Post on 30-Jun-2015
transcript
Advanced Chemistry Development, Inc. (ACD/Labs)
A Fully Automated System for Chromatographic
Method Development Utilizing LC/MS/DAD
Detection
Mike McBrien, Andrey Vazhentsev, and Alexey GalinMarch 15/06
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Outline
The ChallengeInstrument ControlStrategy BuildingMutual Automated Peak MatchingComposite SamplesConclusion
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Method Development Software Today
Users:Design studiesConfigure instruments
Computers Collect, process dataUsers:
Organize datafilesInterpret/extract dataTransfer data to modeling software
Computers model separationUsers:
reconfigure instrument…
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The ChallengeSoftware can do better:
Configuring sessions on the CDSInterpreting dataManaging informationContributing to workflowEliminating unnecessary injections
AutoChrom is designed to further enhance the MethDev workflow.
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Improving on MDS
The next generation of method development:
Instrument controlStreamlining data interpretationEnabling data managementConsistent decision-makingClear, concise reportingEasy redesign of methods
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ACD/AutoChrom
Direct instrument control Automated peak tracking:
UV- and MS-MAPCustom WorkflowsComplete data management:
Data organization by method/wave“Live” peak transferInstrument control
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Managing Instrument Data
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Instrument Control - Configuration
AutoChrom is designed to support virtually any setup that the CDS supports:
Column switchersOvensReservoirs and solvent volumesInjection vials and wells
Even manually-controlled valves can be used; AC prompts the user to change settings.
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Managing Method Development Waves
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Instrument Control – Conditions
Currently, AutoChrom controls only ChemStation PlusConfigured for complete control of the CDS:
TemperatureGradientColumn selectionEtc.
AutoChrom is designed to allow the user to specify virtually any experiment required.
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ACD/AutoChrom 1.0Configuring Console
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Data Management – MethDev Tasks
The MethDev strategy is divided into tasks:
ScreeningScreening/optimization1D optimization2D optimization
Upon completion of a given task, the software waits for instructions or moves to the next task.
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Designing Method Development Approaches
Strategy Builder
Users drop waves into strategy, and double-click
each wave to configure specific requirements.
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Decision-makingDecisions are based on existing LC Simulator algorithms:
Selection modes (screening);Optimization modes (continuous variables).
Suitability can be a function of:Run time;Retention;Resolution.
Users establish their success criteria for each method development wave.
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Data Review
Combination of manual and automated toolsHyphenated detection (MS and/or DAD) aids in component trackingReviewed data is automatically updated to the data management and decision support system
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Data Review (2)
AutoChrom reads peak tables directly post-analysisUsers retain access to hyphenated data throughout the method development sessionAt any time in the future, LC/MS/DAD data can be reviewed.
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Data Interpretation
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Peak Matching
Perhaps the greatest challenge in computer-assisted method development.All peaks must be tracked across all experimentsHumans use spectra, intuition, and test injections.Computers are more limited.
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Matching Peaks for Complex Work
Each component in a separation must be tracked
Manual peak tracking can be tedious in CA MethDev; it’s impossible in
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XTerra RP18 C18 pH 7
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Peak Matching
DAD detectors are ubiquitousMS detection is becoming commonThe challenge:
Develop a tracking system that works for DAD alone, or DAD/MS.
The solution:Mutual Automated Peak matching (MAP)
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UV-Mutual Automated Peak Matching
Data augmentationDeducing the number of componentsKey Set selectionKey Set refinementFinding peaksDetection of “missing” components
Anal. Chim. Acta, 490 (2003) 41-58.
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MS-Mutual Automated Peak Matching
MS offers significant advantages:Greater resolutionS/NOrthogonal signal strength
The MS-MAP algorithm was designed to take advantage of the differences in these detectors
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LC/MS Componentization: IntelliXtract
Combined approach of logic and chemometricsUser-transparent extraction of chromatographic informationIdeally suited to support of chromatographic peak matching2.5 years’ development at ACD/Labs
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Basic Componentization Workflow
Starting list of potential chromatographic peaksFiltering Real Peaks from NoiseGrouping peaks based on retention timesPrimary ion cluster sortingSearching for missing isotope ionsIon cluster analysisIon Cluster Annotation
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MS-MAP: Reconciling Components
IntelliXtract is used primarily as a noise reduction and peak picking toolMatches are based on clustersPerfect co-elution is a problem (today)Solution:
Logic-based reconciliation (if new component arises in run2, then re-examine run1)Orthogonal detection (UV-MAP)
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A note on detectors
DAD and MS are complementary:Intensities don’t correlateFunctional vs. weight difference
UV-MAP and MS-MAP are both useful:
MS-MAP is sensitive to co-elutionUV-MAP is sensitive to S/N
Using UV and MS greatly reduces “misses”
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Information Management
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Data Organization
AutoChrom’s interface is designed specifically for transparent communication of method development workWave data are arranged hierarchically:
MethodTrial
Probe (subsample injected)Detector (one or more datafiles)
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Data HierarchyProject
Wave
Experiment
Subsample
Detector
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Managing Experimental Information
All experimental data for the session are managed, grouped
in waves.
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Managing Experimental Information
All signals are linked to the original injection.
Peak tables are extracted from the original data.
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Linking multiple method development waves
Managing Method Development Sessions
Column ScreeningGradient Optimization
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A Data Explosion
Forced degradation samples:ComplexLow level, unknown impuritiesMultiple samples – one methodLOTS of hyphenated data
Each time a formulation is changed, new impurities may appear – the method
must be redesigned.
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Composite Samples
Sometimes more than one injection represents a single sampleChromatograms must be combined to represent an overall, expected behavior
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Composite Chromatograms
Multiple conditions = different impurities:
HOOHHeatLightAcidBase
The final method should separate all components
for all samples.
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Too Much Data
Each set of chromatographic conditions gives 18 (!) datafiles:
6 subsamples3 detectors
All elution data must be cross-correlated and transferred to method design software
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Example: Managing Stability Data
Each method has had 5 samples injected.
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Example: Managing Stability Data
AutoChrom combines tRs into one chromatogram
per method.
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Reports
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Communicating a MethDev Project
AutoChrom is designed to create a concise summary of an immense amount of dataRigorous method development for composite samples generates GB of instrument filesAutoChrom organizes, interprets, and communicates this dataConcise reports can be created in minutes
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The Math
In one MethDev session:~50 sets of conditions~5-7 samples~10 components~3 detectors
We need to extract/track/organize at least 7500 peaks!
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Speed vs. Organization
The method development console organizes raw data:
Peak data is summarized for entire projectRaw data is indexed but opened only on demandProjects are ~100KRaw data can be more than 4GB
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Compatibility
Instrument control in v1.0 for ChemStation Plus:
Fully automatedUser-interactive
All instruments can be used:Manual input of data still enables considerable time savings in data interpretation and organization
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Conclusion
Streamlining the method development session:
MS/UV supportChemometricsComposite chromatogramsUnprecedented rigor in modeling and reporting
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Acknowledgements
ACD/Labs:Rhiannon JonesAndrey Bogomolov
Johnson and Johnson:Rudy SneyersWilly Janssens
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Supplemental Information
Slides available at www.acdlabs.comBooth#2340Poster 470-67P:
“Managing LC/MS/DAD Data in Forced Degradation Studies”
Presentation 860-6: “Chromatographic Peak Tracking through
Chemometric Analysis of Hyphenated Data”