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Ion-exchange thin-layer chromatography

1965; Elsevier BV; Volume: 12; Issue: 1 Linguagem: Inglês

10.1016/0003-2697(65)90145-4

1096-0309

Erika Randerath, Kurt Randerath,

RNA modifications and cancer

A procedure is described by which small amounts of nucleotides can be eluted quantitatively from PEI-cellulose anion-exchange thin-layer chromatograms. The method comprises direct transfer of the compounds from the layer to a paper wick and subsequent elution from the paper. Spectrophotometric measurements show quantitative recoveries of 5′-CMP, 5′-AMP, 5′-UMP, 5′-GMP, DPN, UDPG, and ATP. Statistical analysis of a number of replicate determinations of 6–10 mμmole samples of these compounds indicates relative standard deviations of 0.9–4.8%. The technique can be used with slight modifications if larger amounts (up to a few hundred micrograms of individual compounds) are to be eluted. It is not restricted to anion-exchange layers, but can be used on plain cellulose and silica gel layers as well. A new one-dimensional separation method on PEI-cellulose thin layers is also described by which an alkaline hydrolyzate of RNA can be resolved into the constituent nucleoside monophosphates. This procedure combined with the quantitative elution procedure should provide a new sensitive technique for determining the base composition of RNA.