Abstract: Isolation,quantitative and qualitative analysis of free and conjugated bile acids might be carried out by combined use of thin layer technique,photo-density meter and micro-computer thin layer scanning.137 samples of human bile were unalyzed by using iso-Octane 10:Ethyl Acetate 5:Acetic Acid glacial 1.5:n-Bntanol 1.5,for free bile acids and Chloroform 65:Methanol 20:Acetic Acid glacial 10:Water 5 for conjugated bile acids.The results showed that the method used in this test is a simple and stable for the analysis of bile acids.