New method for the differentiation of ribonucleotides from deoxyribonucleotides

Abstract

Nucleotides were separated on thin layer of PEI or standard cellulose developed in buffered aqueous ammonium isobutyrate at pH 4.6. In addition to R f values, nucleoside mono-, di-, and triphosphates were further differentiated according to the rate at which a nucleotide forms a stable blue complex with ammonium molybdate. The blue color formation, in turn, depends upon the ease at which a nucleotide liberates its P i; this property appears to be closely related to the nature of the phosphate-phosphate bond, the sugar-phosphate bond, and the base-sugar bond. Differentiation was made between: ( a) ribonucleotide and deoxyribo-nucleotide, ( b) purine derivative and pyrimidine derivative, ( c) 5′-ribonucleotide and 2′,3′-ribonucleotide, ( d) nucleoside monophosphates (e.g., 5′-AMP, 5′-CMP, 5′-UMP, and 5′-GMP), and ( e) nucleoside mono-, di-, and triphosphates (e.g., AMP, ADP, and ATP).