The solubility of drugs in the gastrointestinal tract is very challenging to simulate with artificial media due to the high complexity of human intestinal fluid (HIF). In particular, bile salt composition, pH and buffer capacity are very important characteristics of HIF, since they determine the solubility of drugs in-vivo. In this study, we have measured the concentrations of individual bile salts in human intestinal fluids (n=6) collected from two different locations (duodenum and jejunum) in the fasted state. Total bile salt concentrations ranged from 570 to 5137 μm in the duodenum and from 829 to 5470 μm in the jejunum. The following rank order of relative bile salt concentration in duodenum was observed: taurocholic acid > glycocholate ≥ glycochenodeoxycholate > glycodeoxycholate > taurochenodeoxycholate > taurodeoxycholate. Cholic acid, tauroursodeoxycholate, chenodeoxycholic acid, and deoxycholic acid represented less than 1% of bile salts present in the samples. Ursodeoxycholate could not be detected in HIF. No statistically significant difference between bile salt composition of duodenal and jejunal aspirates was observed. The buffer capacity of HIF was compared with other media commonly used for solubility/dissolution determinations, indicating a relatively low buffer capacity of HIF (4-13 mmol L-1/pH). This low buffer capacity was reflected in the change in pH (between 4 and 9.5) that occurred in HIF after addition of model compounds covering a broad pKa range. Interindividual variability in pH, buffer capacity and bile salt contents of HIF will contribute to differences in the rate and extent of absorption of compounds for which dissolution/solubility is the rate limiting step. The variability observed warrants further research to explore the impact of intraluminal conditions on drug solubility.