1. Automating Drug Metabolism Studies
MetaboLynx
Automating Drug Metabolism Studies
The next slide give a tour through he MetaboLynx software and highlight some of the key features and the benefits those features bring to the analysis of drug metabolites.
For those who want to be able to use the MetaboLynx software ‘live’ there are some basic instructions on how to access the displayed windows within MassLynx.

2. 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)
MetaboLynx has been designed to automate these processes.
It is an option which can be loaded as an extension of MassLynx and will accelerate the processing and reporting of drug metabolism data

3. 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
May need to acquire MS/MS data
Set-up, processing and reporting is time-consuming
NEED AUTOMATED METHOD SET-UP AND PROCESSING
Where can Micromass help?
Often drug metabolites can be predicted by a knowledge of the parent drug structure and of metabolic pathways. If using manual processing this would mean plotting an extracted mass chromatogram for each expected metabolite, integrating those chromatograms and then producing background-subtracted spectra for each detected peak. It is also necessary to check a control or blank sample (matrix with no added drug) analysis to ensure that none of the detected peaks are due to compounds found it the matrix. To do this manually for a large number of compounds is repetitive and time-consuming.
Not all metabolites can be predicted and it is often necessary to search through a data file looking for low level metabolites which are not visible in the TIC as they are hidden by more abundant matrix peaks. This means it is necessary to go through a data file scan-by-scan or chromatogram-by-chromatogram to find these metabolites. This can obviously take many hours.
In addition, once in development, it is necessary to generate product ion spectra to confirm or to identify a metabolite. Setting up and processing MS/MS data is also time-consuming.
Automating these steps can reduce the time taken from days to hours or from hours to minutes.

4. Acquire control & metabolized samples Search expected & unexpected
MetaboLynx Flow Path
LC/MS
Report LC/MS results
Acquire control & metabolized samples
Create MS/MS method
MetaboLynx
Auto-start MS/MS
Search expected & unexpected
Process MS/MS data
Append to report
Control comparison

5. 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

6. MetaboLynx Set-up - Peak Detection

7. MetaboLynx Set-up - Expected Metabolites

8. 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

9. 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

10. MetaboLynx Set-up - Isotope Clusters

11. Isotope Cluster Plot (Br)

12. Data Visualization - MetaboLynx Browser

13. 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

14. MetaboLynx Browser - MS/MS

15. 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)

16. 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