Determination of acid dissociation constants of flavin analogues by capillary zone electrophoresis

Abstract

Acid dissociation constants (pK(a)) of nine kinds of flavin analogues as molecular catalyst candidates were determined by CZE. Although some of the analogues are instable and degradable under the light exposure or in alkaline aqueous solutions, the effective electrophoretic mobility of the flavin analogue of interest has been measured with the residual substance. The pK(a) values of the flavin analogues were analyzed through the changes in the effective electrophoretic mobility with varying pH of the separation buffer. One or two steps pK(a) values were determined by the analysis. One of the degraded species from the flavin analogues, lumichrome, was also detected in the CZE analysis, and its pK(a) values were also determined. While coexisting impurities generated over the storage conditions were found in some analogues, the pK(a) values of the target analogues were successfully determined with the help of the CZE separations. A pressure-assisted CZE was utilized for the determination or the estimation of the pK(a) values of such analogues as possessing carboxylic acid moiety.