Urease-immobilized magnetic microparticles in urine sample preparation for metabolomic analysis by gas chromatography-mass spectrometry
Jazyk angličtina Země Nizozemsko Médium print-electronic
Typ dokumentu časopisecké články
PubMed
31315811
DOI
10.1016/j.chroma.2019.07.009
PII: S0021-9673(19)30677-6
Knihovny.cz E-zdroje
- Klíčová slova
- GC–MS, Immobilized urease, Metabolomics, Urine sample preparation,
- MeSH
- analýza hlavních komponent MeSH
- chromatografie kapalinová metody MeSH
- enzymy imobilizované moč MeSH
- glutaryl-CoA-dehydrogenasa nedostatek metabolismus MeSH
- lidé MeSH
- magnetické jevy * MeSH
- metabolické nemoci mozku metabolismus MeSH
- metabolom MeSH
- metabolomika metody MeSH
- metody pro přípravu analytických vzorků * MeSH
- močovina metabolismus MeSH
- plynová chromatografie s hmotnostně spektrometrickou detekcí metody MeSH
- reprodukovatelnost výsledků MeSH
- studie proveditelnosti MeSH
- tandemová hmotnostní spektrometrie MeSH
- ureasa moč MeSH
- vrozené poruchy metabolismu aminokyselin metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- enzymy imobilizované MeSH
- glutaryl-CoA-dehydrogenasa MeSH
- močovina MeSH
- ureasa MeSH
Urea, as an end product of protein metabolism and an abundant polar compound, significantly complicates the metabolomic analysis of urine by GC-MS. We developed a sample preparation method removing urea from urine samples prior the GC-MS analysis. The method based on urease immobilized on magnetic microparticles was compared with the others that are conventionally used (liquid-liquid extraction, free urease protocol), and samples without any treatment. To study the impact of sample preparation approaches on the quality of analytical data, we employed comprehensive metabolomic analysis (using both GC-MS and LC-MS/MS platforms) of standard material based on human urine. Multivariate statistical analysis has shown that immobilized urease treatment provides similar results to a free urease approach. However, significant alterations in the profiles of metabolites were observed in the samples without any treatment and after the extraction. Compared to other approaches that were tested, the immobilization of urease on microparticles reduces both the number of artifacts and the variability of the metabolites (average CV of extraction 19.7%, no treatment 11.4%, free urease 5.0%, and immobilized urease 2.5%). The method that was developed was applied in a GC-MS metabolomic experiment of glutaric aciduria type I, where both known diagnostically important biomarkers and unknowns, as the most discriminating compounds, were found.
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