Comparison of ethylammonium formate to methanol as a mobile-phase modifier for reversed-phase liquid chromatography.

Ethylammonium formate (EAF), (C2H5NH3+HCO2-), is a room-temperature ionic liquid that has a polarity similar to that of methanol (MeOH) or acetonitrile. The separation at 1 mL/min of a test mixture of vitamins or phenols on a polystyrene-divinylbenzene column using either an EAF- or MeOH-water mobile phase is similar in terms of both resolution and analysis time. Because the viscosity of EAF is higher than that of MeOH, the plate count for phenol at room temperature is lower by about a factor of 1.1-1.4 depending on the flow rate. However, van Deemter plots show that this loss in plate count at 1 mL/min can be recovered and improved from 1500 to 2400 plates by working at a slightly elevated temperature of 55 degrees C. A slower flow rate such as 0.8 mL/min can also substantially improve the plate count as compared to 1-1.5 mL/min. Log P (octanol partition coefficient) versus log k' data for a variety of neutral test solutes are again similar whether EAF or MeOH is used as the organic modifier. Resolution of certain peak pairs such as 2,4-dinitrophenol/2,4,6-trinitrophenol and p-aminobenzoate/benzoate is enhanced using EAF as compared to MeOH. One advantage of EAF is that control of retention of solutes such as water-soluble vitamins under totally aqueous mobile phase conditions is environmentally preferable for quality control applications. In addition, EAF seems to be a milder mobile-phase modifier than MeOH for certain proteins such as lysozyme.