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Related Concept Videos

High-Resolution Mass Spectrometry (HRMS)01:15

High-Resolution Mass Spectrometry (HRMS)

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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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Related Experiment Video

Updated: Mar 15, 2026

Untargeted Metabolomics from Biological Sources Using Ultraperformance Liquid Chromatography-High Resolution Mass Spectrometry UPLC-HRMS
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Investigating Variability in Metabolomics: A Comparative Study of Analytical Platforms and Blood Matrices Using

Giulia Guerra1, Alessio Polymeropoulos2, Elisabetta Venturelli3

  • 1Epidemiology and Prevention Unit, Fondazione IRCCS Istituto Nazionale dei Tumori di Milano, 20133 Milan, Italy.

Molecules (Basel, Switzerland)
|March 14, 2026
PubMed
Summary

Standardizing untargeted metabolomics is crucial for large studies. Monophasic extractions with reversed-phase chromatography offer the best reproducibility and metabolite coverage, minimizing variability in clinical research.

Keywords:
blood matrixdata qualityhuman plasmametabolomicsvalidation

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Last Updated: Mar 15, 2026

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Area of Science:

  • Metabolomics
  • Analytical Chemistry
  • Clinical Research

Background:

  • Untargeted metabolomics studies face significant standardization challenges.
  • Variability in sample preparation and analytical workflows impacts data reproducibility.
  • Need for a robust framework for large-scale clinical and epidemiological studies.

Purpose of the Study:

  • To systematically evaluate factors affecting metabolomics reproducibility.
  • To establish a mechanism-informed framework for improving standardization.
  • To identify optimal protocols for large-scale untargeted metabolomics.

Main Methods:

  • Compared three extraction protocols (IPA, MeOH:ACN, Matyash) using pooled plasma.
  • Evaluated four blood matrices (serum, heparin plasma, EDTA, citrate).
  • Analyzed samples using untargeted metabolomics with HILIC and RP-HPLC columns coupled to mass spectrometry.

Main Results:

  • Monophasic extractions (IPA, MeOH:ACN) yielded >80% features with CV% ≤ 30% for both RP and HILIC.
  • RP chromatography detected >80% metabolites with CV% < 30% across matrices.
  • Serum and heparin plasma showed better reproducibility than EDTA and citrate.

Conclusions:

  • Monophasic extractions combined with RP chromatography maximize reproducibility and metabolite coverage.
  • This standardized workflow minimizes methodological artifacts for robust biological discovery.
  • The proposed framework enhances reliability in large-scale untargeted metabolomics studies.