Fast high-throughput method for the determination of acidity constants by capillary electrophoresis. II. Acidic internal standards
Cabot, JM and Fuguet, E and Rafols, C and Roses, M, Fast high-throughput method for the determination of acidity constants by capillary electrophoresis. II. Acidic internal standards, Journal of Chromatography A, 1217 pp. 8340-8345. ISSN 0021-9673 (2010) [Refereed Article]
A fast method for the determination of acidity constants by CZE has been recently developed. This method is based on the use of an internal standard of pKa similar to that of the analyte. In this paper we establish the reference pKa values of a set of 24 monoprotic neutral acids of varied structure that we propose as internal standards. These compounds cover the most usual working pH range in CZE and facilitate the selection of adequate internal standards for a given determination. The reference pKa values of the acids have been established by the own internal standard method, i.e. from the mobility differences between different acids of similar pKa in the same pH buffers. The determined pKa values have been contrasted to the literature pKa values and confirmed by determination of the pKa values of some acids of the set by the classical CE method. Some systematic deviations of mobilities have been observed in NaOH buffer in reference to the other used buffers, overcoming the use of NaOH in the classical CE method. However, the deviations affect in a similar degree to the test compounds and internal standards allowing thus, the use of NaOH buffer in the internal standard method. This fact demonstrates the better performance of the internal standard method over the classical method to correct mobility deviations, which together with its fastness makes it an interesting method for the routine determination of accurate pKa values of new pharmaceutical drugs and drug precursors.