Retention of ionisable compounds on high-performance liquid chromatography. XV. Estimation of the pH variation of aqueous buffers with the change of the acetonitrile fraction of the mobile phase.
The most commonly used mobile phases in reversed-phase high-performance liquid chromatography (RP-HPLC) are hydro-organic mixtures of an aqueous buffer and an organic modifier. The addition of this organic solvent to buffered aqueous solutions involves a variation of the buffer properties (pH and buffer capacity). In this paper, the pH variation is studied for acetic acid-acetate, phosphoric acid-dihydrogenphosphate-hydrogenphosphate, citric acid-dihydrogencitrate-citrate, and ammonium-ammonia buffers. The proposed equations allow pH estimation of acetonitrile-water buffered mobile phases up to 60% (v/v) of organic modifier and initial aqueous buffer concentrations between 0.001 and 0.1 mol L(-1), from the initial aqueous pH. The estimated pH variation of the mobile phase and the pKa variation of the analytes allow us to predict the degree of ionisation of the analytes and from this and analyte hydrophobicities, to interpret the relative retention and separation of analyte mixtures.
Subirats
Xavier
X
Departament de Química Analítica, Universitat de Barcelona, Diagonal 647, E-08028 Barcelona, Spain.
Bosch
Elisabeth
E
Rosés
Martí
M
D000097:Acetonitriles / Q000737:chemistry*; D002021:Buffers; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D007477:Ions
D000097:Acetonitriles; D002021:Buffers; D007477:Ions; C032159:acetonitrile