Analysis of 3-sulfated and nonsulfated bile acids by one-step solvolysis and high performance liquid chromatography.

Y Hirano,F Nakayama,H Miyazaki,S Higashidate

doi:10.1016/s0022-2275(20)38586-2

Abstract

A facile solvolysis procedure of 3-sulfated bile acid was devised using trifluoroacetic acid, tetrahydrofuran, and methanol. The sulfate esters were completely solvolyzed within only 2 hr by the present method. The clinical utility of the solvolysis procedure and high performance liquid chromatography using immobilized 3 alpha-hydroxysteroid dehydrogenase was demonstrated in the analysis of bile acids in serum of patients with obstructive jaundice. The quantities of 3-sulfated bile acids were calculated from the difference in the amount of bile acids before and after solvolysis. A significantly large proportion of 3-sulfated glycochenodeoxycholic acid, i.e., 21.9 to 31.3% of total glycochenodeoxycholic acid, was found in the serum of patients with obstructive jaundice. Thus, the present method permits simultaneous quantitation of 3-sulfated as well as nonsulfated bile acids in biological samples.

Highlights

Was devised using trifluoroacetic acid, tetrahydrofuran, and MS), and radioimmunoassay (RIA)
This report describes a method of simultaneous determination of 3-sulfated and nonsulfated bile acids using the combination of a novel solvolysis procedure and HPLC with immobilized 3a-HSD, and its clinical application
Tetrahydrofuran (THF) was refluxed for 2 hr over potassium hydroxide and lithium aluminum hydride, since rigorous purification of T H F is very important for later analysis. [24''C]Glycolithocholic acid-3-sulfate was synthesized from [24-'4C]lithocholic acid in our laboratory according to the method of Tserng and Klein [5]. [11,12-3H2]Glycochenodeoxycholicacid-3-sulfate was prepared from [11,12-3H2]chenodeoxycholicacid according to the method of Parmentier and Eyssen [6]

Summary

Introduction

Was devised using trifluoroacetic acid, tetrahydrofuran, and MS), and radioimmunoassay (RIA). The present method permits and glycine- and taurine-conjugated bile acids can simultaneous quantitation of 3-sulfated as well as nonsulfated bile acids in biological samples.-Hirano, Y.,H. This report describes a method of simultaneous determination of 3-sulfated and nonsulfated bile acids using the combination of a novel solvolysis procedure and HPLC with immobilized 3a-HSD, and its clinical application.

Results

Conclusion