カラムスイッチング法を利用した高速液体クロマトグラフィーによる血清中のタンパク結合型胆汁酸の分別定量

概要

論文の詳細を見る
A simple and rapid technique for the fractional determination of bile acids bound with protein in the serum was developed by using high performance liquid chromatography with a dual-column switching system. The serum samples were directly injected onto a first column (hydroxyapatite), which was initially flushed with 1 mM phosphate buffer. Serum proteins were strongly retained on a column of hydroxyapatite, but free bile acids were not retained. The bile acids were adsorbed on a second column (Serumout-25[○!R]) and eluted onto a column of immobilized 3α-hydroxysteroid dehydrogenase (3α-HSD) with an elution solvent (CH_3CN-MeOH-30 mM ammonium acetate, 30 : 30 : 40,v/v/v). Reduced nicotinamideadenine dinucleotide was produced on the immobilized 3α-HSD column and then determined fluorometrically. Subsequently, the hydroxyapatite column was flushed with 20 mM phosphate buffer. Bile acids bound with albumin were eluted and condenced on Serumout-25. The phosphate buffer (400 mM) was finally used for the elution of bile acids bound with globulin from the hydroxyapatite column. Each condenced bile acid was eluted onto the immobilized 3α-HSD column as described above.
社団法人日本薬学会の論文
1989-11-25