血中胆汁酸分画の臨床的意義に関する研究第1編高速液体クロマトグラフィーと酵素螢光測定法を組合わせた血中胆汁酸分画の高速度同時測定法の開発と基礎的検討

概要

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A highly sensitive and simultaneous method for quantitive determination of individual 3α-hydroxy bile acids in serum using high-performance liquid chromatography (HPLC) combined with enzymatic fluorometric measurement was described. The separation of thirteen components of bile acids, free and glycine-or taurine-conjugated bile acids, was satisfactorily attained by HPLC on a μ-Bondapak phenyl column, using 0.3% ammonium carbonate mixed with acetonitrile as an eluent and 3α-hydroxysteroid dehydrogenase as a reagent. A linier relationship was observed between the peak hight and the known amount prepared as standard of each bile acid. The recovery rate of each bile acid added to serum wasapproxymatry 81.7 to 106.6%. The CV values were given in the range of 0.68 to 10.0% by inter-assay and 0.70 to 9.09% by intra-assay. The sensitivity limit of each bile acid by this method was in the range of 6.3-31.0 ng on the column. The concentration of bilirubin ranging 10-40mg/dl did not interfere with this method of determination. The present new method made it possible to separate the components of bile acids simultaneously within comparatively short time and to determine free and conjugated bile acids using only 1 ml of human serum. This method provides more information on the metabolism of bile acids and may be utilized for clinical diagnosis of hepatobiliary diseases.
神戸大学の論文