亚甲蓝为作酶电极电子传递介体的性能研究

研究了亚甲蓝修饰的玻璃电极的催化性能，并以此电极为基础电极研制了介体修饰型多酚氧化酶电极，对多巴胺测定的检出限为５×１０＾－６ｍｏｌ／Ｌ，较空白电极降低了４００倍，而对维生素Ｃ的响应同显著减小，其电流值降低至空白电极的十分之一，因此不仅显著提高了灵敏度，选择性也得到很大的改善。     

修饰酶电极的响应特性

在考虑电极电位、浓度极化等影响因素的情况下, 提出了介体型酶电极响应的数学表达式, 并对吸附法制备的修饰酶电极进行了初步的实验验证和模拟计算, 其实验结果与理论推导是一致的.     

葡萄糖氧化酶介体电极稳定性的研究

本文阐述了介体行丢失速率与电极基线电流大小的关系，以此为依据，讨论了介体液态石蜡中的溶解度对介体丢失的影响，通过向电极表面添加ＤＥＡＥ－葡萄糖来减小带正电荷的介体丢失速率，研究了ＤＥＡＥ－葡萄糖对葡萄糖氧化酶介体电极测试性能的影响。     

漆酶电极的研制及应用

本文报道二茂铁修饰电极为基底的漆酶电极的研制及应用，讨论了电极性能及响应机理，找出了实验最佳条件。并对环境污水中对苯二酚含量进行规定。一结果令人满意。     

介体型酶电极二级催化反应速度常数的测定

采用开路驰豫法，测定了以二茂铁及其衍生物作为电子传递体的介体型酶电极的均相二级催化反应速度常数，结果表明二甲氨基甲基二茂铁，在绝氧条件下对于酶的再生反应具有高的催化活性，是一种性能优良的电子传递体．此外，讨论了开路弛豫法在酶电极研究中的适用条件及各种因素对该方法测定结果的影响．     

壳聚糖固定化醇脱氢酶的制备及其酶学性质研究

报道了醇脱氢酶(ADH)的固定化和酶学性质研究。以壳聚糖作为载体,戊二醛作为交联剂。固定化ADH的最适条件为:以6%戊二醛将壳聚糖交联2 h,与ADH反应2.5 h。对游离和固定化ADH酶学性质的研究表明:酶促反应的最适pH均为8.2,最适温度分别为37℃和40℃,对乙醇的表观米氏常数Km分别为33.9 mmol/L和46.2 mmol/L。与游离酶相比,固定化酶具有良好的操作稳定性。     

过氧化氢酶电极的研制及其性能

生物传感器是一种新兴的测试工具,其中酶电极因具有较常规离子选择电极更短的响应时间,更好的选择性与抗干扰能力强等特性,引起了人们的关注。本文用新鲜合成的三乙基     

聚吡咯修饰乳酸氧化酶电极的研制

根据聚吡咯修饰电极掺杂和去掺杂原理，将乳酸氧化酶固定在玻碳电极表面形成一种新型的乳酸酶电极，该电极灵敏度高，稳定性好，易于制作。本文报道了该电极的研制过程，探讨了影响电极响应的各种因素，找出了最佳实验条件。将此电极用于实际样品中乳酸含量的测定，结果令人满意。     

A New Amperometric Carbon Paste Enzyme Electrode for Ethanol Determination

Abstract

In this study, a new amperometric carbon paste enzyme electrode for determination of ethanol was developed. The carbon paste was prepared by mixing alcohol dehydrogenase, its coenzyme nicotinamide adenine dinucleotide (oxidized form, NAD⁺), poly(vinylferrocene) (PVF) that was used as a mediator, graphite powder and paraffin oil, then the paste was placed into cavity of a glass electrode body. Determination of ethanol was performed by oxidation of nicotinamide adenine dinucleotide (reduced form, NADH) generated enzymatically at +0.7 V. The effects of enzyme, coenzyme and PVF amounts; pH; buffer concentration and temperature were investigated. The linear working range of the enzyme electrode was 4.0×10^?4–4.5×10^?3 M, determination limit was 3.9×10^?4 M and response time was 50 s. The optimum pH, buffer concentration, temperature, and amounts of enzyme, NAD⁺ and PVF for enzyme electrode were found to be 8.5, 0.10 M, 37°C, 2.0, 6.0, and 12.0 mg, respectively. The storage stability of enzyme electrode at +4°C was 7 days. Enzyme electrode was used for determination of ethanol in two different wine samples and results were in good agreement with those obtained by gas chromatography.     

电聚合修饰碳纤维电极及乳酸脱氢酶活性的测定

用电聚合方法将亚甲基绿 (MG)修饰在碳纤维电极 (直径 7μm)上 ,并用该修饰电极测定了乳酸脱氢酶 (LDH)活性。在烟酰胺腺嘌呤二核苷酸 (NAD+ )浓度为 6 .0× 10 - 4mol L ,乳酸浓度为 5 .0× 10 - 3mol/L的pH7.0NaOH KH2 PO4 缓冲介质中 ,电位恒定在 +0 .10V下 ,用电流法测定乳酸脱氢酶活性 ,线性范围为 15～ 2 4 0U/mL ,检测限为 10U/mL ,响应时间为 15s。该修饰微电极稳定性好、灵敏度高、测定干扰小     

A D-Carnitine Dehydrogenase Electrode For The Assessment Of Enantiomeric Purity Of L-Carnitine Preparations

ABSTRACT

The enantiomeric purity of pharmaceutical L-carnitine preparations can be assessed within 60 seconds using a highly selective bienzyme electrode. D-carnitine dehydrogenase from Agmbacterium is highly specific for the non-physiological D-enantiomer and was therefore used as the recognition element. NADH produced in the primary reaction was oxidized by salicylate hydroxylase (EC 1.14.13.1) in an oxygen and salicylate dependent reaction. The consumption of oxygen was monitored with a miniature Clark-electrode. A linear calibration graph from 0.01 mM through 0.6 mM D-carnitine was obtained in phosphate buffer pH 8 comprising 0.5 mM concentrations of the cosubstrates NAD and salicylate. The sensitivity for DL-carnitine was exactly 50% of the respective value for pure D-carnitine, while L-carnitine and ascorbic acid, a common interferent, gave no response at all. Mixtures of both enantiomers containing 1% and 3% D-carnitine, respectively, could be distinguished from each other and from pure (i.e. >98 %) L-carnitine preparations with the new sensor. The biosensor method is faster and less laborious than established HPLC and ¹H-NMR methods since it requires no chemical derivatization. The lower detection limit was 10-fold reduced as compared with a recently published enzymatic assay.     

Amperometric Ethanol Biosensor Based on Integration of Alcohol Dehydrogenase with Meldola's Blue/Ordered Mesoporous Carbon Electrode

An amperometric ethanol biosensor was fabricated by integration of alcohol dehydrogenase (ADH) with meldola's blue (MB)/ordered mesoporous carbon (OMC) composite modified glassy carbon electrode (MB/OMC/GCE). The MB/OMC/GCE was highly sensitive for nicotinamide adenine dinucleotide (NADH) measurement (9.1±0.25 μA/mM) and gave a low detection limit of 0.21±0.02 μM. The ethanol biosensor exhibited a wide linear range up to 6 mM with a lower detection limit of 19.1±0.58 μM as well as a high sensitivity of 34.58±2.43 nA/mM without suffering any interference from some common electroactive compounds.     

甲苯胺蓝修饰石墨电极为基体的乙醇脱氨酶生物传感器

报道了电流型乙醇生物传感器。该传感器以甲苯胺蓝键合修饰浸蜡石墨电极为基 体电极，将醇脱氢酶、烟酰胺腺嘌呤二核苷酸（ＮＡＤ＋）同时固定在蚕丝蛋白膜上，成为无试剂 的醇传感器。在 ｐＨ ８． ５的 Ｔｒｉｓ－ＨＣｌ介质中，该传感器的响应电流与乙醇浓度在 ５． ０× １０－５～ １．１０×１０－３ｍｏｌ／Ｌ范围内有良好的线性关系。响应时间为２０ｓ。本文讨论了影响传感器响应 的各种因素。用该传感器测定了啤酒中乙醇的含量，结果与气相色谱法一致。     

A Bioanode Using Thermostable Alcohol Dehydrogenase for an Ethanol Biofuel Cell Operating at High Temperatures

To extend the range of biofuel cell applications, we wish to increase their maximum operational temperatures. Using a thermostable alcohol dehydrogenase as a biocatalyst, we prepared an enzyme‐immobilized bioanode that can operate at high temperatures. The catalytic current for ethanol oxidation was increased using this electrode at temperatures up to 80 °C.     

Study on the United Glucose Oxidase Electrode Constructed by Composite Electrode

A novel type of united glucose oxidase (GOD) electrode was designed. Glucose oxidase and ferrocene (Fc), which was a mediator, were added into the composite electrode that was constructed by graphite powder, acetylene black, and epoxy resin. These three materials in composite electrode kept constant proportion in weight. And the optimum amounts of GOD and Fc among united enzyme electrode were 5% and 2%, respectively. The glucose was detected linearly in the concentration range 0.01–9.0 mM with a 20-s steady-state response time and 36 nA/mM of the sensitivity at 0.15 V applied potential. And electrode fouling problem and the response current from the interferents were avoided. The response current of the united GOD electrode had no obvious deterioration within 80 days when stored at 4°C in a refrigerator. The detecting results of human serum by the united GOD electrode had good consistency with that by standard enzyme method. The maximum deviation between these two detecting values was 5%. It might be used for detecting the blood sugar in clinical assay.     

聚苯胺葡萄糖氧化酶电极的催化过程

用电化学方法固定在直径为0.5mm铂丝上的聚苯胺(PANI)葡萄糖(GOD)电极对葡萄糖有催化氧化作用.在0～-0.6V(vs.SCE)的电极范围内,在电极的循环伏安曲线上观察到与葡萄糖浓度有关的氧的还原峰和GOD还原态的氧化峰,用此GOD还原态的氧化峰电流可定量检测葡萄糖的浓度。本文提出在PANI电极上存在着酶反应氧化还原电荷直接传递的可能性。     

High‐sensitive Electrochemical Determination of Ethyl Carbamate Using Urethanase and Glutamate Dehydrogenase Modified Electrode

An amperometric biosensor for ethyl carbamate (EC) was developed for the first time through the cascade reactions of urethanase and glutamate dehydrogenase (GLDH). Urethanase decomposes ethyl carbamate to produce ammonia, which converts to L‐glutamate under the catalysis of GLDH in the presence of α‐ketoglutarate and NADH. Then the change of NADH can be detected chronoamperometrically. The two enzymes were entrapped into chitosan/gelatine/γ‐glycidoxy propyl trimethoxy silane sol‐gel and immobilized on the surface of pyrolytic graphite electrode (PGE). The modified electrode was characterized by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). Under the optimized conditions, the amperometric EC biosensor exhibits a linear detection range from 0.5 to 40 μM with a low detection limit of 5.30 nM. The biosensor was successfully used to detect EC in mimic Chinese rice wine samples, and satisfactory recovery and relative standard deviation were achieved.