固定化酵素リアクター及び化学発光検出器を用いた高速液体クロマトグラフィーによるスクロースとグルコースの定量

概要

論文の詳細を見る
Immobilized enzymes were used as column reactors in a high-performace liquid chromatographic system for the specific detection of sucrose and glucose. Invertase (EC 3.2.1.26,Inv) and pyranose oxidase (EC 1.1.3.10,PyOD) were immobilized onto porous glass beads. The saccharides were separated on a TSKgel SCX column (30cm×7.8mm i.d.) with a 0.01M acetate buffer (pH 4.5) as a mobile phase. Inv was capable of hydrolyzing sucrose nearly quantitatively (98%) to fructose and glucose, which was oxidized to glucosone and hydrogen peroxide by PyOD. The produced hydrogen peroxide was monitored chemiluminometrically. The calibration graph was linear from 6.0×10^<-7> to 5.0×10^<-4>M sucrose or glucose. The detection limits for sucrose and glucose were 2.0×10^<-7> (3.4ng) and 3.0×10^<-7>M (2.7ng in a 50 μl injection), respectively.
社団法人日本分析化学会の論文
1993-10-05