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A new approach for cytokinin isolation from Arabidopsis tissues using miniaturized purification: pipette tip solid-phase extraction
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SYSNO ASEP 0385990 Druh ASEP J - Článek v odborném periodiku Zařazení RIV J - Článek v odborném periodiku Poddruh J Článek ve WOS Název A new approach for cytokinin isolation from Arabidopsis tissues using miniaturized purification: pipette tip solid-phase extraction Tvůrce(i) Svačinová, Jana (UEB-Q)
Novák, Ondřej (UEB-Q) RID, ORCID, SAI
Plačková, Lenka (UEB-Q) ORCID, RID
Lenobel, René (UEB-Q) ORCID
Holík, Josef (UEB-Q) RID, ORCID
Strnad, Miroslav (UEB-Q) RID, ORCID
Doležal, Karel (UEB-Q) RID, ORCIDZdroj.dok. Plant Methods. - : BioMed Central
Roč. 8, _ (2012), s. 17Poč.str. 14 s. Jazyk dok. eng - angličtina Země vyd. GB - Velká Británie Klíč. slova Pipette tip solid-phase extraction (PT-SPE) ; Arabidopsis thaliana ; Cytokinins Vědní obor RIV EC - Imunologie CEP TA01010861 GA TA ČR - Technologická agentura ČR KAN200380801 GA AV ČR - Akademie věd CEZ AV0Z50380511 - UEB-Q (2005-2011) UT WOS 000311423700001 DOI 10.1186/1746-4811-8-17 Anotace Background: We have developed a new analytical approach for isolation and quantification of cytokinins (CK) in minute amounts of fresh plant material, which combines a simple one-step purification with ultra-high performance liquid chromatography-fast scanning tandem mass spectrometry. Results: Plant tissue samples (1-5 mg FW) were purified by stop-and-go-microextraction (StageTip purification), which previously has only been applied for clean-up and pre-concentration of peptides. We found that a combination of two reverse phases and one cation-exchange phase, was the best tool, giving a total extraction recovery higher than 80%. The process was completed by a single chromatographic analysis of a wide range of naturally occurring cytokinins (bases, ribosides, O- and N-glucosides, and nucleotides) in 24.5 minutes using an analytical column packed with sub-2- microne particles. In multiple reaction monitoring mode, the detection limits ranged from 0.05 to 5 fmol and the linear ranges for most cytokinins were at least five orders of magnitude. The StageTip purification was validated and optimized using samples of Arabidopsis thaliana seedlings, roots and shoots where eighteen cytokinins were successfully determined. Conclusions: The combination of microextraction with one-step high-throughput purification provides fast, effective and cheap sample preparation prior to qualitative and quantitative measurements. Our procedure can be used after modification also for other phytohormones, depending on selectivity, affinity and capacity of the selected sorbents. Pracoviště Ústav experimentální botaniky Kontakt David Klier, knihovna@ueb.cas.cz, Tel.: 220 390 469 Rok sběru 2013
Počet záznamů: 1