Immobilized Enzymes on Magnetic Beads for Separate Mass Spectrometric Investigation of Human Phase II Metabolite Classes

Ioanna Tsiara, Amelie Riemer, Mario S P Correia, Ana Rodriguez-Mateos, Daniel Globisch

Research output: Contribution to journalArticlepeer-review

1 Citation (Scopus)

Abstract

The human body has evolved to remove xenobiotics through a multistep clearance process. Non-endogenous metabolites are converted through a series of phase I and different phase II enzymes into compounds with higher hydrophilicity. These compounds are important for diverse research fields such as toxicology, nutrition, biomarker discovery, doping control, and microbiome metabolism. One of the challenges in these research fields has been the investigation of the two major phase II modifications, sulfation and glucuronidation, and the corresponding unconjugated aglycon independently. We have now developed a new methodology utilizing an immobilized arylsulfatase and an immobilized β-glucuronidase to magnetic beads for treatment of human urine samples. The enzyme activities remained the same compared to the enzyme in solution. The separate mass spectrometric investigation of each metabolite class in a single sample was successfully applied to obtain the dietary glucuronidation and sulfation profile of 116 compounds. Our new chemical biology strategy provides a new tool for the investigation of metabolites in biological samples with the potential for broad-scale application in metabolomics, nutrition, and microbiome studies.

Original languageEnglish
Pages (from-to)12565-12571
Number of pages7
JournalAnalytical Chemistry
Volume95
Issue number33
DOIs
Publication statusPublished - 22 Aug 2023

Keywords

  • Humans
  • Enzymes, Immobilized
  • Mass Spectrometry
  • Sulfatases
  • Metabolomics
  • Magnetic Phenomena

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