使用固定化吡喃糖氧化酶反应器通过阴离子交换色谱法同时测定血清中的葡萄糖和1-脱氧葡萄糖。

Simultaneous determination of glucose and 1-deoxyglucose in serum by anion-exchange chromatography with an immobilized pyranose oxidase reactor.

作者信息

Kiba N, Goto Y, Furusawa M

机构信息

Department of Applied Chemistry and Biotechnology, Faculty of Engineering, Yamanashi University, Kofu, Japan.

出版信息

J Chromatogr. 1993 Oct 22;620(1):9-13. doi: 10.1016/0378-4347(93)80045-6.

DOI:10.1016/0378-4347(93)80045-6

PMID:8106597

Abstract

A liquid chromatographic system for the determination of glucose and 1-deoxyglucose in serum using an immobilized pyranose oxidase reactor and chemiluminescence detection is described. Separation was achieved on a TSK gel SAX column (2 cm x 4 mm I.D.) with an eluent of 0.1 M sodium hydroxide solution. Serum was diluted 30-fold with 0.1 M NaOH. Sample solution (50 microliters) was injected into the system. The hydrogen peroxide produced was detected by measuring the chemiluminescence emitted on admixing with luminol and potassium hexacyanoferrate (III). The calibration graphs were linear from 1 to 600 microM glucose and from 0.6 to 400 microM 1-deoxyglucose; the detection limits for glucose and 1-deoxyglucose were 0.5 and 0.3 microM, respectively. The sample throughput was 10/h. The immobilized enzyme reactor was stable for at least 2 months.

摘要

描述了一种使用固定化吡喃糖氧化酶反应器和化学发光检测法测定血清中葡萄糖和1-脱氧葡萄糖的液相色谱系统。在TSK凝胶SAX柱（2 cm×4 mm内径）上，以0.1 M氢氧化钠溶液作为洗脱液实现分离。血清用0.1 M NaOH稀释30倍。将50微升样品溶液注入系统。通过测量与鲁米诺和六氰合铁（III）酸钾混合时发出的化学发光来检测产生的过氧化氢。校准曲线在1至600 microM葡萄糖和0.6至400 microM 1-脱氧葡萄糖范围内呈线性；葡萄糖和1-脱氧葡萄糖的检测限分别为0.5和0.3 microM。样品通量为每小时10个。固定化酶反应器至少稳定2个月。