Abstract An improved method of preparation of 5α-anhydrocyprinol from large quantities of carp bile is reported. The following materials were obtained by this method: 5a-anhydrocyprinol, 5α-cholestane-3α, 7α, 12α, 25-tetrol, 5α-cholestane3α, 7α, 12α, 26-tetrol, an unidentified sterol from the neutral lipid fraction, allocholic acid and allochenodeoxycholic acid from the bile acid fraction. Yields of 5α-anhydrocyprinol and bile acids vary in different batches. The identity and purity of these materials were verified by thin-layer chromatography, gas liquid chromatography and gas chromatography-mass spectrometry.