Determination of nucleotide pools in plant tissue by high-performance liquid chromatography

A procedure for the determination of nucleotide pools in plant tissue by HPLC is described. Sample preparation includes the extraction with 0.4 M HClO4, a purification step, which proved to be essential, on a disposable prepacked phenyl-bonded column, neutralization by KOH, and concentration by freeze-drying. The determination of a broad spectrum of ribonucleotides including the ribonucleosides was performed by combining anion-exchange and reversed-phase HPLC. Data are presented for suspension-cultured cells of Nicotiana tabacum and Datura innoxia and for the leaf and root of tobacco.