The ACQUITY Advanced Polymer Chromatography (APC™) System is a breakthrough technology that defines the ultimate in size-based chromatographic separations, delivering more information about your polymers faster than ever before. This means better characterization, improved asset utilization and a superior solution for achieving corporate innovation and sustainability goals.

1. ©2014 Waters Corporation 1
“Innovation in Size Based Separations"

2. ©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

3. ©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

4. ©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

5. ©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

6. ©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

7. ©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

8. ©2014 Waters Corporation 8
High Resolution SEC Characterization
-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
Minutes
2.00 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.

9. ©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

10. ©2014 Waters Corporation 10
Cellulose Based Polymer
– Target Molar Mass 300,000
- Polystyrene Equiv. 100,000

11. ©2014 Waters Corporation 11

12. ©2014 Waters Corporation 12
High Throughput Screening SEC
Dextran 410k
Dextran 47k
Dextran 165kDextran 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
LogDPw
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

13. ©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

14. ©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

15. ©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

16. ©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

17. ©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

18. ©2014 Waters Corporation 18
Selecting the Correct Pore Size
µRIU
0.00
15.00
30.00
Minutes
0.00 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

19. ©2014 Waters Corporation 19
Selecting the Column Length
µRIU
0.00
4.00
8.00
12.00
Minutes
0.00 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

20. ©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?

21. ©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

22. ©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)

23. ©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

24. ©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

25. ©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
Minutes
0.0 1.0 2.0 3.0 4.0 5.0 6.0

26. ©2014 Waters Corporation 26
Choose Appropriate Calibration Kit

27. ©2014 Waters Corporation 27
µRIU
0.0
1.0
2.0
3.0
4.0
5.0
6.0
7.0
Minutes
0.0 1.0 2.0 3.0 4.0 5.0 6.0
µRIU
0.00
4.00
8.00
12.00
16.00
Minutes
2.80 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
LogMolWt
3.0
4.0
5.0
6.0
Elution Volume
2.50 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

28. ©2014 Waters Corporation 28
Data Review

29. ©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

30. ©2014 Waters Corporation 30
Set Calibration Limits V0 and Vt
V0
Vt

31. ©2014 Waters Corporation 31
Alternate Data Processing
How to Process – Single Mode Limits

32. ©2014 Waters Corporation 32
Multi Mode Processing

33. ©2014 Waters Corporation 33
Characterization of a Broad
Epoxy Sample

34. ©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

35. ©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

36. ©2014 Waters Corporation 36
Thank you for your attention.
Questions?