發(fā)布時間：2018-04-18 08:24

  本文選題：銅離子 + 絲網印刷電極　； 參考：《暨南大學》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;然而銅過量易導致阿爾茨海默病,對人體產生危害;目前銅鹽在水產養(yǎng)殖業(yè)中大量使用,易導致水體污染、魚類含銅量超標等危害;構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)的結合產物(MWNT-Cys)組裝在絲網印刷金電極表面,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,蛋白質的活性位點具有許多銅離子結合基序,本研究從銅離子結合基序中選擇常見氨基酸殘基,設計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,利用電化學方法研究寡肽在構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行測量;利用差分脈沖陽極溶出伏安法(DPSVO)對銅濃度進行表征;利用電感耦合等離子體質譜法(ICP-MS)檢測標準銅離子溶液濃度;利用循環(huán)伏安法(CV)、差分脈沖伏安法(DPV)、交流阻抗法(EIS)對寡肽的構象變化進行表征。結果:Cu~(2+)濃度在10nmol/l~1μmol/l、1μmol/l~9μmol/l、9μmol/l~0.9mmol/l范圍內具有良好的線性關系;利用紅外濕法消解法消解魚肉樣品,ICP結果測得銅離子加標回收率為101.00%,相對標準偏差(RSD)為3.5%;利用傳感器對樣品進行三次加標檢測,相對標準偏差(RSD)5%;對銻(Sb)和鉛(Pb)實現半定量檢測,最低檢出濃度可達到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合需要單一的環(huán)境,多離子的共存對寡肽與銅的結合影響較大;KGHGH與金屬離子的結合具有p H值依賴性。結論:所構建的絲網印刷電極傳感器能較準確的定量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.
【學位授予單位】：暨南大學
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：TP212.3;O657.1

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