Abstract: Equol was one of metabolites of daidzin and daidzein by intestinal bacteria decomposition. The disturbance from genistein to equol was reduced by using thin-layer chromatography (TLC) followed by high performance liquid chromatography ( HPLC) . This method solved the problem that equol could hardly be separated effectively by HPLC. Developed in a TLC solvent system of chloroform∶methanol (24∶1) , equol and genistein were observed on Rf value of 0.40 and 0.29, respectively. This new qualitative method for the analysis of urinary equol was more accurate and less disturbed than using only HPLC analysis.