High-Performance Liquid Chromatographic Determination of 5'-and 2'(3')-Mononucleotides in Seaweeds and Fishes

Abstract

Anion exchange chromatography was carried out by stepwise and gradient eluting method, using a high performance liquid chromatograph of Hitachi 655 A-11 type, Hitachi gell 3013 N of anion exchange porous polymer and the authentic samples of nucleic acid related substances. Eluting solution A: 0.045M NH₄Cl·0.0075M KH₂PO₄·0.0075M K₂HPO₄·2% CH₃CN, pH 8.2. Eluting solution B: 0.06M NH₄Cl·0.01M KH2PO4·0.01M K₂HPO₄·5%CH₃CN, pH 82. Eluting solution C: 0.105M NH₄Cl·0.0175M KH₂PO₄·0.0175M K₂HPO₄·6%CH₃CN, pH 3.0. After nucleosides and purine and pyrimidine bases were eluted by eluting solution A, 5'-mononucleotides and 2'(3')-mononucleotides could be eluted separately by the gradient method of eluting solutions A and B. And then, ADP and ATP could be separated continuously by eluting solution C. The nucleotides of 5'-mononucleotides, 2'(3')-mononu-cleotides, ADP, and ATP in several seaweeds and fishes were separated and determined by the above described method. The seaweeds contained 2'(3')-mononucleotides in large quantities and 5'-mononucleotides in small quantities. However, 5'-GMP, 5'-IMP, ATP could be scarcely observed in the seaweeds. The fishes contained a large quantity of 5'-IMP (90-350mg/100g). However, the contents of 5'-mononucleotides except 5'-IMP and 2'(3')-mononucleotides were little in the fishes.