發(fā)布時間：2018-04-18 08:24

  本文選題：銅離子 + 絲網(wǎng)印刷電極　； 參考：《暨南大學(xué)》2016年碩士論文

【摘要】：目的:銅離子存在于多種氧化還原酶的活性位點,對酶的功能發(fā)揮有重要作用;然而銅過量易導(dǎo)致阿爾茨海默病,對人體產(chǎn)生危害;目前銅鹽在水產(chǎn)養(yǎng)殖業(yè)中大量使用,易導(dǎo)致水體污染、魚類含銅量超標(biāo)等危害;構(gòu)建快速檢測銅含量的方法非常必要。本研究利用多壁碳納米管(MWNT)和半胱氨酸(Cys)的結(jié)合產(chǎn)物(MWNT-Cys)組裝在絲網(wǎng)印刷金電極表面,利用差分脈沖陽極溶出伏安法(DPSVO)作為表征方法,構(gòu)建一個快速檢測魚肉樣品中銅離子濃度的傳感器。此外,蛋白質(zhì)的活性位點具有許多銅離子結(jié)合基序,本研究從銅離子結(jié)合基序中選擇常見氨基酸殘基,設(shè)計Lys-Cys-Met-Cys(KCMC)、Lys-Cys-Met-Met-Cys(KCMMC)、Lys-Cys-His-Cys(KCHC)、Lys-Cys-His-His-Cys(KCHHC)、Lys-Gly-His-Gly-His(KGHGH)五條寡肽序列,利用電化學(xué)方法研究寡肽在構(gòu)建檢測銅離子傳感器中的可行性。方法:利用傅里葉變換紅外光譜(FT-IR)對多壁碳納米管(MWNT)的羧基化程度進(jìn)行表征;利用納米粒度及Zeta電位分析儀對碳納米管的粒徑進(jìn)行測量;利用差分脈沖陽極溶出伏安法(DPSVO)對銅濃度進(jìn)行表征;利用電感耦合等離子體質(zhì)譜法(ICP-MS)檢測標(biāo)準(zhǔn)銅離子溶液濃度;利用循環(huán)伏安法(CV)、差分脈沖伏安法(DPV)、交流阻抗法(EIS)對寡肽的構(gòu)象變化進(jìn)行表征。結(jié)果:Cu~(2+)濃度在10nmol/l~1μmol/l、1μmol/l~9μmol/l、9μmol/l~0.9mmol/l范圍內(nèi)具有良好的線性關(guān)系;利用紅外濕法消解法消解魚肉樣品,ICP結(jié)果測得銅離子加標(biāo)回收率為101.00%,相對標(biāo)準(zhǔn)偏差(RSD)為3.5%;利用傳感器對樣品進(jìn)行三次加標(biāo)檢測,相對標(biāo)準(zhǔn)偏差(RSD)5%;對銻(Sb)和鉛(Pb)實現(xiàn)半定量檢測,最低檢出濃度可達(dá)到nmol/l級別;不含巰基(-SH)的寡肽組裝穩(wěn)定,寡肽傳感器對銅的結(jié)合需要單一的環(huán)境,多離子的共存對寡肽與銅的結(jié)合影響較大;KGHGH與金屬離子的結(jié)合具有p H值依賴性。結(jié)論:所構(gòu)建的絲網(wǎng)印刷電極傳感器能較準(zhǔn)確的定量Cu~(2+)的濃度,并且具有制作簡便、檢測快速、可棄式使用等特點;寡肽在結(jié)合金屬離子的過程中,需要單一環(huán)境才能與金屬離子特異性結(jié)合,具有p H值依賴性。
[Abstract]:Objective: copper ions exist in the active sites of many redox enzymes and play an important role in the function of the enzymes. However, excessive copper can lead to Alzheimer's disease and cause harm to human body. At present, copper salt is widely used in aquaculture.It is easy to lead to water pollution and fish copper content exceeds the standard, so it is necessary to establish a method for rapid detection of copper content.In this study, multiwalled carbon nanotubes (MWNTs) and cysteine (CysCys) binding products, MWNT-Cys), were assembled on the surface of silk-screen printed gold electrodes. Differential pulse anodic stripping voltammetry (DPSVOO) was used as a characterization method.A sensor for rapid detection of copper ion concentration in fish samples was constructed.In addition, the active sites of proteins have many copper ion binding motifs. In this study, we selected common amino acid residues from copper ion binding motifs and designed Lys-Cys-Met-Cys-Cys-Met-Cys-Cys-Cys-Met-Cys-Cys-Cys-Cys-His-Cys-Cys-Cis-Cis-Cys-CKCHH KCHHC-Lys-Gly-His-Gly-Hisy-HisKGHGHH5 oligoseptide sequences from copper ion binding motifs.The feasibility of oligoseptide in the construction of copper ion sensor was studied by electrochemical method.Methods: the carboxylation degree of MWNTs was characterized by Fourier transform infrared spectroscopy (FTIR), and the diameter of MWNTs was measured by nano-particle size and Zeta potential analyzer.The copper concentration was characterized by differential pulse anodic stripping voltammetry (DPSVOO), and the concentration of standard copper ion solution was detected by inductively coupled plasma mass spectrometry (ICP-MS).The conformation of oligopeptides was characterized by cyclic voltammetry, differential pulse voltammetry (DPVN) and AC impedance spectroscopy (EIS).Results there was a good linear relationship in the range of 10nmol/l~1 渭 mol / L ~ (1 渭 mol / L ~ (-1) mol/l~9 / L ~ (-1) 渭 mol / L ~ (9) 渭 mol/l~0.9mmol/l), the recovery rate of Cu ~ (2 +) was 101.00% and the relative standard deviation (RSD) was 3.5% by using infrared wet digestion method.Relative standard deviation (RSD5); semi-quantitative detection of antimony, SB) and lead, the lowest detectable concentration can reach nmol/l level; oligoseptides without sulfhydryl group are assembled stably, and oligoseptide sensors require a single environment for the binding of copper.The coexistence of multi-ions has a significant effect on the binding of oligopeptide to copper. The binding of KGHGH to metal ions is pH dependent.Conclusion: the proposed screen-printing electrode sensor can accurately quantify the concentration of Cu~(2, and has the advantages of simple fabrication, rapid detection and disposable use.It needs a single environment to specifically bind to metal ions, which is dependent on pH value.
【學(xué)位授予單位】：暨南大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：TP212.3;O657.1

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