Separation of 2′- and 3′-mononucleotides of plant viral ribonucleic acids by an improved two-dimensional thin-layer chromatography

Abstract

Thin-layer chromatography (TLC) has been used widely in qualitative as well as small-scale quantitative separation of complex mixtures of nucleotides and their derivatives (1–4). The use of DEAE-cellulose, 1 1 Abbreviations: RNA, ribonucleic acid. 2′,3′-UMP, 2′,3′-CMP, 2′,3′-AMP, 2′,3′-GMP, the 2′,3′-monophosphates of uridine, cytidine, adenosine, and guanosine, respectively. MN300G, cellulose powder with CaSO 4·2 1 H 2O as binder. PPO, 2,5-diphenyloxazole. PEI, polyethyleneimine. DEAE, diethylaminoethyl. Sephadex, polyphosphate cellulose, phosphocellulose, silica gel, ECTEOLA-cellulose, PEI-cellulose, and polyamide as layering materials has been reported (1); however, cellulose powder is the most used layering material. The recognized advantages of TLC, namely, convenience, sensitivity, and short developing time as compared with paper chromatography, have been clearly demonstrated in nucleic acid studies (5). Sharper separations of nucleotides, nucleosides, and bases have been observed with cellulose layers (5). This paper describes a new two-dimensional TLC procedure to separate 2′- and 3′-nucleotide isomers of the mononucleotides derived from the ribonucleic acids. This method has been employed successfully in the analysis of base composition of plant viral RNA. Separation of the nucleosides was also achieved with this method.