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Related Concept Videos

High-Resolution Mass Spectrometry (HRMS)01:15

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The resolution of a mass spectrometer depends on the efficiency of separating ions with different ion masses. The mass of an atom is approximated to the sum of the masses of protons and neutrons inside, considering the masses of protons and neutrons as equal. However, the masses of the proton (1.6726 × 10−24 g) and neutron (1.6749 × 10−24 g) are not truly equal. There is a minor error in the expression of atomic masses relative to the simplest atom of hydrogen. For...
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Mass spectrometry is an important technique for the identification of pure compounds. However, it has some limitations for the analysis of complex mixtures, often due to excessive fragmentation making the spectrum too complicated to decipher. Mass spectrometry can be combined with suitable separation methods in sequence, forming hyphenated methods, which are useful in the analysis of complex mixtures.
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Updated: Apr 20, 2026

Untargeted Metabolomics from Biological Sources Using Ultraperformance Liquid Chromatography-High Resolution Mass Spectrometry UPLC-HRMS
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Practical considerations for optimizing untargeted metabolomics with Synapt XS HRMS: A tutorial.

Lars Kaiser1, Oliver Riester1, Hans-Peter Deigner2

  • 1Institute of Precision Medicine, Furtwangen University, Jakob-Kienzle-Strasse 17, Villingen-Schwenningen, 78054, Germany.

Journal of Chromatography. A
|April 18, 2026
PubMed
Summary
This summary is machine-generated.

This tutorial details untargeted metabolomics using Synapt XS HRMS, focusing on optimizing settings for labile metabolites and transferable protocols for diverse biological samples. It ensures sensitive and reproducible analyses across various mass spectrometers.

Keywords:
High-resolution mass spectrometryIon heatingLC-MS protocolsSynapt XSTraveling wave ion mobilityUntargeted metabolomics

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Area of Science:

  • Metabolomics
  • Analytical Chemistry
  • Biochemistry

Background:

  • Metabolomics is crucial for understanding organismal metabolism.
  • Liquid chromatography-high resolution mass spectrometry (LC-HRMS) is a dominant technique for global metabolomics.
  • Transferring protocols between different LC-HRMS instruments is challenging due to hardware and ion mobility variations.

Purpose of the Study:

  • To provide a tutorial for untargeted metabolomics using the Synapt XS HRMS.
  • To address challenges in transferring protocols, especially for labile polar metabolites affected by traveling wave ion mobility spectrometry (TWIMS).
  • To offer broadly applicable methods for sample preparation, chromatographic separation, data analysis, and quality control.

Main Methods:

  • Step-by-step protocols for sample preparation and chromatographic separation of polar and non-polar compounds.
  • Adjustment of TWIMS parameters to mitigate ion heating and fragmentation of labile metabolites.
  • Utilizing Progenesis QI for data analysis, including spectral library creation and fragment database integration.

Main Results:

  • The Synapt XS HRMS enables sensitive and reproducible untargeted metabolomics across diverse biological matrices.
  • Optimized TWIMS parameters improve the analysis of labile polar metabolites.
  • Developed protocols are largely transferable to other high-performance mass spectrometers.

Conclusions:

  • The tutorial provides a framework for sensitive and reproducible metabolomics analyses.
  • Methods are broadly applicable to modern mass spectrometers from various manufacturers.
  • Specific focus on Synapt XS HRMS settings enhances analysis of labile metabolites.