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流动注射法结合固定化谷氨酸氧化酶和谷氨酸脱氢酶反应器测定人血清和大鼠脑样本中的谷氨酸。

Flow injection determination of glutamate in human serum and rat brain samples with immobilized glutamate oxidase and glutamate dehydrogenase reactors.

作者信息

Stalikas C D, Karayannis M I, Tzouwara-Karayanni S M

机构信息

Department of Chemistry, University of Ioannina, Greece.

出版信息

Eur J Clin Chem Clin Biochem. 1994 Oct;32(10):767-72. doi: 10.1515/cclm.1994.32.10.767.

Abstract

Two methods are proposed for the determination of regional concentrations of glutamate in the rat brain as well as in human serum. Glutamate oxidase was immobilized on non-porous glass beads and glutamate dehydrogenase was immobilized on glass derivatives. These supports were employed for the construction of Single Bead String Reactors and Packed Bed Reactors, respectively, which in turn were linked to Flow Injection Analysis systems with either photometric or fluorometric detection. Analytical working curves are linear in the range 1-200 mumol/l for packed bed reactors and 10-500 mmol/l for single bead string reactors. The samples were pretreated depending on their origin and the applied measuring system. Optimal dilution factors were established for the two techniques. Optimal dilution ratios were established and the influence of several added substances was investigated. Recovery and method comparison studies including high performance liquid chromatography verified the accuracy of the proposed methods. Results from within-day and between-day measurements gave relative standard deviations of 4.7 and 5.9% for serum samples and 2.5 and 4.0% for brain samples, respectively.

摘要

本文提出了两种测定大鼠脑和人血清中谷氨酸区域浓度的方法。谷氨酸氧化酶固定在无孔玻璃珠上,谷氨酸脱氢酶固定在玻璃衍生物上。这些载体分别用于构建单珠串反应器和填充床反应器,进而与具有光度或荧光检测的流动注射分析系统相连。填充床反应器的分析工作曲线在1-200μmol/l范围内呈线性,单珠串反应器的分析工作曲线在10-500mmol/l范围内呈线性。根据样品来源和应用的测量系统对样品进行预处理。确定了两种技术的最佳稀释因子。确定了最佳稀释比例,并研究了几种添加物质的影响。包括高效液相色谱在内的回收率和方法比较研究验证了所提方法的准确性。日内和日间测量结果表明,血清样品的相对标准偏差分别为4.7%和5.9%,脑样品的相对标准偏差分别为2.5%和4.0%。

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