1. Raman Spectroscopy: a Valuable New Tool for Polymorph Analysis

2. 2 Overview Raman spectroscopy –Rapidly growing technique New solution for polymorph screening – Multi-well format – Automated data collection and analysis Result of a year-long collaboration with a major pharmaceutical company

3. 3 Polymorphism Important issue for pharmaceutical industry –Physical properties Bioavailability, stability, dissolution, manufacturability –Development and manufacturing –Regulatory –Intellectual property

4. 4 Current Industry Status Polymorph characterization is moving to earlier stages in drug development More exhaustive screening –Parallel crystallization experiments Temperature Solvent selection Salt selection Recrystallization rate Increased analytical workload More data to analyze Demand for more automated analytical tools

5. 5 Multi-Well Recrystallization Solvent 1Solvent 2Solvent 3Solvent 4 Solvent 5 Solvent 6Solvent 7Solvent 8Solvent 9 Solvent 10Solvent 11Solvent 12 Acid A Acid B Acid G Acid C Acid D Acid F Acid H Acid E

6. 6 Common Techniques used in Polymorph Characterization X-ray Powder Diffraction (XRD) Differential Scanning Calorimetry (DSC) Thermal Gravimetric Analysis (TGA) Molecular information is lacking

7. 7 How Do We Fill the Molecular Information Gap? Raman spectroscopy –Measures shifts in wavelength of scattered light –Shifts are characteristic of compound and its molecular environment High information content –Molecular identification –Differentiates between: Polymorphs Solvates Hydrates Amorphic vs. crystalline forms

8. 8 Additional Benefits of Raman Spectroscopy Raman is well-suited to high throughput screening Non destructive Fast data acquisition (< 1 sec/sample) Little or no sample prep Flexible sampling –Microscope slides, glass vials,…etc. –Measure directly in Microtiter™ plates (48, 96, 384 well) –In situ monitoring Highly sensitive – Sample size flexibility (bulk or single crystal)

9. 9 5-Methyl-2-[(2-nitrophenyl)amino]-3-thiophenecarbonitrile also known as “ROY” * Raman Spectroscopy and Polymorphism N Yu, L.; Stephenson, G. A.; Mitchell, C. A.; Bunnell, C. A.; Snorek, S. V.; Bowyer, J. J.; Borchardt, T. B.; Stowell, J. G; Byrn, S. R. J. Am. Chem. Soc. 2000, 122, 585. * Yellow Polymorph Orange Polymorph Red Polymorph

10. 10 Raman Spectroscopy – The Past

11. 11 The Thermo Electron Solution The Nicolet™ Almega™ XR –Walk up instrument –Analytical tool –Designed for ease of use –Highest reliability –Highly automated –Regulatory compliant

12. 12 Raman Spectroscopy - Nicolet Almega XR Completely integrated system Laser safety Class I Micro analysis –Single crystals –96-well plates –High quality visual imaging –Polarized light Extensive automation No maintenance Reliable results Regulatory compliant Comprehensive software

13. 13 No Maintenance System alignment –Traditionally high maintenance –Thermo solved this problem –Patented autoalignment technique Added benefit – maximum performance

14. 14 Trusted Results Automated system calibration –Integrated on-board standards –Unattended operation –Scheduled during off hours

15. 15 Regulatory Compliance System validation –ValPro™ instrument qualification package Comprehensive qualification tools – DQ, IQ, OQ Traceable standard Digital signatures

16. 16 OMNIC™ software suite –Automates all aspects of instrument control –True push-button operation –Minimal training time –Automated data analysis Comprehensive Software

17. 17 OMNIC Array Automation Software Most advanced solution for polymorph characterization Developed and tested with pharmaceutical partner –Full year collaboration in a true drug development environment Automates extraction of useful information –Data collection –Data processing –Data analysis LIMS compatible Robotics ready

18. 18 Easy Experimental Set Up Pre-defined templates –24, 48, 96, 384, 1536-well microtiter plates –Capillary arrays –Micro-arrays –Custom arrays

19. 19 Point and click navigation Specify which wells to sample Specify data collection method for each well Easy Experimental Set Up

20. 20 Easy Data Collection

21. 21 Easily Understandable Results Graphical presentation of data analysis –Point and click navigation and retrieval of spectral information –Photo documentation of each measurement

22. 22 Overcoming the Challenge of Automated Data Analysis Eliminate background interference Facilitate automated spectrum to spectrum comparisons

23. 23 Elimination of Background Fluorescence Norris derivative –Eliminates fluorescence interference while revealing spectral similarities –Permits batch elimination of interference

24. 24 Example of Applying Norris Derivative

25. 25 Results Extraction From Entire Data Set The challenge –Compare all spectra with all the others in the array to identify known classes and potential new forms Cluster analysis algorithm performs a cross correlation between all members in a data set –Assigns spectra to groups based on similarity –Identifies outliers –Results are processed and presented visually –Immediately grasp results of entire experiment Rapid analysis of large and complex data sets

26. 26 Yellow Cluster Green Cluster Blue Cluster Red Cluster Purple Outliers Visual Display of Grouped Wells Cluster analysis displays wells grouped together by spectral similarity Each line represents a single well

27. 27 Identifies Known Polymorphs and New Forms Color-codes wells into four groups –Colored red, green, blue and yellow Twenty wells do not fall into any of the four groups –Colored purple –Potentially new forms Reports results to LIMS Manual spectral comparison takes skilled analyst 4 hours per 96-well plate Array Automation takes 5 minutes

28. 28 Apply Norris derivative to eliminate fluorescence interference Cluster analysis Rapid Data Processing with Array Automation Polymorphs identified by color coding

29. 29 Summary Raman fills the critical molecular information gap The Thermo Raman tool is particularly well suited for the pharmaceutical analytical laboratory –High degree of automation –High reliability –Automated data analysis –Complete instrument qualification tools Thermo has developed an advanced solution for polymorph screening Thermo is committed to partnering with you