Band profiles of reacting acido-basic compounds with water-methanol eluents at different SWpHs and ionic strengths in reversed-phase liquid chromatography.

Overloaded band profiles of aniline (25 microL injection of a 30.5mM solution) eluted with a methanol-aqueous buffer solution (30/70, v/v) were recorded at the exit of a 150 mm x 4.6mm column packed with 3.5 microm XBridge-C(18)porous particles. The SWpHs of the mobile phase was adjusted with phosphate (SWpHs approximately 2.7 and SWpHs approximately 7.5) or acetate buffers (SWpHs approximately 5.3) of different concentrations (11, 56, and 278 mM). The elution times and profiles of the bands observed at low ionic strength were successfully accounted for using the extended Debye-Hückel theory to estimate the activity coefficients of the ions in the bulk phase and a simple non-competitive Langmuir adsorption model, the adsorption of pure aniline or pure anilinium onto XBridge-C(18) being described by a Langmuir isotherm. The band profiles were calculated using this adsorption model and the equilibrium-dispersive model of chromatography. The calculated and the experimental band profiles are in excellent agreement at all buffer pH and ionic strength. This demonstrates that the elution times and the band profiles are controlled by the chromatographic dilution process and by the reaction of aniline with the buffer solution.