Study on the development of the electrochemical screening for endocrine disruptors

内分泌干​​扰物电化学筛选研究进展

• 批准号: 12440207
• 负责人: TANAKA Shunitz
• 金额: $ 8.06万
• 依托单位: HOKKAIDO UNVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12440207/
• 关键词: endocrine disruptors; electrochemical screening; estrogen receptor; electroactive labeled ligand; daunomycin; 17β-estradiol; bisphenol A; electrochemical decomposition; ノニルフェノール

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.

在对蛋白质与其配体（如亲和素和生物素）的相互作用进行基础研究后，开发了一种新的内分泌干扰化学物质（EDC）电化学筛选方法。为了评价EDC与雌激素受体(EK)的结合能力，制备了用道诺霉素标记的17β-雌二醇作为电活性化合物。与柔红霉素相比，制备的标记雌二醇(LE)的电化学灵敏度较高。通过 LE 电极响应的降低观察到 LE 和 ER 之间的相互作用。发现这种减少是由于 LE 与 ER 的特异性结合。 LE 和 17β-雌二醇之间对 ER 上限制结合位点的竞争反应导致 LE 峰值电流增加。然后可以从峰值电流的增加得到针对LE的竞争者的浓度。 1×10^-8M 17 β-雌二醇的相对标准偏差为约10％(n＝7)。还通过与 17β-雌二醇-ER 相互作用进行比较来评估 EDC 和 ER 之间的结合亲和力。双酚A、对壬基苯酚和p,p,p'-DDT 被用作EDC 的测试化合物。所有测试化合物都表现出一定的与 ER 结合的能力。这种电化学结合测定说明了一种评估 EDC 与 ER 结合能力的新方法，无需对结合和游离 LE 进行分离程序。使用硫酸化 β-环糊精通过毛细管电泳开发了用于 EDC 分离分析的迁移系统。此外，分别使用SnO_2/Ti和碳纤维电极研究了水中EDCs的电化学分解和去除。

项目成果

H.Kuramitz: "Electrochemical sensing of avidin-biotin interaction using redox markers"Electroanalysis. 12. 1299-1303 (2000)

H.Kuramitz：“使用氧化还原标记物对亲和素-生物素相互作用进行电化学传感”电化学分析。

K.Sugawara: "Voltammetry detection of lectin using sugar labeled with electroactive substance"Anal. Sci.. 17. 21-25 (2001)

K.Sukawara：“使用电活性物质标记的糖进行凝集素的伏安法检测”Anal。

K. Sugawara: "Voltammetric behaviors of Lectin-Sugar binding using Au electrode modified with Galactosamine"Chemistry. Letters. 214-215 (2000)

K. Sukawara：“使用半乳糖胺修饰的金电极进行凝集素-糖结合的伏安行为”化学。

Y.Itoh: "High-Performance Liquid Chromatographic Separation of Potential Hydroxylated Metabolites of Estradiol 17-Sulfate by Female Rat Liver Microsomes"Anal.Sci.. 17. 659-661 (2001)

Y.Itoh：“通过雌性大鼠肝微粒体对雌二醇 17-硫酸盐的潜在羟基化代谢物进行高效液相色谱分离”Anal.Sci.. 17. 659-661 (2001)

K.Sugawara,: "Voltammetric behaviors of lectin-sugar binding using Au electrode modified with galactosamine,"Chemistry Letters,. 2000. 214-215 (2000)

K.Sukawara，：“使用半乳糖胺修饰的金电极进行凝集素-糖结合的伏安行为”，《化学快报》。