Bilirubin, the yellow-orange tetrapyrrole pigment of jaundice, is essentially insoluble in pure water, but is much more soluble in solutions of bile salts such as sodium taurocholate. The biophysical chemistry of bilirubin in bile salt solutions is affected by changes in the pH of the solution in the range 5-9, suggesting that interactions with bile salt molecules and micelles may alter the acidity of the pigment. We have examined this possibility by determining the apparent pKa values for a series of carboxyl 13C-enriched model compounds, including the bilirubin analog mesobilirubin XIIIalpha, in solutions of sodium taurocholate and sodium taurodeoxycholate. Apparent pKa values were determined by 13C NMR titrations in dimethyl sulfoxide-water mixtures. The results show that the acidity of all compounds is decreased, or pKa increased, in micellar bile salt solution relative to pure water and that the effect is greatest for the larger, less water-soluble compounds. We have proposed a model to explain these results and discussed the implications of these findings for the biophysical chemistry of bilirubin in bile.