| Project/Area Number |
12440207
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
分離・精製・検出法
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| Research Institution | HOKKAIDO UNVERSITY |
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Principal Investigator |
TANAKA Shunitz Hokkaido Univ., Graduate School of Environmental Earth Sci., Prof., 大学院・地球環境科学研究科, 教授 (30142194)
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| Co-Investigator(Kenkyū-buntansha) |
ITOH Shinji Hokkaido College of Pharmacy, Faulty of Pharmacy, Asso. Prof., 薬学部, 講師 (80137447)
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| Project Period (FY) |
2000 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥12,600,000 (Direct Cost: ¥12,600,000)
Fiscal Year 2001: ¥5,300,000 (Direct Cost: ¥5,300,000)
Fiscal Year 2000: ¥7,300,000 (Direct Cost: ¥7,300,000)
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| Keywords | endocrine disruptors / electrochemical screening / estrogen receptor / electroactive labeled ligand / daunomycin / 17β-estradiol / bisphenol A / electrochemical decomposition / ノニルフェノール |
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| Research Abstract |
A new electrochemical screening method for endocrine disrupting chemicals(EDCs) was developed after the fundamental investigation of the interaction of a protein and its ligand, such as avidin and biotin. To evaluate the binding ability of EDCs to the estrogen receptor(EK), 17 β-estradiol labeled with daunomycin as an electroactive compound was prepared. The electrochemical sensitivity of the prepared labeled estradiol(LE) was high, as compared with daunomycin. The interaction between LE and ER was observed by the decrease in the electrode response of LE. This decrease was found to be due to the specific binding of LE with ER. The competitive reaction between LE and 17β-estradiol for the limiting binding sites on ER produced increase in the peak current of LE. Then the concentration of competitors against LE can be obtained from the increase in the peak current. The relative standard deviation at 1 x 10^<-8> M 17 β-estradiol was about 10 % (n=7). The binding affinity between EDCs and ER were also evaluated by comparison with 17β-estradiol-ER interaction. Bisphenol A, p-nonylphenol and p,p,p'-DDT were used as a test compound for EDCs. All test compounds demonstrated some ability to bind with ER.
This electrochemical binding assay illustrates a new method for evaluating the binding capacity of EDCs to ER without the need for a separation procedure for the bound and free LE.
The development of a migrating system for the separation analysis of EDCs was performed by capillary electrophoresis using sulfated β-cyclodextrin. Moreover, the electrochemical decomposition and removal of EDCs in water was investigated using SnO_2/Ti and carbon fiber electrode, respectively.
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