黃素輔酶在氧化鋅納米棒光電極上的光致電化學(xué)響應(yīng)及其應(yīng)用的研究
發(fā)布時(shí)間：2018-12-27 10:21
【摘要】：黃素輔酶,包括黃素單核苷酸(FMN)和黃素腺嘌呤二核苷酸(FAD)以及它們的前體核黃素(RF),是黃素蛋白(主要為氧化還原酶)的輔基,它可以接受2個(gè)氫而還原為FMNH2或FADH2,參與人體內(nèi)多種氧化還原反應(yīng),促進(jìn)糖、氨基酸、脂和蛋白質(zhì)的代謝。因此,黃素輔酶及其相關(guān)蛋白質(zhì)的測(cè)定具有重要的意義。如果黃素輔酶能作為電子給體,與光敏納米材料組成一種新的光致電化學(xué)(PEC)反應(yīng)系統(tǒng),借助于這個(gè)PEC分析的平臺(tái),就能建立測(cè)定黃素輔酶的新方法,是非常有意義的工作。本文利用四電極雙通道電化學(xué)系統(tǒng),在電化學(xué)還原黃素輔酶后,使其與光激發(fā)下的氧化鋅納米棒光電極(ZnONRP)發(fā)生PEC反應(yīng),構(gòu)建了一種納米界面光電極與黃素輔酶的PEC反應(yīng)通道,通過光電流的測(cè)量實(shí)現(xiàn)了對(duì)黃素輔酶的直接檢測(cè)。本文主要做了以下三個(gè)方面的研究:1.用電化學(xué)沉積法制備了ZnONRP,優(yōu)化了制備方法,并通過掃描電鏡(SEM)和X射線衍射譜(XRD)對(duì)電極的修飾結(jié)果進(jìn)行了表征。以制備的ZnONRP作為光敏界面,由第二工作電極還原RF為RFH2,使其與ZnONRP發(fā)生PEC反應(yīng),建立了以RF為電子供體,ZnONRP為電子受體的PEC反應(yīng)系統(tǒng)。基于ZnONRP對(duì)RF的光致電化學(xué)響應(yīng),通過檢測(cè)光電流實(shí)現(xiàn)了對(duì)RF的定量分析。文中討論了光電化學(xué)敏感界面對(duì)RF的響應(yīng)機(jī)理,討論了偏壓、電解液pH值、光照強(qiáng)度以及工作電極與第二工作電極之間的距離對(duì)PEC反應(yīng)和測(cè)定RF的影響。在優(yōu)化的實(shí)驗(yàn)條件下,光電流強(qiáng)度與1.00?10-11?1.00?10-6 mol/L RF濃度的對(duì)數(shù)值成正比,檢測(cè)限為6.0?10-12 mol/L(S/N=3),靈敏度為195.6 nA/logC(μmol/L)。對(duì)實(shí)際樣品測(cè)定的相對(duì)標(biāo)準(zhǔn)偏差(RSD)小于6.25%,回收率為99.0%?104%,常見生化物質(zhì)對(duì)RF的光電流響應(yīng)無干擾。2.用FMN和FAD代替RF,利用黃素輔酶在ZnONRP上的PEC反應(yīng),創(chuàng)建了黃素輔酶的PEC分析法。在優(yōu)化的實(shí)驗(yàn)條件下,光電流強(qiáng)度與FMN或FAD濃度對(duì)數(shù)成正比,在1.00?10-11?1.00?10-6 mol/L的濃度范圍內(nèi),光電流強(qiáng)度與FMN和FAD濃度的對(duì)數(shù)值成正比,線性相關(guān)系數(shù)R2分別為0.996和0.998,檢出限分別為8.00?10-12和5.00?10-12 mol/L(S/N=3)。3.FAD是葡萄糖氧化酶(GOD)的輔酶,葡萄糖是GOD的底物,用葡萄糖還原GOD中的FAD,再利用殼聚糖的電荷傳遞功能,通過直接電子轉(zhuǎn)移和ZnONRP-FAD光致電化學(xué)系統(tǒng),建立了葡萄糖的PEC分析法。在優(yōu)化了殼聚糖的濃度、GOD負(fù)載量和電解液pH后,光電流強(qiáng)度與葡萄糖濃度在0.60~2.80mmol/L濃度范圍內(nèi)呈指數(shù)關(guān)系,相關(guān)系數(shù)R2=0.991,檢出限為0.20 mmol/L。將該方法用于對(duì)人血清中葡萄糖含量的檢測(cè),RSD為0.86%,回收率為98.5%~103.3%。
[Abstract]:Flavin coenzymes, including Flavin mononucleotide (FMN) and Flavin adenine dinucleotide (FAD), as well as their precursor riboflavin (RF), are the counits of the Flavin protein (mainly redox enzyme). It can accept two hydrogen and reduce to FMNH2 or FADH2, in human body to participate in a variety of redox reactions, promoting the metabolism of sugar, amino acids, lipids and proteins. Therefore, the determination of Flavin coenzyme and its related proteins is of great significance. If Flavin coenzyme can be used as electron donor to form a new photochemical (PEC) reaction system with Guang Min nanomaterials, with the help of this PEC analysis platform, a new method for the determination of Flavin coenzyme can be established, which is of great significance. In this paper, a four-electrode two-channel electrochemical system was used to make PEC reaction with photoinduced photoelectrode (ZnONRP) of zinc oxide nanorods after electrochemical reduction of Flavin coenzyme. A PEC reaction channel between photoelectrode and Flavin coenzyme was constructed, and the direct detection of Flavin coenzyme was realized by photocurrent measurement. This paper mainly does the following three aspects of research: 1. The preparation method of ZnONRP, was optimized by electrochemical deposition. The modified results of the electrode were characterized by scanning electron microscopy (SEM) and X-ray diffraction (XRD). Using the prepared ZnONRP as Guang Min interface, the second working electrode was used to reduce RF to RFH2, to induce PEC reaction with ZnONRP. A PEC reaction system with RF as electron donor and ZnONRP as electron receptor was established. Based on the photochemical response of ZnONRP to RF, the quantitative analysis of RF is realized by detecting photocurrent. The response mechanism of photochemically sensitive interface to RF is discussed. The effects of bias voltage, pH value of electrolyte, illumination intensity and the distance between the working electrode and the second working electrode on the PEC reaction and the determination of RF are discussed. Under the optimized experimental conditions, the photocurrent intensity is proportional to the logarithmic value of the concentration of 1.00 ~ (-11) ~ (-1) ~ (-1) ~ (10 ~ (-6) mol/L RF, and the detection limit is 6.0 ~ 10 ~ (-12) mol/L (S/N=3). The sensitivity is 195.6 nA/logC (渭 mol/L). The relative standard deviation (RSD) for the actual samples was less than 6.25, the recovery was 99.0 and 104, and the common biochemical substances had no interference with the photocurrent response of RF. 2. Using FMN and FAD instead of RF, the PEC analysis method of Flavin coenzyme was established by using PEC reaction of Flavin coenzyme on ZnONRP. Under the optimized experimental conditions, the photocurrent intensity is in direct proportion to the logarithm of FMN or FAD concentration. In the range of 1.00 ~ (-11) ~ (-1) ~ (-1) ~ (-6) mol/L, the optical current intensity is directly proportional to the logarithmic values of FMN and FAD concentrations. The linear correlation coefficient R2 was 0.996 and 0.998, and the detection limits were 8.00 ~ (-12) and 5.00 ~ (-12) mol/L (S/N=3), respectively. 3.FAD was the coenzyme of glucose oxidase (GOD), and glucose was the substrate of GOD. The FAD, in GOD was reduced with glucose and the charge transfer function of chitosan was utilized. The PEC analysis of glucose was established by direct electron transfer and ZnONRP-FAD photochemical system. When the concentration of chitosan, GOD loading and electrolyte pH were optimized, there was an exponential relationship between the photocurrent intensity and glucose concentration in the range of 0.60~2.80mmol/L concentration, the correlation coefficient was R2N 0.991and the detection limit was 0.20 mmol/L.. The method was applied to the determination of glucose in human serum. The RSD was 0.86 and the recovery rate was 98.5 and 103.3%.
【學(xué)位授予單位】：青島科技大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：O657.1
本文編號(hào)：2392916
[Abstract]:Flavin coenzymes, including Flavin mononucleotide (FMN) and Flavin adenine dinucleotide (FAD), as well as their precursor riboflavin (RF), are the counits of the Flavin protein (mainly redox enzyme). It can accept two hydrogen and reduce to FMNH2 or FADH2, in human body to participate in a variety of redox reactions, promoting the metabolism of sugar, amino acids, lipids and proteins. Therefore, the determination of Flavin coenzyme and its related proteins is of great significance. If Flavin coenzyme can be used as electron donor to form a new photochemical (PEC) reaction system with Guang Min nanomaterials, with the help of this PEC analysis platform, a new method for the determination of Flavin coenzyme can be established, which is of great significance. In this paper, a four-electrode two-channel electrochemical system was used to make PEC reaction with photoinduced photoelectrode (ZnONRP) of zinc oxide nanorods after electrochemical reduction of Flavin coenzyme. A PEC reaction channel between photoelectrode and Flavin coenzyme was constructed, and the direct detection of Flavin coenzyme was realized by photocurrent measurement. This paper mainly does the following three aspects of research: 1. The preparation method of ZnONRP, was optimized by electrochemical deposition. The modified results of the electrode were characterized by scanning electron microscopy (SEM) and X-ray diffraction (XRD). Using the prepared ZnONRP as Guang Min interface, the second working electrode was used to reduce RF to RFH2, to induce PEC reaction with ZnONRP. A PEC reaction system with RF as electron donor and ZnONRP as electron receptor was established. Based on the photochemical response of ZnONRP to RF, the quantitative analysis of RF is realized by detecting photocurrent. The response mechanism of photochemically sensitive interface to RF is discussed. The effects of bias voltage, pH value of electrolyte, illumination intensity and the distance between the working electrode and the second working electrode on the PEC reaction and the determination of RF are discussed. Under the optimized experimental conditions, the photocurrent intensity is proportional to the logarithmic value of the concentration of 1.00 ~ (-11) ~ (-1) ~ (-1) ~ (10 ~ (-6) mol/L RF, and the detection limit is 6.0 ~ 10 ~ (-12) mol/L (S/N=3). The sensitivity is 195.6 nA/logC (渭 mol/L). The relative standard deviation (RSD) for the actual samples was less than 6.25, the recovery was 99.0 and 104, and the common biochemical substances had no interference with the photocurrent response of RF. 2. Using FMN and FAD instead of RF, the PEC analysis method of Flavin coenzyme was established by using PEC reaction of Flavin coenzyme on ZnONRP. Under the optimized experimental conditions, the photocurrent intensity is in direct proportion to the logarithm of FMN or FAD concentration. In the range of 1.00 ~ (-11) ~ (-1) ~ (-1) ~ (-6) mol/L, the optical current intensity is directly proportional to the logarithmic values of FMN and FAD concentrations. The linear correlation coefficient R2 was 0.996 and 0.998, and the detection limits were 8.00 ~ (-12) and 5.00 ~ (-12) mol/L (S/N=3), respectively. 3.FAD was the coenzyme of glucose oxidase (GOD), and glucose was the substrate of GOD. The FAD, in GOD was reduced with glucose and the charge transfer function of chitosan was utilized. The PEC analysis of glucose was established by direct electron transfer and ZnONRP-FAD photochemical system. When the concentration of chitosan, GOD loading and electrolyte pH were optimized, there was an exponential relationship between the photocurrent intensity and glucose concentration in the range of 0.60~2.80mmol/L concentration, the correlation coefficient was R2N 0.991and the detection limit was 0.20 mmol/L.. The method was applied to the determination of glucose in human serum. The RSD was 0.86 and the recovery rate was 98.5 and 103.3%.
【學(xué)位授予單位】：青島科技大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：O657.1
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