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Related Experiment Video

Updated: Apr 12, 2026

Untargeted Metabolomics from Biological Sources Using Ultraperformance Liquid Chromatography-High Resolution Mass Spectrometry UPLC-HRMS
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Solid phase extraction methodology for UPLC-MS based metabolic profiling of urine samples.

Filippos Michopoulos1,2, Helen Gika3, Dimitrios Palachanis3

  • 1Department of Chemistry, Aristotle University of Thessaloniki, Thessaloniki, Greece.

Electrophoresis
|May 13, 2015
PubMed
Summary

Solid-phase extraction (SPE) optimizes sample preparation for untargeted metabolomics. Different SPE phases effectively fractionate, concentrate, and desalt urine samples for enhanced ultra-performance LC-TOF-MS analysis.

Keywords:
Metabolic profilingMetabolomicsSample preparationSolid phase extractionUrine analysis

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Area of Science:

  • Analytical Chemistry
  • Metabolomics
  • Biochemistry

Background:

  • Untargeted metabolic phenotyping (metabolomics/metabonomics) typically minimizes sample preparation.
  • However, desalting, preconcentration, and metabolite fractionation are crucial for specific analyses.
  • Solid-phase extraction (SPE) offers a simple, automatable solution for sample preparation prior to ultra-performance LC-TOF-MS.

Purpose of the Study:

  • To investigate the properties of various SPE phases for extracting metabolites from urine.
  • To evaluate the suitability of different SPE materials for sample fractionation, concentration, and desalting.
  • To determine optimal SPE methods for subsequent HILIC and RPLC analyses.

Main Methods:

  • Investigated a range of SPE phases including alkyl-modified silica (C8, C18-OH, C18) and polymeric materials.
  • Evaluated metabolite extraction efficiency from urine samples using different SPE phases.
  • Assessed the performance of SPE for sample fractionation into polar and nonpolar constituents.

Main Results:

  • The C18 SPE phase effectively fractionated urine into polar and nonpolar fractions suitable for HILIC and RPLC, respectively.
  • Polymeric SPE materials demonstrated superior performance for sample concentration and desalting.
  • SPE phases exhibited distinct capabilities in metabolite extraction and sample manipulation.

Conclusions:

  • SPE is a valuable technique for optimizing urine sample preparation in metabolomics studies.
  • The choice of SPE phase (C18 vs. polymeric) depends on the specific analytical goal: fractionation or concentration/desalting.
  • Optimized SPE protocols enhance the subsequent analysis of polar and nonpolar metabolites by LC-MS.