Skip to main content

Lund University Publications

LUND UNIVERSITY LIBRARIES

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

Zborníková, Eva ; Knejzlík, Zdeněk ; Hauryliuk, Vasili LU orcid ; Krásný, Libor and Rejman, Dominik (2019) In Talanta 205. p.1-10
Abstract

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... (More)

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.

(Less)
Please use this url to cite or link to this publication:
author
; ; ; and
publishing date
type
Contribution to journal
publication status
published
keywords
Escherichia coli, HILIC, HPLC-MS, Nucleotide, ppGpp, Stringent response
in
Talanta
volume
205
article number
120161
pages
1 - 10
publisher
Elsevier
external identifiers
  • pmid:31450400
  • scopus:85069720759
ISSN
0039-9140
DOI
10.1016/j.talanta.2019.120161
language
English
LU publication?
no
additional info
Funding Information: This work was supported by Czech ministry of health (Grant No. 17-29680A to D.R. and L.K.) Czech Science Foundation grant number (grant No. 15-11711S to D.R.), EU and Czech ministry of Education and Sport via JPIAMR (grant No. 8F19006 to D.R. and V.H.), Swedish Research council (grant 2017–03783 to VH) and Charles University , Department of Analytical Chemistry – Specific University Research No. SVV260440 to E.Z. Funding Information: This work was supported by Czech ministry of health (Grant No.17-29680A to D.R. and L.K.) Czech Science Foundation grant number (grant No. 15-11711S to D.R.), EU and Czech ministry of Education and Sport via JPIAMR (grant No. 8F19006 to D.R. and V.H.), Swedish Research council (grant 2017?03783 to VH) and Charles University, Department of Analytical Chemistry ? Specific University Research No. SVV260440 to E.Z. Publisher Copyright: © 2019 The Authors Copyright: Copyright 2019 Elsevier B.V., All rights reserved.
id
0c593c6d-c013-4758-9d5d-fe13948b1e39
date added to LUP
2021-09-24 20:33:17
date last changed
2024-09-08 23:57:04
@article{0c593c6d-c013-4758-9d5d-fe13948b1e39,
  abstract     = {{<p>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 &gt;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.</p>}},
  author       = {{Zborníková, Eva and Knejzlík, Zdeněk and Hauryliuk, Vasili and Krásný, Libor and Rejman, Dominik}},
  issn         = {{0039-9140}},
  keywords     = {{Escherichia coli; HILIC; HPLC-MS; Nucleotide; ppGpp; Stringent response}},
  language     = {{eng}},
  pages        = {{1--10}},
  publisher    = {{Elsevier}},
  series       = {{Talanta}},
  title        = {{Analysis of nucleotide pools in bacteria using HPLC-MS in HILIC mode}},
  url          = {{http://dx.doi.org/10.1016/j.talanta.2019.120161}},
  doi          = {{10.1016/j.talanta.2019.120161}},
  volume       = {{205}},
  year         = {{2019}},
}