A novel switchable water stationary phase for supercritical fluid chromatography

A novel pH switchable water stationary phase is presented for use in supercritical fluid chromatography (SFC). By adding NH₄OH to the water coating and system hydration, changes in CO₂ pressure and temperature allow a wide range of stationary phase pH conditions (∼3-9) to be achieved, which impact analyte retention properties. For example, 100 atm and 50 °C produces an acidic water stationary phase (pH near 4.0) where octanoic acid readily elutes while the base caffeine does not. Conversely, at 80 atm and 120 °C a basic water stationary phase (pH near 8.0) is obtained and the opposite occurs. Further, under constant pressure and temperature conditions, simply adding or removing NH₄OH from the system is also found to readily allow switching between the basic and acidic water stationary phase modes and demonstrates control over ionizable analyte elution. For instance, hexanoic acid elution is near 40 times more delayed on a basic water stationary phase and, as such, it can be eluted at later points in time as desired by removing the NH₄OH and switching to an acidic stationary phase. Experiments indicate that stationary phase pH switching occurs uniformly across the 15 m column length within about 18 s and that analyte retention times are very reproducible upon performing a switch (1.4% RSD; n = 3). Results demonstrate the selectivity factor between acidic and neutral analytes can be reversed and increased about 35 times, while in other trials resolution also similarly increased near 40-fold. By rapidly switching the stationary phase pH back and forth between acidic and basic modes, the selectivity between ionizable analytes could also be increased as desired. Various applications with the system show that it can vastly increase the separation between target analytes and matrix components as required by the dynamics of a particular separation.