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RESEARCH PRODUCT

Determination of total ribonucleotide pool in plant materials by high-pH anion-exchange high-performance liquid chromatography following extraction with potassium hydroxide

Christophe RiondetGérard AlcarazSylvain Morel

subject

RibonucleotidePotassium CompoundsNicotiana tabacumIon chromatographyBiochemistryHigh-performance liquid chromatographyAnalytical Chemistrychemistry.chemical_compoundSpecies SpecificityHydroxidesSample preparationAnion Exchange ResinsPotassium hydroxideChromatographybiologyIon exchangeOrganic ChemistryExtraction (chemistry)General MedicineHydrogen-Ion ConcentrationPlantsReference StandardsRibonucleotidesChromatography Ion Exchangebiology.organism_classificationchemistry

description

A new, improved method that only requires a potassium hydroxide extraction procedure is presented for the analysis of a full nucleotide pool in plant materials. Quantification was performed by high-pH anion-exchange chromatography (HPAEC) with UV detection after a potassium hydroxide extraction, and allowed the quantification of 13 linear ribonucleotides in a single run. The method has been validated by comparison of six extraction methods and also by measurement of the intracellular nucleotide levels of three plant species (cell cultures and leaves). The evolution of the nucleotide pool of Nicotiana tabacum cell culture during growth has also been measured, and showed an increase in the pool until the fifth day, where the growth rate reaches a maximum, after which a decrease was observed.

yearjournalcountryeditionlanguage
2005-07-09Journal of Chromatography A
https://doi.org/10.1016/j.chroma.2005.04.036