An acid dissociation constant of tetrabromophenolphthalein ethyl ester (TBPE) was determined through the measurement of electrophoretic mobility by capillary zone electrophoresis (CZE). Although TBPE is degradable in acidic pH region and it gradually degraded at pH conditions around and below its pKa values in the time scale of the CZE measurement, equilibrium species of interest were detected as a peak-shaped signal with tailing of the degraded species. Changes in electrophoretic mobility of the equilibrium species of TBPE were analyzed at its detectable pH range in the presence of phenol red as a mobility standard. An acid dissociation constant of 3.47 ± 0.06 (pKa, I = 0.01, 25 C, standard error) was determined for TBPE.
2Here, we report a novel method consisting of capillary electrophoretic separation followed by two-color laser-induced fluorescence detection with postcolumn derivatization.
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