本文選題：脫氧核糖核酸 + 懸汞電極　； 參考：《河北大學(xué)》2009年碩士論文

【摘要】：脫氧核糖核酸(DNA)是染色體的主要化學(xué)成分,同時也是組成基因的材料。脫氧核糖核酸的含量測定是生命科學(xué)、臨床檢驗、生化研究、環(huán)境科學(xué)等研究領(lǐng)域的重要課題。本文應(yīng)用懸汞電極(HMDE)和活化玻碳電極(AGCE),利用三電極電化學(xué)系統(tǒng),針對脫氧核糖核酸進(jìn)行了電化學(xué)檢測方法的研究,研究了對硝基苯酚與DNA的相互作用并對以對硝基苯酚為電化學(xué)探針測定DNA的分析方法進(jìn)行了研究。取得了以下研究結(jié)果： 1.闡述了DNA測定對電分析化學(xué)的挑戰(zhàn),并對DNA分析測定的研究現(xiàn)狀進(jìn)行了評述。 2.研究了Britton-Robinson緩沖溶液中,在懸汞電極(HMDE)上以孔雀石綠為電化學(xué)探針采用差分脈沖溶出伏安法間接測定DNA的方法,并對其測定條件(pH值、支持電解質(zhì)、富集電位、富集時間等)進(jìn)行了優(yōu)化。在最佳條件下,峰電流的降低值與DNA濃度在4.0～120μg mL-1范圍內(nèi)呈良好的線性關(guān)系,檢測限(36)為1.43μg mL-1,相對標(biāo)準(zhǔn)偏差(R.S.D.%)為2.65%-4.33%。此法應(yīng)用于模擬DNA樣品的測定,平均回收率為96.0%-106.5%。 3.研究了在氫氧化鈉溶液中活化玻碳電極的制備方法并探討了對DNA的電催化機(jī)理。結(jié)果表明,在Britton-Robinson緩沖溶液中,在活化玻碳電極(ACCE)上用線性掃描伏安法測定DNA有更好的檢出限和線性范圍,并且活化玻碳電極有很好的穩(wěn)定性,適于連續(xù)多次的測定。在最佳條件下,峰電流強(qiáng)度與DNA濃度在2.5μgmL-1～200μg mL-1范圍內(nèi)呈線性關(guān)系,檢測限(36)為0.184μg mL-1,相對標(biāo)準(zhǔn)偏差(R.S.D.%)為2.24%～4.43%。此法應(yīng)用于DNA樣品的測定,平均回收率為97.4%～104.9%。 4.研究了在玻碳電極上對硝基苯酚(p-NP)與DNA的相互作用,求解得出反應(yīng)的結(jié)合比為2：3和結(jié)合常數(shù)為7.4×106。并對結(jié)合條件進(jìn)行了優(yōu)化,發(fā)現(xiàn)在最佳條件下p-NP峰電流的降低值與DNA濃度成良好的線性關(guān)系,據(jù)此建立了以p-NP為電化學(xué)探針測定DNA的分析方法,實現(xiàn)了對DNA樣品的測定。在最佳條件下,峰電流的降低值與DNA濃度在1.0～50μg mL-1范圍內(nèi)呈良好的線性關(guān)系,檢測限(36)為0.21μg mL-1,相對標(biāo)準(zhǔn)偏差(R.S.D.)為2.43%～4.16%。將此法應(yīng)用于DNA樣品的測定,平均回收率為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%.
【學(xué)位授予單位】：河北大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2009
【分類號】：R341

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