High Performance Liquid Chromatography for Selective Separation of Nucleotides and Oligonucleotides on Nucleic Acid Base-Immobilized Silica Gel

Abstract

Nucleotides and oligonucleotides could be separated by using silica-based supports on which immobilized were thymine (Si-Thy), uraci (Si-Ura), cytosine (Si-Cyt), adenine (Si-Ade) and guanine (Si-Gua) derivatives. The chromatographic conditions were as follows: Column: Si-Thy, Si-Ura, Si-Cyt, Si-Ade and Si-Gua; sample: nucleosides, nucleotides and oligonucleotides; column length: 150mm×4.6mm id.; eluent: 0.1M triethylammonium acetate (TEAA) aqueous solution (pH 6.8±0.2) containing acetonitrile; flow rate: 1.0ml/min; injection volume: 4-10μl; temperature: 35°C; detector: UV (254nm). These resins showed the ability of the selective separation of nucleotides and oligonucleotides by the specific interaction between complementary bases.