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Simulated Moving BedChromatography in thePharmaceutical Industry
Ron Bates
Bristol-Myers SquibbApril 19, 2004
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Outline
Short Biography
What is Bristol-Myers Squibb
Chromatography
Batch vs continuous HPLC, LC, SMB, P-CAC
Simulated Moving Bed Chromatography Introduction
Theory (brief) Operation
Applications in the Pharmaceutical Industry
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B.S. Chemical Engineering, RPI, 1993
Ph.D. Biochemical Engineering, University ofMaryland, Baltimore County, 1999 Focus: ion-exchange chromatography
Pfizer, Groton, CT, 1999-2003 Focus: small molecule chromatography, HPLC, LC, SFC, SMB,
FLASH, extraction, crystallization, precipitation
Bristol-Myers Squibb, Syracuse, NY, 2003-present Focus: protein chromatography
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Bristol-Myers Squibb
Top-ten pharmaceutical company Products in numerous therapeutic areas
Cardiovascular & Metabolic Diseases Mental HealthPravachol, Coumadin Abilify
Headache and Migrane Infectious DiseasesExcedrin Reyataz, Sustiva
OncologyErbitux, Taxol
Strong pipeline focused in 10 therapeutic areas Oncology, Cardiovascular, Infectious Diseases, Inflammation, etc.
Sites around the world U.S. Research/Manufacturing sites
MA, NY, NJ, CT, IL, Puerto Rico
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Bristol-Myers SquibbSyracuse, NY
Clinical and Commercial Manufacturing Plant
Small-molecule pilot plants Process development and optimization
Clinical manufacturing Penicillin-based products
Last US-based Penicillin manufacturer
Bio-synthetic products
Biotechnology Development, Manufacturing, Analytical Biosciences, Quality
Control / Assurance
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Bristol-Myers SquibbSyracuse, NY - Biotechnology
Two lead protein therapeutics Abatacept: commericial in 2005
Commercial-scale manufacturing
Commercial launch out of Syracuse Facility BLA filing Dec. 2004
LEA29Y: Phase III clinical trials in 2005 Development for next generation process
Clinical production in 2004
Expansion in analytical and quality groupsto support processes
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Batch
vs.
Continuous Chromatography
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Discrete starting and ending points
Example: 10 minute HPLC cycle
Types: GC, HPLC, FLASH, FPLC, LC, etc.
Can be run in many modes: Linear, overloaded, frontal, etc.
Batch Chromatography
0 2 4 6 8 10 12time
Concentration
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Batch Chromatography
(Raffinate)
Feed
Desorbent
Desorbent
Effluentto Waste
Load
Elution
Elution
Effluent
to Waste
Desorbent Elution
(Extract)
(To Waste)
Strong
Solvent
Regeneration
Reference: Linda Wang, Perdue University
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Batch Chromatography
Empty zone
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Continuous Chromatography
Feed is loaded onto column and product iscollected continuously
Annular (P-CAC)
Preparative continuous annular chromatography
Countercurrent
Simulated moving bed chromatography (SMB)
Feed
column
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P-CAC
Reference: Genetic Engineering News, Oct. 1, 1999
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P-CAC
Reference: Genetic Engineering News, Oct. 1, 1999
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P-CAC
Reference: Genetic Engineering News, Oct. 1, 1999
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P-CAC
Reference: Genetic Engineering News, Oct. 1, 1999
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Simulated Moving Bed
Chromatography (SMB)
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What is SMB
SMB is Simulated Moving Bed Chromatography.
SMB is continuous countercurrent chromatography. The feed is pumpedinto the system and two (or more) product streams are continuouslycollected.
SMB has been used for the production of millions of tons of bulkcommodities (p-xylene, high fructose corn syrup, etc...) for the past fourdecades.
Due to improvements in column and equipment technology, SMB has
recently been used in the pharmaceutical industry (Sandoz, SmithKline,UCB, Pfizer). HPLC costs: $100/kg to $5000/kg
SMB costs: $50/kg to $200/kg
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SMB versus HPLC
Advantages of SMB: Lower solvent utilization (up to 10 times less than batch HPLC) Generally can use less expensive, larger stationary phases Able to get high recovery and high purity Sometimes better productivity Lower labor and QC costs Only partial separation of solutes is required to obtain high purity. Higher yield than batch 10% more than batch. High throughput 5 to 10 fold increase. Lower solvent consumption An order of magnitude lower. Continuous process.
Disadvantage of SMB: Binary separation only Complexity
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Commercial Applications of SMB Hydrocarbons
Sugars
Agrochemicals
Antibiotics
Peptides Chiral Drugs
Gaining tremendous momentum FDA approves of the technology Chiral resin manufacturers sell resins specifically made for SMB
Proteins? Useful as polishing step?
SEC: remove aggregated form of product
Multicomponent separations more difficult than traditional uses 8, 12, even 16 zone systems being examined
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Basic Principle
Mobile Phase
Feed
Continuous Countercurrent Chromatography
stationary column
A sample is injected in the centre of a stationary column
The two components move at different speeds and are separated
If we now move the column from right to left, at a speed halfway
between that of the solutes, they now move in different directions ...
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Basic Principle
Mobile Phase
Feedcolumn
The two solutes now move in different directions relative to a stationaryobserver. If the column is very long, the bands will continue to separate.
Continuous Countercurrent Chromatography
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Basic Principle
Mobile Phase
Feedcolumn
If we continue to add sample at the center, the components will continueto separate
Continuous Countercurrent Chromatography
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This is clearly a continuous system, but there are problems.
The column needs to be of infinite length, the actual moving of solids isvery difficult and some way to introduce and remove the sample and theproducts are needed.
We solve this by cutting the column into small segments and simulatingthe moving of them
Basic Principle
Mobile Phase
Feedcolumn
Continuous Countercurrent Chromatography
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The feed and solvent inlets are now placed between the segments
and are moved each time a segment is moved from one end to the other
Basic Principle
Mobile Phase
Feedcolumn
Continuous Countercurrent Chromatography
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Products are removed by bleeding off a carefully calculated flow
at suitable exit points. This changes the velocity of the bands in
the column and forces the products to move toward the ports
This ensures that the column segments are clean before they are moved
and that the solvent can be recycled directly back through the system
Mobile PhaseBasic Principle
Mobile Phase
Feedcolumn
Continuous Countercurrent Chromatography
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True Moving Bed
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Binary Separation in a True Moving Bed
Desorbent
Desorbent
Raffinate
Extract
Feed
Feed
Extract Raffinate
Time : t
Time : t + t
Reference: Linda Wang, Perdue University
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Binary Separation in a True Moving Bed
Desorbent
Raffinate
Extract
Feed
ExtractRaffinate
Time : t + 3t
Feed
Time : t + 2t
Desorbent
Reference: Linda Wang, Perdue University
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Binary Separation in a True Moving Bed
Desorbent
Desorbent
Raffinate
Extract
Feed
Feed
Extract Raffinate
Time : t + 4t
Time : t + 5t
Reference: Linda Wang, Perdue University
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TMB to SMB
Since its very difficult to move solids, true
countercurrent chromatography does notexist.
Instead, the bed is broken into manyfractions and their movement is simulatedby changing the inlet and outlet ports
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Simplified SMB - 1Feed
Solvent
Extract Raffinate
FeedSolvent
Extract Raffinate
The system is started.....
A frontal elution separation
occurs in Section 3.
1 2 3 4
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Simplified SMB - 2FeedSolvent
Extract Raffinate
FeedSolvent
Extract Raffinate
The separation continues.....
Eventually the front of
pure product 1 reaches the
outlet. It is distributed
between the final Sectionand the product port
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FeedSolvent
Extract Raffinate
Simplified SMB - 3
FeedSolvent
Extract Raffinate
Finally, the mixed product
reaches the outlet. To avoid
collecting impure material, it
is necessary to move the
columns 1 position upstream.
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FeedSolvent
Extract Raffinate
Simplified SMB - 4
FeedSolvent
Extract Raffinate
The frontal separation
continues; at the same time,
the slow moving product starts
to separate from the tail of the
mixed product band in Section 2
Eventually the fast moving
product again reaches theoutlet and more pure product
is collected.
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FeedSolvent
ExtractRaffinate
Simplified SMB - 5When the mixed band reaches
the end of Section 3 its tail has
left Section 2 (if the separation
has been correctly designed) and
only pure product 2 remains inSection 2.
FeedSolvent
ExtractRaffinate
To avoid collecting impure
raffinate, the columns aremoved once more. Now, the
pure component 2 is in Section 1.
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FeedSolvent
ExtractRaffinate
Simplified SMB - 6
FeedSolvent
ExtractRaffinate
The second component is now
collected at the Extract port while
the separation continues in Sections
2 and 3.
The faster component reaches the
Raffinate port and is again collected;
note that the outlet concentrations areneither constant nor concurrent.
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FeedSolvent
ExtractRaffinate
Simplified SMB - 7
FeedSolvent
ExtractRaffinate
Eventually, the mixed zone
reaches the raffinate port and
the columns are again switched.
This simplified system is now
in a steady state mode and will
continue to cycle.
Switch
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The moving of the bed is simulated by moving the pointsof feed and mobile phase addition, as well as the points
of raffinate and extract removal while keeping thecolumn positions fixed.
Time = 0Extract
Feed Raffinate
Mobile
Phase
Feed
Raffinate
Time = 1
Mobile
Phase
Extract
Packed
Column
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The zones are made up of one or more columns
Six-column SMB System
Eight-column SMB system
I II III IV I II III IV
I II III IV I II III IV
SMB Configurations
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Li uid
RAFFINATE
ELUENT
FEED
EXTRACT
t0
Li uid
RAFFINATE
ELUENT
FEED
EXTRACT
t0 + T / 2
SMB Operation
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Li uid
t0 + 1 T
EXTRACT
FEED
RAFFINATE
ELUENT
Li uid
RAFFINATE
FEED
ELUENT
EXTRACT
t0 + 1 T + T / 2
SMB Operation
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Theory Governing Equations
For another day
Maybe
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Theory Working Equations / Definitions
k1 = capacity factor = (tr-t0) / t0
= k2/ k1
Rs = 2* (tr1-tr2) / (w1-w2)
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SMB Method Development
1. Start with linear batch experiments
2. Increase either mass or volume of load to overload thecolumn
3. Calculate isotherm
4. Determine resistance to mass transfer (if important)
5. Calculate necessary flow rates
6. Optimize (either on-the-fly or with a proven model)
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Linear Chromatography
0 2 4 6 8 10 12time
Concentration
tr1
tr2
t0
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Batch Chromatography Experiments
Feed concentration As concentrated as possible to minimize disruption to
Zone III velocity
Need to run batch experiments at appropriate
concentrations and solvents Desorbent composition
Solubility of products
Strength
Trade-off between time and mobile phase utilization
Sorbent Capacity, selectivity, resolving power
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Feed Concentration
Feed concentration: Consider two systems
A: Concentrated feed
B: Dilute feed
Run batch experiments to examine effect ofconcentration
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Desorbent composition
Multiple trade-offs: Solubility of products and effectiveness of the
solvent Not always complimentary
Often solubility dictates solvent composition Speed
Low k = high throughput More wear and tear on equipment Larger system needed
Large k = low throughput Less wear and tear Smaller system acceptable
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Choice of Sorbent
Capacity: higher = better?
Selectivity: higher = better?
Resolving power: higher Rs
= better?
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Linear Chromatography
0 2 4 6 8 10 12time
Concentration
tr1
tr2
t0
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Volume Overloading
time
Absorbance
B t h Ch t h t SMB
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Batch Chromatography to SMBInitial Operating Conditions
Determine optimal feed concentration,stationary phase and mobile phasecomposition (highest with lowest
capacity factors)
Calculate isotherm and mass transferresistances
Either use software package or rules ofthumb to generate initial SMB flow rates
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Zone velocities vI = vRecycle + vD vII = vI - vX vIII = vII + vF vRecycle = vIII - vRaff
Solvent Mass Balances Flow Rates
IIIIII IVvI vII vIII
vRecycle
vX vRaffvFvD
Overall Mass Balance vD + vF = vX + vRaff
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Flow rates Commercial SMB design models available
Given batch results from 5-10 column experiments Flow rate, feed concentrations, retention times Solubility data
Predict zone velocities, productivities Problems:
Usually assumes simple adsorption model and lumped masstransfer coefficients
Often difficult to interpret overloaded chromatograms
Rules of Thumb
Educated guesses based upon batch results fromlinear and overloaded experiments VII and VIII ratio (based upon retention times) VI to flush back-side of slowest component from zone I Feed concentration and flow rate based upon solubility data and
solvent mass balance
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Period
The period is the time a column stays inone zone also called switching time.
Changing the period has the effect of
changing all 4 zones simultaneously, thuseither speeding up or slowing down thesolutes
Example of switching time
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Li uid
RAFFINATE
ELUENT
FEED
EXTRACT
t0
Example of switching time
Liquid
t0 + 1 T
EXTRACT
FEED
RAFFINATE
ELUENT
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SMB Optimization
Independent variables:
Flow rates
Recycle, Desorbent, Raffinate, Extract, Feed
Period (switching time) Thats it.
Procedure:
Get the system bound, manipulate the flowrates to maximize throughput at requiredpurity
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SMB Optimization
IIIIII IVvI vII vIII
vRecycle
vX vRaffvFvD
Questions:
What is the effect of increasing the Zone I flow rate?
How would one accomplish this?
Zone II? Zone III?
What if the system is underutilized (i.e., more feed canbe added to the system) how would one do this
without affecting the other zone flow rates?
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Two component SMB System
FeedDesorbent
Extract Raffinate
Conc.
I II III IV
Bed Position
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SMB Optimization
IIIIII IVvI vII vIII
vRecycle
vX vRaffvFvD
Questions:
Extract contains too much of the weakly adsorbedspecies what do you do?
If situation was reversed?
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Two component SMB System
FeedDesorbent
Extract Raffinate
Conc.
I II III IV
Bed Position
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SMB Optimization
IIIIII IVvI vII vIII
vRecycle
vX vRaffvFvD
Questions:
Extract contains too much of the weakly adsorbedspecies what do you do?
If situation was reversed?
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Two component SMB System
FeedDesorbent
Extract Raffinate
Conc.
I II III IV
Bed Position
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Examples of SMB
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Two component SMB System
M lti t S t
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Multi-component System
0 10 20 30 4000.10.20.30.40.50.60.70.8
Time [min]
Ci/CF,i
Sulfuric AcidGlucoseXyloseAcetic Acid
Single-component pulse data
Reference: Linda Wang, Perdue University
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Multi-Component SMB System
Desorbent
Extract(2, 3)
Feed
(1, 2, 3)
Raffinate(1)
I II III IV
1 Fast Solute2 Intermediate Solute3 Slow Solute
Concentration
Bed Position
Reference: Linda Wang, Perdue University
Complete Separation in Tandem SMB
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Complete Separation in Tandem SMB
Column Number
0 5 10 15 200
0.5
1
Ci/CF,i
Des. Ext. Feed Raf.Sulfuric AcidGlucose
Acetic Acid
0 5 10 15 200
0.5
1
Ci/CF,i
Des. Ext. Feed Raf.
Reference: Linda Wang, Perdue University
Profiles of a Parallel SMB
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Profiles of a Parallel SMB
Glucose yield: 94% Glucose purity: 99%
0 5 10 15 200
0.2
0.4
0.6
0.8
1
1.2
Column Number
Ci/CF
,i
E1D1
B(o) F
R1 D2
E2 B(i)
R2
I
*
*
II III IV V VI VII VIII IXSulfuric Acid
Glucose
Acetic Acid
Reference: Linda Wang, Perdue University
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Other Questions?