Oral presentation at ACS2010 National Meeting on Aug. 22, 2010.

1. ACS 2010 National Meeting
Division of Analytical Chemistry
GPC-IR to Characterize Macromolecular
Excipients in Pharmaceutical Formulations
Ming Zhou, William Carson,
Sidney Bourne & Tom Kearney
Spectra Analysis, Inc.
August 22, 2010
Contact: ZhouM@Spectra-Analysis.com
Tel. 508-281-6276
1

2. OUTLINE
 GPC-IR Hyphenated Technology: Instrumentation
 Excipient Characterization: Copovidone PVP/VAc
 Excipient Degradation from HME Process:
HPMCAS, Eudragit L100-55 (PEA/MAA)
 Summary: GPC-IR Applications in Pharma Formulations
Q&A
2

3. Hyphenated Technologies &
Major Applications
LC-MS LC-IR
Separation
Liquid Chromatography
Detection & Mass Infra Red
Data Analysis Spectroscopy Spectroscopy
Applications Small Molecules, Proteins Polymers
Pharma API’s Polymeric Excipients

4. LC-IR Hyphenation

5. What is Direct Deposition FTIR?
Separated Dot Depositing on Disk Separated Dots from HPLC-IR Continuous Polymer Tracks (GPC-IR)

6. Direct Deposition FTIR
& Data Processing
ZnSe Disk

7. Features of DiscovIR-LC
 High Quality Solid Phase Transmission IR Spectra
 Real-Time On-line Detection
 Microgram Sensitivity
 Compatible with all LC Solvents and Gradients
• e.g. Water, ACN, Methanol, THF, Chloroform, HFIP
 Compatible with all GPC/SEC Solvents
 Fully Automated Operation: No Fractionation
 Multi-Sample Processing: 10 Hr ZnSe Disk Time

9. Compositional Drift Analysis of
Copolymer Poly(A-B) by GPC-IR
A
A/B Ratio B
Absorption
High MW Low MW Molar Mass
Ratio 10/8 12/12 2/4 Total 24/24
A% 56% 50% 33% 50%

10. Compositional Heterogeneity of
Copolymer Poly(A-B)
GPC-IR GPC
Absorption
IR Bulk 50% (NMR)
(MS)
High MW Low MW Molar Mass
Ratio 10/8 12/12 2/4 Total 24/24
A% 56% 50% 33% 50%

11. GPC-IR Spectrum of Copovidone
Excipient - VP/VAc Copolymer
Peak 1680 cm-1 from VP comonomer
Peak 1740 cm-1 from VAc comonomer

12. Excipient Compositional Drift
w/ MWD Vs. Bulk Average
GPC-IR Chromatogram Overlay with Comonomer IR Peak Ratios
Copovidone
Bulk Average
(Molecular Weight Distribution)
Abs. Peak Ratio: AVA / AVP = (k1*b*MVA) / (k2*b*MVP) = k (MVA / MVP) ~ Comonomer Ratio

13. Excipient Compositional Drift
w/ MWD Vs. Bulk Average
.6
Copovidone: sample A 50
molecular weight
.5
% acetate comonomer
distribution
max. IR absorbance
45
.4
.3 Bulk Average comonomer composition 40
40% VAc distribution
.2
35
.1
0 30
106 105 104 103 102 Molecular Weight

14. Copovidone MW Distributions from
Different Suppliers (Manf. Processes)
.6
Copovidone: sample A
sample B
.5
sample C
max. IR absorbance
.4
.3
.2
.1
0
Molecular Weight
106 105 104 103 102
Copovidone A gave clear tablets while Copovidone C led to cloudy ones.

15. Copovidone Compositional Drifts
from Different Manf. Processes
.6
Copovidone: sample A
50
sample B
.5
% acetate comonomer
sample C
45
.4
Molecular Weight
max. IR absorbance
Distribution Comonomer Composition
.3
Distribution
40
Bulk 40% VAc
.2
35
.1
0 30
Molecular Weight
106 105 104 103 102
Copovidone A gave clear tablets while Copovidone C led to cloudy ones.

16. Excipient Characterization
by LC-IR in Pharma Formulations
 Copolymer Compositional Analysis with MW Distributions
• Comonomer Ratio Drift (Functional Groups) vs. Bulk Average
• Excipient Lot-to-Lot Variations: QbD Studies
 Excipient Performance & Functional Group Correlations
• Hydrophobic/Hydrophilic Ratio Drift vs. Phase Separations
• Effects on Excipient Dissolution Behavior
Reference
(1) Chemical Heterogeneity on Dissolution of HPMC,
EU J. of Pharma Sci., P392 (2009), A. Viriden et al.
(2) Comp Drift Effect on Dissolution of PMMA/MAA,
Materials Letters, P1144 (2009), E. Manias et al.
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17. Excipient Degradation from
Hot Melt Extrusion Process
 Hot Melt Extrusion Process: To Make Solid Dispersions
for Low Solubility Drugs to Improve Bioavailability
 Degradation Issues
• Excipient & API Degradation at High Temp. (100-200C)
• Discoloration / Residues
• Degradant / API Interactions
 Process Variables
• Temperature
• Time (Screw Speed)
• Torque
• Screw / Die Designs
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18. Excipient HPMCAS Degradation
in Hot Melt Extrusion Process
Unprocessed
Processed at 160C
Degradant
Processed at 220C

19. Degradant from HPMCAS (220C)
in Hot Melt Extrusion Process
IR Database Search Result: Succinic Acid

20. HPMCAS Degradation
in Hot Melt Extrusion Process
-C=O
OH
Functional Group Ratio Changes from High Temp Process (Sample C)

21. GPC-IR Analysis of HPMCAS
Degradation in HME Process
 Detected Degradants: Succinic Acid & Derivatives
 Detected Functionality Ratio Change: Hydroxyl Vs. Carbonyl
 Help Understand Excipient Degradation Mechanism
 Study Excipient / API Interactions
 Define Safe Process Window: QbD
 Excipient Blends with Plasticizers and Additives
HOOC-CH2-CH2-C=O
CH3-C=O
Fig. A Schematic Structure of HPMC-AS

22. Eudragit L100-55 Characterization
by GPC-IR (Chromatograph + IR Spectra)

23. Eudragit L100-55 Compositional Drift at
Different Elution Times (Red 8’ & Blue 10’)
COOEt
1735
COOH
1705
CH2 CH3
Areas: L R
Acid / Ester Co-Monomer Ratio ~ Acid / Ester Peak Area Ratio = [(L+R)-2L] / (2L)

24. IR Spectra of L100-55 Samples at
Polymer Peak Center (Elution Time ~9.4’)
S0 – Green Ref COOEt
S1 – Pink 130C 1735
S2 – Blue 160C
S3 – Black 190C
COOH
1705
NCE?
1805 cm-1
CO-OH
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25. Excipient L100-55 Crosslinked from
COOH to Anhydride at Higher Temp
COOEt
1735 S0 – Green Ref
S1 – Pink 130C
S2 – Blue 160C
COOH S3 – Black 190C
1705
NCE?
1805 cm-1
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26. Summary: Eudragit L100-55
Degradation & Stability from HME
Sample # Extrusion Screw Sample Sample Degradant Polymer
Temp. Speed Color in THF Formed Change
(~0.5%)
S0 Not White Clear None None
Processed Solution
S1 130 C 250 rpm Off Clear Trace
White Solution Anhydrides
S2 160 C 250 rpm Off Clear Anhydrides Acid/Ester
White Solution Ratio
Decreased
S3 190 C 250 rpm Brownish Some Anhydrides Acid/Ester
Residue Ratio
Decreased
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27. Common Polymeric Excipients
Cellulose Derivatives
• HydroxyPropyl Methoxy Cellulose (Hypromellose): HPMC
• HPMC Acetate Succinate: HPMC-AS
• HPMC Phthalate: HPMC-P
• HydroxyPropyl Cellulose: HPC
 Copovidone: PolyVinyl Pyrrolidone / Vinyl Acetate – PVP/VAc
 SoluPlus Terpolymer: PEG / PVAc / PVCap
 Methacrylic or Methacrylate Copolymers: Eudragit
 Polyethylene Oxide: PEO (MW > 20K) or PEG (MW < 20K), PEG/PPG
 PLGA Copolymers: Biodegadable
 Excipient Combinations with Plasticizers and Additives
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28. GPC-IR Applications for Excipient
Analysis in Drug Formulations
Excipient Formulation Develop. Formulated Drugs
Manufacturing Drug Manufacturing Shelf Life Stability
• Process Control • Incoming QC • Stressed
• Lot-to-lot • Excipient Functionality Degradation
Variations • Formulation
• CoA Development • De-Formulate
• QbD Excipient Blends
• Novel Excipient
R&D • Process Degradation
(Hot Melt Extrusion)
• Define Safe Process
Window / QbD
• Process Monitoring
• Trouble Shooting • Trouble Shooting • Trouble-Shoot
Problem Drugs in
the Market
Users: Excipient Pharma Co. Pharma Co.
Manufacturers HME Service Providers Generic Drug Co.

29. Excipient QbD Space
GPC-IR-Performance
Slide from USP International Excipient Workshop (July 2009)
Performance
GPC
IR GPC-IR

30. Excipient Analysis with GPC-IR
in Pharceutical Formulations
 Polymeric Excipient Characterization
 Compositional Variations with MWD: Functional Group Ratios
 Lot-to-Lot, Supplier-to-Supplier Variations
 Degradation Analysis in Thermal Process (HME)
 Detect Degradants (Low MW)
 Polymer Structural Changes:
• Cross-Linking (New Chemical Entity)
• Functional Group Changes

31. GPC-IR & HPLC-IR Applications
 Excipient Characterization, Functionality & Degradation Analysis
 Copolymer Compositional Analysis across MW Distribution
 Polyolefin Copolymer Branching Analysis by High Temp GPC-IR
 Polymer Blend Ratio Analysis across MW Distribution
 Polymer Additive & Impurity Analysis
 De-Formulation for Polymers and Additives: Competitive Analysis
 Process Control & Optimization
 Excipients, Plastics, Rubbers, Films, Fibers, Foams & Composites
 Reactive Polymer Analysis for Coating, Adhesive, Sealant & Elastomer
 Isomer Analysis for Chemicals, Forensics & Pharmaceuticals
 General Analytical Capability: Trouble Shooting
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32. GPC-IR Operating Conditions
& Sample Preparation
 GPC Chromatograph: Agilent® 1200
• GPC Column Temperature: Ambient
• Solvent: THF at 1.0 ml/min
• Column: Jordi Gel DVB Mixed Bed– 250 x 10 mm
• Sample Injections: 100 ml at ~0.5% weight / volume THF
 IR Detection
• DiscovIR-LC® solvent-removing direct-deposition solid phase FTIR
• Cyclone Temperature: 150oC
• ZnSe Disk Temperature: -10 ~ -15oC
 Sample Preparation:
• 0.050 g excipient solid samples were dissolved in 10 ml THF in ~1
hr and filtered with 0.45 mm PTFE syringe filter before GPC injection

33. Acid/Ester Ratio Changes with
Elution Time (MWD) and Processing Temp.
S0 – Green Ref.
S1 – Pink 130C
S2 – Blue 160C
S3 – Black 190C
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34. HPMCAS Grade-to-Grade
Difference (LF/MF/HF) by GPC-IR
-C-O-C-
1060
HOOC-CH2-CH2-C=O
AS A
C=O Acetyl
1740 1235
CH3-C=O
HP
C/HP M
CH3
OH OCH3
1372
3470 2830

35. HPMCAS Grade-to-Grade
Difference (HF/MF/LF) by GPC-IR
Acetyl / C=O (total AS): Peak 1235cm-1 /1740 cm-1 Ratios
HF—0.8
MF—0.5
LF—0.4

36. IR Band Identifications of HPMCAS
Excipients for Ratio Drift Analysis
S A
HP HOOC-CH2-CH2-C=O CH3-C=O
C O
M M M M
o
O O
M CH3 M CH3 M
CH3-C=O A
Groups HP M C A AS Notes
CH3 1372 HP
OCH3 2830 M
OH 3470 (Unsub. OH & HP OH) OH
COCH3 1235 A
Total C=O 1740 AS
CH2 2935 2935 2935 2935 CH2
C-O-C 1060 BackBone
(BB)