A method for the quantitative determination of three main bile acids, cholic acid (CA), ursodesoxycholic acid (UDCA) and chenodesoxycholic acid (CDCA), in bear gall, drainage from bear gall and bear gallstone is described. Experimental conditions: TLC Scanner CS-910, fluorescence scanning, lambda ex 470 nm and lambda em 550 nm for CA; lambda ex 380 nm and lambda em 450 nm for UDCA and CDCA. The results showed that the contents of UDCA and CDCA in bear gall drainage were higher than those in bear gall. The method is simple, rapid and sensitive. The reproducibility is good. The average recovery is 98.4%, CV is 1.4%.