Raman Spectroscopy: a Valuable New Tool for Polymorph Analysis

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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

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Polymorphism

• Important issue for pharmaceutical industry–Physical properties

• Bioavailability, stability, dissolution, manufacturability

–Development and manufacturing

–Regulatory

–Intellectual property

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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

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Multi-Well Recrystallization

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Acid A

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Common Techniques used in Polymorph Characterization

• X-ray Powder Diffraction (XRD)

• Differential Scanning Calorimetry (DSC)

• Thermal Gravimetric Analysis (TGA)

• Molecular information is lacking

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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

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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)

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5-Methyl-2-[(2-nitrophenyl)amino]-3-thiophenecarbonitrile also known as “ROY” *

Raman Spectroscopy and Polymorphism

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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.

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Yellow Polymorph

Orange Polymorph

Red Polymorph

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Raman Spectroscopy – The Past

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The Thermo Electron Solution

• The Nicolet™ Almega™ XR–Walk up instrument

–Analytical tool

–Designed for ease of use

–Highest reliability

–Highly automated

–Regulatory compliant

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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

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No Maintenance

• System alignment–Traditionally high maintenance

–Thermo solved this problem

–Patented autoalignment technique• Added benefit – maximum performance

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Trusted Results

• Automated system calibration– Integrated on-board standards

– Unattended operation

– Scheduled during off hours

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Regulatory Compliance

• System validation

–ValPro™ instrument qualification package

• Comprehensive qualification tools

– DQ, IQ, OQ

• Traceable standard

• Digital signatures

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• OMNIC™ software suite–Automates all aspects of instrument control

–True push-button operation

–Minimal training time

–Automated data analysis

Comprehensive Software

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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

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Easy Experimental Set Up

• Pre-defined templates–24, 48, 96, 384, 1536-well

microtiter plates

–Capillary arrays

–Micro-arrays

–Custom arrays

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• Point and click navigation

• Specify which wells to sample

• Specify data collection method for each well

Easy Experimental Set Up

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Easy Data Collection

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Easily Understandable Results

• Graphical presentation of data analysis– Point and click navigation and retrieval of spectral information

– Photo documentation of each measurement

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Overcoming the Challenge of Automated Data Analysis

• Eliminate background interference

• Facilitate automated spectrum to spectrum comparisons

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Raman shift (cm-1)

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Elimination of Background Fluorescence

• Norris derivative – Eliminates fluorescence

interference while revealing spectral similarities

– Permits batch elimination of interference

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Example of Applying Norris Derivative

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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

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YellowCluster

Green Cluster

BlueClusterRed

Cluster

Purple Outliers

Visual Display of Grouped Wells

• Cluster analysis displays wells grouped together by spectral similarity

• Each line represents a single well

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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

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Apply Norris derivative to eliminate fluorescence interference

Cluster analysis

Rapid Data Processing with Array Automation

Polymorphs identified by color coding

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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