Separation of arabinosyl, ribosyl, and deoxyribosyl purine nucleotides by thin-layer chromatography.

Studies on the cellular metabolism of endogenous nucleosides and structurally related analogs are facilitated by the thin-layer chromatographic (TLC) separation of nucleotides, Complex mixtures of ribosyl and deoxyribosyl nucleotides can be separated by ascending two-dimensional TLC on PEI-cellulose anion-exchange layers”“. Nucleoside mono-, diand triphosphates are first separated in one dimension by stepwise development in solvents of increasing ionic strength4e5. Ribosyl and deoxyribosyl nucleotides then are resolved in the other dimension using borate-containing solvent systems4-“. Nucleosides and bases present in cell extracts interfere with resolution of nucleotides, however, and must be removed by an initial development of the chromatogram. We report here a TLC system which resolves arabinosyl as well as ribosyl and deoxyribosyl nucleotides. Nucleosides and bases do not interfere with resolution of nucleotides and no prior development is required. The system has been used routinely to separate purine nucleotides in our studies of the cellular metabolism of 9-p-Darabinofuranosyladenine, an active antiviral drug.