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

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 tox...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 2023-08, Vol.95 (33), p.12565-12571
Main Authors: Tsiara, Ioanna, Riemer, Amelie, Correia, Mario S. P., Rodriguez-Mateos, Ana, Globisch, Daniel
Format: Article
Language:English
Subjects:
Analytical chemistry
Arylsulfatase
Beads
Biological properties
Biological samples
Biomarkers
Chemical compounds
Diet
Enzymatic activity
Enzymes
Immobilized enzymes
Metabolism
Metabolites
Metabolomics
Microbiomes
Spectrometry
Sulfation
Toxicology
Xenobiotics
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Summary: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.
ISSN:0003-2700
1520-6882
1520-6882
DOI:10.1021/acs.analchem.3c02988