Development of a System Containing a Flow-Through Electrochemiluminescent Detector and Its Application to High-Sensitive Determination

流通式电化学发光检测器系统的研制及其在高灵敏测定中的应用

• 批准号: 11650835
• 负责人: KUSU Fumiyo
• 金额: $ 2.11万
• 依托单位: Tokyo University of Pharmacy and Life Science
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11650835/
• 关键词: Electrochemiluminescence; Flow-through cell; High-sensitive determination; Luminol; Hydrogen peroxide; Glucose; Uric acid; Immobilized enzyme

A system containing a flow-through electrochemiluminescent detector was developed for high-sensitive determination of biologically important substances. The main contents of the results are as follows :1. Construction of the electrochemiluminescent detection systemThe system developed was fabricated from a carrier solution reservoir, degasser, pump, injector of test solution, electrochemiluminescent cell and phototube. The experimental conditions, such as the composition of carrier solution, the size and length of Teflon tube for the flow line and the cell design, were optimized to give reproducible data with high sensitivity.2. High-sensitive determination of hydrogen peroxideDetermination of hydrogen peroxide was carried out using luminol, an electrochemiluminescent reagent. Hydrogen peroxide amount ranging from 59 fmol to 59 nmol could be determined using 5.0 μM luminol in a phosphate buffer (pH 7.6) at a flow rate of 0.4 ml/min and an applied potential of 1.1 V vs. Ag/AgCl.3. Analysis of serumAs examples of analysis of biological samples, glucose and uric acid in serum were determined. Glucose oxidase or uricase were immobilized on Chitopearl filled in an enzyme reactor tube located between the injector and detector. The present system should be applicable to a simple method for determining biologically important substances with high sensitivity.

开发了一种包含流通式电化学发光检测器的系统，用于高灵敏度测定生物重要物质。研究结果的主要内容如下：1。电致发光检测系统的构建该系统由载液池、脱气器、泵、试液进样器、电致发光池和光电管组成。对实验条件如载液组成、流动线所用聚四氟乙烯管的尺寸和长度以及池的设计进行了优化，得到了重复性好、灵敏度高的数据.高灵敏度测定过氧化氢采用鲁米诺电致发光法测定过氧化氢。使用5.0 μM鲁米诺的磷酸盐缓冲液（pH 7.6），流速为0.4 ml/min，外加电位为1.1 V（相对于Ag/AgCl），可测定59 fmol至59 nmol范围内的过氧化氢量。3.血清分析作为生物样品分析的实例，测定血清中的葡萄糖和尿酸。将葡萄糖氧化酶或尿酸酶固定在填充在位于进样器和检测器之间的酶反应管中的壳聚糖上。本系统应适用于一个简单的方法，用于确定具有高灵敏度的生物重要物质。

项目成果

K.Arai,T.Sato,H.Tamanouchi,M.Ohtomo,K.Takamura,T.Sueoka,and F.Kusu: "Electrolysis of 2-Methyl-1,4-naphthoquinone and 3,5-Di-tert-butyl-1,2-benzoquinone Using the Flow-Cell System with a Column Electrode"Electrochim.Acta.,. 45. 3029-3034 (2000)

K.Arai、T.Sato、H.Tamanouchi、M.Ohtomo、K.Takamura、T.Sueoka 和 F.Kusu：“2-甲基-1,4-萘醌和 3,5-二叔-的电解”

K.Arai, T.Sato, H.Tamanouchi, M.Ohtomo, K.Takamura, T.Sueoka, and F.Kusu: "Electrolysis of 2-Methyl-1,4-naphthoquinone and 3,5-Di-tert-butyl-1,2-benzoquinone Using the Flow-Cell System with a Column Electrode"Electrochim.Acta. 45. 3029-3034 (2000)

K.Arai、T.Sato、H.Tamanouchi、M.Ohtomo、K.Takamura、T.Sueoka 和 F.Kusu：“2-甲基-1,4-萘醌和 3,5-二叔-的电解”

K. Arai,K. Takahashi,F. Kusu: "An Electrochemiluminescence flow-through cell and its applications to sensitive immunoassay using N-(aminobutyl)-N-ethylisoluminol"Anal.Chem.. 71. 2237-2240 (1999)

K.荒井，K.

K.Arai,T.Sato,H.Tamanouchi,M.Ohtomo,K.Takamura,T.Sucoka,and F.Kusu: "Electrolysis of 2-Methyl-1,4-naphthoquinone and 3,5-Di-tert-butyl-1,2-benzoquinone Using the Flow-Cell System with a Column Electrode"Electrochim.Acta.,. 45. 3029-3034 (2000)

K.Arai、T.Sato、H.Tamanouchi、M.Ohtomo、K.Takamura、T.Sucoka 和 F.Kusu：“2-甲基-1,4-萘醌和 3,5-二叔-的电解”