Analysis of nucleotide pools in bacteria using HPLC-MS in HILIC mode

0507420 - ÚOCHB 2020 RIV NL eng J - Článek v odborném periodiku
Zborníková, Eva - Knejzlík, Zdeněk - Hauryliuk, V. - Krásný, Libor - Rejman, Dominik
Analysis of nucleotide pools in bacteria using HPLC-MS in HILIC mode.
Talanta. Roč. 205, Dec 1 (2019), č. článku 120161. ISSN 0039-9140. E-ISSN 1873-3573
Grant CEP: GA MZd(CZ) NV17-29680A; GA ČR GA15-11711S; GA MŠMT(CZ) 8F19006
Institucionální podpora: RVO:61388963 ; RVO:61388971
Klíčová slova: nucleotide * HPLC-MS * HILIC * ppGpp * stringent response * Escherichia coli
Obor OECD: Analytical chemistry; Microbiology (MBU-M)
Impakt faktor: 5.339, rok: 2019
Způsob publikování: Open access
https://www.sciencedirect.com/science/article/pii/S0039914019307878?via%3Dihub

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.
Trvalý link: http://hdl.handle.net/11104/0298412