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Tandem mass spectrometry identification and LC-MS quantification of intact cytokinin nucleotides in K-562 human leukemia cells
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SYSNO ASEP 0351018 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Tandem mass spectrometry identification and LC-MS quantification of intact cytokinin nucleotides in K-562 human leukemia cells Author(s) Béres, Tibor (UEB-Q) RID
Zatloukal, Marek (UEB-Q) ORCID
Voller, Jiří (UEB-Q) RID, ORCID
Niemann, P. (DE)
Gahsche, M.C. (DE)
Tarkowski, Petr (UEB-Q)
Novák, Ondřej (UEB-Q) RID, ORCID, SAI
Hanuš, Jan (UEB-Q)
Strnad, Miroslav (UEB-Q) RID, ORCID
Doležal, Karel (UEB-Q) RID, ORCIDSource Title Analytical and Bioanalytical Chemistry. - : Springer - ISSN 1618-2642
Roč. 398, č. 5 (2010), s. 2071-2080Number of pages 10 s. Language eng - English Country DE - Germany Keywords Cytokinins ; Nucleotides ; HPLC Subject RIV EB - Genetics ; Molecular Biology R&D Projects 1M06030 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) GD522/08/H003 GA ČR - Czech Science Foundation (CSF) GA522/08/0920 GA ČR - Czech Science Foundation (CSF) CEZ AV0Z50380511 - UEB-Q (2005-2011) UT WOS 000283244500028 DOI 10.1007/s00216-010-4126-5 Annotation We describe here a new reversed phase high performance liquid chromatography with mass spectrometry detection method for quantifying intact cytokinin nucleotides in human K-562 leukemia cells. Tandem mass spectrometry (MS/MS) was used to identify the intracellular metabolites (cytokinin mono- di- and tri-phosphorylated nucleotides) in riboside-treated cells. For the protein precipitation and sample preparation, a trichloroacetic acid extraction method is used. Samples are then back-extracted with diethyleter, lyophilized, reconstituted and injected into the LC system. Analytes were quantified in negative selected ion monitoring (SIM) mode using a single quadrupole mass spectrometer. The method was validated in terms of retention time stabilities, limits of detection (LOD), linearity, recovery and analytical accuracy. The developed method was linear in the range of 1-1000 pmol for all studied compounds. The limits of detection for the analytes vary from 0.2 to 0.6 pmol. Workplace Institute of Experimental Botany Contact David Klier, knihovna@ueb.cas.cz, Tel.: 220 390 469 Year of Publishing 2011
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