MetaboLynx

Automating Drug Metabolism Studies

What is MetaboLynx?

• MetaboLynx is the application manager for automated processing and reporting of metabolism data

• It is an extension of MassLynx designed specifically for this application

• MetaboLynx accelerates the processing and reporting of drug metabolism data

• It can help to carry out data manipulation in hours (minutes) where it would previously have taken days (hours)

What is required?

• Many metabolites can be predicted• Processing is time-consuming and repetitive

• NEED AUTOMATED PROCESSING• Not all metabolites can be predicted• Need to search data for unknowns• They may be hidden by abundant endogenous peaks• Data processing is extremely labour-intensive

• NEED AUTOMATED PROCESSING• May need to acquire MS/MS data• Set-up, processing and reporting is time-consuming

• NEED AUTOMATED METHOD SET-UP AND PROCESSING

MetaboLynxMetaboLynx

Acquire control & metabolized samples

Acquire control & metabolized samples

LC/MS LC/MS

Control comparisonControl comparison

Search expected & unexpected

Search expected & unexpected

Report LC/MS resultsReport LC/MS results

Auto-start MS/MSAuto-start MS/MS

MetaboLynx Flow PathMetaboLynx Flow Path

Append to reportAppend to report

Create MS/MS methodCreate MS/MS method

Process MS/MS dataProcess MS/MS data

MetaboLynx Set-up

• Flexible parameters - discovery or development• Batch or single sample analysis• Expected metabolites menu• Unexpected metabolites• Control sample• Search data using MS, PDA or analog

• radioactivity detection, fluorescence, etc.• Exact mass measurement and elemental composition• Isotope cluster search

• chlorine, bromine, isotopic labeling• Automated MS/MS set-up

MetaboLynx Set-up - Peak Detection

MetaboLynx Set-up - Expected Metabolites

Unexpected Metabolites

• Small peaks in complex matrices• Requires intensive data mining• Greatest time-saving through automation• Using chromatogram peak detection• User defined step size and thresholds

Control Comparison

• Direct subtraction not feasible• Acquire control and experimental sample• Process control and experimental sample• Identify peaks by mass, RT and response• Specify windows for RT and response• Filter out matching peaks• Utility depends on control sample

MetaboLynx Set-up - Isotope Clusters

Isotope Cluster Plot (Br)

Data Visualization - MetaboLynx Browser

Automated LC-MS/MS

• Creates and starts LC-MS/MS acquisition • builds MS/MS method based on browser report• creates new sample list• creates new file name(s) and sample text• automatically starts MS/MS (product ion) acquisition • can choose full automation or manual intervention• processes MS/MS data• adds data to LC-MS report file and browser

MetaboLynx Browser - MS/MS

Instrument Compatibility

• ZMD / ZQ (single quad)• LC-MS

• Quattro micro / LC / Ultima (triple quad)• LC-MS/MS• processing of precursor and neutral loss data

• LCT (oa-TOF)• exact mass LC-MS

• Q-TOF micro / II / Ultima• exact mass LC-MS and LC-MS/MS

• GCT (GC-TOF)

MetaboLynx - Summary

• Automation of metabolism studies• Flexibility for metabolite screening or low level detection• Speed

• sample throughput in discovery• data mining in development

• Data visualisation and reporting• Not just metabolism

• Impurity studies• Further developments on-going