Počet záznamů: 1

Tandem mass spectrometry identification and LC-MS quantification of intact cytokinin nucleotides in K-562 human leukemia cells

  1. 1.
    0351018 - UEB-Q 2011 RIV DE eng J - Článek v odborném periodiku
    Béres, Tibor - Zatloukal, Marek - Voller, Jiří - Niemann, P. - Gahsche, M.C. - Tarkowski, Petr - Novák, Ondřej - Hanuš, Jan - Strnad, Miroslav - Doležal, Karel
    Tandem mass spectrometry identification and LC-MS quantification of intact cytokinin nucleotides in K-562 human leukemia cells.
    Analytical and Bioanalytical Chemistry. Roč. 398, č. 5 (2010), s. 2071-2080 ISSN 1618-2642
    Grant CEP: GA MŠk 1M06030; GA ČR GD522/08/H003; GA ČR(CZ) GA522/08/0920
    Výzkumný záměr: CEZ:AV0Z50380511
    Klíčová slova: Cytokinins * Nucleotides * HPLC
    Kód oboru RIV: EB - Genetika a molekulární biologie
    Impakt faktor: 3.841, rok: 2010

    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.
    Trvalý link: http://hdl.handle.net/11104/0190862