The concentrations of 3 beta-hydroxy-5-cholestenoic acid, 3 beta,7 alpha-dihydroxy-5-cholestenoic acid, and 7 alpha-hydroxy-3-oxo-4-cholestenoic acid were determined in plasma from patients with different liver diseases and compared with those of unconjugated and conjugated C24 bile acids. The levels of the cholestenoic acids were similar in patients with extrahepatic cholestasis and in controls (median concentration 153 and 162 ng/ml, respectively), whereas significantly elevated levels were found in plasma from patients with primary biliary cirrhosis (median concentration 298 ng/ml) and alcoholic liver cirrhosis (median concentration 262 ng/ml). As expected, conjugated C24 bile acids were elevated in most patients whereas the corresponding unconjugated compounds were low in cholestasis and elevated in alcoholic liver cirrhosis. The levels of the individual C27 acids were usually positively correlated to each other and also to the levels of conjugated C24 bile acids in plasma from patients with liver cirrhosis. In contrast, there was no correlation between the levels of C27 acids and conjugated bile acids in patients with extrahepatic cholestasis. The levels of unconjugated C24 bile acids were not correlated to C27 acids or conjugated bile acids in any of the groups. The results indicate that there is a close metabolic relationship between the individual C27 acids, that they do not participate in an enterohepatic circulation, and that the liver is important for their elimination/metabolism.