LIQUID-CHROMATOGRAPHIC SEPARATION OF HEXOPYRANOSYLATED CYTOSINE NUCLEOSIDES FROM THEIR DEGRADATION PRODUCTS

Development of a liquid chromatographic method which can separate each of a series of hexopyranosylated cytosine nucleosides from their degr adation products formed at acid, neutral and basic pH is described. Bo th silica-based reverse-phase and polymer columns were examined. Influ ence of the mobile phase pH, ion-pairing agent, concentration of the b uffer and type and concentration of organic modifier were systematical ly investigated. The concentration of the ion-pairing agent and the bu ffer were found to have a major effect on selectivity. Samples were fi nally analyzed on a poly(styrene-divinylbenzene), PLRP-S 100 Angstrom (8 mu m) 250 x 4.6 mm I.D. column at 60 degrees C and with a mobile ph ase consisting of acetonitrile-sodium octanesulphonate (pH 2.5; 0.02 M )-potassium phosphate buffer (pH 2.5; 0.2 M)-water (X:25:50:25 -X, v/v , where X is variable). (C) 1997 Elsevier Science B.V.