Poster - Waters Corporation · purification process. Efficiently managing this burden requires...
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Introduction Traditionally, drug discovery laboratories have screened large libraries with minimal to no purity requirements. Today, laboratories screen libraries that are smaller and more focused with specific purity limits, typically 85% or greater. This places a greater burden on the overall purification process. Efficiently managing this burden requires purification strategies that dovetail into the overall "synthesis-to-screen" process. This poster illustrates how a compound library is taken through a purification process, using the new Waters ® AutoPurify ™ Software. This software allows for automation from the initial QC, through the purification, to the fraction reanalysis. Intelligent decisions at each step increase the overall quality and throughput of the process. For example, using the target’s purity from analytical results, the appropriate purification method, if necessary, is selected for that sample based on the user- defined purification strategy. Additionally, the selected purification method can be a narrow gradient, identified based on the analytical retention time. This provides optimal target separation from closely eluting impurities and thus improving the resulting fraction purity. Additional tools allow for simple data importing/exporting using barcoded samples and fractions. Furthermore, data archiving using a relational data allows for easy search and retrieval capabilities of historical data to aid in maintaining a patent compliant position regarding newly synthesized chemical entities. System: Waters AutoPurification ™ System: 2525 Binary Gradient Module, 2767 Sample Manager, Column Fluidics Organizer, 2996 Photodiode Array Detector, ZQ ™ Mass Detector, MassLynx ™ and FractionLynx ™ Software, XTerra ® C 18 5 μm Column; Analytical: 2.1 x 50 mm, Preparative: 19 x 50 mm Samples: Samples were prepared by mixing various drug- like test compounds. Total compound concentration per sample was about 10 mg/mL PosterREPRINT Paul M. Lefebvre 1 , Andrew Brailsford 2 , David Brindle 2 , Clare North 2 , Ronan Cleary 1 , Warren B. Potts III 1 , Brian W. Smith 1 1 Waters Corporation, 34 Maple St., Milford, MA 01757; 2 Waters Corporation, 2 Floats Road, Wythenshawe, Manchester, UK, M23 9LZ Presented at PITTCON 2003, Orlando, FL, 9th - 14th, March 2003 COMPOUND PURIFICATION WORKFLOW MANAGEMENT AND OPTIMIZATION Waters FractionLynx ™ MS-based AutoPurification ™ System
Transcript of Poster - Waters Corporation · purification process. Efficiently managing this burden requires...
Introduction
Traditionally, drug discovery laboratories have screenedlarge libraries with minimal to no purity requirements.Today, laboratories screen libraries that are smaller andmore focused with specific purity limits, typically 85%or greater. This places a greater burden on the overallpurification process. Efficiently managing this burdenrequires purification strategies that dovetail into theoverall "synthesis-to-screen" process.
This poster illustrates how a compound library is takenthrough a purification process, using the new Waters®
AutoPurify™ Software. This software allows forautomation from the initial QC, through the purification,to the fraction reanalysis. Intelligent decisions at eachstep increase the overall quality and throughput of theprocess. For example, using the target’s purity fromanalytical results, the appropriate purification method, ifnecessary, is selected for that sample based on the user-defined purification strategy. Additionally, the selectedpurification method can be a narrow gradient, identified
based on the analytical retention time. This providesoptimal target separation from closely eluting impuritiesand thus improving the resulting fraction purity.
Additional tools allow for simple dataimporting/exporting using barcoded samples andfractions. Furthermore, data archiving using arelational data allows for easy search and retrievalcapabilities of historical data to aid in maintaining apatent compliant position regarding newlysynthesized chemical entities.
System: Waters AutoPurification™ System: 2525Binary Gradient Module, 2767 Sample Manager,Column Fluidics Organizer, 2996 Photodiode ArrayDetector, ZQ™ Mass Detector, MassLynx™ andFractionLynx™ Software, XTerra® C18 5 µm Column;Analytical: 2.1 x 50 mm, Preparative: 19 x 50 mm
Samples: Samples were prepared by mixing variousdrug- like test compounds. Total compoundconcentration per sample was about 10 mg/mL
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Paul M. Lefebvre1, Andrew Brailsford2, David Brindle2, Clare North2, Ronan Cleary1, Warren B. Potts III1, Brian W. Smith1
1Waters Corporation, 34 Maple St., Milford, MA 01757; 2Waters Corporation, 2 Floats Road, Wythenshawe, Manchester, UK, M23 9LZ
Presented at PITTCON 2003, Orlando, FL, 9th - 14th, March 2003
COMPOUND PURIFICATIONWORKFLOW MANAGEMENT
AND OPTIMIZATION
Waters FractionLynx™ MS-based AutoPurification™ System
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Gradient: Total Flow: Analytical: 0.5 mL/min;Preparative: 41 mL/minSolvent; A: Water with 0.1% formic acid; B:Acetonitrile with 0.1% formic acid
End1095 – 58.25 – 8.5
958 – 8.255 – 951 – 8
50 – 1Composition (%B)Time (Minutes)
Narrow end - 957.5 – 8
End1095 – 58.25 – 8.5
958 – 8.25
Narrow gradient1 – 7.55 - Narrow start0 – 1Composition (%B)Time (Minutes)
Generic: Focused Preparative
95637-106350.16-7
50.137.25-637.224.34-524.311.43-411.450-3
Ending %BStarting %BRetention Time
The analytical retention time determines whichnarrow purification method is run.
The resulting narrow and generic gradient profilesare shown at right.
GenericNarrow ANarrow BNarrow CNarrow DNarrow ENarrow F
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Benefit of Narrow Gradients
Mixture of 4 closely eluting compounds - Target M/Z = 283.2
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011703s42 1: Scan ES+ 277.9835.82e7
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011703s42 1: Scan ES+ 279.9356.53e7
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011703s42 1: Scan ES+ 282.8958.33e7
4.05
011703s42 1: Scan ES+ 268.858.36e6
3.88
011703s42 1: Scan ES+ TIC
1.48e84.104.06
0.59 3.88
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Generic Purification Gradient
• Generic Purification Gradient - Inadequate resolution and co-elution
• Narrow Purification Gradient- Baseline separation for the earlier
eluting impurity- Co-eluting impurity over 50% baseline
resolved
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011703s43 1: Scan ES+ 277.9833.85e7
4.71
011703s43 1: Scan ES+ 279.9354.44e7
3.203.29
011703s43 1: Scan ES+ 282.8953.96e7
2.862.81
2.97
011703s43 1: Scan ES+ 268.852.75e6
1.90
9.20
011703s43 1: Scan ES+ TIC
5.83e73.20
2.852.78
0.50
1.89
4.713.29
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Preparing for Intelligent Purification
AutoPurify™ Method Windows
• Option for single or multiple targets• Targets can be mass- or UV-based• Single or multiple chromatograms used to assess purity • Pass/Tentative/Fail thresholds defined
Target found and its purity is greater thanrequirement – No need to purify
Target found but its purity was betweenthe fail and pass threshold – Run a narrowgradient since retention time is known
Target found but its purity was below thefail threshold – Not worth purifying
Target was found but a peak to assignpurity could not be found – Run a genericgradient with an MS trigger
Target was not detected – No need to purify
Explanation
None
Narrow
None
Generic 1
None
Strategy
Ideal synthesisPassFound
Synthesis was OK, but not goodenough to meet the purityrequirement
TentativeFound
Very poor synthesis, only a smallamount of sample made
FailFound
Synthesis worked, but the qualityis unknown – not UV active
Not FoundFound
Failed SynthesisN/ANot Found
Possible ScenarioPurityTarget
Example Purification Strategy
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Strategy View
Analytical Results
Three different graphical summary views of the sample data • Classic view
- Displays target as Found/Not Found • Purity view
- Displays results after analytical interrogation- Combined view of Found/Not Found and Pass/Tentative/Fail
• Strategy view- Displays the samples that will and will not be purified
Purity View
FoundNot Found
Classic View
Found + PassFound + TentativeFound + FailFound + No PeakNot Found
To be purifiedNot to be purified
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Automated Stages or Manual Review
• Level of process automation or manual review is flexible- From fully automated with no manualintervention to user controlled steps
- Sample information is carried through theprocess regardless of the automation level
Reanalysis Results
• Collected fractions were automaticallyreanalyzed to reassess the purity of each tube
• The new results are interrogated with the sameparameters used on the original data
• Using the purity summary view allows for asimple display of the samples/fractions whichstill do not meet the purity threshold
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OpenLynx™ Global SERVER
• Sample all summary reports can be imported intoa relational database
• Allows for easy storage and retrieval ofarchived results
• Example query shows all the results for aspecific sample- Analytical - Preparative- Reanalysis
• Results are shown in a similar graphicalsummary, along with the other samples fromthat batch
Export List
Tab delimited text files can be exported with a user-defined set offields, allowing for simple process integration and population ofcorporate LIMS system.
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Summary of Benefits
• Increased throughput- Only purifying necessary samples
- Reduced solvent consumption- Reduced disposal cost
• Increased fraction quality- Narrow gradients
• Fully automated- Extent of manual intervention is flexible
• Easier process integration- Supports the use of barcodes on samples
and fractions- File import and export to interface with
complementary equipment- Databases- Liquid handlers
• Improved data handling- Data entered once
- More efficient- Less chance of error
- Results querying and archiving- Patent filing
• Can be done on a single system orinterchangeable between systems
Acknowledgements
• Schering AG, GmbH- Marion Wenz, Marcus Koppitz, Alexandra
Kappfer, Olaf Prien• GNF
- John Isbell and team• GSK, Stevenage
- Keith Brinded
• Waters Corporation- Darcy Shave- Mike Jackson- Tom Wheat- Simon Wilkins
Waters, AutoPurify, AutoPurification, ZQ, MassLynx, FractionLynx, OpenLynx and XTerra aretrademarks of Waters Corporation.All other trademarks are the property of their respective owners.©2003 Waters Corporation Printed in the U.S.A. March 2003 720000696EN LW-PDF
WATERS CORPORATION34 Maple St.Milford, MA 01757 U.S.A.T: 508 478 2000F: 508 872 1990www.waters.com
