A NEW ANALYTICAL METHOD OF INDIVIDUAL BILE ACIDS USING HIGH PERFORMANCE LIQUID CHROMATOGRAPHY AND IMMOBILIZED 3α-HYDROXYSTEROID DEHYDROGENASE IN COLUMN FORM

Abstract

Abstract We studied the use of immobilized enzyme, covalently bound to aminopropyl-CPG, to the analysis on free and conjugated individual bile acids. A microcolumn with the immobilized 3α-hydroxysteroid dehydrogenase was prepared and used in a system of high performance liquid chromatography (HPLC). Reduced NAD+ produced from bile acid by this enzyme was fluorimetrically determined with fluorometer. As the result, the free, glycine- and taurine-conjugated individual bile acids which were separated by the HPLC were determined on high sensitivity and specificity. Clinical trials were done for the serum of patients with hepatobiliary disease.