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Preparation of DNA-hydroxyapatite Modified Electrod and Its Application in Biological Analysis

Author: ZhangNuo
Tutor: WeiQin
School: Jinan University
Course: Analytical Chemistry
Keywords: CT DNA Trimethyl ammonium phenyl porphyrin Vitamin B12 Chloromycetin Modified electrode
CLC: O657.1
Type: Master's thesis
Year: 2010
Downloads: 63
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Abstract


DNA is the most important molecules in living cells, which contain all the genetic information of a specific cell, in a very wide field of molecular biology and biomedical research. DNA biosensor is a research hotspot of today's bio-sensing technology. Research DNA with DNA-modified electrodes interaction with other substances, not only overcome the solution electrochemical method, background signal is difficult to extract the disadvantage of a weak signal, and greatly reduced the amount of DNA to improve the sensitivity of the method, the sample in less consumption get a variety of effects parameters. The study DNA and other small molecule interactions is a major issue in the biochemistry and molecular biology, disease diagnosis, environmental testing, pharmaceutical research and drug design plays an important role. Water-soluble cationic porphyrin has excellent biological activity can be used as potential anti-cancer, anti-bacterial drugs and nucleic acid structure and dynamics of the probe, and its interaction with DNA to become a hot research topic. Porphyrins and DNA binding mode also helps to seek the relationship between the biological activity of the porphyrin compounds and DNA binding mode. Study of interaction of DNA with drug molecules, to help people understand the impact of some of the drug molecules in vivo replication and transcription of DNA, and the resulting species variation, the size of a variety of compounds with affinity for DNA and chemical nucleic acid enzyme The mechanism of action and other information. Surface electrochemical method of DNA synthesis with excellent biocompatibility and unique adsorption of hydroxyapatite (HAP), and prepared DNA-nano hydroxyapatite modified electrode (dsDNA / HAp / GCE) Interaction with other small molecules. The paper mainly includes the following aspects: The first part outlines the significance and application of DNA modified electrode preparation method Research and DNA modified electrode. The content and features of this thesis. The second section describes the Preparation and Characterization of HAp. HAP [Ca10 (PO4) 6 (OH) 2] is commonly used in bone repair material, its rules of three-dimensional chemical structure and unique multi-adsorption sites make it much attention in the field of catalysis, protein separation. In this paper, the co-precipitation method and microemulsion method to prepare the nano-HAP, and analyzed the synthesis of HAp in the microemulsion medium. The third part of the DNA and water-soluble cationic porphyrin - the trimethylammonium porphyrin (TAPP) interaction and DNA damage was detected. Using drop coating method has excellent biocompatibility and unique adsorption HAP formation of nano-thin film modified glassy carbon electrode. The electrochemical experiments results show that the nano-HAP film can effectively double-stranded DNA adsorbed on its surface. By cyclic voltammetry and electrochemical impedance spectroscopy system to study the interaction between the DNA and fixed in the HAp film TAPP and the modified electrode for the detection of DNA damage and determination of porphyrins. The experimental results show that the adsorption reaction control in the reaction process of the electrodes within the range of 50 ~ 250 mV · s-1 scanning speed based surface; in the range of pH value of 3.7 to 9.1, with the pH value increases TAPP redox peaks occurrence of positive shift, suggesting that TAPP electrode redox process not only e-participation, but also protons involved in DNA modification; pH = 7.0 phosphate buffer solution (PBS), with the ionic strength of the solution increases, TAPP DNA-modified electrode apparent formal potential constantly shift, indicating that the interaction between TAPP and DNA intercalation of small grooves, both can generate supramolecular compounds. According to the Langmuir formula obtained between TAPP and DNA binding constant of 1.48 x 105 mol L-1. The fourth part of the vitamin B12 interaction with DNA. By cyclic voltammetry study of vitamin B12 DNA in pH = 4.9 of HAc-NaAc interactions electrochemical behavior and concluded more convincing than the solution electrochemical method. The experimental results show that the presence of DNA can lead to the reduction peak current of vitamin B12 reduce. Modified electrode electrochemical parameters by measuring the bare glassy carbon electrode and DNA to prove that vitamin B12 DNA in the experimental conditions combine to generate a non-electrically active supramolecular compound, and a series of equations to obtain the supramolecular compound the composition is 1: 1, the binding constant beta = 5.35 × 105 mol · L-1. The fifth part of the DNA modified electrode electrocatalytic activity of chloramphenicol. The use-dsDNA / HAp / GCE on chloramphenicol electric catalytic role in the establishment of an electrochemical method for quantitative analysis of chloramphenicol. PH = 6.0 of KH2PO4-Na2HPO4 buffer solution and 0.1 mol · L-1 KCl solution, the concentration of chloramphenicol and the peak current in the range of 2.80 × 10-7 ~ 3.60 × 10-6 mol · L-1 showed a good linear relationship between the linear regression equation and the linear correlation coefficient were: ip (μA) = 5.865 .4784 c (umol L-1), r = 0.9942, the detection limit of 1.35 × 10-7 mol L-1. Chloramphenicol eye drops using the method for quantitative analysis, 8, parallel to the sample analysis result of a relative standard deviation of less than 3%, fully meet the requirements of the quantitative analysis.

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CLC: > Mathematical sciences and chemical > Chemistry > Analytical Chemistry > Instrument analysis ( physics and physical chemistry ) > Electrochemical analysis
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