Untargeted Metabolomics Workflow

Untargeted metabolomics workflow

Identify putative biomarkers, understand system-wide metabolism, and detect unknown metabolites in routine applications. To ensure comprehensive metabolome coverage, our separation technologies and high resolution-accurate mass (HRAM) Thermo Scientific Orbitrap mass spectrometers provide reproducibility, sensitivity, and the ability to measure a broad range of metabolites, from very hydrophilic to hydrophobic.

 

Enhance your discovery analysis with the Thermo Scientific AcquireX Intelligent Data Acquisition, which optimizes MS2 interrogation of relevant compounds for greater coverage and more confident annotation than traditional data-dependent analysis. Our specially designed metabolomics software transforms large datasets into meaningful insight by efficiently analyzing rich HRAM Orbitrap mass spectrometry data.

What you get from our untargeted metabolomics platform

When combined with advanced separation techniques, industry-leading Orbitrap mass spectrometers deliver high-throughput and quantitative capabilities, expanding the scope of understanding of metabolites involved in cell metabolism and biological pathways and in putative biomarker discovery.

Confident identification and quantitation

With resolutions up to 1 million, Orbitrap mass analyzers enhance identification and quantitation confidence through high specificity, enabling precise mass assignments, particularly for metabolites with similar masses and in isotope-labeling studies.

The very high resolving power of Orbitrap mass analyzers resolves isobaric metabolites.

Sensitivity

Metabolomics, a quantitative science, requires detection of vastly differing concentrations of endogenous metabolites for a comprehensive understanding of their biological states. Orbitrap mass spectrometers enable quantitation down to low-femtomole concentrations, providing up to five orders of linear dynamic range with precise coefficients of variation (CV).

Sensitivity of Orbitrap mass analyzer: (A) Quantitation of citric acid was achieved at low-femtomole sensitivity using full scan MS. (B) Shows the amount of endogenous citric acid obtained from each plate containing ~3 × 106 cells.

Reproducibility

Thanks to its sensitivity and selectivity, Orbitrap mass spectrometry has become the preferred method for metabolomics research. To derive meaningful insights from the extensive datasets typical in metabolomics studies, Orbitrap mass spectrometers deliver accurate and reproducible data from run to run, eliminating the need for internal calibration.

Full scan MS R = 70,000; Peak width = 3.6 s, mean 14.5 scans; D3-hippuronic acid, theoretical.

Throughput

Enhanced coverage is achieved by utilizing both positive and negative ionization modes on the mass spectrometer. For higher throughput and more efficient and productive analysis, polarity switching can be performed in a single run on Orbitrap mass spectrometers.

Base peak chromatograms of positive and negative ion modes of bovine heart lipid extract (1 μg on column) in a single run.

Untargeted metabolomics workflow

Comprehensive coverage of the metabolome enabled by orthogonal separations and deep analysis with Orbitrap mass spectrometers

 

The Thermo Scientific untargeted metabolomics workflow enables you to:

  • Annotate and identify biomarkers
  • Understand cellular metabolism
  • Identify trends between samples and populations
  • Overcome the technical limitations associated with studying the most challenging samples

Our separation technologies and high-resolution, accurate-mass (HRAM) Orbitrap mass spectrometers are proven to be highly reproducible, sensitive, and capable of measuring a wide range of metabolites, from very hydrophilic to hydrophobic, for comprehensive coverage. With specially designed metabolomics software, you can examine the rich HRAM Orbitrap mass spectrometry data more efficiently and convert these large data sets into meaningful insights.

 

Acquisition and interpretation of mass spectra to characterize and identify unknowns is a formidable task in untargeted metabolomics. Sample complexities, background interferences, compound structural diversity, and lack of robust data processing tools can make it difficult to set up analytical methods and interpret data. Overcoming these bottlenecks requires advances in both data acquisition and computational tools. Extract meaningful data faster and with higher confidence with the novel, automated AcquireX Intelligent Data Acquisition Workflow that includes various characterization capabilities using intelligent MSn and a multitude of mass spectral data-processing tools along with the Orbitrap mass spectrometer.

What customers are saying about Orbitrap IQ-X Tribrid Mass Spectrometer

“Progress has been made to putatively annotate metabolites using MS or MS/MS data using established UHPLC/MS assays through the metabolomics database and mass spectral library searching workflows.

 

The Orbitrap Tribrid MS delivers complementary information with multiple dissociation techniques and robust MSn data required to deduce structural information and increase confidence in metabolite annotation. With built-in intelligent data acquisition, AcquireX, we now collect more informative data, and not just more data, we can maximize metabolome coverage and increase confidence in the identification on unknowns applying intelligent-DDA approaches.”

 

Warwick Dunn, PhD, Professor in Analytical and Clinical Metabolomics, Phenome Centre Birmingham, School of Biosciences and Institute of Metabolism and Systems Research, University of Birmingham, UK

What customers are saying about the Orbitrap Astral Mass Spectrometer

“While intelligent DDA and rolling exclusion lists have been extremely helpful for common metabolites, the approach presents challenges for rare chemicals that only occur in a subset of samples. The capability to generate MS/MS data for nearly all compounds in a single, short run on an Orbitrap Astral MS is a huge step forward in metabolomics. It will be particularly important for characterizing exposure chemicals, such as drugs and certain food components.”

 

Dr Gary J. Patti,  PhD, Professor of Chemistry, Michael and Tana Powell, Washington University School of Medicine in St. Louis

What customers are saying about the Orbitrap Excedion Pro Mass Spectrometer

“Mass spectrometers with extended dynamic range offer a significant advantage by enabling the simultaneous analysis of both trace-level and highly-abundant analytes without signal saturation or the loss of low-intensity peaks. This perspective will benefit the analysis of biological, environmental, and industrial samples with a broad range of analyte concentrations. By improving isotopic and adduct distribution analysis and enhancing detection limits, such advanced mass spectrometers can significantly improve quantitative accuracy, making them indispensable for high-precision analytical applications.”

 

Oliver Fiehn, PhD, Director, West Coast Metabolomics Center Davis Genome Center

For Research Use Only. Not for use in diagnostic procedures.