Retention behavior of ionizable isomers in reversed-phase liquid chromatography:  a comparative study of porous...

The retention behaviors of 36 positional isomers of ionizable substituted benzene compounds have been compared on two different packing materials:  porous graphitic carbon (PGC) and octadecyl bonded silica (ODS) using 35% aqueous acetonitrile as the mobile phase. The effect of the mobile phase pH on the solute retention was studied over a range of pH values from pH 2.0 to 7.0. The retention as a function of pH was modeled using equations based on solute ionization. With PGC, the theoretical equations fitted the observed retention data for each class of solute, indicating that the retention mechanism was uniform over the whole pH range. However, with ODS, only the acidic solutes showed agreement with the theoretical model; for the amine-containing compounds, serious deviations from the theory were observed, suggesting that strongly acidic silanols gave added retention at low mobile phase pH. Overall, PGC demonstrated a higher selectivity toward positional isomers than ODS. This was attributed to the greater steric discriminating ability arising from the flat surface of the PGC compared with the more fluid nature of the ODS bonded phase.