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©2018 Waters Corporation 1 COMPANY CONFIDENTIAL
Metabolite Identification
Webinar Content Needs
Presented by:
Steve Preece
Eric van Beelen
Diana Perrier
©2018 Waters Corporation 2 COMPANY CONFIDENTIAL
That’s the benefit of participating in the Live Session!
– Post your questions into the Q&A facility on the left.
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©2018 Waters Corporation 3 COMPANY CONFIDENTIAL
Metabolite Identification
Webinar Content Needs
Presented by:
Steve Preece
Eric van Beelen
Diana Perrier
©2018 Waters Corporation 4 COMPANY CONFIDENTIAL
Ion Mobility – the Basics (Steve Preece)
– What is IMS and how does it work
– Collision Cross Section (CCS)
Metabolite Identification (Eric van Beelen)
– Challenges in Metabolite Identification
– How Ion Mobility addresses these challenges
– Advanced Data Mining options
Q&A
Outline
©2018 Waters Corporation 5 COMPANY CONFIDENTIAL
Ion Mobility
RESISTANCE Determined by size (mass) and shape
DRIVING FORCE Determined by mass and charge
N2
Drift Tube
Electric field
IONS GAS
Ions separated based on their mass, charge and shape
©2018 Waters Corporation 6 COMPANY CONFIDENTIAL
LC-IMS-MS
©2018 Waters Corporation 7 COMPANY CONFIDENTIAL
IMS
-9 -8 -7 -6
TOF MS LC
-5 -4 -3 -2 -1 0 1 2 3 4 n
10n seconds
LC-IMS-MS
+ +
-
-
+ GAS
Field
ION
SOURCE
COLLISION
CELL
CHROMATOGRAPHIC
SEPARATION
ION MOBILITY
SEPARATION
TOF MASS
SEPARATION
©2018 Waters Corporation 8 COMPANY CONFIDENTIAL
LC-IM-MS
LC
time
MS
IMS
drift time drift time drift time
©2018 Waters Corporation 9 COMPANY CONFIDENTIAL
Data Independent Acquisitions
V
V
L
H
Precursor Ions (MS)
Fragment Ions (‘MS/MS’)
MSE
©2018 Waters Corporation 10 COMPANY CONFIDENTIAL
Single sample injection
Alternating low and high energy spectra
– Precursor and fragment ions
– MW information
– Structural information
– Confirmatory ions
All of the data all of the time
– No data dependent switching where information can be missed
MSE
Data Independent Acquisitions
precursors
fragments
©2018 Waters Corporation 11 COMPANY CONFIDENTIAL
C + B + A
Low energy
High energy
Precursors
Fragments
RT
CCS
Clarity
HDMSE
DIA With ION MOBILITY
©2018 Waters Corporation 12 COMPANY CONFIDENTIAL
Verapamil in PPT Plasma MSE
617.3076
©2018 Waters Corporation 13 COMPANY CONFIDENTIAL
Verapamil in PPT Plasma HDMSE
Verapamil
518.2235
441.2758
289.1923
Unlimited Product ion
HDMSE
617.3076
©2018 Waters Corporation 14 COMPANY CONFIDENTIAL
IMS drift time CCS
Additional identification point
Matrix independent
System independent
The ability to see more
Resolution of isomers
Cleaner spectra
IMS
Orthogonal Separation Collision Cross Section
©2018 Waters Corporation 15 COMPANY CONFIDENTIAL
Differentiating characteristic of an ion
—Chemical structure
(mass, size)
—Dimensional information
(shape)
Physicochemical property of an ion
Aiding confirmation/identification
What is CCS?
©2018 Waters Corporation 16 COMPANY CONFIDENTIAL
Ion Mobility – the Basics (Steve Preece)
– What is IMS and how does it work
– Collision Cross Section (CCS)
Metabolite Identification (Eric van Beelen)
– Challenges in Metabolite Identification
– How Ion Mobility addresses these challenges
– Advanced Data Mining options
Q&A
Outline
©2018 Waters Corporation 17 COMPANY CONFIDENTIAL
Metabolite Identification in Drug Development
--
Metabolic Stability
--
Metabolic Biological Profiling
--
Metabolic Quantification
Activity Study metabolites (new analogues)
Metabolite Tox Study
Metabolite IP -- --
--
Market
©2018 Waters Corporation 18 COMPANY CONFIDENTIAL
Metabolite Identification Challenges - Technical
©2018 Waters Corporation 19 COMPANY CONFIDENTIAL
Metabolite Identification Challenges - Workflow
https://www.bioanalysis-zone.com/2018/07/30/in-the-zone-biotransformation_inz_biot_waters/
©2018 Waters Corporation 20 COMPANY CONFIDENTIAL
Make
Test Analyze
Design
The Discovery LC-MS Workflow for Metabolite Identification
©2018 Waters Corporation 21 COMPANY CONFIDENTIAL
Ion Mobility for Spectral Clarity in Metabolite Identification
Kirk, Jayne, Russell Mortishire-Smith and Mark Wrona. Integrating Ion Mobility into Routine Metabolite Identification
Studies using the Vion IMS QTof Mass Spectrometer. Waters Application Note 720006121EN. 2017.
©2018 Waters Corporation 22 COMPANY CONFIDENTIAL
Ion Mobility to Resolve Co-Eluting Metabolites
Kirk, Jayne, Russell Mortishire-Smith
and Mark Wrona. Integrating Ion
Mobility into Routine Metabolite
Identification Studies using the Vion
IMS QTof Mass Spectrometer. Waters
Application Note 720006121EN. 2017.
©2018 Waters Corporation 23 COMPANY CONFIDENTIAL
Ion Mobility to Resolve Co-Eluting Metabolites
Kirk, Jayne, Russell Mortishire-Smith and Mark Wrona. Integrating Ion Mobility into Routine Metabolite Identification
Studies using the Vion IMS QTof Mass Spectrometer. Waters Application Note 720006121EN. 2017.
©2018 Waters Corporation 24 COMPANY CONFIDENTIAL
Ion Mobility for Condition Independent Metabolite Tracking
Kirk, Jayne, Russell Mortishire-Smith and Mark Wrona. Integrating Ion Mobility into Routine Metabolite Identification
Studies using the Vion IMS QTof Mass Spectrometer. Waters Application Note 720006121EN. 2017.
©2018 Waters Corporation 25 COMPANY CONFIDENTIAL
UNIFI Metabolite Identification Application
Small Molecule Example
©2018 Waters Corporation 26 COMPANY CONFIDENTIAL
Biotransformation Localization Step
Wrona, Mark, Sherri Naughton, Yun Alelyunas, Russell Mortishire-Smith and Jayne Kirk. Metabolite Identification:
Working with Large Preclinical Multispecies Data Sets. Waters Application Note 720006106EN. 2017.
©2018 Waters Corporation 27 COMPANY CONFIDENTIAL
Trendplot Step
Wrona, Mark, Sherri Naughton, Yun Alelyunas, Russell Mortishire-Smith and Jayne Kirk. Metabolite Identification:
Working with Large Preclinical Multispecies Data Sets. Waters Application Note 720006106EN. 2017.
©2018 Waters Corporation 28 COMPANY CONFIDENTIAL
Reporting
Wrona, Mark, Sherri Naughton, Yun Alelyunas, Russell Mortishire-Smith and Jayne Kirk. Metabolite Identification:
Working with Large Preclinical Multispecies Data Sets. Waters Application Note 720006106EN. 2017.
©2018 Waters Corporation 29 COMPANY CONFIDENTIAL
UNIFI with VION IMS QTof is a great tool for identifying the metabolic pathways of your drug.
Ion Mobility data is beneficial to the metabolic profiling
– Cleaner spectra
– CCS values
The powerful reporting features of the UNIFI Scientific Information System allow easy and
comprehensive presentation of your results
The compliant ready software (UNIFI) ensures compliance accross different stages of the drug
development process
Possible to work with third party applications which have the potential to add significant value to the
native capabilities of the UNIFI Scientific Information System.
UNIFI SCIENTIFIC INFORMATION SYSTEM
©2018 Waters Corporation 30 COMPANY CONFIDENTIAL
Technical superiority of Vion and IMS acquired data, now combined with the gold
standard for third party data processing software
UNIFI API with Mass-Metasite and WebMetabase
©2018 Waters Corporation 31 COMPANY CONFIDENTIAL
The Discovery LC/MS Workflow for Metabolite Identification
Make
Test Analyze
Design
©2018 Waters Corporation 32 COMPANY CONFIDENTIAL
What are Mass-MetaSite and WebMetabase?
Mass-MetaSite
– engine that processes LC-MS data to identify
drug metabolites in biological samples
WebMetabase
– processes MMS data from multiple compounds
and conditions for databasing and visualization
Together, MMS and WMB capture all in vitro and in vivo biotransformations into a
database, enabling data sharing, mining and new chemistry design.
©2018 Waters Corporation 33 COMPANY CONFIDENTIAL
The Vion/UNIFI/Mass-MetaSite and WebMetabase Workflow
©2018 Waters Corporation 34 COMPANY CONFIDENTIAL
Increased Confidence with Improved Data Quality
J Kirk, I Zamora, A Riera, T Radchenko, A Escola, Y Alelyunas, R Mortishire-Smith and M Wrona. Characterising
the Catabolism of Peptides Using Ion Mobility Enabled High Resolution Mass Spectrometry with Mass-MetaSite
Integration for Data Processing. Poster presented at IMSC, 2018.
©2018 Waters Corporation 35 COMPANY CONFIDENTIAL
UNIFI API with Mass-MetaSite and WebMetabase
Peptide Example
J Kirk, I Zamora, A Riera, T Radchenko, A Escola, Y Alelyunas, R Mortishire-Smith and M Wrona. Characterising
the Catabolism of Peptides Using Ion Mobility Enabled High Resolution Mass Spectrometry with Mass-MetaSite
Integration for Data Processing. Poster presented at IMSC, 2018.
©2018 Waters Corporation 36 COMPANY CONFIDENTIAL
UNIFI workflows for regulated, compliant,
network-based vial to file environments.
Mass-MetaSite and WebMetabase for high
throughput, batch processing, cloud-based
flexible environments.
Waters Met ID Solutions for Both Development and Discovery
©2018 Waters Corporation 37 COMPANY CONFIDENTIAL
What Does This All Mean for the DMPK Lab?
Time Automation Capacity
• Better spectral quality by IMS means increased confidence in identification, structural elucidation, distinguishing co-eluting metabolites
• CCS values means another separation dimension acquired at the same time that is matrix and condition independent
• Integration of ion mobility data with software means a less manual workflow, allowing scientists to work on value added projects
©2018 Waters Corporation 38 COMPANY CONFIDENTIAL
Upcoming Events Where You Can Learn More
Russell Mortishire-Smith, PhD (Waters)
Technological advancements in
screening platforms for metabolite data
access and evaluation
Richard Clayton, PhD (Covance)
The Application of Ion Mobility Mass
Spectrometry in DMPK Workflows
©2018 Waters Corporation 39 COMPANY CONFIDENTIAL
Additional Resources
dmpk.waters.com
Posters
Application Notes Tech Briefs
Presentations
©2018 Waters Corporation 40 COMPANY CONFIDENTIAL
Thank you for your attention! Q&A
That’s the benefit of participating in the Live Session!
– Post your questions into the Q&A facility on the left.
– Refresh the Q&A window from time to time to view what it may contains.
– We will answer your questions live.
Eric van Beelen
Steve Preece
