Determination of pKa Values of Oxocholanoic Acids by Potentiometric Titration

The pK a and the maximum solubility values of cholic, deoxycholic salts and their oxo-derivatives have been measured by the method of potentiometric titration. In the monomer phase (under the critical micellar concentration, CMC), the bile salts have different pK a values, as a result of their structural differences (the number of hydroxyl and oxo groups in the steroid skeleton) and different hydration properties of the acid anions. In the micellar phase (above the CMC), the bile salts have higher pK a values than in the monomer phase (under the CMC). This increase in the pK a values is greater in more hydrophobic bile salts (cholate and deoxycholate), than in less hydrophobic oxo derivatives, which can be explained by the different aggregation numbers of these bile salts. The oxo-derivatives are more likely to form dimeric micelles, where the carboxylic groups are situated on the two sides of the micelle, not causing any electrostatic repulsion. In the more hydrophobic bile salts, aggregation numbers are higher, which causes electrostatic repulsion of the nearby situated carboxylic anions and consequential protonation of these anions (which leads to the stabilization of the micelle). The maximum solubility values are higher for the oxo-derivatives. If the steroid skeleton of the bile salt is more hydrophobic, the capacity to solubilize the unionized bile acid is higher, i.e. a smaller amount of the bile acid anion is needed for the solubilization of the bile acid monomer. The oxo-derivatives are less hydrophobic, but alongside their hydrophobicity, the structure of the micelle determines the solubilization capacities.