innovation in size based polymer separations
TRANSCRIPT
©2014 Waters Corporation 1
“Innovation in Size Based Separations"
©2014 Waters Corporation 2
Current Status on Method Development for GPC
GPC needs method development – However, there are challenges that prevent routine method
development o Time constraints due to lengthy analysis times o Columns/ systems often dedicated to one solvent o Screening of conditions often not a viable option
– And now there are new challenges that make method development even more critical o Complex polymer chemistry o Many more process situations o Faster reactions – in process monitoring a challenge
©2014 Waters Corporation 3
Advanced Polymer Chromatography A Definition
Application technique for the size based separation of polymers in
solution using columns packed with sub-3µm, rigid, high-pore-
volume, hybrid particles combined with a robust, fully optimized low
dispersion ACQUITY system
©2014 Waters Corporation 4
The ACQUITY® Advanced Polymer Chromatography™ (APC™) System
Precise solvent
management
Low system dispersion
Compatibility with
challenging solvents
Rigid, solvent-resilient columns
Versatile column
management
Stable refractive
index detection
Flexible and robust data processing
Wide range of APC
standards
©2014 Waters Corporation 5
Benefits of APC Approach to the SEC Experiment
Speed of Analysis
Routine bracketed
calibration
Method conditions screening capability
Flexibility
Innovative SEC column choices
Fast system equilibration
Automated solvent
exchange
Resolution
Separation of highly complex polymer blends
Accurate reliable
data
©2014 Waters Corporation 6
A Systematic Approach to Developing APC Methods
Speed of Analysis: Increased Data Points Method Development Factors
Flexibility: Column Selection Automated Solvent Change
Maximize Resolution: Data Processing and Reporting
©2014 Waters Corporation 7
A Systematic Approach to Developing APC Methods
Speed of Analysis: Increased Data Points Method Development Factors
Flexibility: Column Selection Automated Solvent Change
Maximize Resolution: Data Processing and Reporting
©2014 Waters Corporation 8
High Resolution SEC Characterization
µ
-2.00
-1.50
-1.00
-0.50
0.00
0.50
1.00
1.50
2.00
2.50
3.00
3.50
4.00
4.50
5.00
Minutes2.00 2.20 2.40 2.60 2.80 3.00 3.20 3.40 3.60 3.80 4.00 4.20 4.40 4.60 4.80 5.00 5.20 5.40 5.60 5.80 6.00
Use multiple small pore columns in a bank to improve low MW resolution
of the Liquid Epoxy Resin
6.00 min.
©2014 Waters Corporation 9
4 samples including replicate bracketed calibration in 90 min
4 samples including replicate bracketed calibration • 90 min. with APC • 1 Day with Traditional GPC
Sample Test Set with APC Sample Test Set with GPC
©2014 Waters Corporation 10
Cellulose Based Polymer – Target Molar Mass 300,000 - Polystyrene Equiv. 100,000
©2014 Waters Corporation 11
©2014 Waters Corporation 12
High Throughput Screening SEC
Dextran 410k Dextran 47k Dextran 165k Dextran 1.75m
µRIU
0
2
4
6
8
10
12
14
Minutes 0.20 0.22 0.24 0.26 0.28 0.30 0.32 0.34 0.36 0.38 0.40 0.42
GPC Calibration Plot
Log
DP
w
2.4
2.6
2.8
3.0
3.2
3.4
3.6
Use a single short column to
achieve very fast elution times for your polymer in
screening applications
©2014 Waters Corporation 13
A Systematic Approach to Developing APC Methods
Speed of Analysis: Increased Data Points Method Development Factors
Flexibility: Column Selection Automated Solvent Change
Maximize Resolution: Data Processing and Reporting
©2014 Waters Corporation 14
Column Selection Criteria
Surface chemistry – Aqueous or organic soluble polymer
Particle size
– smaller particles for higher resolution – larger particles to avoid shear degradation of very high MW components
Pore size
– depends on molecular weight range of sample – avoid exclusion of sample – maximize pore volume in required separation region
Column Length
– compromise between resolution and analysis time
©2014 Waters Corporation 15
Surface Chemistry
Two surface chemistries available – Designed for use with aqueous soluble polymers – Temperature rated up to 45C – pH range of 1 – 9
– Designed for use with organic soluble polymers – Temperature rated up to 90C – pH range of 1 - 11
AQ
XT
©2014 Waters Corporation 16
BEH Particle Resistance to Swelling
ACQUITY APC Columns are based on our BEH particle technology – Rigid pore structure – Excellent mechanical
stability – Excellent solvent
compatibility
But I have to select columns that are packed in my
chromatographic solvent to avoid bed swelling
Columns are shipped dry. Just equilibrate
with 20 column volumes of your chromatographic
solvent
©2014 Waters Corporation 17
Efficiency: Impact of Particle Size on SEC Separations
Data courtesy of Miroslav Janco, The Dow Chemical Co.
Column: ACQUITY BEH C18 (130Å, ~0.7 mL/g), 4.6x150mm; Sample: Polystyrene (Mp=11.6K, Đ=1.03); Mobile phase: THF; Detection: UV
©2014 Waters Corporation 18
Selecting the Correct Pore Size µR
IU
0.00
15.00
30.00
Minutes0.00 1.50 3.00 4.50 6.00
Selecting a column bank with a pore size range to cover the
MW distribution of the polymer is critical for getting an accurate measurements
2 Columns in series 45A + 125A 3 Columns in series 45A + 125A + 450A
©2014 Waters Corporation 19
Selecting the Column Length
µRIU
0.00
4.00
8.00
12.00
Minutes0.00 3.00 6.00
Selecting a column bank with increasing length will improve the resolution of
the separation
3 Columns in Series 30mm 75mm 150mm
©2014 Waters Corporation 20
Solvent Selection Criteria
Solubility – Need a solvent that can entirely dissolve your polymer to get real
measurement
Viscosity – What is the impact on the separation conditions? – Can it be managed?
Additives – How to identify and eliminate surface interactions to get purely SEC
mode separations
System/Detection Considerations – The solvent is compatible with your polymer, but what about the
chromatographic system?
©2014 Waters Corporation 21
Solvent Selection and Refractive Index
Ascending order of Refractive Index
Solvent Refractive Index
Hexafloroisopropanol 1.275 Methanol 1.329 Water 1.33 Acetonitrile 1.344 Acetone 1.359 Ethanol 1.361 Ethyl Acetate 1.37 Hexane 1.373 Methyl ethyl Ketone 1.379 Isopropanol 1.38 Heptane 1.387 Isooctane 1.404 Tetrahydrofuran 1.408 Dichloromethane 1.424 Dimethylformamide 1.428 Dimethylacetamide 1.438 Chloroform 1.443 N-methylpyrrolidone 1.468 Dimethyl sulfoxide * 1.477 Toluene 1.496
Test Polymer
©2014 Waters Corporation 22
Polysilane RI Detector Response with Toluene, THF and Ethyl Acetate
Solvent :Toluene (negative detector signal)
Solvent: Toluene (negative signal – detector polarity switch)
Solvent :THF (positive detector signal)
Solvent :Ethyl Acetate (positive detector signal)
©2014 Waters Corporation 23
Fully compatible with the ACQUITY APC supported solvents – 6 solvent lines
Enables automated solvent
switching on the ACQUITY APC System – Solvent resilient ACQUITY APC
hybrid particles not susceptible to swelling with solvent switching
Optional Solvent Select Valve
©2014 Waters Corporation 24
A Systematic Approach to Developing APC Methods
Speed of Analysis: Increased Data Points Method Development Factors
Flexibility: Column Selection Automated Solvent Change5
Maximize Resolution: Data Processing and Reporting
©2014 Waters Corporation 25
Epoxy 1009
ACQUITY APC XT 45A,125A,450A in Series 30mm 75mm 150mm
Use a range of columns in a bank to characterize broad samples and their trace oligomeric species
1 x 450Å (20,000-400,000) 150mm length 1 x 125Å (1,000-30,000) 150mm length
1 X 45Å (200-5,000) 150mm length
µRIU
0.0
1.0
2.0
3.0
4.0
5.0
6.0
7.0
Minutes0.0 1.0 2.0 3.0 4.0 5.0 6.0
©2014 Waters Corporation 26
Choose Appropriate Calibration Kit
©2014 Waters Corporation 27
µRIU
0.0
1.0
2.0
3.0
4.0
5.0
6.0
7.0
Minutes0.0 1.0 2.0 3.0 4.0 5.0 6.0
µR
IU
0.00
4.00
8.00
12.00
16.00
Minutes2.80 3.50 4.20 4.90 5.60 6.30
V1 = 1,800,000 + 277,000 + 34,800 + 3,470 V2 = 1,210,000 + 130,000 + 17,600 + 474 V3 = 552,000 + 66,000 + 9,130 + 266
Log M
ol W
t
3.0
4.0
5.0
6.0
Elution Volume2.50 3.00 3.50 4.00 4.50 5.00
ACQUITY APC Calibration Standards: Polystyrene
9 min
High MW Standards MW: 266 - 1,800,000 Columns: 45A + 125A + 450A 150 mm each THF at 0.8 ml/min 10 ul RI Detector
©2014 Waters Corporation 28
Data Review
©2014 Waters Corporation 29
Calibration Using Narrow Standards Polynomial – All data points fitted with one function of the form; Linear (1st order): Log M = A + B(V) Quadratic (2nd order): Log M = A + B(V) + C(V2) Cubic (3rd order): Log M + A + B(V) + C(V2) + D(V3) Fifth Order: Log M + A + B(V) + C(V2) + D(V3) + E(V4) + F(V5) Cubic Spline Sets of points fitted with a series of Cubic equations Point to Point Points fitted with a series of linear equations
Extrapolation
©2014 Waters Corporation 30
Set Calibration Limits V0 and Vt
V0
Vt
©2014 Waters Corporation 31
Alternate Data Processing How to Process – Single Mode Limits
©2014 Waters Corporation 32
Multi Mode Processing
©2014 Waters Corporation 33
Characterization of a Broad Epoxy Sample
©2014 Waters Corporation 34
Summary
APC method development follows the same workflow as GPC. A number of factors must be considered when developing a
robust and appropriate test method including; – Goal of analysis – Polymer properties – Solvent selection – Detector response
Factors such as solvent selection and use of solvent additives can be easily screened and scaled on a single column set.
Selection of calibration standards can be made to match column pore size.
Calibration can be easily added to all test sets Use of analysis specific processing can improve data consistency
and direct data reporting
©2014 Waters Corporation 35
The ACQUITY® Advanced Polymer Chromatography™ (APC™) System Innovation in Size Based Separations
Precise solvent
management
Low system dispersion
Compatibility with
challenging solvents
Rigid, solvent-resilient columns
Versatile column
management
Stable refractive
index detection
Flexible and robust data processing
Wide range of APC
standards
©2014 Waters Corporation 36
Thank you for your attention.
Questions?