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Related Concept Videos

Mass Spectrometry: Complex Analysis01:21

Mass Spectrometry: Complex Analysis

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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.
GC–MS is a powerful hyphenated method commonly used in forensics and environmental...
922

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Proteomic Profile of EPS-Urine through FASP Digestion and Data-Independent Analysis
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Profiling Urinary Sulfate Metabolites With Mass Spectrometry.

Christopher C J Fitzgerald1, Rikard Hedman1, Dimanthi R Uduwela1

  • 1Research School of Chemistry, Australian National University, Acton, ACT, Australia.

Frontiers in Molecular Biosciences
|March 14, 2022
PubMed
Summary
This summary is machine-generated.

This study introduces a new workflow for analyzing urinary sulfated metabolites using UHPLC-HRMS/MS. The method identified novel steroid metabolites in horses and assessed sulfatase enzyme activity in human urine.

Keywords:
anti-dopingmass spectrometrymetabolomicssteroidsulfatasesulfate estersulfation

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Area of Science:

  • Metabolomics
  • Analytical Chemistry
  • Biochemistry

Background:

  • Urinary phase II sulfate metabolites are crucial for understanding biological compound metabolism.
  • Existing methods for profiling sulfated metabolites can be limited.
  • A need exists for rapid, untargeted analysis of these compounds.

Purpose of the Study:

  • To develop and validate a novel workflow for untargeted metabolic profiling of urinary sulfated metabolites.
  • To apply the workflow to identify equine steroid metabolites and assess sulfatase enzyme activity.

Main Methods:

  • Ultra-high-performance liquid chromatography-high resolution tandem mass spectrometry (UHPLC-HRMS/MS) with data-dependent acquisition (DDA).
  • Automated script-based data processing pipeline with k-means clustering for sulfate identification.
  • Application to equine urine post-testosterone propionate administration and human urine with different sulfatase enzymes.

Main Results:

  • Detected ten elevated sulfated steroid metabolites in horse urine after testosterone propionate administration.
  • Identified a novel equine metabolite: 5α-androstane-3β,17α-diol 3-sulfate.
  • Demonstrated higher selectivity of Pseudomonas aeruginosa arylsulfatases (PaS) for human urinary sulfated metabolites compared to Helix pomatia arylsulfatase (HpS).

Conclusions:

  • The novel UHPLC-HRMS/MS workflow enables rapid, systematic, and untargeted metabolic profiling of urinary sulfated metabolites.
  • The method successfully identified new equine steroid metabolites and differentiated sulfatase enzyme activities.
  • This approach offers a valuable tool for metabolomic research involving sulfated compounds in biological matrices.