This presentation will provide an introduction to the technology and benefits of Waters ACQUITY Ultra Performance Convergence Chromatography system (UPC2) and illustrate how it can impact the analysis of chemicals and polymers.

1. ©2015 Waters Corporation 1
Enhancing Separations Performance for
Chemicals and Polymers
Tim J Jenkins Ph.D
Director, Business Development

2. ©2015 Waters Corporation 2
Rebirth of Polymer Science

3. ©2015 Waters Corporation 3
Renewed Focus on Polymer Analysis
Ion mobility enabled HDMS
Comprehensive 2D
Separations
Thermal Analysis
& Rheology
APC : Polymer MWD

4. ©2015 Waters Corporation 4
Evolution of Separation Technology
Gas Chromatography Convergence Chromatography
GC
Capillary GC
HPLC
UPLC
SFC
UPC2
Liquid Chromatography

5. ©2015 Waters Corporation 5
Definitions
Convergence Chromatography is a category of separation science
that provides orthogonal and increased separation power,
compared to liquid or gas chromatography, to solve separation
challenges.
UltraPerformance Convergence Chromatography [UPC2] is a
holistically designed chromatographic system that utilizes
liquid CO2 as a mobile phase to leverage the chromatographic
principles and selectivity of normal phase chromatography
while providing the ease-of-use of reversed-phase LC.

6. ©2015 Waters Corporation 6
Driving UPC2 performance
–Selectivity that can be addressed
–Management of supercritical fluids
–Innovative chemistries

7. ©2015 Waters Corporation 7
What Drives UPC2 Performance?
- Addressable Selectivity
Solvent
Pentane, Hexane, Heptane
Xylene
Toluene
Diethyl ether
Dichloromethane
Chloroform
Acetone
Dioxane
THF
MTBE
Ethyl acetate
DMSO
Acetonitrile
Isopropanol
Ethanol
Methanol
Stationary Phase
Silica / BEH
2-ethylpyridine
Cyano
Aminopropyl
Diol
Amide
PFP
Phenyl
C18 < C8
Reversed-
phase
Selectivity
Space
Normal Phase
Selectivity
Space

8. ©2015 Waters Corporation 8
Convergence
Chromatography
Selectivity Space
What Drives UPC2 Performance?
- Addressable Selectivity
Solvent
Pentane, Hexane,
Heptane
Xylene
Toluene
Diethyl ether
Dichloromethane
Chloroform
Acetone
Dioxane
THF
MTBE
Ethyl acetate
DMSO
Acetonitrile
Isopropanol
Ethanol
Methanol
Stationary Phase
Silica / BEH
2-ethylpyridine
Cyano
Aminopropyl
Diol
Amide
PFP
Phenyl
C18 < C8
WeakStrong
Supercritical
CO2
Organic
Modifier

9. ©2015 Waters Corporation 9
What Drives UPC2 Performance?
- Managing Supercritical Fluids
Historical challenges for SFC
 Lack of robustness
– Shifting retention times
– Low accuracy for partial loop injections
– Unstable modifier delivery at low percentages of co-solvent (< 5%)
 Lack of instrument performance
– Insufficient instrumentation reliability (pumping system, injection
mechanism, backpressure regulator)
– Large system dispersion and dwell volumes prevented adoption of
smaller particles and high throughput analysis
 Low sensitivity
– High detector and pump noise
– Refractive index effects of CO2

10. ©2015 Waters Corporation 10
What Drives UPC2 Performance?
- Innovative Chemistries
ACQUITY UPC2 BEH 2-EP
• Good retention, peak shape and selectivity
• Lipids, steroids, pesticides
ACQUITY UPC2 BEH
• Heightened interaction with polar groups such as phospholipids
• OLED’s, polymer additives, pesticides
ACQUITY UPC2 CSH Fluoro-Phenyl
• Good retention of weak bases
• Alternate elution for acidic and neutral compounds
• Vitamin D metabolites, steroids, natural products
ACQUITY UPC2 HSS C18 SB
• Reversed-phase-like selectivity
• Fat soluble vitamins, lipids (Free fatty acids)
Scalable to larger particle sizes
ACQUITY UPC2/Viridis BEH
(1.7, 3.5 and 5 µm)
ACQUITY UPC2/Viridis BEH 2-EP
(1.7, 3.5 and 5 µm)
ACQUITY UPC2/Viridis CSH Fluoro-
phenyl (1.7, 3.5 and 5 µm)
ACQUITY UPC2 HSS C18 SB
(1.8 and 3.5 µm)

11. ©2015 Waters Corporation 11
 Built upon proven UPLC Technology
– Quantifiable increase in productivity
– Ultra-low dispersion enable the use of small
diameter particles
 Exceptional increase in available selectivity
– Solve routine & complex separation challenges
 2012 Pittcon Editors Gold Award
 2013 Green Innovation Award
 2013 Best New Separation Product
 2013 R&D 100 Award

12. ©2015 Waters Corporation 12
ACQUITY UPC2 Flow Path and
Components
Inject
valve
Auxiliary
Inject valve
Column Manager
PDA detector
Back Pressure Regulator
(Dynamic and Static)
Waste Modifier CO2
Supply CO2
Pump
Modifier
Pump
mixer
Thermo-electric
heat exchanger
Make-up
Pump
Mass Spec
Splitter

13. ©2015 Waters Corporation 13
UPC2 Configuration for MS
UV Detector
Make-Up
Pump
Convergence
Manager
MS

14. ©2015 Waters Corporation 14
The Key Benefits of UPC²
 Simplify the workflow with UPC2
– Combine multiple techniques (LC & GC into CC)
– Access robust normal phase separations
– Eliminate solvent exchange steps for organic extracts
 Deal with compound Similarity challenges
– Chiral Separations (enantiomers & diastereomers)
– Positional isomers (differ in location of functional groups)
 Deliver Orthogonal separations
– Different relative retention helps ensure full characterization
– Check method specificity by comparison to a second
procedure
– Reveal “hidden” impurity or degradation peaks
– Increase confidence in characterization of complex samples
SIMPLICITY
SIMILARITY
ORTHOGONALITY

15. ©2015 Waters Corporation 15
Oligomeric Materials

16. ©2015 Waters Corporation 16
UPC2 Polymer Separations:-
Polystyrene
AU
0.00
0.20
0.40
0.60
0.80
AU
0.00
0.20
0.40
0.60
0.80
AU
0.00
0.02
0.04
0.06
0.08
0.10
0.12
Minutes
0.00 0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00
Column: Waters ACQUITY UPLC HSS
Cyano 1.8 um, 3x100 mm. Gradient: 10
%B to 45 %B over 3 min, hold for 30 s,
back to 10 % B in 30 s
Flow rate: 1.7 mL/min, ABPR: 2800 psi
PS-1000
PS-2500
PS-1300

17. ©2015 Waters Corporation 17
UPC2 Polymer Separations:-
PMMA
AU
-0.002
0.000
0.002
0.004
0.006
0.008
0.010
0.012
0.014
Minutes
1.00 2.00 3.00 4.00 5.00 6.00
AU
0.000
0.002
0.004
0.006
0.008
Minutes
2.00 4.00 6.00 8.00 10.00 12.00 14.00
Column: Waters ACQUITY UPLC
HSS Cyano 1.8 um, 3x100 mm.
Flow rate: 2.0 mL/min, ABPR: 2000
psi. Gradient: 5 %B to 30 %B over
15 min, hold for 1 min, back to 5
%B in 0.1 min
PMMA-2500
PMMA-4250
Column: Waters ACQUITY UPLC
HSS Cyano 1.8 um, 3x100 mm.
Flow rate: 2.0 mL/min, ABPR:
2000 psi Gradient: 5 %B to 25
%B over 5 min, then to 30 %B
over 1 min, back to 5 %B in
0.1 min.

18. ©2015 Waters Corporation 18
UPC2 Polymer Separations:-
Triton X-100
 Non-ionic surfactant: used in cosmetics, industrial materials,
and as detergents
 Composition requires monitoring
– Differences in ethoxy chain length affect viscosity, solubility, polarity,
and other characteristics of the product
NP-HPLC
HT-GC
SFC

19. ©2015 Waters Corporation 19
UPC2 Polymer Separations:-
Triton X-100
System: ACQUITY UPC2 with UPC2PDA
Column: ACQUITY UPC2 BEH 2.1mm x 50mm, 1.7µm
Mobile Phase: A: CO2
B: Methanol
Wash Solvents: 70:30 Methanol:Isopropanol
Separation Mode:Gradient starting at 2% B to 35% over 1.25 minutes, back to
2% B in 5 s
Flow Rate: 2.0 mL/min
ABPR: 1500 psi
Column Temp.: 40°C
Injection Volume:1.0 µL
Run Time: 2 minutes
Detection: PDA 3D Channel: PDA, 210-400nm;
PDA 2D Channel: 222nm @ 4.8nm Resolution
(Compensated 380-480nm)
CDS: Empower® 3 CDS

20. ©2015 Waters Corporation 20
Peak1-0.271
Peak2-0.455
Peak3-0.574
Peak4-0.662
Peak5-0.733
Peak6-0.792
Peak7-0.841
Peak8-0.884
Peak9-0.921
Peak10-0.954
Peak11-0.985
Peak12-1.013
Peak13-1.041
Peak14-1.067
Peak15-1.093
Peak16-1.119
Peak17-1.158
Peak18-1.188
Peak19-1.217
Peak20-1.248
AU
0.00
0.10
0.20
0.30
0.40
0.50
0.60
0.70
0.80
0.90
1.00
1.10
1.20
1.30
Minutes
0.10 0.15 0.20 0.25 0.30 0.35 0.40 0.45 0.50 0.55 0.60 0.65 0.70 0.75 0.80 0.85 0.90 0.95 1.00 1.05 1.10 1.15 1.20 1.25 1.30 1.35 1.40 1.45 1.50
Triton X 100
UPC2 Polymer Separations:-
Triton X-100
• CO2 and Methanol gradient @ 40°C
• 75s to elute all components
• Approx. 20 oligomers separated and detected
1.5 mins

21. ©2015 Waters Corporation 21
ColumnName: SampleName:tritonh/e65deg3-20gr 20mniDateAcquired:8/16/20128:44:13AMEDTInstrumentMethodId:8156InjectionId:8174
AU
0.000
0.010
0.020
0.030
0.040
0.050
0.060
0.070
0.080
0.090
0.100
UPC2 Polymer Separations:-
Triton X-100
 Optimise separation for resolution (20 mins run time)
 See significant fine structure
 Ability to detect and monitor minor components and by-products

22. ©2015 Waters Corporation 22
UPC2 Polymer Separations:-
Condensation co-polymer synthesis
Bisphenol A and formaldehyde condensation co-polymer
Formaldehyde
Bisphenol A
Resin “Trimer”

23. ©2015 Waters Corporation 23
UPC2 Polymer Separations:-
Condensation co-polymer synthesis
AU
0.00
0.05
0.10
0.15
0.20
0.25
0.30
0.35
0.40
0.45
Minutes
0.00 0.50 1.00 1.50 2.00 2.50 3.00
Unknown
m/z 227
Dimer
m/z 467
Trimer
m/z 707
UPC2 with UV and MS detection

24. ©2015 Waters Corporation 24
UPC2 Polymer Separations:-
Condensation co-polymer synthesis
AU
0.00
0.05
0.10
0.15
0.20
0.25
0.30
0.35
Minutes
0.00 0.50 1.00 1.50 2.00 2.50 3.00 3.50
Combined- SQ1: MSScan1: 200.00-2000.00ES-, Centroid, CV=Tune
227.0
271.0
Intensity
0
50000
100000
150000
200000
250000
300000
350000
400000
450000
500000
m/z
200.00 250.00 300.00 350.00 400.00
Sample
Mass spectrum
Bisphenol-A
standard
Mass spectrum

25. ©2015 Waters Corporation 25
UPC2 Polymer Separations:-
Poly [Phenylglycidyl ether–co-formaldehyde]
 Potentially complex mixture of oligomers depending on where
linkage occurs between the units.
 3 Potential dimers are shown below
Phenylglycidyl ether
+ Formaldehyde
Dimers
(1)
(2)
(3)

26. ©2015 Waters Corporation 26
UPC2 Polymer Separations:-
Poly [Phenylglycidyl ether–co-formaldehyde]
Dimer1
Trimer1
Dimer2
Dimer3
Trimer2
Trimer3
Trimer5Trimer4
Trimer6
Trimer7
UPC2 with UV detection

27. ©2015 Waters Corporation 27
UPC2 Polymer Separations:-
Poly [Phenylglycidyl ether–co-formaldehyde]
Intensity
0.0
5.0x107
1.0x108
1.5x108
2.0x108
Minutes
0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 11.00 12.00 13.00 14.00 15.00 16.00 17.00
Dimers
m/z 312
Trimers
m/z 474 Tetramers
m/z 636
m/z
404 m/z
402
m/z 404
m/z 402
UPC2 with MS detection

28. ©2015 Waters Corporation 28
Extractable Materials

29. ©2015 Waters Corporation 29
“Hot Topic”
– Extractables & Leachables
Focus from ..
 Packaging and Coatings Industries
 Consumer Product Manufacturers
 Cosmetics and Personal care Producers
Focus on …
 Extractables
 Leachables
 Intentionally added substances (IAS)
 Non-intentionally added substances (NIAS)
Focus because …
 Regulation

30. ©2015 Waters Corporation 30
Typical extractables, IAS and NIAS
 Impurities in starting materials
 Chemical additives, plasticizers, antioxidants and contaminants
present in individual polymers
 Monomers & oligomers from incomplete polymerization
 Volatile compounds from secondary packaging; inks, adhesives
 Residual compounds from the surfaces of the molding
equipment, antistatics etc

31. ©2015 Waters Corporation 31
Controlled extraction study
 4 material types
– High Density Polyethylene bottle (HDPE)
– Low Density Polyethylene container (LDPE)
– Ethylene Vinyl Acetate plasma bag (EVA)
– Polyvinyl Chloride blister pack (PVC)
 3 different extraction solvents /
techniques
– Water (Conventional oven)
– Isopropanol (with Microwave and ASFE)
– Hexane (with Microwave and Soxhlet)
 3 separation techniques
– GC
– UPLC
– UPC2 Apps Note 720004509en.pdf
Apps Note 720004490en.pdf

32. ©2015 Waters Corporation 32
Irgafos 168
5-chloro-2-hydroxy-4-
methylbenzophenone (5-Cl-2-
OH-4-methyl BP)
Diphenyl phthalate Uvitex OB
Irganox 245
Irganox 1076
4-hydroxy-2-octyloxybenzophenone (4-OH-2-octyloxy BP)
Tinuvin 328Irganox 1010
Lowinox 44B25
Irganox 1330
BHT
Naugard 445
Tinuvin P
Structures of Polymer Additives
Investigated

33. ©2015 Waters Corporation 33
4 min separation by UPC2 vs.
9.5 min by UPLC
UPC2
BHT
5-Cl-2-OH-4-methylBP
TinuvinP
Tinuvin328
Irgafos168
2-OH-4-octyloxyBP
Irganox1076
Diphenylphthalate
UvitexOB
Naugard445
Irganox1330
Irganox1010
Irganox245
Lowinox44B25
AU
0.00
0.05
0.10
0.15
0.20
0.25
0.30
0.35
0.40
0.45
0.50
Minutes
0.00 0.20 0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40 2.60 2.80 3.00 3.20 3.40 3.60 3.80 4.00
5-Cl-2-OH-4-methylBP
UPLC
TinuvinP
Diphenylphthalate
BHT
Irganox245
Lowinox44B25
4-OH-2-octyloxyBP
UvitexOB
Naugard445
Tinuvin328
Irganox1076
Irganox1330
Irganox1010
Irgafos168
AU
-0.15
-0.10
-0.05
0.00
0.05
0.10
0.15
0.20
0.25
0.30
0.35
0.40
0.45
0.50
0.55
0.60
Minutes
0.00 0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00 5.50 6.00 6.50 7.00 7.50 8.00 8.50 9.00 9.50 10.00
Chromatographic separations

34. ©2015 Waters Corporation 34
Irgafos168
Irganox1076
Irganox1010
AU
-0.008
-0.006
-0.004
-0.002
0.000
0.002
0.004
0.006
0.008
0.010
0.012
0.014
0.016
0.018
0.020
0.022
0.024
0.026
0.028
0.030
Minutes
0.00 0.20 0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 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
UV chromastogram of LDPE SFE
extract analysed by UPC2

35. ©2015 Waters Corporation 35
Intensity
0.0
5.0x105
1.0x106
1.5x106
2.0x106
2.5x106
3.0x106
3.5x106
Minutes
0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00
Combined - SQ 1: MS Scan 1: 200.00-1200.00 ES+, Centroid, CV=Tune
475.5
476.5 531.6
548.6
549.6
553.6
554.6 569.5
Intensity
0.0
2.0x106
4.0x106
6.0x106
8.0x106
1.0x107
1.2x107
1.4x107
1.6x107
1.8x107
2.0x107
m/z
450.00 460.00 470.00 480.00 490.00 500.00 510.00 520.00 530.00 540.00 550.00 560.00 570.00 580.00 590.00 600.00
Confirmation of identity using MS
Intensity
0.0
2.0x106
4.0x106
6.0x106
8.0x106
1.0x107
Minutes
0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00
Combined - SQ 1: MS Scan 1: 200.00-1200.00 ES+, Centroid, CV=Tune
475.5
476.5 531.6
548.6
549.6
553.6
554.6
572.6
Intensity
0.0
2.0x106
4.0x106
6.0x106
8.0x106
1.0x107
1.2x107
1.4x107
1.6x107
1.8x107
2.0x107
2.2x107
m/z
450.00 460.00 470.00 480.00 490.00 500.00 510.00 520.00 530.00 540.00 550.00 560.00 570.00 580.00 590.00 600.00
Peak in LDPE extract
Peak from
Irganox 1076 std

36. ©2015 Waters Corporation 36
Workflow Benefit of ACQUITY UPC2 for
the Analysis of Polymer Extracts
Non-Polar Solvent
Extraction
Polar Solvent
Extraction
Inject direct
on GC
Evaporate and reconstitute
in a more polar solvent for
LC injection
Inject direct
on LC
Back-extract with a non-
polar solvent for GC injection
Polar or Non-Polar
Extraction
Inject direct
On UPC2

37. ©2015 Waters Corporation 37
Analytical Scale SFE (MV-10 ASFE)
Advantages of SFE
 Little to No Residual Solvents
 Superior Yield and Purity
 Lower Operating Costs
 Processes Thermolabile Compounds
 Safe
 Scalable
 Variable solvent power (Tunable)
 Gas-like mass transfer
 Zero surface tension
 SC-CO2 has high affinity with organic solvents

38. ©2015 Waters Corporation 38
Analytical Supercritical Fluid
Extraction
Low IPA SFE
High IPA SFE
Soxhlet
Microwave
Column Name: 2-EPSampleName: LDPEhigh IPA SFEDate Acquired: 9/6/2012 9:28:52 PMEDT Instrument Method Id: 1953 Injection Id: 2310
AU
-0.005
0.000
0.005
0.010
0.015
Column Name: 2-EPSampleName: LDPElow IPA SFEDate Acquired: 9/6/2012 8:41:45 PMEDT Instrument Method Id: 1953 Injection Id: 2266
AU
-0.006
-0.004
-0.002
0.000
0.002
0.004
0.006
Column Name: 2-EPSampleName: LDPEIPA sox Date Acquired: 9/6/2012 6:38:07 PMEDT Instrument Method Id: 1953 Injection Id: 2151
AU
-0.005
0.000
0.005
0.010
0.015
0.020
Column Name: 2-EPSampleName: LDPEIPA mw Date Acquired: 9/6/2012 5:50:59 PMEDT Instrument Method Id: 1953 Injection Id: 2107
AU
-0.005
0.000
0.005
0.010
Minutes
0.00 0.20 0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 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

39. ©2015 Waters Corporation 39
SIMPLICITY
SIMILARITY
ORTHOGONALITY

40. ©2015 Waters Corporation 40
 Expanding selectivity with
Convergence Chromatography (CC)
– Principles of CC
 Understanding the technology and
the instrument
 Getting started with CC
 Method development
 Applications
 [Click Here] for Web Overview
 Order a Paper Copy [Here]
Available Now
Convergence Chromatography Primer