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Development of precise micro sensor of urinary protein and glucose using ion associate formation with anionic dye

利用与阴离子染料形成离子缔合物开发尿蛋白和葡萄糖的精确微传感器

基本信息

批准号：
17550087
负责人：
SAKAI Tadao
金额：
$ 2.42万
依托单位：
Aichi Institute of Technology
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2005
资助国家：
日本
起止时间：
2005 至 2007
项目状态：
已结题

项目摘要

1) Tetrabromophenol blue (HTBPB) reacted with bovine serum albumin (BSA) to form a blue ion associate at pH 3.2 in the presence of Triton X-100. BSA could be determined in the range of 0-20 ppm by measurring the absorbance at 625nm. The LOD was 0.16 ppm BSA. Moreovere, a visual method based on the color changes was developed after the collection of TBPB-binding BSA on the membrane filter in the absence of TX-100. In the visual method, the standard color calibration chart on a series of MF could be made in the range of 0-9 ppm BSA.2) A sensitive, rapid and accurate determination of protein in patient urine was carried out by flow injection analysis (FIA) using tetrabromophenolphthalein ethyl ester (TBPEH) and Triton X-100 at pH 3.0. The detection system was based on the ion association formation between human serum albumin (HAS) and TBPEH in the micelle formed by Triton X-100. The calibration graph was linear in the range of 0.15-2mg/dL. The LOD (3S/N) was 0.05mg/dL at 610nm. The sample … More throughput was 30 per hour3) A new sequential injection (SI) system with spectrophotometric detections has been developed for successive determination of protein and glucose. The protein assay is based on ion-association of protein with tetrabromophenolphthe\alein ethyl ester (TBPE) in the presence of Triton X-100 at pH 3.2. The blue product is monitored for absorbance at 607nm. For glucose, hydrogen peroxide, generated by the oxidation of glucose in the presence of glucose oxidase immobilized on glass beads packed in a minicolumn, is monitored using iron-catalyzed oxidation reaction of p-anisidine to form a red colored product (520nm). The SI procedure takes advantage in performing the protein assay during the incubation period for glucose oxidation. Linear ranges were up to 10mg/dL human serum albumin with a limit of detection (3S/N) of 0.3mg/dL, and up to 12.5mg/dL glucose with LOD of 0.08mg/dL Sample throughput for the whole asay of both protein and glucose is 6/h. The automated system has been demonstrated for the successive assay of protein and glucose in urine samples taken from diabetic disease patients. This developed SI system is an alternative automation for screening for diabetic diagnosis. Less

1)在Triton X-100存在下，四溴酚蓝（HTBPB）与牛血清白蛋白（BSA）在pH 3.2的条件下形成蓝色离子缔合物。在625 nm处测定吸光度，可测定0-20 ppm范围内的BSA。LOD为0.16 ppm BSA。此外，在不存在TX-100的情况下，在膜过滤器上收集TBPB结合BSA后，开发了基于颜色变化的目视方法。2）采用流动注射分析法，以四溴酚酞乙酯（TBPEH）和Triton X-100为显色剂，在pH3.0条件下，对病人尿中蛋白质进行了灵敏、快速、准确的测定。该检测体系基于人血清白蛋白（HAS）与TBPEH在Triton X-100形成的胶束中形成离子缔合物。线性范围为0.15- 2 mg/dL。在610 nm处的LOD（3S/N）为0.05 mg/dL。样品 ...更多信息 3）建立了一种新的连续进样分光光度法测定蛋白质和葡萄糖的方法。蛋白质测定基于在pH 3.2下在Triton X-100存在下蛋白质与四溴苯酚酞菁乙酯（TBPE）的离子缔合。监测蓝色产物在607 nm处的吸光度。对于葡萄糖，使用铁催化的对茴香胺氧化反应监测葡萄糖在微型柱中固定在玻璃珠上的葡萄糖氧化酶存在下氧化产生的过氧化氢，以形成红色产物（520 nm）。SI程序在葡萄糖氧化孵育期间进行蛋白质测定中具有优势。线性范围：人血清白蛋白10 mg/dL，检出限（3S/N）0.3mg/dL;葡萄糖12.5mg/dL，检出限（LOD）0.08mg/dL。该自动化系统已被证明用于连续测定取自糖尿病患者的尿样中的蛋白质和葡萄糖。这个开发的SI系统是一个替代的自动化筛查糖尿病诊断。少