2007 Fiscal Year Final Research Report Summary
Development of precise micro sensor of urinary protein and glucose using ion associate formation with anionic dye
| Project/Area Number |
17550087
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Analytical chemistry
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| Research Institution | Aichi Institute of Technology |
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Principal Investigator |
SAKAI Tadao Aichi Institute of Technology, Department of Applied Chemistry, Faculty of Engineering, Professor (30076038)
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| Co-Investigator(Kenkyū-buntansha) |
TESHIMA Norio Aichi Institute of Technology, Department of Applied Chemistry, Faculty of Engineering, Associate Professor (30292501)
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| Project Period (FY) |
2005 – 2007
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| Keywords | protein / visual detection / flow injection analysis / anionic dye / sequential injection analysis / glucose / diabetic disease / automated screening |
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| Research Abstract |
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 hour
3) 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
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