Presenter: Mark Sartain, Ph.D., Scientist, Agilent
A major challenge facing translational metabolomics is the successful implementation and harmonization of targeted
methodologies to measure large population cohorts and to improve inter-laboratory precision enough to enable
cross-laboratory comparisons of measurements. Come join Agilent and learn about:
A highly curated and proven robust plasma lipidomics LC/TQ method that balances depth of coverage with sample
throughput across large population cohorts
Performance results from an interlaboratory multiday study to assess variance and demonstrate transferability
in method implementation
METLIN: How to Analyze a Million Molecular Standards
Dr. Gary Siuzdak
Sr. Director of Scripps Center for Metabolomics and Professor of Chemistry, Molecular and Computational Biology
Analyzing over one million molecular standards is a challenging endeavor, especially when it needs to be accomplished
at multiple collisional energies and in both positive and negative ionization modes. I will describe some of the details
of how this was performed and why we have pursued not only tandem mass spectrometry (MS2) data acquisition but also a
newly created (and only) neutral loss database that is also embedded within METLIN. The primary reason behind this effort
is to create high confidence molecular identification of known molecules, and preliminary characterization of novel,
unknown molecules (unknowns). METLIN is moving to become a truly comprehensive database with data on ~1% of PubChem’s 93
million compounds, essentially a number that can be characterized as all the currently known chemical space.
Applications of GC×GC are as broad and extensive as for GC, testifying to the interest in evaluating the technique
against one-dimensional methods. Metabolomics should be where GC×GC excels simply because it is—or should be—the
ultimate in untargeted profiling of volatile compounds in samples. As an approach to investigate cancers, COVID-19,
and other disease states, breath analysis has been an emerging, active field, although the studies are still few in
terms of the populations surveyed, typically ranging from 10 to 200 subjects. Here, I will talk about some of those
breath studies and the inherent importance of employing standardized approaches at every stage.
Innovations and Applicationis using Widely-Targeted Metabolomics
Dr. Shih-Chieh Chu
Metware Biotechnology Inc.
The number of metabolites that we can identify today is only a fraction of what is predicted. Metware Biotechnology focuses on
developing innovative metabolomics processes and database to improve metabolite identification and quantification. This talk will
describe Widely-Targeted Metabolomics process with Metware's metabolite database development and how it applies to life science
and biomedical research.
Employing new Technology for Bridging the Gap between Untargeted and Targeted Metabolomics
Dr. Maryam Goudarzi - Sr. Global Market Development & Marketing Manager, Small Molecule Omics
When performing metabolite identification from biological matrices, several strategies can be applied. On HRMS systems,
untargeted analysis is performed using information dependent acquisition/data dependent acquisition (IDA/DDA). While this
approach enables the identification of both expected and unknown metabolites, it suffers from gaps in MS/MS coverage or
poor-quality MS/MS. In previous work, we showed that activation of the Zeno trap increased the MS/MS signal and peak area
for polar metabolite fragments by up to 14x, leading to improved quantification and identification. Here, we will show a
wider array of biological sample types to investigate using the Zeno trap with SWATH acquisition for data independent
acquisition (DIA) to address current challenges in metabolomics analysis.
Quantitative Metabolic Pathway-specific Metabolomics using MRM based Techniques
Dr. Jun Han - Adjunct Assistant Professor, Metabolomics group leader and senior scientist, Genome BC Proteomics Centre, University of Victoria
Precision metabolomics necessitates high-sensitivity and reliable quantitation of endogenous metabolites in biological
samples. Multiple-reaction monitoring mass spectrometry (MRM/MS) in combination with front-end LC separations has been
the gold standard techniques in this regard. However, successful measurements of all known metabolites in specific
metabolic pathways are often complicated by diversified structures of different metabolites and their wide concentration
ranges. With the development and validation of dozens of different yet complementary LC-MRM/MS methods, with or without
pre-analytical chemical derivatization, pathway-specific detection and precise/accurate quantitation of hundreds of
endogenous metabolites involved in major metabolic pathways have been achieved. Examples for high-precision and high-accuracy
metabolite analysis of central carbon metabolism, fatty acid metabolism and cholesterol synthesis/metabolism will be presented.
Intelligence-driven Metabolomics Workflows: Hardware and Software Innovations for Confident Differential Analysis,
Unknown Annotation, and Biomarker Discover
Introduction
The field of Metabolomics has been advancing at an impressive rate, inspiring key analytical innovations designed
to keep pace with the biological needs of a study. Despite these advances, challenges remain around compound annotation
and identification, differential analysis as well biological interpretation. Here we highlight key hardware and software
innovations designed to address these study bottlenecks. The unique fragmentation capabilities available on the Orbitrap
IQ-X ™ Tribrid MS, such as UVPD, HCD, and CID, are used to distinguish isobaric structural isomers in the absence of
chromatographic separation. Improved AcquireX workflows are shown to streamline structural library generation, leading
to information-rich fragmentation of more experimentally relevant compounds, and most importantly, Compound Discoverer™
3.3 software algorithm improvements and enhanced features, efficiently translating raw data from intelligence-driven
data acquisition modes into meaningful results. This updated software version boasts a new peak detection algorithm,
optimized for large data sets, that includes peak quality thresholding to improve detection and relative quantification.
The hardware improvements of HRAM Orbitrap technology and CD 3.3 data processing software innovations provide confident
differential analysis, unknown annotation, and biomarker discovery.
Conclusion
ThermoFisher Scientific is committed to advancing the field of metabolomics through intelligence-driven data acquisition,
streamlined software processing strategies, and key hardware innovations that keep pace and push beyond the current
analytical demands of the field.
Senior Manager, Omics Business Development, Waters Corporation
As a sponsor of this year’s MANA lunch Seminar, Waters will discuss the SELECT SERIES MRTs performance for both imaging
and LC-MS workflows in metabolomics and lipidomics applications. This novel time-of-flight technology combines
unprecedented resolution and mass accuracy with the benefits of TOF, producing consistent mass resolution over a broad
mass range and varying scan speeds.
Distinguishing the fine isotope structure of small molecules and resolving nominally isobaric interferences is now
possible—without compromising on scan speeds compatible with the best chromatography or tissue imaging experiments. The
results are unambiguous compound identification for even the most complex samples.
Translational Mass Spectrometry Imaging: Doctor did you get it all?
Speaker: Martin Kaufmann, PhD
Research Associate, Department of Medicine, Queen's University
Mass spectrometry imaging methods such as DESI (desorption electrospray ionization mass spectrometry) enable metabolomic
profiling of tissue sections that may one day serve as ancillary tools for tissue diagnosis in the pathology lab. Another
technique ‘I-knife’ based on rapid evaporative ionization mass spectrometry (REIMS) has been proposed as an intraoperative
tool to help guide surgeons during tumor removal to reduce the need for re-operation due to remaining cancer cells on the
periphery, known as positive margins. Unlike DESI, the spatial location of mass spectra cannot be determined by I-knife
in its current form, which limits the ability to validate intraoperative data, or in future, inform the surgeon on the
location a potential positive margin so that more tissue can be removed. The presentation will focus on the creation of ‘3D
mass spectrometry imaging’ by combining I-knife with a spatio-temporal navigated cautery, and the application of this
technology in the operating theatre to breast cancer surgery. Implications for real-time intraoperative margin assessment,
and anticipated improvements to the ‘patient journey’ will be discussed.
