基于DNA-甲苯胺蓝生物聚合离子膜的多环有机物电化学传感器的研究
Study on Electrochemical Biosensor for Polycyclic Organic Compounds Based on DNA-Toluidine Blue Bio-polyion Complex Membrane
摘 要
利用DNA与小分子之间的相互作用,以DNA/壳聚糖生物聚合离子膜固定电活性小分子,制备了DNA-甲苯胺蓝/壳聚糖聚合离子复合膜修饰电极,并利用多环有机物与染料分子对DNA特异结合的竞争关系,构筑了多环有机物非试剂添加型DNA电化学生物传感器。以盐酸四环素为模式分子,利用循环伏安法和方波伏安法研究了该修饰电极的电化学特性以及该电极对盐酸四环素的电化学响应,结果表明,DNA和甲苯胺蓝成功地固定在电极表面,电极表面的甲苯胺蓝保持了很好的电化学活性。利用紫外-可见分光光度法研究了电极对盐酸四环素响应的作用机理。该传感器的线性范围为2.5~100 μmol·L-1。
DNA聚合离子膜
多环有机物
甲苯胺蓝
DNA electrochemical biosensor
DNA polyion complex membrane
Polycyclic organic compound
Toluidine blue
Abstract
Based on the interaction between DNA and small molecules, a DNA/chitosan (CTS) bio-polyion complex membrane was used for immobilization of DNA binding electro-activators. A DNA-toluidine blue (TB)/CTS bio-polyion complex membrane modified electrode was developed for a reagent-less DNA electrochemical biosensor of polycyclic organic compounds (POCs), which based on the specific and competitive binding between POCs and dye molecules to DNA. Using tetracycline (TC) as a model analyte of polycyclic organic compounds, the electrochemical characteristic of the modified electrode and the response to the TC were studied by cyclic voltammetry and square wave voltammetry. It was found that DNA and TB were successfully co-immobilized on the electrode, and the embedded TB maintained its electro-active properties. The mechanism by which the modified electrode responded to TC was investigated by using UV-vis spectrophotometry. Linearity range of the biosensor found was 2.5-100 μmol·L-1.
中图分类号 O657.14
所属栏目 试验与研究
基金项目 中科院环境化学与生态毒理学国家重点实验室开放课题项目(KF2012-06);教育部留学回国人员科研启动基金(20071108);辽宁省教育科学“十二五”规划课题(JG11DB140)
收稿日期 2013/9/10
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备注顾婷婷(1973-),女,辽宁鞍山人,教授,博士,主要研究方向为生物电化学传感器。
引用该论文: GU Ting-ting,XIA Hong-qi. Study on Electrochemical Biosensor for Polycyclic Organic Compounds Based on DNA-Toluidine Blue Bio-polyion Complex Membrane[J]. Physical Testing and Chemical Analysis part B:Chemical Analysis, 2014, 50(9): 1070～1075
顾婷婷,夏洪齐. 基于DNA-甲苯胺蓝生物聚合离子膜的多环有机物电化学传感器的研究[J]. 理化检验－化学分册, 2014, 50(9): 1070～1075
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参考文献
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【3】CARLO M D, MARCELLO M D, GIULIANI M, et al. Detection of benzo(a)pyrene photodegradation products using DNA electrochemical sensors［J］. Biosensors and Bioelectronics, 2012,13(1):270-276．
【4】WONG E L S, GOODING J J. The electrochemical monitoring of the perturbation of charge transfer through DNA by cisplatin［J］. Journal of American Chemical Society, 2007,129:8950-8951．
【5】WANG Li-rong, QU Na, GUO Liang-hong. Electrochemical displacement method for the investigation of the binding interaction of polycyclic organic compounds with DNA［J］. Analytical Chemistry, 2008,80:3910-3914．
【6】GU Ting-ting, HASEBE Y. Novel amperometric assay for drug-DNA interaction based on an inhibitory effect on an electrocatalytic activity of DNA-Cu(Ⅱ) complex［J］. Biosensors and Bioelectronics, 2012,33:222-227．
【7】MATSUMOTO Y, TERUI N, TANAKA S. Accumulated detection of ethidium bromide using a UV-irradiated DNA film modified electrode and its application for electrochemical detection of an environmental pollutant［J］. Journal of Electroanalytical Chemistry, 2007,610:193-198．
【8】HASEBE Y, GU Ting-ting. DNA-Cu(Ⅱ) complex as a novel electrocatalyst for a hydrogen peroxide sensor［J］. Journal of Electroanalytical Chemistry, 2005,576:177-181．
【9】LI Na, ZHAO Hong-wen, YUAN Ruo, et al. An amperometric immunosensor with a DNA polyion complex membrane/gold nanoparticles-backbone for antibody immobilisation［J］. Electrochimica Acta, 2008,54:235-241．
【10】WU Wei, YI Ping, HE Ping, et al. Nanosilver-doped DNA polyion complex membrane for electrochemical immunoassay of carcinoembryonic antigen using nanogold-labeled secondary antibodies［J］. Analytica Chimica Acta, 2010,673:126-132．
【11】CHI Zhen-xing, LIU Ru-tao, SUN Ya-jing, et al. Investigation on the toxic interaction of toluidine blue with calf thymus DNA［J］. Journal of Hazardous Materials, 2010,175:274-278．
【12】PAUL P, KUMAR G S. Spectroscopic studies on the binding interaction of phenothiazinium dyes toluidine blue O, azure A and azure B to DNA［J］. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, 2013,107:303-310．
【13】GU Ting-ting, XIA Hong-qi, WANG Jian-li, et al. Peroxidase and toluidine blue-incorporated DNA bio-polyion complex film as biosensing layer for hydrogen peroxide determination［J］. Materials Science Forum, 2013,750:84-87．
【14】PINGARRN J M, YEZ-SEDEO P, GONZ-LEZ-CORTSA. Gold nanoparticle-based electrochemical biosensors［J］. Electrochimica Acta, 2008,53:5848-5866．
【15】LIANG Ru-ping, CHEN Yun-xia, QIU Jiang-dong. A sensitive amperometric immunosensor for hepatitis B surface antigen based on biocompatible redox-active chitosan-toluidine blue/gold nanoparticles composite film［J］. Anal Methods, 2011,3:1338-1343．
【16】LAVIRON E J. General expression of the linear potential sweep voltammogram in the case of diffusionless electrochemical systems［J］. Electroanal Chem and Interfacial Electrochemistry, 1979,101:19-28．
【17】费丹,王兴明,黎泓波,等.亚甲基蓝与鲱鱼精DNA相互作用的光谱法研究［J］.化学学报, 2008,66(4):443-448．
【18】WU Li-dong, LU Xian-bo, JIN Jing, et al. Electrochemical DNA biosensor for screening of chlorinated benzene pollutants［J］. Biosensors and Bioelectronics, 2011,26:4040-4045．
【19】KAMITORI S, TAKUSAGAWA F. Crystal structure of the 2: 1 complex between d (GAAGCTTC) and the anticancer drug actinomycin D［J］. Journal of Molecular Biology, 1992,225:445-456．
【20】KLUMP H, HERZOG K. Scanning calorimetry as an analytical tool to characterize DNAs［J］. Berichte der Bunsengesellschaft für physikalische Chemie, 1984,88:20-24．
【21】QU Feng, LI Na-qiang, JIANG Yu-yang. Electrochemical studies of NiTMpyP and interaction with DNA［J］. Talanta, 1998,45(5):787-793．
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