β Scribed by A. Floridi; C.A. Palmerini; C. Fini
No coin nor oath required. For personal study only.
A single-column high-performance liquid chromatographic system for the simultaneous separation of bases, nucleosides and nucleoside mono-and polyphosphates has been developed, in which a strong porous anion-exchange resin (Aminex A-14) is used. The chromatographic run, carried out at 55" and at alkaline pH by using a linear gradient both of ionic strength and pH, takes less than 225 min. The quantitative application of the described procedure to the analysis of cell nucleotide pools is reported_
The nucleotide pattern of cell extracts is of interest not only for defining the metabolic stage of the cell, but also for studying the compartmentation and the normal or altered metabolism of these compounds. Various chromatographic techniques can be employed to analyse bases, nucleosides and nucleotides (two-dimensional paper chromatography, paper electrophoresis, ion-exchange paper chromatography, thin-layer chromatography), but the best method, at present, for detecting, identifying and quantitatively determining such compounds is ion-exchange chromatography. In this field, the classic approach by Cohn', the application of a gradient elution system2, the development of an ultraviolet elIluent monitoring device3 and the use of new efficient packing materials --have allowed the development of highperformance liquid chromatography (HPLC) ion-exchange systems. The analysis of nucleotides from cell extracts can be performed either with pellicular ion-exchange resin&'-" or by reversed-phase chromatographyl~. The HPLC separation of bases or nucIeosides-17 and of nucleosides in the presence of their bases18-20 with porous ion-exchange resins has also been described. More complex mixtures, in which bases, nucleosides and nucleotide monophosphates are simultaneously present, have been separated with using ,either Cation21 or anion-exchange?chromatography.
Good results, however, for the simultaneous analysis of high resolution by bases, nucleosides '204 A. FLORIDI, C. A. PALMERINI, C. FINI
π SIMILAR VOLUMES
Nucleotides, nucleosides, and purine bases were extracted from human endomyocardial biopsies, freeze-clamped rat hearts, and porcine coronary sinus plasma. Perchloric acid extracts were neutralized with Freon-trioctylamine and analyzed at 250 nm by reverse-phase ion-pairing high-performance liquid c
The chromatographic behaviour of some natural and modified pyrimidine bases and nucleosides on an octadecyl stationary phase was studied. The retention and selectivity parameters of the separation of the compounds studied were derived on the basis of solvophobic theory. The mechanism of base and nuc