固定化酵素を組み合わせた高速液体クロマトグラフィーによる簡易高感度血中胆汁酸分画測定法の開発とその臨床応用

概要

論文の詳細を見る
We have developed a highly sensitive and simultaneous method for the determination 6f individual 3α-hydroxy bile acids in serum using high-performance liquid chromatography (H. P. L. C.) combined with immobilized enzyme. Amberlite XAD-7 was used for the extraction of bile acids. Samples were analized on the column (W-260) made of copolymer which was tolerable even in severe alkalinized condition. Immobilized enzyme colume was performed of cellulose particles fixed with 3α-hydroxy steroid dehydrogenase according to the procedure described by S. C. March. The eluent of H. P. L. C. which consists of ammonium phosphate solution and acetonitrile was flowed into the immobilized enzyme column with β-NAD solution (pH 9.5). Enzymatic reaction was measured by fluorometer through the immobilized enzyme column. The linear correlated standard curves were obtained. The recovery ratios and reproducibility of intra and inter assays were satisfactory. The sensitivity limit of each bile acid was within 0.08 μmol/l.
神戸大学の論文