High-performance liquid chromatography of tRNAs on novel stationary phases

Rapid separation of a group of tRNAs was carried out on novel siliceous bonded stationary phases with aqueous eluents by using gradient elution with increasing or decreasing salt gradient, as usual in electrostatic interaction chromatography or hydrophobic interaction chromatography, respectively. The stationary phases consist of microparticulate macroporous silica with surface-bound polar moieties, containing weak cationic and/or hydrophobic binding sites. Depending on the nature of the binding sites, the stationary phases exhibit different retention behavior and selectivity for tRNAs. Aqueous phosphate solutions were used as the eluent, and in many cases isocratic elution was sufficient to separate seven tRNAs. Addition of magnesium ions or n-decylbetaine to the eluent resulted in lower retention, the latter causing a greater increase in the eluent strength. The optimum pH range of the eluent was 5.5-6.5.