功能碳黑修饰的丝网印刷碳糊电极葡萄糖生物传感器的特性与机理

从碳黑表面引发苯乙烯磺酸钠的原子转移自由基聚合制备了聚(苯乙烯磺酸钠)改性碳黑(CB-g-PSS),并分别以掺杂和沉积两种不同方式修饰电极的生物敏感膜,再在生物敏感膜上吸附固定葡萄糖氧化酶,制作了两种葡萄糖氧化酶传感器,得到了不同的效果.实验结果表明,将CB-g-PSS与成膜材料掺杂制作的生物传感器与无修饰传感器相比,响应灵敏度下降了1/3;将CB-g-PSS沉积修饰丝网印刷碳糊电极制作的传感器与无修饰传感器相比,响应灵敏度提高了2倍,且对1.1～33.3 mmoL/L的葡萄糖待测样本,RSD均＜7%,稳定性良好,有较高的应用价值.通过实验分析了CB-g-PSS以不同方式修饰电极的工作机理,结果表明,选择正确的修饰方式,能够发挥CB-g-PSS的导电效应及纳米效应,使其有利于酶的固定,提高响应灵敏度并改善酶促反应动力学特性.     

亲水金和憎水二氧化硅纳米颗粒对葡萄糖生物传感器响应灵敏度的增强作用

用亲水金、憎水二氧化硅纳米颗粒固定葡萄糖氧化酶(GOD),采用聚乙烯醇缩丁醛(PVB)为辅助固酶膜基质来制备葡萄糖生物传感器,并考察了亲水金、憎水二氧化硅纳米颗粒对酶电极电流响应的影响.实验表明,引入纳米粒子可显著增强电极响应灵敏度.并对两种不同性质纳米颗粒所起作用的可能机理进行讨论,从理论和实验上证明了纳米颗粒对固定酶的作用.为制备有实用价值的葡萄糖生物传感器提供了可供参考的实验和理论依据.     

金属纳米颗粒的制备及其在酶生物传感器中的应用研究

近年来,金属纳米颗粒的制备研究引起了人们的广泛兴趣.与相应的块体材料相比,金属纳米颗粒具有独特的化学和物理性质,可应用于电学、催化、磁性材料、光催化、生物染色剂、药物输送等许多领域.其中,传感器是纳米颗粒最有前途的应用领域之一.传感器的微型化是传感器发展的主要研究方向,将纳米颗粒用于传感器的研究将促进这一目标的实现.本论文利用纳米颗粒材料的独特效应来提高葡萄糖传感器的响应电流.将自制的银金、铂以及二氧化硅和铂复合纳米颗粒用于固定化酶,使酶电极的电流响应值得到了大幅度的提高,从而为纳米增强的新型葡萄糖生物传感器的研究、制备和应用提供了可供参考的实验和理论依据,并为传感器的小型化开辟了一条新途径.     

亲水金和憎水二氧化硅纳米颗粒对葡萄糖生物传感器响应灵 …

用亲水金、憎水二氧化硅纳米颗粒固定葡萄糖氧化酶（ＧＯＤ）,采有聚乙烯醇缩丁醛（ＰＶＢ）为辅助固酶膜基质来制备葡萄糖生物传感器,并考察了亲水金、憎水二氧化硅纳米颗粒对酶电极电流响应的影响。实验表明,引入纳米粒子可显著增强电极响应灵敏度,并对两种不同性质纳米所起作用的可能机理进行讨论,从理论和实验上辛 明了纳米颗粒对固定酶的作用。为制备有实用价值的葡萄糖生物传感器提供了可供参考的实验和理论依据。     

基于纳米氧化铜修饰的异戊巴比妥的新型电化学传感器的制备与应用

在纳米氧化铜修饰的玻碳电极表面电聚合一种能够快速检测尿液中异戊巴比妥(AMB)的分子印迹敏感膜,研究了该敏感膜的最佳成膜条件及最佳工作条件.通过扫描电子显微镜(SEM)、循环伏安(CV)和电化学交流阻抗法(EIS)研究了印迹膜的表面形貌及性能.电化学实验结果表明,纳米氧化铜能提高传感器对AMB的灵敏度.在最佳实验条件下,铁氰化钾分子探针的差分脉冲伏安(DPV)峰电流响应值与AMB的浓度在1.0×10-7~1.4×10-4mol/L范围内呈现良好的线性关系(线性相关系数R=0.9966);检出限为2.1×10-9mol/L(S/N=3).此印迹传感器可用于尿液中AMB的检测,加标回收率为94.00%~104.67%.     

基于磁性分子印迹聚合物的电化学传感器的研究进展

分子印迹聚合物与磁性纳米材料结合,制备成磁性分子印迹纳米敏感膜,这样做不仅可以发挥分子印迹聚合材料的优势,而且磁性纳米粒子可有效提高电化学传感器的灵敏度、稳定性以及生物相容性等。近年来将磁性分子印迹纳米敏感膜应用于电化学传感器制备成的磁性分子印迹电化学传感器得到了较快的发展。本文就近5年来磁性分子印迹电化学传感器敏感膜所用的磁性材料、敏感膜制备方法以及磁性分子印迹电化学传感器在环境、食品以及临床方面的应用进行了综述总结。     

高分子对酶、抗体、DNA的修饰、固定化及其生物医学应用

为发展适于生物医用的生物功能高分子材料,本实验室近年来研究了可溶性高分子对L-天冬酰胺酶的修饰、纳米磁性高分子微粒对酶或抗体的固定化、亚微米高分子微球固定化碱性磷酸酶及其在DNA检测中的应用、高分子微球固定化酶的合成与性能、酶在导电高分子膜上的固定化及生物传感器制备等. 本文对此进行简要总结.     

镍纳米粒子修饰碳糊电极直接电催化鸟嘌呤的研究

将镍纳米粒子与石蜡、石墨按照一定比例混合制备镍纳米粒子修饰碳糊电极,采用循环伏安法(CV)对修饰碳糊电极进行电化学表征,在0.1 mol/L B-R缓冲溶液(pH4.5)中研究了鸟嘌呤在该修饰电极上的电化学行为。结果表明,与裸碳糊电极相比,以掺杂法制备的镍纳米粒子修饰电极能够明显降低鸟嘌呤的过电位,增大其氧化电流,很好地催化氧化鸟嘌呤。在优化的实验条件下,鸟嘌呤在该修饰电极上的氧化峰电流与其浓度在1.0×10-5~5.0×10-4mol/L范围内呈良好的线性关系,检出限(3σ)为7.5×10-6mol/L。     

葡萄糖氧化酶在纳米金修饰的丝网印刷电极上的直接电子传递及应用研究

制作了一次性使用的无介质丝网印刷传感器。研究了葡萄糖氧化酶在纳米金修饰的丝网印刷碳电极(GOD/Au/SPCE)上的电化学行为,在0.1mol/L和pH5.0的磷酸盐缓冲体系中,吸附在金胶纳米粒子上的葡萄糖氧化酶能保持其生物活性并催化溶解氧的还原,还原催化电流随葡萄糖溶液的加入而降低,线性范围为3.0×10-5～4.0×10-4mol·L-1,检出限为1.0×10-5mol·L-1。方法简便快速,可用于葡萄糖的测定。     

高分子对酶,抗体DNA的修饰,固定化及其生物医学应用

为发展适于生物医用的生物功能高分子材料，本实验室近年来研究了可溶性高分子对Ｌ－天冬酰胺酶的修饰，纳米磁性高分子微粒对酶或抗体的固定化，亚微米高分子微球固定化碱性磷酸酶及其在ＤＮＡ检测中的应用，高分子微球固定化酶的合成与性能，酶在导电高分子膜上的固定化及生物传感器制备等，本文对此进行简要总结。     

A Novel DNA Probe Based on Molecularly Imprinted Polymer Modified Electrode for the Electrochemical Monitoring of DNA

A DNA probe that was based on methylene blue (MB) imprinted polyvinyl pyridine polymer (MIP) modified carbon paste electrodes were developed for the first time for electrochemical monitoring of DNA. Probes were built up by adsorbing MB onto modified electrodes prior to DNA immobilization. It was shown that DNA strongly immobilizes on MIP modified electrodes when MB was adsorbed in advance of DNA immobilization. The performance of the MB imprinted polymer modified carbon paste electrodes (MIP‐CPE) to rebind the template molecule (MB) were compared to those of control polymer modified (non‐imprinted polymer NIP‐CPE) and bare (CPE) electrodes. Electrochemical signal resulting from the oxidation of guanine moiety of the immobilized probe DNA was high enough on the constructed platform, implicating that probes of this kind could be favorably used for DNA analysis. These probes exhibited high selectivity for its complementary DNA sequences (target). HBV‐DNA hybridization was studied to evaluate the selectivity of the probes for complementary, non‐complementary and mismatch sequences. The detection limit of the probe for the target DNA was 8.72 µg/mL (1.38 µM), which was better than those attained by some earlier DNA sensor studies.     

A Novel and Selective Methylene Blue Imprinted Polymer Modified Carbon Paste Electrode

In this study, methylene blue (MB) imprinted microbeads were synthesized and characterized by Fourier transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM).Differential pulse voltammetric responses of carbon paste electrodes modified with MB imprinted polymer were used to evaluate the adsorption and selectivity features of the polymer. For selectivity studies two basic dyestuffs (thionine blue and toluidine blue) which have similar structure to MB were chosen. Comparison of the voltammetric responses obtained with pure carbon paste and carbon paste modified with either imprinted or nonimprinted polymer electrodes revealed that MB imprinted polymer presented a higher selectivity to the template molecule MB in contrast to structurally similar molecules, thionine blue and toluidine blue.     

A glucose electrode based on carbon paste chemically modified with a ferrocene-containing siloxane polymer and glucose oxidase,coated with a poly(ester-sulfonic acid) cation-exchanger

A carbon-paste chemically modified with glucose oxidase and a ferrocene-containing siloxane polymer was further modified by coating the electrode surface with a poly(ester-sulfonic acid) cation-exchanger, Eastman AQ-29D. The polymer is obtained as a homogeneous aqueous dispersion at pH 5–6; when dried, the polymer coating is not water-soluble. The coating was shown not to be detrimental to the enzyme activity but to prevent electrochemically active anionic interferents such as ascorbate and urate from reaching the electrode surface. The polymer coating also prevented glucose oxidase from leaking out of the carbon paste into the contacting solution and protected the electrode surface from fouling agents present in urine and bovine serum albumin. Uncoated electrodes lost some 10-2-15% of their original response to glucose after storage in buffer for three weeks whereas the response of the coated electrodes remained constant. Calibration curves for glucose were strictly linear up to about 5 mM for uncoated and up to 20 mM for coated electrodes. The response current to glucose was not decreased after coating.     

Amperometric detection of glucose using self-catalytic carbon paste electrodes

The analytical detection of d-glucose by means of self-catalytic carbon paste electrodes is described. In the construction of these electrodes, carbon powder has been modified with the redox liquid n-butylferrocene, which simultaneously serves both to help bind the paste together whilst also acting as a mediator in the enzymatic oxidation of d-glucose by glucose oxidase. The sensor then functions by monitoring the electrochemical oxidation of the constituent n-butylferrocene itself. Through testing in model glucose solutions, the electrodes were found to yield a linear response over a d-glucose concentration range of 2-20 mM. They were also successfully employed in the determination of d-glucose levels in a spiked blood sample, giving a detection limit of 0.8 mM (based on the 3sigma criterion).     

Association interaction and voltammetric determination of 1-aminopyrene and 1-hydroxypyrene at cyclodextrin and DNA based electrochemical sensors

The aim of this work was voltammetric determination of 1-aminopyrene and 1-hydroxypyrene using carbon paste electrodes modified with cyclodextrin derivatives and double stranded deoxyribonucleic acid (dsDNA). The detection schemes based on a preconcentration and differential pulse voltammetric (DPV) determination at beta-cyclodextrin and gamma-cyclodextrin modified carbon paste electrode (beta-CD/CPE, gamma-CD/CPE), neutral beta-cyclodextrin polymer and carboxymethyl-beta-cyclodextrin polymer modified screen-printed electrode (beta-CDP/SPE, beta-CDPA/SPE) and dsDNA modified screen-printed electrode (DNA/SPE) are proposed for the trace determination of studied analytes within the concentration range from 2 x 10(-8) to 4 x 10(-7) mol dm(-3) and from 2 x 10(-7) to 4 x 10(-6) mol dm(-3) with the limits of quantification down to 10(-8) mol dm(-3). Depending on pH, 1-aminopyrene interacts with both surface attached CD and DNA by electrostatic bonds and supramolecular complexation while 1-hydroxypyrene associates with the CD hosts via complexation. The 1-aminopyrene interaction with dsDNA was confirmed by fluorimetric measurements in the solution phase using a competing DNA-TO-PRO-3 dye complex. In addition, the effect of temperature on this association was investigated using an electrically heated DNA-modified carbon paste electrode (DNA/CPE).     

生物功能电极(Ⅴ)─葡萄糖氧化酶在环糊精聚合物中的固定化

将葡萄糖氧化酶(GOD)固定在α-环糊精聚合物中,而电子传递体分子被包含在环糊精腔穴中。固定化酶膜的FTIR测定表明,GOD与环糊精聚合物发生共价连接。制备了含电子传递体的不同GOD酶电极并比较了它们的性能。含四硫代富瓦烯的酶电极具有良好的电流响应特性,可望成为第二代葡萄糖酶电极的新构型。     

Dopamine and Glucose Sensors Based on Glassy Carbon Electrodes Modified with Melanic Polymers

This work deals with the study of polymers electrogenerated from different catechols at glassy carbon electrodes and the analytical applications of the resulting modified electrodes for dopamine quantification and glucose biosensing. The electropolymerization was performed from a 3.0×10^?3 M catechol solution (catechol, dopamine, norepinephrine, epinephrine or L ‐dopa in a 0.050 M phosphate buffer pH 7.40) by applying 1.00 V for 60 min. The properties of the polymers are very dependent on the nature of the catechol, L ‐dopa being the best. Glassy carbon electrodes modified with melanic polymers electrogenerated from L ‐dopa and norepinephrine were found to be suitable for dopamine determinations in flow systems, although the behavior was highly dependent on the nature of the monomer. Detection limits of 5.0 nM dopamine and interferences of 9.0 and 2.6% for 5.0×10^?4 M ascorbic acid and 5.0×10^?5 M dopac, respectively, were obtained at the glassy carbon electrode modified with a melanin‐type polymer generated from L ‐dopa (using 1.0×10^?3 M AA in the measurement solution). The advantages of using a melanin‐type polymer generated from dopamine to improve the selectivity of glucose biosensors based on carbon paste electrodes containing Pt and glucose oxidase (GOx) are also discussed. The resulting bioelectrodes combines the high sensitivity of metallized electrodes with the selectivity given by the polymeric layer. They exhibit excellent performance for glucose with a rapid response (around 10 seconds per sample), a wide linear range (up to 2.5×10^?2 M glucose), low detection limits (143 μM) and a highly reproducible response (R.S.D of 4.9%). The bioelectrodes are highly stable and almost free from the interference of large excess of easily oxidizable compounds found in biological fluids, such as ascorbic acid (AA), uric acid (UA) and acetaminophen.     

Ordered Mesoporous Carbon Paste Electrodes for Electrochemical Sensing and Biosensing

Surface renewable ordered mesoporous carbon paste electrodes (OMCPE) were prepared by mechanical mixing ordered mesoporous carbon (OMC) and mineral oil. Electrochemical behavior of the composite electrode was evaluated and compared with the conventional graphite paste (GPE) and carbon nanotubes paste (CNTPE) electrodes. The OMCPE provided improved electron transfer kinetics and catalytic capabilities in connection with oxidation and/or reduction of different redox systems, such as ferricyanide and some biological species, e. g. ascorbic acid (AA), uric acid (UA), β‐nicotinamide adenine dinucleotide (NADH), dopamine (DA), epinephrine (EP), acetaminophenol (AP) and hydrogen peroxide. The substantial decrease in the over voltage of the hydrogen peroxide oxidation along with the facile incorporation of glucose oxidase (GOD) into the composite matrix allowed us successfully to fabricate a sensitive and selective glucose biosensor. A linear response up to 15 mM glucose was obtained for the OMCPE modified with 10% GOD (w/w) with a detection limit of 0.072 mM. In addition, we also successfully applied the OMCPE to the anodic stripping voltammetric analysis of heavy metal ions with improved sensitivities in comparison with CNTPE and GPE. The excellent experimental results implicate that the new developed paste electrode holds great promise in the design of electrochemical devices, such as sensors and biosensors.     

Electroanalytical Study of Prussian Blue Modified Glassy Carbon Paste Electrodes

Two types of glassy carbon (GC) powder (i.e., Sigradur K and Sigradur G) have been mixed with mineral oil to obtain glassy carbon paste electrodes (GCPE's). The electrochemical behavior of such electrodes at different percentages of glassy carbon has been evaluated with respect to the electrochemistry of ferricyanide as revealed with cyclic voltammetry and the best paste composition was chosen. GC was then modified with Prussian Blue (PB), mixed at different percentages with unmodified GC and with a fixed amount of mineral oil in order to obtain PB modified glassy carbon paste electrodes (PB‐GCPE's). PB‐GCPE's with different percentages of GC modified with PB (PB‐GC) were compared and the dependence on the amount of PB on their performances was evaluated by studying the parameters of cyclic voltammetry (i.e., current peak, ΔE_p, anodic and cathodic current ratio, charge density) and the amperometric response to H₂O₂. Data interpretation based on the GC surface area is presented. GCPE's with a selected amount of PB‐GC were then tested as H₂O₂ probes and all the analytical parameters together with the dependence on pH were evaluated. Some preliminary experiments with these electrodes assembled as glucose, lysine and lactate biosensors are also reported.     

Highly Sensitive and Selective Glucose Biosensing at Carbon Paste Electrodes Modified with Electrogenerated Magnetite Nanoparticles and Glucose Oxidase

This work reports the advantages of carbon paste electrodes modified with electrogenerated magnetite nanoparticles. The nanoparticles present catalytic activity towards hydrogen peroxide reduction. The incorporation of glucose oxidase (GOx) and magnetite in a carbon paste matrix have made possible the development of an efficient glucose biosensor. The effect of the amount of GOx and magnetite present in the composite on the response of the biosensor was critically evaluated. The biosensors demonstrated to be highly selective, with negligible interference of ascorbic acid and uric acid. The proposed biosensor was challenged with human blood serum demonstrating an excellent correlation with the spectrophotometric method.