Gallbladder bile of the bullfrog, Rana cetesbeiana was found to contain a number of minor bile acids and bile alcohols in unconjugated forms. The following bile acids and bile alcohols were identified by combined gas-liquid chromatography-mass spectrometry: cholic acid, allocholic acid, 3 alpha,7 alpha,12 alpha-trihydroxy-5 alpha-cholestan-26-oic acid, 3 alpha,7 alpha,12 alpha-trihydroxy-24-methyl-5 beta-cholestan-26-oic acid, 3 alpha,7 alpha,12 alpha,24-tetrahydroxy-5 alpha-cholestan-26-oic acid, 3 alpha,7 alpha,12 alpha,24-tetrahydroxy-5 beta-cholestan-26-oic acid, 3 alpha,7 alpha,12 alpha,26-tetrahydroxy-5 alpha-cholestan-27-oic acid, 3 alpha,7 alpha,12 alpha,26-tetrahydroxy-5 beta-cholestan-27-oic acid, 3 alpha,7 alpha,12 alpha-trihydroxy-27-nor-5 alpha-cholestan-24-one, and 3 alpha,7 alpha,12 alpha-trihydroxy-27-nor-5 beta-cholestan-24-one. These minor constituents of the bile may represent intermediates in the biosynthetic pathway of bile acids and bile alcohols in the bullfrog.