Development Of Chemiluminometric μ-Flow-Through Detectors

化学发光μ流过式检测器的开发

• 批准号: 13555231
• 负责人: KIBA Nobutoshi
• 金额: $ 3.39万
• 依托单位: University of Yamanashi
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13555231/
• 关键词: FLOW SYSTEM; MICROFLUID SYSTEM; CHEMILUMINESCENCE; DETECTOR; MINIATURIZED; HYDROGEN PEROXIDE; HIGH THROUGHPUT; ENZYME; 化学発光検出; 検出器; 小型化; 過酸化水素; ハイスループット分析; 固定化酵素; マイクロTAS; 化学発光; 科学発光

A MICROMACHINED FLOW CELL (OVERALL SIZE; 25x25x1 mm^3) WAS DESIGNED FOR HIGH THROUGHPUT ANALYSIS. THE FLOW CELL CONSISTED OF A SILICON BOTTOM CHIP (0.5mm THICK) WHICH HAD A SPIRAL CHANNEL (20 TURNS), TWO PIERCED HOLES (1mm DIAMETER), AND A GLASS TOP PLATE (0.5mm THICK). THE CHANNEL (50cm LENGTH, 150μm WIDTH, 20μm DEPTH, 150μm SPACING BETWEEN CHANNELS) WAS PRODUCED BY ECHING. TW0 HOLES SERVE AS THE FLUID INLET (CENTER) AND OUTLET (SIDE). THE CHIP WAS PLACED IN FRONT OF A WINDOW OF A PHOTOMULTIPLIER TUBE. ALSO, TRANSPARENT FUSED SILICA CAPILLARY (50μm I. D., 375μm O. D.) WHICH IS COATED WITH TEFLON AF WAS USED FOR MORE RAPID ANALYSIS. THE CHIP AND CAPILLARY WERE USED AS FLOW CELLS IN A SINGLE-LINE CHEMILUMINOMETRIC FLOW INJECTION SYSTEM. PEROXIDASE (EC 1.11.1.7) WAS COVALENTLY IMMOBILIZED ON THE INNER SURFACE OF THE CHANNEL. THE SAMPLE VOLUME FOR ONE MEASUREMENT WAS 0.2μL. THE MAXIMUM SAMPLING RATE WAS 315/h AT A CARRIER SOLUTION FLOW RATE OF 10μl/min. A CALIBRATION GRAPH FOR H_20_2 WAS LINEAR FOR 5 nM -5μM. THE SYSTEM WITH μ-FLOW-THROUGH DETECTOR WAS APPLIED TO THE DETERMINATIONS OF H_20_2 IN ENVIROMENTAL SAMPLES AND OF QXIDASE SUBSTRATES IN BIOCHEMICAL SAMPLES.

微机械流动池（总尺寸；25x25x1毫米^3）被设计用于高通量分析。流动电池由一个硅底芯片（0.5毫米厚）组成，它有一个螺旋通道（20圈），两个直径1毫米的穿孔，和一个玻璃顶板（0.5毫米厚）。该通道（长50cm，宽150μm，深20μm，通道间距150μm）由ECHING公司生产。两个孔作为流体入口（中心）和出口（侧面）。该芯片被放置在光电倍增管的窗口前。此外，透明熔融石英毛细管（50μm内径，375μm外径）表面涂有聚四氟乙烯，用于更快的分析。在单线化学发光流动注射系统中，采用芯片和毛细管作为流动细胞。过氧化物酶（ec 1.11.1.7）被共价固定在通道的内表面。每次测定的进样量为0.2μl。在载体溶液流速为10μl/min时，最大采样率为315次/h。H_20_2在5 nM ~ 5μ m范围内呈线性关系。将该系统应用于环境样品中h_20_2和生化样品中q氧化酶底物的测定。

项目成果

Nobutoshi Kiba: "Flow-through Chemiluminescence Sensor Using Immobilized Oxidases for the Selective Determination of L-Glutamate in a Flow-Injection System."Analytical Sciences. 17. 929-933 (2001)

Nobutoshi Kiba：“使用固定化氧化酶的流通式化学发光传感器在流动注射系统中选择性测定 L-谷氨酸。”分析科学。

Kazue Tani: "Retention Behavior of Monosaccharides and Disaccharides on Titania."Chromatographia. 57. 409-412 (2003)

Kazue Tani：“单糖和二糖在二氧化钛上的保留行为。”色谱法。

Nobutoshi Kiba: "Chemiluminometric Sensor for Simultaneous Determination of L-Glutamate and L-Lysine with Immobilized Oxidases in a Flow …"Analytical Chemistry. 74. 1269-1274 (2002)

Nobutoshi Kiba：“用固定化氧化酶在流动中同时测定 L-谷氨酸和 L-赖氨酸的化学发光传感器……”分析化学。74。1269-1274 (2002)

Nobutoshi Kiba: "Simultaneous Determination of Choline and Acetylcholine Based on a Trienzyme Chemiluminometric Biosensor in a Single Line Flow Injection System."Analytical Sciences. 19. 1647-1651 (2003)

Nobutoshi Kiba：“基于单线流动注射系统中的三酶化学发光生物传感器同时测定胆碱和乙酰胆碱。”分析科学。

Kazue Tani: "Evaluation of Titania as an Ion-Exchanger and as a Ligand-Exchanger in HPLC."Chromatographia. 55. 33-37 (2002)

Kazue Tani：“二氧化钛作为离子交换剂和配体交换剂在 HPLC 中的评估。”色谱法。