电化学测定脱氧核糖核酸的研究

发布时间：2018-06-21 06:54

本文选题：脱氧核糖核酸 + 悬汞电极　；参考：《河北大学》2009年硕士论文

【摘要】：脱氧核糖核酸(DNA)是染色体的主要化学成分,同时也是组成基因的材料。脱氧核糖核酸的含量测定是生命科学、临床检验、生化研究、环境科学等研究领域的重要课题。本文应用悬汞电极(HMDE)和活化玻碳电极(AGCE),利用三电极电化学系统,针对脱氧核糖核酸进行了电化学检测方法的研究,研究了对硝基苯酚与DNA的相互作用并对以对硝基苯酚为电化学探针测定DNA的分析方法进行了研究。取得了以下研究结果： 1.阐述了DNA测定对电分析化学的挑战,并对DNA分析测定的研究现状进行了评述。 2.研究了Britton-Robinson缓冲溶液中,在悬汞电极(HMDE)上以孔雀石绿为电化学探针采用差分脉冲溶出伏安法间接测定DNA的方法,并对其测定条件(pH值、支持电解质、富集电位、富集时间等)进行了优化。在最佳条件下,峰电流的降低值与DNA浓度在4.0～120μg mL-1范围内呈良好的线性关系,检测限(36)为1.43μg mL-1,相对标准偏差(R.S.D.%)为2.65%-4.33%。此法应用于模拟DNA样品的测定,平均回收率为96.0%-106.5%。 3.研究了在氢氧化钠溶液中活化玻碳电极的制备方法并探讨了对DNA的电催化机理。结果表明,在Britton-Robinson缓冲溶液中,在活化玻碳电极(ACCE)上用线性扫描伏安法测定DNA有更好的检出限和线性范围,并且活化玻碳电极有很好的稳定性,适于连续多次的测定。在最佳条件下,峰电流强度与DNA浓度在2.5μgmL-1～200μg mL-1范围内呈线性关系,检测限(36)为0.184μg mL-1,相对标准偏差(R.S.D.%)为2.24%～4.43%。此法应用于DNA样品的测定,平均回收率为97.4%～104.9%。 4.研究了在玻碳电极上对硝基苯酚(p-NP)与DNA的相互作用,求解得出反应的结合比为2：3和结合常数为7.4×106。并对结合条件进行了优化,发现在最佳条件下p-NP峰电流的降低值与DNA浓度成良好的线性关系,据此建立了以p-NP为电化学探针测定DNA的分析方法,实现了对DNA样品的测定。在最佳条件下,峰电流的降低值与DNA浓度在1.0～50μg mL-1范围内呈良好的线性关系,检测限(36)为0.21μg mL-1,相对标准偏差(R.S.D.)为2.43%～4.16%。将此法应用于DNA样品的测定,平均回收率为96.4%～104.9%。
[Abstract]:Deoxyribonucleic acid (DNA) is the main chemical component of chromosomes and the material of genes. Deoxyribonucleic acid content determination is an important subject in life science, clinical examination, biochemical research, environmental science and so on. In this paper, a three-electrode electrochemical system was used to study the electrochemical detection of DNA by using HMDE) and activated glassy carbon electrode (AGCEE). The interaction between p-nitrophenol and DNA was studied and the analytical method of DNA determination with p-nitrophenol as electrochemical probe was studied. The results are as follows: 1. In this paper, the challenge of DNA determination to electroanalytical chemistry is reviewed, and the research status of DNA analysis is reviewed. 2. A method for indirect determination of DNA by differential pulse stripping voltammetry with malachite green as an electrochemical probe in Britton-Robinson buffer solution was studied. The pH value, supporting electrolyte, and enrichment potential were determined by differential pulse stripping voltammetry. The enrichment time was optimized. Under the optimum conditions, the decrease of peak current showed a good linear relationship with the concentration of DNA in the range of 4.0 ~ 120 渭 g mL ~ (-1). The detection limit of 36) was 1.43 渭 g mL ~ (-1), and the relative standard deviation was 2.65-4.33. The method was applied to the determination of simulated DNA samples. The average recovery was 96.0- 106.5. The preparation method of activated glassy carbon electrode in sodium hydroxide solution and the electrocatalytic mechanism of DNA were studied. The results show that in Britton-Robinson buffer solution, the detection limit and linear range of DNA determination by linear scanning voltammetry on activated glassy carbon electrode (ACCEE) are better, and the activated glassy carbon electrode has good stability and is suitable for continuous determination. Under the optimum conditions, the peak current intensity was linearly correlated with DNA concentration in the range of 2.5 渭 gm-1 ~ (-1) ~ 200 渭 g 路mL ~ (-1), the detection limit was 36) was 0.184 渭 g mL ~ (-1), and the relative standard deviation was 2.24 渭 m ~ (-1) and 4.43 渭 g 路mL ~ (-1), respectively. The method was applied to the determination of DNA samples, and the average recovery was 97.40.104.9. The interaction between p-nitrophenol p-NP) and DNA on glassy carbon electrode was studied. The binding ratio of the reaction was 2:3 and the binding constant was 7.4 脳 10 ~ 6. The binding conditions were optimized, and it was found that the decrease of p-NP peak current had a good linear relationship with the concentration of DNA. Based on this, a method for the determination of DNA with p-NP as electrochemical probe was established, and the determination of DNA samples was realized. Under the optimum conditions, the decrease of peak current showed a good linear relationship with the concentration of DNA in the range of 1.0 渭 g mL ~ (-1), the detection limit of 36 was 0.21 渭 g mL ~ (-1), and the relative standard deviation was R. S. D. 2.43 and 4.16. The method was applied to the determination of DNA samples and the average recovery was 96. 4% 104.9%.
【学位授予单位】：河北大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R341

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