Abstract
Many trials have been made with HPLC, on bile acids determination,and introduced, the enzymatic method uslng immobilized 3α-hydroxysteroid dehydrogenase (3α-HSD) as an effective method to detect of bile acidsand its conlugates, for post column reac-tion technique,in this paper.
DuPont Zorbax C8 were used as the LC column which was housed in column oven to keep at 55°, and used the alcohol-phosphate buffer mixture as the mobile phase.
The enzyme were immobilized on aminated porous glass beads with glutaraldehyde, and packed in stainless steel column. The reaction reagent contaming NAD and DTT (dithio threitol) in phosphate buffer was mixied just before the immobilied enzyme column.
The fluorescence intensity of the NADH thus produced were measured, in the presence of bile acids as substrate.
Some results are described on some conditions for separation and determination in this paper.
