polyrmc, tulane center for polymer reaction monitoring and characterization recently acquired lab...
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PolyRMC, Tulane Center for Polymer Reaction Monitoring and Characterization
Recently acquired lab space
Aerial view of Tulane campus
Motto:
Value and impact based on scientific and technical excellence, integrity, and relevance
Mission statement:
To be the world’s premier center for R&D polymerization reaction monitoring
http://tulane.edu/sse/polyRMC PolyRMC is a non-profit entity
Founded in Summer 2007
Wayne F. Reed, Founding DirectorAlina M. Alb, Associate Director for ResearchMichael F. Drenski, Associate Director for Instrumentation Alex Reed, Assistant Director for Operations
Tightly focused but broadly applicable
:
Through PolyRMC personnel’s many years of industrial collaboration the process of dealing with confidentiality, intellectual property rights, and all other legal issues has been streamlined to produce rapid agreements on desirable terms for industries.
Polymer ‘born characterized’
‘on-command’ polymers
Monitoring and control
• new polymeric materials • medical applications• nanotechnology • new high performance materials
Resins, Paints, Coatings
Industrial R&D and problem solving
Fundamental and applied research
Accelerate R&D of new materials
Advanced characterization
Multi-detector SEC analysis:Multi-angle Light Scattering,
Viscometer, RI, UV detectors
Fast Turn-around polymer characterization
• Reduce bottlenecks and lengthy turn around time in workflows.
• The services can be used to prioritize and complement each industry’s own in-house characterization efforts.
• Set standards of quality control and product reproducibility leading to higher efficiency, and establish means of characterizing and improving new products.
R&D, product development, and problem solving in the polymer/pharmaceutical/ natural products industries
• failure to meet product grade specifications; • inconsistency of product quality;• product instabilities, such as precipitation, degradation, or phase separation;• Defects in the products, such as colloids, particulates, and undesirable colors;
Some of PolyRMC’s Initiatives
Development of natural products
• Release of proteins, polysaccharides, and other components during extraction, including enzymatic activation/deactivation processes.
• Monitoring chemical and physical changes when natural products are modified by chemical, thermal, radiative, or enzymatic treatments.
• Determining the types of micro- and nanostructures that can be formed from natural products.
• Measurement of encapsulation and time-release properties of natural products used with pharmacological, food, and other substances.
◦ Deep expertise broadly applicable to applied polymer issues; creativity; advanced instrumentation base; entrepreneurial energy
◦ Adapting our approaches to the many complex processing aspects of natural products; extraction, enzymatic modifications, chemical tailoring, encapsulation, etc.
◦ A track record of success in solving problems:
- gelatin/oligosaccharide phase separation
- aggregation, degradation, micro-gelation, dry powder dissolution, polymerization - characterization of natural products; xanthan, pectin, gum arabic, alginates;
- multi-detector SEC characterization; - origin and detection of polymer product anomalies; - determination of physical/chemical processes in production of copolymers; - characterization of water soluble polymers for water purification, paints, cosmetics, food;
PolyRMC Expertise
e.g.
• Deep and powerful expertise in highly focused but broadly applicable areas
• Ability to conceptualize problems in general, far-reaching terms
• Complete, state-of-the-art instrumentation and skills within a ‘clean’ university environment
• Ready access to many online resources
• Chance to outsource research and problem solving without the overhead investment
• PolyRMC is used to dealing with industrial partners and their concerns for IP rights, confidentiality, etc.
Advantages
Types of reactions that we monitor• Synthetic polymerization reactions: free radical, ‘living’,
polycondensation, homogeneous and emulsion phase, in batch, semi-batch, and continuous reactors
• Postpolymerization reactions: hydrolysis, PEGylation, ‘click’ reactions, grafting, amination, etc.
• Modifications of natural products, especially polysaccharides
• Poplypeptide synthesis reactions
• Oligonucleotide synthesis reactions
• Polymer degradation reactions due to enzymes, chemical agents, heat, radiation, acids, bases, etc.
• Protein aggregation and other instabilities
• Phase separation, microgelation
• Kinetics of interacting components in complex solutions
• Dissolution of dry powders, emulsions, pastes, etc.
• Release of encapsulated and associated agents
• Production or hydrolysis of polymers amidst bacterial populations
The types of quantities and events that we monitor during these reactions Evolution of polymer molecular weight
Reaction kinetics; e.g. polymerization, degradation, aggregation rates
Particle size distributions
Degree of reaction completion
Tracing residual monomers and other reagents
Monomeric and comonomeric conversion
Reactivity ratios
Composition drift and distribution
Intrinsic viscosity
Unusual or unexpected events during reactions; onset of turbulence, microgelation
Attainment of desired properties, such as stimuli responsiveness; ability to encapsulate drugs or other agents, micellization or other supramolecular structuration, solubility changes, ability to interact or not interact with specific agents, etc.
Equilibrium characterization of polymer solutions
Non-equilibrium characterization; PolyRMC
methods • ACOMP (Automatic Continuous Monitoring of Polymerization reactions); Monitor synthetic reactions, polypeptide synthesis, polymer modifications, etc.
• Heterogeneous Time Dependent Static Light Scattering (HTDSLS); Characterize co-existing populations of polymer and colloids; e.g. bacteria and polymers
• Simultaneous Multiple Sample Light Scattering (SMSLS); high throughput screening of protein aggregation, solution stability in general.
• Multi-detector Size Exclusion Chromatography (SEC), a standard method• Automatic Continuous Mixing (ACM), characterize complex, multicomponent solutions along selected composition gradients. A PolyRMC method.
Methods and techniques used for reaction monitoring and characterization
Achtung!
Many biological polymers in aqueous solutions are inherently unstable and can aggregate, form microgels, precipitate, or otherwise degrade in time.
• The time for such instabilities may be seconds, hours, days, even months or longer.
• It is hence imperative to know if such a solution is in equilibrium, or at least in a long lived metastable state, before making equilibrium measurements, such as chromatographic determinations, or single scattering or other measurements.
• This is why we have developed a number of methods, briefly outlined below, SMSLS, ACOMP, ACM, and HTDSLS, for monitoring the kinetics and characteristics of non-equilibrium processes.
• Unfortunately, many researchers spend a lot of time making measurements on kinetically unstable systems, leading to irreproducible results and confusion.
Do not use equilibrium characterization methods to characterize non-equilibrium systems.
Multi-detector size exclusion chromatography; to be used when the polymer solution is in equilibrium
Example of state-of-the art multi-detector Size Exclusion Chromatography
viscosifyingemulsifying
bulk
Analyzed gum arabic SEC data
Determining the molecular origins of how a natural product works to emulsify and thicken alimentary
products.
Dextran has a small population of very high mass chains causing separation:
Mn=1,600g/mole
Mw=12,500g/mole
SEC: Origin of oligosaccharide/gelatin phase separation
It’s the oligosaccharide, not the gelatin!
- Seen in SEC light scattering
RI
& L
S90
o (ar
b. u
nits
)Mn controls the sensation of sweetness, and determines commodity price, but Mw controls phase separation. This approach provides a means of screening this highly variable natural product.
Monitoring polymer degradation processes
Light Scattering and Degradation
Signatures for time dependent light scattering enzymatic degradation of linear molecules with different numbers of strands
time [104 sec]
Degradation by laminarinase
Beta glucan is a mixture of double and triple strands
New signatures for time-dependent light scattering degradation of branched polymers; determination of polymer architecture, kinetics, modes of cleavage
sidechain strippingrandom chain cleavagerandom backbone cleavage
Proteoglycan ‘monomer’
glycosaminoglycan sidechains
protein backbone
- sidechain stripping, backbone intact
- random sidechain degradation, backbone intact
- sidechain stripping and backbone degradation
Simultaneous Multiple Sample Light Scattering (SMSLS); when high throughput and/or long term solution stability screening is important
Simultaneous Multiple Sample Light ScatteringSMSLS: High throughput screening
A single instrument can monitor stability and reactions of many different samples for hours, days, months, automatically, and with a single computer; e.g. Monitor protein aggregation.
A typical SMSLS prototype with both flow and batch cells
SMSLS scheme for automatic, continuous monitoring of protein aggregation
NxM protein samples under any desired set of T, pH, concentration, agitation, ionic strength
Mw vs. time for all NxMsamples, continuously and simultaneously on the computer screen
NxM protein samples under any desired set of T, pH, concentration, agitation, ionic strength
Mw vs. time for all NxMsamples, continuously and simultaneously on the computer screen
Note, for aggregating systems that become turbid M =1
M= # of series cells
N= # of parallel cell banks
PolyRMC will run assays on systems determined by pharmaceutical sector colleagues, e.g. protein solution stability under a matrix of conditions.
If SMSLS proves useful for a given pharmaceutical sector collaborator, PolyRMC, or associated entity, will build and deliver a turn-key, customized SMSLS instrument and associated software for the collaborating company.
Developing and delivering complete SMSLS systems to Pharmaceutical companies;
How technology transfer will work
PolyRMC also provides an as-needed access service to SMSLS assays, and related problem solving, in cases where the company might not need an instrument of its own.
Online monitoring/characterization of aggregation processes
Gelatin aggregation
Aggregation process for gelatin solutions at different temperatures, monitored by ACOMP
Therapeutic protein aggregation monitored by SMSLS
Map
p. /
Map
p., t
=0
t (h)
-at ionic strength:1.56 – 50mM
Protein aggregation
All solutions are unstable over time
Ranked methods for monitoring and quantifying protein aggregation
Most important aspect of aggregation is change in Mass
1. Static Light Scattering: Absolute, model-independent change in Mw at the slightest change
SMSLS: for high throughput
2. Dynamic Light Scattering: Runner-up. Model dependent, sensitive to <D>z, only indirectly sensitive to Mass.
3. Low angle Mie scattering/diffraction: e.g. Master Sizer. Misses the boat. Reports aggregation only after very advanced. Gives ‘size’ not mass.
4. Fluorescence. Indirect, insensitive, but better than nothing.
0
2 1010
4 1010
6 1010
8 1010
1 1011
0 2000 4000 6000 8000 1 104 1.2 104
y = 6.2873e+09 + 8.9157e+06x R= 0.99696
1/v
1/c
1/v=1/vmax
+Km
/cvmax
Vmax
=1.6x10-10 M/s
Kmax
=0.00142 cm3/g
Michaelis-Menten-Henri Enzyme kinetics
Rapid determination of enzyme kinetics
Hyaluronate degradation by hyaluronidase
Enzymatic degradation monitored by SMSLS
Dissolution of polymers and time release studies
* Small population of aggregates present in dry powder* Aggregates dissolve in time
Dissolution of a polyelectrolyte
Polystyrene sulfonate
Dissolution of dry polysaccharides
Origin of poor dissolution due to formation of aggregates
0
0.05
0.1
0.15
0.2
0.25
0 1 2 3 4 5 6 7 8
Rele
sed
am
ou
nt
dru
g(m
g)
Time (h)
(2)
(1)
(b)
OCH2CHCH2NHCH(CH3)2.HCl
OH (a)
Ethylene Glycol DimethacrylateN-isopropylacrylamide
AcrylonitrileCN
Monitoring drug release by nanohydrogels
The release of propanolol, PPL from core-shell p(AN-c-NIPAM) 1” and amidoximated p(AN-c-NIPAM) “2” was continuously monitored by UV detection with ACM.
Poly(acrylonitrile-co-Nisopropylacrylamide), p(AN-c-NIPAM) core-shell hydrogel nanoparticles were synthesized by microemulsion polymerization and their feasibility as drug carrier was investigated.
Monitoring heterogeneous solutions of polymers and colloids; e.g. proteins amidst bacteria.
Heterogeneous Time Dependent Static Light Scattering (HTDSLS)
Determine large particle densities amid polymer chains; e.g. spherulites, microgels, bacteria, crystallites, etc.
Follow evolution of large particles; e.g. in biotechnology reactors where bacteria/polymers co-exist. e.g. xanthan productions, degradation of polysaccharides, other fermentation reactions
Permits useful characterization of polymers in solutions which, up until now, would be considered far too contaminated with dust and other scatterers.
HTDSLS: Use flow to create countable scattering peaks from colloidal particles, while simultaneously monitoring the background scattering due to co-existing polymers
Applications of Heterogeneous Time Dependent Static Light Scattering (HTDSLS)
HTDSLS: Good data from a classically intractable case of high particulate contamination:
0
500
1000
1500
I(m
V)
0 50 100 150 200 250 300 350
t(s)
[PVP] F(1/s) . 1.5 mg/ml 0.4561BL 856mV...1 mg/ml 0.3205BL 630mV.0.75 mg/ml 0.3596BL 513mV....0.25 mg/ml 0.4328BL 213mV...water 0.4333BL 25mV av 0.40 + 14%
0.0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1.0 1.1 1.21.4x10-6
1.6x10-6
1.8x10-6
2.0x10-6
2.2x10-6
2.4x10-6
2.6x10-6
2.8x10-6
3.0x10-6
3.2x10-6
3.4x10-6
Kc/
I
sin 2̂(/2) + 100*c
Mw=6.1x105 g/mol A2=3.34x10-4mL-mol/g2, Rg=460 A
Schimanowsky, Strelitzki Mullin, Reed, Macromolecules 32, 7055, 1999
5200, 2 micron latex spheres/mL
Heterogeneous time dependent static light scattering (HTDSLS)
Co-existing E. Coli and PVP polymers in solution
Schimanowsky, Strelitzki Mullin, Reed, Macromolecules 32, 7055, 1999
Automatic Continuous Online Monitoring of Polymerization reactions (ACOMP)
Fundamental studies of polymerization kinetics and mechanisms
Optimization of reactions at bench and pilot plant levels
Full scale, feedback control of industrial reactors
Automatic Continuous Online Monitoring of Polymerization reactions: ACOMP
Continuously extract and dilute viscous reactor liquid producing a stream through the detectors so dilute that detector signals are dominated by the properties of single polymers, not their interactions.
Principle of ACOMP
ACOMP ‘front-end’: Extraction/dilution/conditioning
ACOMP ‘back-end’: Detector train
UV detector
Refractive indexdetector
Viscometer
Light scattering
Solvent
Reactor
Recent ACOMP advances Copolymerization
Predictive control
Heterogeneous phase; emulsion and inverse emulsion
Living-type polymerization
Continuous reactors
About ACOMP- Monitor important characteristics of
polymerization reactions while they are occurring
- Develop new polymeric materials, understand kinetics and mechanisms.
- Optimize reactions at bench and pilot plant level.
- Full feedback control of large scale reactors:Increased energy efficiencyMore efficient use of non-renewable resources, plant and personnel timeLess emissions and pollution
Stem the flight of manufacturing overseas: Jobs. ACOMP lab. unit
Left: polymer Mw and r vs. conversion; Right: particle size distribution and specific surface area
Raw data and analysis for free radical polymerization of MMA in emulsion at 70C.
- first simultaneous online monitoring of both polymer and particle properties
A. M Alb, W. F Reed, Macromolecules, 41, 2008
Emulsion Polymerization: Example of raw data and analysis
Summary: PolyRMC works with many pharmaceutical, synthetic, and natural product polymers, with a particular emphasis on monitoring processes in solutions of these in order to
• Better understand the processes and mechanisms involved in producing such polymers
• Quantitatively control the factors responsible for the reactions
• Monitor processes for completion, unusual events, specific thresholds of product stimuli responsiveness, etc.
• Produce products that consistently meet or exceed specifications.
• These capabilities can be used in the discovery, development, formulation, and quality control stages