QUANTITATIVE DETERMINATION OF BILE SALTS IN BILE USING THIN-LAYER CHROMATOGRAPHY AND 3ALPHA-HYDROXYSTEROID DEHYDROGENASE

A method is described for the quantitative determination of bile salts in human bile using 3α-hydroxysteroid dehydrogenase and the spectrophotometric determination of NADH, The optimum conditions for the reaction were assessed and found to be pH 10, a temperature of 26° and a reaction time of 40 min. Other components of bile, including cholesterol, did not interfere with the reaction. With this procedure it is possible to determine bile salt concentrations ranging from 10-80 mg/100 ml. The method is reproducible and can be extended to include individual bile salts if these are first separated by thin-layer chromatography. © 1969.