Improved pH measurement of mobile phases in reversed-phase liquid chromatography

Abstract

Mobile phase pH is a critically important parameter in reversed-phase liquid chromatographic (RPLC) separations involving analytes that display acidic or basic properties in the pH range used for the mobile phase. The main problem in measuring mobile phase pH lies in the fact that RPLC mobile phases are typically aqueous–organic mixtures. In addition to experimental difficulties, the pH values refer to different aqueous–organic compositions that cannot be correctly compared. Given this situation, the unified pH (^w_abspH, also termed as ) based on the absolute chemical potential of the solvated proton has been proposed as a rigorous way of characterising mobile phase acidity that is fully inter-comparable between mobile phases of any composition. Here we report the ^w_abspH values of 78 reversed-phase liquid chromatography-mass spectrometry mobile phases that were carefully measured by potential differences in a symmetric cell with two glass electrode half-cells and almost ideal ionic liquid triethylamylammonium bis((trifluoromethyl)sulfonyl)imide [N₂₂₂₅][NTf₂] salt bridge with multiple overlapping measurements. The system of altogether 300 Δ^w_abspH values was anchored to the pH value of standard pH 7.00 aqueous buffer solution. The consistency standard deviation of the whole set of measurements was 0.09 pH units. In addition to the differential potentiometric reference method, simpler measurement methods that use double junction reference or double junction combined electrodes were tested and were found suitable for routine laboratories if high accuracy is not required.