Differentiation of isomeric metabolites of carbamazepine based on acid-base properties; Experimental vs theoretical approach

Abstract

Metabolism of carbamazepine is complex and leads to the three isomeric derivatives whose occurrence is dependent on the type of sample material. Their unambiguous differentiation is overall important. In this work, the qualitative analysis of 2-hydroxycarbamazepine, 3-hydroxycrbamazepine and carbamazepine-10,11-epoxide was attempted for the first time using capillary zone electrophoresis, based on the models linking electrophoretic mobility with pKa value determining the acidity of the hydroxyl groups. For this purpose, pKa values ​​were determined using electrophoretic and theoretical methods, and then the compliance of the obtained mobility models with the measured values ​​was analyzed. Despite the slight difference in acidity ​​(0.3-0.4 pH unit), the obtained results prove that the correct identification of the metabolites under consideration, and reliable prediction of the selectivity of their separation, are possible on the basis of experimentally determined pKa values, even with highly simplified methods assuming the lack of certain data. However, it is important to choose the optimal pH value, which should be close to pKa. On the other hand, worse results were obtained for the theoretically determined mobilities, which differed significantly from the experimental values. An attempt was also made to explain the acidity of hydroxycarbamazepines and the associated thermodynamic parameters - deprotonation enthalpy and entropy, with respect to their structure. The lack of intramolecular hydrogen bonds and the significant contribution of entropic effects stabilizing the protonated form seems to be significant. The higher pKa value for CBZ-2-OH probably results from the stronger effect of the energetically unfavorable organization of solvent dipoles due to ionization.