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Urease-immobilized magnetic microparticles in urine sample preparation for metabolomic analysis by gas chromatography-mass spectrometry

J. Jáčová, M. Jořenek, K. Pospíšková, L. Najdekr, L. Zajoncová, D. Friedecký, T. Adam,

. 2019 ; 1605 (-) : 360355. [pub] 20190710

Language English Country Netherlands

Document type Journal Article

Persistent link   https://www.medvik.cz/link/bmc20006139

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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$a Jáčová, Jaroslava $u Laboratory of Metabolomics, Institute of Molecular and Translational Medicine, Faculty of Medicine and Dentistry, Palacký University Olomouc, Hněvotínská 5, 779 00, Olomouc, Czech Republic; Laboratory for Inherited Metabolic Disorders, Department of Clinical Biochemistry, University Hospital Olomouc, I.P. Pavlova 6, 775 20, Olomouc, Czech Republic. Electronic address: jaroslava.jacova@gmail.com.
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$a Urease-immobilized magnetic microparticles in urine sample preparation for metabolomic analysis by gas chromatography-mass spectrometry / $c J. Jáčová, M. Jořenek, K. Pospíšková, L. Najdekr, L. Zajoncová, D. Friedecký, T. Adam,
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$a 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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$a Jořenek, Miroslav $u Department of Biochemistry, Faculty of Science, Palacký University Olomouc, Šlechtitelů 27, 783 71, Olomouc, Czech Republic. Electronic address: MiraCRXJorenek@seznam.cz.
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$a Pospíšková, Kristýna $u Regional Center of Advanced Technologies and Materials, Palacký University Olomouc, Šlechtitelů 27, 783 71, Olomouc, Czech Republic. Electronic address: kristyna.pospiskova@upol.cz.
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$a Najdekr, Lukáš $u Laboratory of Metabolomics, Institute of Molecular and Translational Medicine, Faculty of Medicine and Dentistry, Palacký University Olomouc, Hněvotínská 5, 779 00, Olomouc, Czech Republic; Laboratory for Inherited Metabolic Disorders, Department of Clinical Biochemistry, University Hospital Olomouc, I.P. Pavlova 6, 775 20, Olomouc, Czech Republic. Electronic address: lukas.najdekr@gmail.com.
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$a Zajoncová, Ludmila $u Department of Biochemistry, Faculty of Science, Palacký University Olomouc, Šlechtitelů 27, 783 71, Olomouc, Czech Republic. Electronic address: ludmila.zajoncova@upol.cz.
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$a Friedecký, David $u Laboratory of Metabolomics, Institute of Molecular and Translational Medicine, Faculty of Medicine and Dentistry, Palacký University Olomouc, Hněvotínská 5, 779 00, Olomouc, Czech Republic; Laboratory for Inherited Metabolic Disorders, Department of Clinical Biochemistry, University Hospital Olomouc, I.P. Pavlova 6, 775 20, Olomouc, Czech Republic. Electronic address: david.friedecky@upol.cz.
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$a Adam, Tomáš $u Laboratory of Metabolomics, Institute of Molecular and Translational Medicine, Faculty of Medicine and Dentistry, Palacký University Olomouc, Hněvotínská 5, 779 00, Olomouc, Czech Republic; Laboratory for Inherited Metabolic Disorders, Department of Clinical Biochemistry, University Hospital Olomouc, I.P. Pavlova 6, 775 20, Olomouc, Czech Republic. Electronic address: tomasadam@gmail.com.
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