Separatory determination of bile acid glucuronides in human bile.

概要

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A precise and sensitive method for the separation and determination of bile acid glucuronides in human bile is described. Bile acids in human bile were hydrolyzed by cholylglycine hydrolase and separated into the unconjugated, glucuronide, and sulfate fractions by ion-exchange chromatography on a newly developed lipophilic gel, piperidinohydroxypropyl Sephadex LH-20. Bile acid glucuronides were then hydrolyzed by the β-glucuronidase preparation from E. coli, and the free bile acids were determined by the enzymatic method. This new assay system requires only 1 to 10μl of human bile and is applicable to the quantitation of bile acid glucuronides in bile with satisfactory precision and reproducibility.Using this method, the bile acid glucuronide levels in gallbladder bile were measured in clinical cases. The amounts of glucuronides relative to total bile acids were 1.1±0.4% and 1.8±0.6% in patients with and without cholecystolithiasis, respectively. The composition ratio of glucuronides in lithocholates was larger than in total bile acids, while that in cholates was smaller.
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