• Je něco špatně v tomto záznamu ?

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

Jazyk angličtina Země Nizozemsko

Typ dokumentu časopisecké články

Perzistentní odkaz   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.

Citace poskytuje Crossref.org

000      
00000naa a2200000 a 4500
001      
bmc20006139
003      
CZ-PrNML
005      
20200518132349.0
007      
ta
008      
200511s2019 ne f 000 0|eng||
009      
AR
024    7_
$a 10.1016/j.chroma.2019.07.009 $2 doi
035    __
$a (PubMed)31315811
040    __
$a ABA008 $b cze $d ABA008 $e AACR2
041    0_
$a eng
044    __
$a ne
100    1_
$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.
245    10
$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,
520    9_
$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.
650    _2
$a vrozené poruchy metabolismu aminokyselin $x metabolismus $7 D000592
650    12
$a metody pro přípravu analytických vzorků $7 D053000
650    _2
$a metabolické nemoci mozku $x metabolismus $7 D001928
650    _2
$a chromatografie kapalinová $x metody $7 D002853
650    _2
$a enzymy imobilizované $x moč $7 D004800
650    _2
$a studie proveditelnosti $7 D005240
650    _2
$a plynová chromatografie s hmotnostně spektrometrickou detekcí $x metody $7 D008401
650    _2
$a glutaryl-CoA-dehydrogenasa $x nedostatek $x metabolismus $7 D050770
650    _2
$a lidé $7 D006801
650    12
$a magnetické jevy $7 D060328
650    _2
$a metabolom $7 D055442
650    _2
$a metabolomika $x metody $7 D055432
650    _2
$a analýza hlavních komponent $7 D025341
650    _2
$a reprodukovatelnost výsledků $7 D015203
650    _2
$a tandemová hmotnostní spektrometrie $7 D053719
650    _2
$a močovina $x metabolismus $7 D014508
650    _2
$a ureasa $x moč $7 D014510
655    _2
$a časopisecké články $7 D016428
700    1_
$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.
700    1_
$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.
700    1_
$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.
700    1_
$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.
700    1_
$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.
700    1_
$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.
773    0_
$w MED00004962 $t Journal of chromatography. A $x 1873-3778 $g Roč. 1605, č. - (2019), s. 360355
856    41
$u https://pubmed.ncbi.nlm.nih.gov/31315811 $y Pubmed
910    __
$a ABA008 $b sig $c sign $y a $z 0
990    __
$a 20200511 $b ABA008
991    __
$a 20200518132348 $b ABA008
999    __
$a ok $b bmc $g 1524997 $s 1096195
BAS    __
$a 3
BAS    __
$a PreBMC
BMC    __
$a 2019 $b 1605 $c - $d 360355 $e 20190710 $i 1873-3778 $m Journal of chromatography. A, Including electrophoresis and other separation methods $n J Chromatogr A $x MED00004962
LZP    __
$a Pubmed-20200511

Najít záznam

Citační ukazatele

Možnosti archivace