Retention of ionizable compounds on high-performance liquid chromatography. VII. Characterization of the retention of ionic solutes in a C18 column by mass spectrometry with electrospray ionization.
The elution of ions from a C18 column with mobile phases containing methanol (60%, v/v) and aqueous buffers is studied by mass spectrometry. It is demonstrated that the anions are excluded from the stationary phase by the ionized silanols. However, the ionized silanols interact strongly with cations, which are retained in the column. These cations are later eluted from the column by ion exchange with the cations present in the pH buffered mobile phase. The size of the ions, the mobile phase cation concentration and the mobile phase pH are the main parameters that affect elution of the retained cations. It is also demonstrated that there are at least two different types of ionizable silanols, with different acidities, that contribute to the retention of cations. An estimate of the pKa values of these two groups of silanols in 60% methanol is given.
Departament de Química Analítica, Universitat de Barcelona, Spain. marti@apolo.qui.ub.es
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Rosés
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Oumada
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Bosch
D002021:Buffers; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002852:Chromatography, Ion Exchange / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D007477:Ions; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
D002021:Buffers; D007477:Ions