<Originals>Measurement of blood bile acid levels by high-performance liquid chromatography and its application to bile acid tolerance test

概要

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Concentrations of the free and conjugated bile acid fractions in blood were measured with a fully automated analyzing system consisting of a high-performance liquid chromatography, an immobilized enzyme (3α-HSD) column, and an automated pretreatment apparatus. This system was also applied to the exogenous bile acid tolerance test (chenodeoxycholic acid 500 mg). Three mixtures of acetonitrile and triammonium phosphate buffer with different mixing ratios and pHs were used as the solvent of the mobile phase, and passed through the separation column with a NAD solution as the reaction fluid by the step-gradient method. Reproducibility in repeated determinations, day-to-day variation, recovery rate, linearity, and correlation with colorimetric analysis were all satisfactory. The blood bile acid levels showed marked differences with pathologic conditions. The values reached a plateau at about 60 minutes after the administration in patients with liver cirrhosis while they attained a peak after 120 minutes in these with chronic hepatitis and acute hepatitis, after 90 minutes in healthy individuals. The present technique enables accurate and rapid measurement by a simple procedure. Requiring only minute amounts of samples (20-100 μl), it is also applicable to children. The method may become in clinical use with the establishment of characteristic concentration profiles in each pathologic condition.
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