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

Béres, Tibor; Zatloukal, Marek; Voller, Ji?í; Niemann, Percy; Gahsche, Marie Christin; Tarkowski, Petr; Novák, Ond?ej; Hanu?, Jan; Strnad, Miroslav; Dole?al, Karel (2010). Tandem mass spectrometry identification and LC-MS quantification of intact cytokinin nucleotides in K-562 human leukemia cells. Analytical and bioanalytical chemistry, 398(5), pp. 2071-80. Heidelberg: Springer 10.1007/s00216-010-4126-5

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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 was used to identify the intracellular metabolites (cytokinin monophosphorylated, diphosphorylated, and triphosphorylated 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 diethyl ether, lyophilized, reconstituted, and injected into the LC system. Analytes were quantified in negative selected ion monitoring mode using a single quadrupole mass spectrometer. The method was validated in terms of retention time stabilities, limits of detection, linearity, recovery, and analytical accuracy. The developed method was linear in the range of 1-1,000 pmol for all studied compounds. The limits of detection for the analytes vary from 0.2 to 0.6 pmol.

Item Type:

Journal Article (Original Article)

Division/Institute:

04 Faculty of Medicine > Department of Head Organs and Neurology (DKNS) > Clinic of Neurology

UniBE Contributor:

Niemann, Petra

ISSN:

1618-2642

Publisher:

Springer

Language:

English

Submitter:

Factscience Import

Date Deposited:

04 Oct 2013 14:14

Last Modified:

17 Mar 2015 19:17

Publisher DOI:

10.1007/s00216-010-4126-5

PubMed ID:

20821313

Web of Science ID:

000283244500028

URI:

https://boris.unibe.ch/id/eprint/3379 (FactScience: 207051)

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