Separation of oligodeoxyribonucleotides, prepared by chemical synthesis, was carried out using reversed 'phase ion-pair chromatography with LiChrospher 100 RP-18e. By this technique, the peak shape and the separation of peaks were highly improved, compared to the results obtained by conventional reversed phase chromatography with the same column. The chain length and the concentration of the tetraalkylammonium ion-pair reagents were found to be responsible for their retention behaviors and tetrabutylammonium phosphate showed marked enhancement of the peak resolution. In the separation with this reagent, a good linear relationship between the elution volumes and the base numbers of oligodeoxyribonucleotides was obtained. This implies that ion-pair chromatography can be used for the separation of oligodeoxyribonucleotides according to their base numbers.
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