Tauroursodeoxycholate (TUDC), a relatively hydrophilic bile salt, reduces disruption of cholesterol-rich membranes by more hydrophobic bile salts such as taurocholate (TC), taurochenodeoxycholate (TCDC), or taurodeoxycholate (TDC). We examined the interactions of these bile salts in adsorption to large unilamellar vesicles to determine whether TUDC may stabilize membranes by preventing adsorption of more toxic bile salts. Fractional adsorption was quantified by rapid ultrafiltration. Adsorption coefficient Ai was defined for each bile salt i as ([bound i]/[free i])/[lecithin]. Affinity of different bile salts for lecithin vesicles varied with their relative hydrophobicity, increasing in the order TUDC < TC << TCDC < or = TDC. Ai of each bile salt fell with its accumulation on membranes, reaching a minimum at bound bile salt/lecithin mole ratio (B:L) between 0.05 and 0.1, then increasing with formation of higher-affinity mixed micelles. Inclusion of cholesterol in vesicles reduced Ai of all bile salts. In heterologous binding studies at submicellar concentrations, Ai of each bile salt varied with total B:L but was independent of the specific bile salts present on the membrane. Addition of TUDC to TDC reduced binding of TDC to membranes only slightly and lowered the threshold TDC concentration associated with transition to mixed micelles. However, above this threshold, TUDC markedly altered the adsorption of TDC to lecithin-containing phases. We conclude that TUDC does not directly stabilize membranes; rather, reduced permeabilization and dissolution of cholesterol-rich membranes after addition of TUDC to TDC may result from effects on the formation and structure of simple and mixed micelles.