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

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下反应形成蓝色离子缔合物。通过测量625nm处的吸光度，可以在0- 20ppm范围内测定BSA。LOD为0.16 ppm BSA。此外，在没有TX-100的情况下，开发了一种基于tbpb结合的BSA在膜过滤器上收集后颜色变化的可视化方法。目测法可在0-9 ppm bsa范围内对一系列MF进行标准色校正图。2)采用流动注射分析法（FIA），以四溴酚酞乙酯（TBPEH）和Triton X-100为检测试剂，在pH 3.0条件下对患者尿液中的蛋白质进行了灵敏、快速、准确的测定。该检测系统基于Triton X-100形成的胶束中人血清白蛋白（HAS）与TBPEH之间的离子缔合形成。校正曲线在0.15 ~ 2mg/dL范围内呈线性关系。610nm处LOD （3S/N）为0.05mg/dL。采用分光光度法连续测定蛋白质和葡萄糖，建立了一种新的连续注射（SI）系统。蛋白质测定是基于蛋白质在Triton X-100存在下，pH值为3.2时与四溴酚烯丙烯乙酯（TBPE）的离子结合。在607nm处监测蓝色产品的吸光度。对于葡萄糖，过氧化氢是在葡萄糖氧化酶存在下由葡萄糖氧化产生的，葡萄糖氧化酶固定在装在小柱中的玻璃珠上，用铁催化对茴香胺的氧化反应来监测，形成红色的产物（520nm）。SI程序在葡萄糖氧化的潜伏期进行蛋白质测定时具有优势。人血清白蛋白在10mg/dL范围内线性检测，检出限（3S/N）为0.3mg/dL；葡萄糖在12.5mg/dL范围内线性检测，LOD为0.08mg/dL。该自动化系统已被证明用于糖尿病患者尿液样本中蛋白质和葡萄糖的连续测定。该开发的SI系统是糖尿病诊断筛选的另一种自动化方法。少

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