Transcript Array Automation Polymorph Analysis

Raman Spectroscopy: a Valuable New
Tool for Polymorph Analysis
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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Acid A
Acid B
Acid C
Acid D
Acid E
Acid F
Acid G
Acid H
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Solvent 12
Solvent 11
Solvent 10
Solvent 9
Solvent 8
Solvent 7
Solvent 6
Solvent 5
Solvent 4
Solvent 3
Solvent 2
Solvent 1
Multi-Well Recrystallization
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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Raman Spectroscopy and Polymorphism
5-Methyl-2-[(2-nitrophenyl)amino]-3-thiophenecarbonitrile
also known as “ROY” *
Red Polymorph
Orange Polymorph
Yellow Polymorph
* 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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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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Comprehensive Software
• OMNIC™ software suite
–Automates all aspects of instrument control
–True push-button operation
–Minimal training time
–Automated data analysis
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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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Easy Experimental Set Up
• Point and click navigation
• Specify which wells to sample
• Specify data collection method for each well
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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
2 800 0
2 600 0
2 400 0
2 200 0
2 000 0
Raman intensity
1 800 0
1 600 0
1 400 0
1 200 0
1 000 0
8 000
6 000
4 000
2 000
3 000
2 500
2 000
Ra ma n s h ift (c m-1)
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1 500
1 000
5 00
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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Visual Display of Grouped Wells
• Cluster analysis displays wells grouped together by
spectral similarity
• Each line represents a single well
Green
Cluster
Yellow
Cluster
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Red
Cluster
Purple
Blue
Outliers
Cluster
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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Rapid Data Processing with Array Automation
Apply Norris derivative to eliminate fluorescence interference
Polymorphs identified by color coding
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Cluster analysis
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
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