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Analysis of nucleotide pools in bacteria using HPLC-MS in HILIC mode
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SYSNO ASEP 0507420 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Analysis of nucleotide pools in bacteria using HPLC-MS in HILIC mode Author(s) Zborníková, Eva (UOCHB-X) RID, ORCID
Knejzlík, Zdeněk (UOCHB-X)
Hauryliuk, V. (EE)
Krásný, Libor (MBU-M) RID, ORCID
Rejman, Dominik (UOCHB-X) RID, ORCIDArticle number 120161 Source Title Talanta. - : Elsevier - ISSN 0039-9140
Roč. 205, Dec 1 (2019)Number of pages 10 s. Language eng - English Country NL - Netherlands Keywords nucleotide ; HPLC-MS ; HILIC ; ppGpp ; stringent response ; Escherichia coli Subject RIV CB - Analytical Chemistry, Separation OECD category Analytical chemistry Subject RIV - cooperation Institute of Microbiology - Microbiology, Virology R&D Projects NV17-29680A GA MZd - Ministry of Health (MZ) GA15-11711S GA ČR - Czech Science Foundation (CSF) 8F19006 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) Method of publishing Open access Institutional support UOCHB-X - RVO:61388963 ; MBU-M - RVO:61388971 UT WOS 000485856500085 EID SCOPUS 85069720759 DOI 10.1016/j.talanta.2019.120161 Annotation Nucleotides, nucleosides and their derivatives are present in all cells at varying concentrations that change with the nutritional, and energetic status of the cell. Precise measurement of the concentrations of these molecules is instrumental for understanding their regulatory effects. Such measurement is challenging due to the inherent instability of these molecules and, despite many decades of research, the reported values differ widely. Here, we present a comprehensive and easy-to-use approach for determination of the intracellular concentrations of >25 target molecular species. The approach uses rapid filtration and cold acidic extraction followed by high performance liquid chromatography (HPLC) in the hydrophilic interaction liquid chromatography (HILIC) mode using zwitterionic columns coupled with UV and MS detectors. The method reliably detects and quantifies all the analytes expected to be observed in the bacterial cell and paves the way for future studies correlating their concentrations with biological effects. Workplace Institute of Organic Chemistry and Biochemistry Contact asep@uochb.cas.cz ; Kateřina Šperková, Tel.: 232 002 584 ; Jana Procházková, Tel.: 220 183 418 Year of Publishing 2020 Electronic address https://www.sciencedirect.com/science/article/pii/S0039914019307878?via%3Dihub
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