Bossi Alessandra, Castelletti Laura, Piletsky Sergey A, Turner Anthony P F, Righetti Pier Giorgio
Department of Science and Technology, University of Verona, Verona, Italy.
Electrophoresis. 2003 Oct;24(19-20):3356-63. doi: 10.1002/elps.200305588.
Extending the previous preliminary study on the construction of a capillary electrophoresis (CE)/sensor for the detection of reducing analytes, we focus the interest on the simultaneous detection of redox active species, which are important indicators of the oxidative damage in tissues, of food preservation, and of pollution. The CE/sensor was built by modifying the detector-portion of the capillary with the redox-sensitive polymer polyaniline (PANI). The analyte is detected by monitoring the changes in optical absorption of the PANI film. The CE/sensor was tested, with good results, with ascorbic acid, glutathione (GSH), as well as with compounds with very close similarity (ascorbic and isoascorbic acid). The kinetics of oxidation and reduction of PANI were evaluated. Further a PANI/CE-biological sensor was developed by coupling an enzyme, glucose oxidase (GOD), to the PANI-modified portion of the capillary. The stability of the immobilized GOD and the sensitivity of the CE/biosensor were studied, by using glucose as test analyte in concentrations within the physiological range. The results indicate that the CE/biosensor had good stability (more than 75% of original activity retained after 30 operational days), manufacturing reproducibility and a sensing range convenient for monitoring physiological glucose (1-24 mM).
在先前关于构建用于检测还原性分析物的毛细管电泳(CE)/传感器的初步研究基础上,我们将兴趣集中在同时检测氧化还原活性物质上,这些物质是组织氧化损伤、食品保鲜和污染的重要指标。通过用对氧化还原敏感的聚合物聚苯胺(PANI)修饰毛细管的检测部分来构建CE/传感器。通过监测聚苯胺膜的光吸收变化来检测分析物。用抗坏血酸、谷胱甘肽(GSH)以及结构非常相似的化合物(抗坏血酸和异抗坏血酸)对CE/传感器进行了测试,结果良好。评估了聚苯胺的氧化和还原动力学。此外,通过将葡萄糖氧化酶(GOD)与毛细管的聚苯胺修饰部分偶联,开发了一种聚苯胺/CE生物传感器。以葡萄糖作为生理范围内浓度的测试分析物,研究了固定化葡萄糖氧化酶的稳定性和CE/生物传感器的灵敏度。结果表明,CE/生物传感器具有良好的稳定性(30个操作日后保留了超过75%的原始活性)、制造重现性以及适用于监测生理葡萄糖(1-24 mM)的传感范围。