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

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61989592%3A15310%2F10%3A10217256" target="_blank" >RIV/61989592:15310/10:10217256 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/61389030:_____/10:00351018

Výsledek na webu

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DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

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

Popis výsledku v původním jazyce

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.

Název v anglickém jazyce

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

Popis výsledku anglicky

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.

Druh

J_x - Nezařazeno - Článek v odborném periodiku (Jimp, Jsc a Jost)

CEP obor

CB - Analytická chemie, separace

OECD FORD obor

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Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)<br>Z - Vyzkumny zamer (s odkazem do CEZ)

Rok uplatnění

2010

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Analytical and Bioanalytical Chemistry

ISSN

1618-2642

e-ISSN

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Svazek periodika

398

Číslo periodika v rámci svazku

5

Stát vydavatele periodika

DE - Spolková republika Německo

Počet stran výsledku

10

Strana od-do

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Kód UT WoS článku

000283244500028

EID výsledku v databázi Scopus

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