Lipopolysaccharide (LPS) was assayed in 78 samples of human common-duct bile, obtained at endoscopic retrograde cholangiography. The LPS was assayed by a chromogenic limulus amebocyte lysate (LAL) test, after dilution of bile samples in heparinized plasma and inactivation of inhibitors. The assay was not influenced by other biliary constituents, as demonstrated by the recovery of standards. Bile pigments did not influence the results. The LAL test was positive in 60 of the samples, 59 of which had a positive culture for gram-negative bacteria or Candida sp. The levels of LPS were significantly correlated to the total number of bacteria (n = 16, R = 0.55, p less than 0.05). The median LPS level was 35,250 ng/l and showed a very large variation (140 ng/l to 27.8 mg/l). In four of the samples gram-negative bacteria were present, but no LPS could be detected. The study demonstrates the presence of LPS in great quantities in human bile and supports the feasibility of using the LAL test on bile samples. The presence of LPS (within the detection limit) appears to be associated with local microbial colonization.