (solid markers) and CV (open markers) as a function of the shear stress
up to 10 L h-1 of injected phase
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1.6 Pa 3.2 Pa
4.7 Pa 9.5 Pa
SINGLE PASS SYSTEM (9 mm insert) Dispersed phase Sunflower oil; continuous phase 2% Tween 20
10 L h-1 of injected phase
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Drop diameter (solid markers) and CV (open markers) as a function of the shear stress
SINGLE PASS SYSTEM (9 mm insert) Dispersed phase 4% PCL in DCM; continuous phase 1% PVA
up to 10 L h-1 of injected phase
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1.6 Pa 3.2 Pa
9.5 Pa 19 Pa
SINGLE PASS SYSTEM (9 mm insert) Dispersed phase 4% PCL in DCM; continuous phase 1% PVA
10 L h-1 of injected phase
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ENCAPSULATED YEAST CELLS W/O emulsion
Aim to produce pH sensitive particles and
exploit the pH to deliver the active ingredient (e.g. cells)
Morelli, Serena, R. G. Holdich, and Marijana M. Dragosavac. "Microparticles for cell encapsulation and colonic delivery produced by membrane emulsification." Journal of Membrane Science 524 (2017): 377-388.
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Intestine delivery of probiotic vRelease of probiotic into the small intestine- colon area, exploiting the pH 7- 8 existing
Yeast species Saccharomyces boulardii
has shown probiotic activity
7- 8
microparticle
cell
3rd Latin-America Symposium on Microencapsulation
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Eudragit S100 coating process
Coated microparticles resulted smaller in size than the uncoated ones - less able to swell
Oil in oil (o1/ o2) solvent evaporation method*:
*Zhang, Lin, et al. "Eudragit® S100 coated calcium pectinate microspheres of curcumin for colon targeting." Journal of microencapsulation 28.7 (2011), 659-667.
Eudragit S100 polymer
H2O
H2O
H2O H2O
H2O
H2O
H2O
H2O
H2O
- Soluble at pH above 7 - Insoluble in water
Uncoated microparticles in water
Coated microparticles in water
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Influence of the cell concentration on emulsion characteristic
The amount of cell encapsulated does not affect the resulting droplet size and uniformity
3.14 x 107 cells mL-1 9.42 x 107 cells mL-1
1.88 x 108 cells mL-1 3.14 x 108 cells mL-1
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Microparticles dissolution time and pH
Eudragit coated particles survived the acidic environment and released the content at pH 7-8
(a) (b) (c) (d)
(a) Uncoated microparticle in water (b) O1/O2 emulsion during the coating process after 0 minutes and
(c) 4 hours, (d) microparticle re-suspended in water after washing in hexane and drying at room temperature.
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Cell viability determination
The yeast released from the particles survived to the encapsulation process and stomach pH
Glucose consumption with time Confocal analysis using fluorescent dyes
% 𝐺𝐺𝐺𝐺𝐺𝐺𝐺𝐺𝐺𝐺𝐺𝐺𝐺𝐺 𝐺𝐺𝐺𝐺𝑐𝑐𝐺𝐺𝐺𝐺𝑢𝑢𝐺𝐺𝑢𝑢 =(𝐺𝐺𝐺𝐺𝐺𝐺𝑖𝑖 − 𝐺𝐺𝐺𝐺𝐺𝐺𝑐𝑐 ) ∗ 100
𝐺𝐺𝐺𝐺𝐺𝐺𝑖𝑖
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CONCLUSION
• Metal membranes and single pass system provide high throughput up to 10 L h-1
• Process can be scaled up • Uniform particles 10 – 250 µm
Homogeniser Membrane emulsification
• Conventional formulation needs to be adjusted to work
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44
44
Micro / Nano – Materials Engineering Group,
Prof. Richard G. Holdich Dr Goran T. Vladisavljevic
Visiting researchers: Larissa Consoli Brazil,
Emma Piaccentini, CNR Italy, Alessandra Imbrognio, CNR Italy
Miguel Angel Suarez Valdes, UNIDAV, Spain
Students: Marianna Petronela Vamanu,
Serena Morelli, Seyitan Odunola, Ryan Barnfield,
Alix Barton,
Dr Konstantin Loponov
ACKNOWLEDGEMENTS
Funding:
Innovate UK Micropore Technologies
Ltd. UK
For the membranes and equipment:
Marijana Dragosavac
Microchannel Array Engineering Team, National Food Research Institute, Tsukuba, Japan.
Prof. Mitsutoshi Nakajima
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