About: Analysis of cytokinin nucleotides by capillary zone electrophoresis with diode array and mass spectrometric detection in a recombinant enzyme in vitro reaction     Goto   Sponge   NotDistinct   Permalink

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Description
  • A capillary zone electrophoresis (CZE) method for separation of adenosine and N-6-isopentenyladenosine (cytokinin) nucleotides was developed, optimized and validated. Aqueous solutions of several amino acids were evaluated as the background electrolyte constituents. Separation of six nucleotides in less than 20 min with high theoretical plate number (up to 400 000 for isopentenyladenosine triphosphate) was achieved using a 100 mM sarcosine/ammonia buffer at pH 10.0. The detection limits of the CZE-UV method are in the low micromolar range (0.69-1.27 mu mol L-1). Good repeatability of migration times (within 1.3%), peak areas (within 1.8%) and linearity (R-2 }0.999) was achieved over the concentration range 5-1000 mu mol L-1. The method was used to assay the activity of the recombinant Arabidopsis thaliana isopentenyltransferase 1 (AtIPT1). Baseline separation of isopentenylated nucleotides by CE-ESI-MS using a volatile buffer (30 mM ammonium formate; pH 10.0) was accomplished. The identities of the reaction products - isopentenyladenosine di- and triphosphate were confirmed by HPLC-QqTOE-MS. Dephosphorylation of ATP was observed as a parallel reaction
  • A capillary zone electrophoresis (CZE) method for separation of adenosine and N-6-isopentenyladenosine (cytokinin) nucleotides was developed, optimized and validated. Aqueous solutions of several amino acids were evaluated as the background electrolyte constituents. Separation of six nucleotides in less than 20 min with high theoretical plate number (up to 400 000 for isopentenyladenosine triphosphate) was achieved using a 100 mM sarcosine/ammonia buffer at pH 10.0. The detection limits of the CZE-UV method are in the low micromolar range (0.69-1.27 mu mol L-1). Good repeatability of migration times (within 1.3%), peak areas (within 1.8%) and linearity (R-2 }0.999) was achieved over the concentration range 5-1000 mu mol L-1. The method was used to assay the activity of the recombinant Arabidopsis thaliana isopentenyltransferase 1 (AtIPT1). Baseline separation of isopentenylated nucleotides by CE-ESI-MS using a volatile buffer (30 mM ammonium formate; pH 10.0) was accomplished. The identities of the reaction products - isopentenyladenosine di- and triphosphate were confirmed by HPLC-QqTOE-MS. Dephosphorylation of ATP was observed as a parallel reaction (en)
Title
  • Analysis of cytokinin nucleotides by capillary zone electrophoresis with diode array and mass spectrometric detection in a recombinant enzyme in vitro reaction
  • Analysis of cytokinin nucleotides by capillary zone electrophoresis with diode array and mass spectrometric detection in a recombinant enzyme in vitro reaction (en)
skos:prefLabel
  • Analysis of cytokinin nucleotides by capillary zone electrophoresis with diode array and mass spectrometric detection in a recombinant enzyme in vitro reaction
  • Analysis of cytokinin nucleotides by capillary zone electrophoresis with diode array and mass spectrometric detection in a recombinant enzyme in vitro reaction (en)
skos:notation
  • RIV/61989592:15310/12:33142716!RIV13-MSM-15310___
http://linked.open...avai/predkladatel
http://linked.open...avai/riv/aktivita
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  • P(ED0007/01/01), P(GA522/08/0920), P(LC06034), Z(AV0Z50380511), Z(MSM6198959216)
http://linked.open...iv/cisloPeriodika
  • NOV
http://linked.open...vai/riv/dodaniDat
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  • 122103
http://linked.open...ai/riv/idVysledku
  • RIV/61989592:15310/12:33142716
http://linked.open...riv/jazykVysledku
http://linked.open.../riv/klicovaSlova
  • Cytokinin biosynthesis; Isopentenyltransferase; Capillary electrophoresis; Cytokinin nucleotides (en)
http://linked.open.../riv/klicoveSlovo
http://linked.open...odStatuVydavatele
  • NL - Nizozemsko
http://linked.open...ontrolniKodProRIV
  • [981CD75F0D80]
http://linked.open...i/riv/nazevZdroje
  • Analytica Chimica Acta
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http://linked.open...v/svazekPeriodika
  • 751
http://linked.open...iv/tvurceVysledku
  • Doležal, Karel
  • Strnad, Miroslav
  • Spíchal, Lukáš
  • Maier, Vítězslav
  • Gemrotová, Markéta
  • Béres, Tibor
  • Dessoy, Marco A
  • Friedecký, David
  • Ganzera, Markus
  • Tarkowski, Petr
  • Wessjohann, Ludger A
http://linked.open...ain/vavai/riv/wos
  • 000311013100021
http://linked.open...n/vavai/riv/zamer
issn
  • 0003-2670
number of pages
http://bibframe.org/vocab/doi
  • 10.1016/j.aca.2012.08.049
http://localhost/t...ganizacniJednotka
  • 15310
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