基于二氧化锆和碳纳米管协同作用的过氧化氢生物传感器

过氧化氢作为一种重要的化工产品在纺织行业、化工行业、造纸工业、环保行业、电子行业、食品卫生行业及其他领域得到广泛的应用~([1]).     

基于纳米金自组装膜固定辣根过氧化物酶的生物传感研究

将辣根过氧化物酶(HRP)通过纳米技术和自组装技术固定于电极表面,制得了酶修饰电极.纳米金与HRP形成了静电复合物并高效地保持了HRP的生物活性,以对苯二酚作为电子媒介体,差示脉冲伏安法(DPV)研究生物酶电极测定H2O2的线性范围为5.0×10-6～1.0×10-3 mol/L,检测限为2.5×10-6 mol/L,线性方程为△I=0.34765+4.05553CH2O2(mM).酶电极的表观米氏常数(K(app))为0.0675 mmol/L.实验同时证明该生物酶电极具有良好的稳定性和使用寿命.     

流动注射化学发光植物组织传感器测定草酸盐

草酸是尿道结石主要成因之一 .统计结果表明 ,90 %以上结石均含草酸钙 .回肠病、口角性肠胃炎及脂肪吸收不良均可引起尿液中草酸的增加 .草酸盐的测定在临床上具有重要意义 .微量草酸测定的传统方法是变色酸比色法或偶氮化合物比色法[1 ] .测定草酸盐的方法还有原子吸收光谱法[2 ] 、高效液相色谱法[3] 、离子色谱法[4] 、光度法[5] 和草酸氧化酶法[6] 等 .这些方法或操作复杂、耗时 ,或灵敏度低 ,使其应用受到限制 .本文提出一种测定草酸盐的新方法 ,其原理为草酸 +Ｏ2草酸氧化酶 ＣＯ2 +Ｈ2 Ｏ2 ,Ｌｕｍｉｎｏｌ+Ｈ2 Ｏ2 ｈｖ　　将含有草…     

基于石墨烯-壳聚糖-辣根过氧化物酶的H2O2生物传感器的研制

制备了石墨烯-壳聚糖(GR-CS)纳米复合材料,并将之与辣根过氧化物酶(HRP)混合,构建了基于石墨烯-壳聚糖-辣根过氧化物酶的生物传感器(GR-CS-HRP/GC)。探针及循环伏安研究表明,该界面具有优异的电子传导能力、较大的比表面积和良好的生物相容性,对H2O2的还原显示出较好的电催化活性,在工作电位为-0.2 V,0.05 mol/L的磷酸盐缓冲盐溶液(PBS,pH 6.8)中,该酶传感器对过氧化氢响应灵敏度高,检测范围宽,测定H2O2的线性范围为5.0×10-7～2×10-3mol/L(相关系数为0.998)。检出限为2.0×10-7mol/L(S/N=3)。并且表现出良好的稳定性和高选择性。该电极用于实际样品中H2O2的测定,结果令人满意。     

基于聚乙烯醇离子液体负载HRP修饰石墨烯/纳米金复合膜的过氧化氢生物传感器

以石墨烯/纳米金修饰玻碳电极为基底, 用聚乙烯醇与离子液体复合物将辣根过氧化物酶固定于电极表面, 制备了过氧化氢生物传感器. 结果表明, 在0.1 mol/L HAc-NaAc+0.1mol/L KCl(pH=6.5)中, H₂O₂的氧化峰电流与其浓度在9.55×10^-6~6.01×10^-3 mol/L间呈良好线性关系, 检出限(3S/N)为3.3×10^-7 mol/L. 用标准加入法做回收实验, 回收率在93.4%~100.5%之间. 该传感器对H₂O₂具有较高的灵敏度和较低的检测限, 稳定性和重现性良好, 使用寿命较长, 且制作成本低, 可多次重复使用.     

基于拮抗作用检测除草剂的类囊体膜生物传感器研究

利用除草剂对植物类囊体束缚酶分解过氧化氢的拮抗作用,研制了一种快速检测痕量除草剂的电化学生物传感器.将植物类囊体用聚乙烯醇-苯乙烯吡啶(PVA-SbQ)光敏聚合剂在紫外光诱导下产生大分子网状结构进行包埋,制成生物敏感膜,并固定在铂电极表面.根据加入除草剂时类囊体膜束缚酶分解过氧化氢活性的变化,对除草剂进行测定.在含有1×10^-3mol/LNaCl,5×10^-3mol/LMgCl₂和0.01mol/LH₂O₂的Tris-HCl缓冲溶液(pH=7.4)中,基于测量0.65V处H₂O₂氧化电流的变化,可以对下列浓度的除草剂进行定量检测:百草枯3×10^-9～1.5×10^-7mol/L,敌草龙1×10^-8～3×10^-7mol/L,扑草净4×10^-8～3×10^-6mol/L,阿特拉津1×10^-7～5×10^-6mol/L,莠灭净1×10^-7～5×10^-6mol/L.利用PVA-SbQ光聚合膜固定类囊体,能够使酶的活性在低温下保持数月.     

Hydrogen peroxide biosensor based on electrodeposition of zinc oxide nanoflowers onto carbon nanotubes film electrode

A new amperometric biosensor for hydrogen peroxide was developed based on adsorption of horseradish peroxidase at the glassy carbon electrode modified with zinc oxide nanoflowers produced by electrodeposition onto multi-walled carbon nanotubes （MWNTs） film. The morphology of the MWNTs/nano-ZnO electrode has been investigated by scanning electron microscopy （SEM）, and the electrochemical performance of the electrode has also been studied by amperometric method. The resulting electrode offered an excellent detection for hydrogen peroxide at -0.11 V with a linear response range of 9.9×10^-7 to 2.9×10^-3 mol/L with a correlation coefficient of 0.991, and response time 〈5 s. The biosensor displays rapid response and expanded linear response range, and excellent stability.     

基于等离子体聚合膜固定酶的H2O2生物传感器

以玻碳电极为基础电极,用微波等离子体技术聚合沉积聚乙二胺等离子体膜,使之形成带氨基功能团的表面,再通过戊二醛交联共价固定辣根过氧化物酶,制得H2O2生物传感器.探讨了等离子体聚合膜的形成条件(如放电功率、单体流速、单体气压和聚合时间),讨论了工作电位、介体浓度和pH值对传感器响应的影响.此外,用红外光谱对等离子体聚合膜进行了表征.该传感器在5×10-7～1.1×10-3mol/LH2O2浓度范围内有线性响应,最低检测限为0.3μmol/L.将此传感器用于实际试样回收率的测定,结果良好.     

基于纳米金和硫堇固定酶的过氧化氢生物传感器

在铂电极上自组装一层纳米金(GNs), 构建负电荷的界面, 然后通过金-硫、金-氮共价键合作用和静电吸附作用自组装一层阳离子电子媒介体硫堇(Thio). 再以同样的作用自组装一层GNs和辣根过氧化酶(HRP)的混合物, 最后在电极最外层滴加一层疏水性聚合物壳聚糖(Chit), 由此制备了一种新型的过氧化氢生物传感器. 研究了工作电位、检测底液pH、温度对响应电流的影响, 以及GNs和HRP之间的相互作用, 探讨了传感器的表面形态、交流阻抗、重现性和稳定性. 该传感器的酶催化反应活化能为12.4 kJ/mol, 表观米氏常数为6.5×10^－4 mo/L, 在优化的实验条件下, 所研制的传感器对H₂O₂的线性范围为5.6×10^－5～2.6×10^－3 mol/L, 检出限为1.5×10^－5 mol/L. 应用此方法制备了HRP和葡萄糖氧化酶(GOD)双酶体系葡萄糖生物传感器, 并应用于实验样品葡萄糖含量的测定.     

基于Hemin/多壁碳纳米管纳米复合物构建的过氧化氢生物传感器的研究

利用卟啉(Hemin)具有模拟酶的功能,与多壁碳纳米管(MWCNTs)构建了一种新型的过氧化氢(H2O2)生物传感器。首先,利用Hemin与MWCNTs之间的π-π键作用,在超声分散下制备Hemin/MWCNTs纳米复合物;采用滴涂技术并在nafion的作用下将其固载在电极表面,制得该H2O2生物传感器(nafion/Hemin/MWCNTs/GCE)。采用紫外-可见分光光度法(UV-Vis)对合成的纳米复合物进行了分析;采用扫描电镜(SEM)对电极的表面形貌进行了表征;采用循环伏安法和计时电流法考察了该修饰电极的电化学行为;并对传感器的行为进行了详细的研究。在最优条件下,此修饰电极对H2O2具有明显的催化作用,电流与H2O2的浓度在6.0×10-7~1.8×10-3 mol/L范围内呈现良好的线性关系,检出限达2.0×10-7 mol/L。此传感器制作简单,具有较高的灵敏度和良好的稳定性及重现性。     

A peroxidase-tetrathiafulvalene biosensor based on self-assembled monolayer modified Au electrodes for the flow-injection determination of hydrogen peroxide

The construction and performance under flow-injection conditions of an integrated amperometric biosensor for hydrogen peroxide is reported. The design of the bioelectrode is based on a mercaptopropionic acid (MPA) self-assembled monolayer (SAM) modified gold disk electrode on which horseradish peroxidase (HRP, 24.3 U) was immobilized by cross-linking with glutaraldehyde together with the mediator tetrathiafulvalene (TTF, 1 μmol), which was entrapped in the three-dimensional aggregate formed.

The amperometric biosensor allows the obtention of reproducible flow injection amperometric responses at an applied potential of 0.00 V in 0.05 mol L⁻¹ phosphate buffer, pH 7.0 (flow rate: 1.40 mL min⁻¹, injection volume: 150 μL), with a range of linearity for hydrogen peroxide within the 2.0 × 10⁻⁷–1.0 × 10⁻⁴ mol L⁻¹ concentration range (slope: (2.33 ± 0.02) × 10⁻² A mol⁻¹ L, r = 0.999). A detection limit of 6.9 × 10⁻⁸ mol L⁻¹ was obtained together with a R.S.D. (n = 50) of 2.7% for a hydrogen peroxide concentration level of 5.0 × 10⁻⁵ mol L⁻¹. The immobilization method showed a good reproducibility with a R.S.D. of 5.3% for five different electrodes. Moreover, the useful lifetime of one single biosensor was estimated in 13 days.

The SAM-based biosensor was applied for the determination of hydrogen peroxide in rainwater and in a hair dye. The results obtained were validated by comparison with those obtained with a spectrophotometric reference method. In addition, the recovery of hydrogen peroxide in sterilised milk was tested.     

Amperometric biosensors based on alumina nanoparticles-chitosan-horseradish peroxidase nanobiocomposites for the determination of phenolic compounds

A horseradish peroxidase (HRP) biosensor based on alumina (Al(2)O(3)) nanoparticles-chitosan (CHIT) nanocomposites was developed for the detection of phenolic compounds. UV-Vis spectra and Fourier transform infrared spectra showed that HRP retained its original structure on the Al(2)O(3)/CHIT film. The surface morphologies of the composite films were characterized by scanning electron microscopy. Cyclic voltammetry and amperometry were used to study the proposed electrochemical biosensor. Optimization of the experimental parameters was performed with regard to pH, applied electrode potential and the concentration of hydrogen peroxide. The linear range, sensitivity and detection limit of the biosensor were investigated for eight phenolic compounds. In particular, the linearity of the biosensor for the detection of hydroquinone was obtained from 5 × 10(-9) M to 7 × 10(-5) M with a detection limit of 1 nM (based on the S/N = 3). The optimized biosensor for hydroquinone determination displayed a high sensitivity of 518.4 nA μM(-1) with a response time of ～5 s.     

Enhanced peroxidase activity of hemoglobin in a DNA membrane and its application to an unmediated hydrogen peroxide biosensor.

Hemoglobin can exhibit not only a direct electron transfer reacting after being entrapped in a DNA membrane, but also a greatly enhanced peroxidase activity toward the reduction of hydrogen peroxide. Based on the direct electrochemical property and nice enzymatic activity of the protein in a DNA membrane, a reagentless hydrogen peroxide biosensor was prepared. The peak current related to hydrogen peroxide was linearly proportional to its concentration in the range of 1.9 x 10(-6)-6.8 x 10(-4) mol L(-1). The detection limit was 1 x 10(-6) mol/L.     

Using novel polysaccharide-silica hybrid material to construct an amperometric biosensor for hydrogen peroxide

A new type of sol-gel organic-inorganic hybrid material was developed and used for the fabrication of an amperometric hydrogen peroxide biosensor. This material was prepared from natural chitosan and recently introduced completely water-soluble precursor, tetrakis(2-hydroxyethyl) orthosilicates (THEOS), by the sol-gel process without the addition of organic solvents and catalysts. The gelation time for the sol-gel transition and dynamic rheological properties of the resultant gel matrix could be modulated by the amount of added THEOS. The structure of the hybrid gel was made up of a network and spherical particles, as confirmed by SEM observation. By electrochemical experiments, it was found that such a hybrid gel matrix could retain the native biocatalytic activity of the entrapped horseradish peroxidase and provide a fast amperometric response to hydrogen peroxide. The linear range for the determination of hydrogen peroxide was found to be from 1.0 x 10(-6) to 2.5 x 10(-4) mol/L with a detection limit of 4.0 x 10(-7) mol/L. The apparent Michaelis-Menten constant was determined to be 2.198 mmol/L. To improve the analytical characteristics of the fabricated biosensor, the effects of applied potential and pH value on the steady-state current were studied. Under the optimized experimental conditions, the fabricated biosensor was found to have good analytical performance, reproducibility, and storage stability.     

Multilayer assembly of Prussian blue nanoclusters and enzyme-immobilized poly(toluidine blue) films and its application in glucose biosensor construction

A multilayered glucose biosensor via sequential deposition of Prussian blue (PB) nanoclusters and enzyme-immobilized poly(toluidine blue) films was constructed on a bare Au electrode using electrochemical methods. The whole configuration of the present biosensor can be considered as an integration of several independent hydrogen peroxide sensing elements. In each sensing element, the poly(toluidine blue) film functioned as both the supporting matrix for the glucose oxidase immobilization and the inhibitor for the diffusion of interferences, such as ascorbic acid and uric acid. Meanwhile, the deposited Prussian blue nanocluster layers acts as a catalyst for the electrochemical reduction of hydrogen peroxide formed from enzymatic reaction. Performance of the whole multilayer configuration can be tailored by artificially arranging the sensing elements assembled on the electrode. Under optimal conditions, the biosensors exhibit a linear relationship in the range of 1 x 10(-4) to 1 x 10(-2) mol/L with the detection limit down to 10(-5) mol/L. A rapid response for glucose could be achieved in less than 3 s. For 1 mM glucose, 0.5 mM acetaminophen, 0.2 mM uric acid, and 0.1 mM ascorbic acid have no obvious interferences (<5%) for glucose detection at an optimized detection potential. The present multilayered glucose biosensor with a high selectivity and sensitivity is promising for practical applications.     

Amperometric sensor for hydrogen peroxide based on electric wire composed of horseradish peroxidase and toluidine blue-multiwalled carbon nanotubes nanocomposite

A kind of nanocomposites with good dispersion in water was prepared through noncovalent adsorption of toluidine blue (Tb) on multiwalled carbon nanotubes (MWCNT) for electric communication between horseradish peroxidase (HRP) and electrode. The nanocomposites could be conveniently cast on electrode surface. With the aid of chitosan, HRP was then immobilized on the nanostructure to form a reagentless amperometric sensor for hydrogen peroxide. UV-vis spectroscopy and electrochemical impedance spectroscopy were used to characterize the adsorption of Tb on MWCNT. The presence of both Tb as mediator of electron transfer and MWCNT as conductor enhanced greatly the enzymatic response to the reduction of hydrogen peroxide. The novel biosensor exhibited fast response towards hydrogen peroxide with a detection limit of 1.7x10(-6)M and the linear range extended up to 4x10(-4)M without the interference of ascorbic acid and uric acid. The Michaelis-Menten constant (K'(m)) of the immobilized HRP was evaluated to be 0.16mM.     

Amperometric Biosensor for Hydrogen Peroxide Based on Electrodeposited Sub-micrometer Gold Modified Glassy Carbon Electrode

IntroductionAmperometricbiosensorofhydrogenperoxideisofpracticalimportancebecauseofitswideapplicationsinchemical,biological,clinical,environmentalandmanyotherfields.Forimprovementofsensor抯quality,vari-ouskindsofchemicalmodificationmethodshavebeendevelopedforreducingredoxoverpotentialsofH2O2atelectrodesurfaces,increasingthedetectionsensitivity,linearrange,stabilityandlivetime.Ithasbeenshownthattheuseofsub-micrometersizedmetalparticlessuchasPt-blackcansignificantlyimprovethequalityofthebiosens…     

毛细管电泳电化学法检测过氧化氢

以金微盘电极和离子液体修饰单壁碳纳米管糊微盘电极分别作为毛细管电泳电化学检测器,试验了两种电极对过氧化氢的响应情况,将金微盘电极与毛细管电泳联用,对过氧化氢进行了定性和定量检测.探讨了分离电压、缓冲溶液pH值和工作电位等条件对H2O2检测的影响.实验结果表明,峰电流与H2O2浓度在1.0×10-6～1.0×10-5mo...     

溶胶-凝胶法固定胆固醇氧化酶偶联普鲁士蓝化学修饰电极的研究

用溶胶-凝胶法将胆固醇氧化酶固定在普鲁士蓝修饰的玻碳电极表面,制成了一种新型胆固醇传感器,实现了低电位下对胆固醇的间接测定,胆固醇的测定范围伏安法为5×10^-7～8×10^-5mol/L,安培法为5×10^-6～5×10^-4mol/L.伏安法检出限为1.2×10^-7mol/L,是目前所见灵敏度最高的胆固醇传感器之一,该传感器对胆固醇的测定可避免常规电化学传感器测定中由于样品中大量存在的易氧化物质所带来的干扰,该传感器的寿命长,使用次数在300次以上.     

Biosensor for H2O2 Response Based on Horseradish Peroxidase: Effect of Different Mediators Adsorbed on Silica Gel Modified with Niobium Oxide

Reagentless biosensors sensitive to hydrogen peroxide have been developed and compared. These biosensors are comprised of a carbon paste electrode modified with horseradish peroxidase (HRP) and one phenothiazine (methylene blue), one phenoxazine (meldola's blue) or one phenazine (phenazine methosulfate) dye adsorbed on silica gel modified with niobium oxide (SN). The enzyme was immobilized onto the graphite powder by cross‐linking with glutaraldehyde and mixing with one of the electron transfer mediators (dyes) adsorbed on SN. The amperometric response was based on the electrocatalytic properties of the dye to mediate electrons, which were generated in the enzymatic reaction of hydrogen peroxide under catalysis of HRP. The dependence on the biosensor response in terms of pH, buffer, HRP amounts and applied potential was investigated. The best results were found with a biosensor containing methylene blue dye showing an excellent operational stability (around 92% of the activity was maintained after 300 determinations). The proposed biosensor also presented good sensitivity (32.87 nA cm^{?2 μ}mol^?1 L) allowing hydrogen peroxide quantification at levels down to 0.52×10^?6 mol L^?1 an optimum response at pH 6.8 and at a potential of ?50 mV (vs. SCE) and showing a wide linear response range (from 1 to 700 μmol L^?1 for hydrogen peroxide).