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Statements

Subject Item
n2:RIV%2F61989592%3A15310%2F12%3A33142888%21RIV13-MSM-15310___
rdf:type
n13:Vysledek skos:Concept
rdfs:seeAlso
http://www.plantmethods.com/content/8/1/17
dcterms:description
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. 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.
dcterms:title
A new approach for cytokinin isolation from Arabidopsis tissues using miniaturized purification: pipette tip solid-phase extraction A new approach for cytokinin isolation from Arabidopsis tissues using miniaturized purification: pipette tip solid-phase extraction
skos:prefLabel
A new approach for cytokinin isolation from Arabidopsis tissues using miniaturized purification: pipette tip solid-phase extraction A new approach for cytokinin isolation from Arabidopsis tissues using miniaturized purification: pipette tip solid-phase extraction
skos:notation
RIV/61989592:15310/12:33142888!RIV13-MSM-15310___
n13:predkladatel
n21:orjk%3A15310
n3:aktivita
n14:S n14:P n14:Z
n3:aktivity
P(ED0007/01/01), P(GA522/09/1576), P(KAN200380801), P(TA01010861), S, Z(AV0Z50380511)
n3:cisloPeriodika
17
n3:dodaniDat
n4:2013
n3:domaciTvurceVysledku
n5:9896821 n5:4712048 n5:4577213 n5:5991803 n5:3686957 n5:4521463
n3:druhVysledku
n16:J
n3:duvernostUdaju
n20:S
n3:entitaPredkladatele
n22:predkladatel
n3:idSjednocenehoVysledku
120341
n3:idVysledku
RIV/61989592:15310/12:33142888
n3:jazykVysledku
n8:eng
n3:klicovaSlova
Tandem mass spectrometry (MS/MS); performance liquid chromatography (UHPLC); Ultra-high; StageTip; Pipette tip solid-phase extraction (PT-SPE); Arabidopsis thaliana; Cytokinins
n3:klicoveSlovo
n9:Ultra-high n9:StageTip n9:performance%20liquid%20chromatography%20%28UHPLC%29 n9:Tandem%20mass%20spectrometry%20%28MS%2FMS%29 n9:Pipette%20tip%20solid-phase%20extraction%20%28PT-SPE%29 n9:Cytokinins n9:Arabidopsis%20thaliana
n3:kodStatuVydavatele
GB - Spojené království Velké Británie a Severního Irska
n3:kontrolniKodProRIV
[A74F8D73D7A1]
n3:nazevZdroje
Plant Methods
n3:obor
n7:EC
n3:pocetDomacichTvurcuVysledku
6
n3:pocetTvurcuVysledku
7
n3:projekt
n10:ED0007%2F01%2F01 n10:GA522%2F09%2F1576 n10:TA01010861 n10:KAN200380801
n3:rokUplatneniVysledku
n4:2012
n3:svazekPeriodika
8
n3:tvurceVysledku
Svačinová, Jana Doležal, Karel Novák, Ondřej Holík, Josef Plačková, Lenka Lenobel, René Strnad, Miroslav
n3:wos
000311423700001
n3:zamer
n17:AV0Z50380511
s:issn
1746-4811
s:numberOfPages
28
n19:doi
10.1186/1746-4811-8-17
n15:organizacniJednotka
15310