High-Performance Liquid Chromatography of Bile Acids Effect of Hydroxyl Groups at C-3, 6 7 and 12 on Bile Acid Mobility

Abstract

The high-performance liquid chromatographic characteristics of a number of bile acids with hydroxyl groups at C-3, 6, 7 and 12 positions are reported. Using Nova-pak C₁₈ reversed-phase columns and mobile phase consisting of acetonitrile/water/methanol/acetic acid mixtures, it was found; that compounds with β -hydroxyl groups were eluted much earlier than those with α-hydroxyl groups. Introduction of a hydroxyl group caused a marked increase in polarity of the bile acid, however, the effect was not well defined in bile acids with vicinal glycol system at C-6,7. The retention volumes of the various bile acids were reproducible and can be useful for characterization of bile acids in biological fluids.