Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
SHR Neuro Krebs Kardio Lipid Stoffw Microb
Schittmayer, M; Birner-Gruenberger, R; Zamboni, N.
Quantification of cellular folate species by LC-MS after stabilization by derivatization.
Anal Chem. 2018;
Doi: 10.1021/acs.analchem.8b00650
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- Führende Autor*innen der Med Uni Graz
- Schittmayer-Schantl Matthias
- Co-Autor*innen der Med Uni Graz
- Birner-Grünberger Ruth
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- Abstract:
- Folate cofactors play a key role in one-carbon metabolism. Analysis of individual folate species is hampered by the low chemical stability and high inter-convertibility of folates which can lead to severe experimental bias. Here we present a complete workflow that employs simultaneous extraction and stabilization of folates by derivatization. We perform reductive methylation employing stable isotope labelled reagents to retain information on position and redox state of one carbon units as well as the redox state of the pteridine ring. The derivatives are analyzed by a targeted LC(HILIC)-MS/MS method without the need for deconjugation, thereby also preserving the glutamation state of folates. The presented method does not only improve analyte coverage and sensitivity as compared to other published methods, it also greatly simplifies sample handling and storage. Finally, we report differences in the response of bacterial and mammalian systems to pharmacological inhibition of dihydrofolate reductase.