Measurement with Amperometric Sensors

Amperometric with macroelectrodes is done using approved technical equipment. There are numerous commercially available devices with equal design. The electrochemical cell works with three electrodes (Chap. 2, Sect. 2.4), i.e. working, auxiliary and reference electrodes. They are driven by a potentiostat designed as an analogue electronic drcuit Digital potential control instead of analogue control circuitry did not stand the test of time, since it does 

In commercially availaljle potentiostats, specific additional electronic modules are provided for picoampere measurements. For classic investigations. 

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The type of molecular recognition reaction determines the form of the transducer used (Table 5.3). Enzymatic reactions often involve an electron transfer. This electrical activity can be measured with amperometric, potentiometric or conductometric sensors. If the bioreaction includes the generation of H+ or OH ions, then a pH sensitive dye in combination with an optical device can be used. For antibody-antigen binding, the mass change on the surface of the transducer can be detected with a piezoelectric device. Exothermic or endothermic reactions can be followed with a temperature sensor. 

D. Preparation and Measurement of Nitric Oxide with Amperometric Sensors.245... 

D. PREPARATION AND MEASUREMENT OF NITRIC OXIDE WITH AMPEROMETRIC SENSORS... 

With amperometric sensors, the electrode is polarized by a predetermined potential, and the resulting electrolysis current is measured. Under certain conditions, saturation is achieved, and the limiting current is proportional to the actual concentration. Instead of the transducer principle of energy conversion (with potentiometry)y we now have the current-limiting transduction. 

Agents for chemical bleaching rely on different types of peroxides. Potentiometric or amperometric biosensors that detect the highly specific and sensitive reaction of enzymes like katalases with their corresponding substrates can be used for on-line measurement [84]. The sensors can be manufactured with simple technologies at moderate cost, but their stability is not sufficient for integration in household appliances. 

In Chapters 6 and 7, we discussed potentiometric and amperometric sensors, respectively. The third basic electrochemical parameter that can yield sensory information is the conductance of the electrochemical cell (Fig. 8.1). Conductance is the reciprocal of resistance. It is related to current and potential through the generalized form of Ohm s law (C.l). If the measurement is done with AC signal conductance (G) becomes frequency-dependent conductance G(co) and the resistance R becomes impedance Z( (o). 

In this chapter, the development of an amperometric sensor will be explained and discussed. The principle of the analysis method will be based on the results described in Chapter4 this means that use will be made of the oxidation reaction of hydrogen peroxide in the prewave, and that the concentration will be determined using the rate equation. In addition to measurement of the electrical current response, temperature and pH will therefore also be measured. Accordingly, it is interesting to start with an investigation of the temperature influence. 

Immobilization of bioactive material on/in the electrode allows combining bio-reaction selectivity with sensitivity of electrochemical detection. Irrespective of reaction in the biosensor, the electrochemical response is measured, in particular, as current at the given potential (amperometric sensor) or electrode potential (potentiometric sensor). 

Acetylcholineesterase A 3 mm diameter, Pt disc electrode, either polished or black, was immersed for 20 min in a solution of avidin (lOOpg/mL). After washing with PBS solution of pH 7.4, the electrode was immersed for 20 min in a biotin-labeled ChO solution (lOOpg/mL). After treatment 10 times, the surface of the sensor was further modified with ChE in the same manner. The amperometric response of the sensor was measured with a three-electrode cell at 0.6 V versus Ag/ AgCl in 0.1 M phosphate buffer of pH 6.8. The response was linear from 1 pM to 1 mm of acetylcholine. [102]... 

Amperometric sensors — A class of electrochemical sensors based on amperometry. A - diffusion-limited current is measured which is proportional to the concentration of an electrochemically active analyte. Preferred technique for - biosensors with or without immobilized enzymes (biocatalytic sensors). The diffusion layer thickness must be kept constant, either by continuous stirring or by means of an external diffusion barrier. Alternatively, micro electrodes can be... 

Enzyme sensors can measure analytes that are the substrates of enzymatic reactions. Thermometric sensors can measure the heat produced by the enzyme reaction [31], while optical or electrochemical transducers measure a product produced or cofactor consumed in the reaction. For example, several urea sensors are based on the hydrolysis of urea by urease producing ammonia, which can be detected by an ammonium ion-selective ISE or ISFET [48] or a conductometric device [49]. Amperometric enzyme sensors are based on the measurement of an electroactive product or cofactor [50] an example is the glucose oxidase-based sensor for glucose, the most commercially successful biosensor. Enzymes are incorporated in amperometric sensors in functionalised monolayers [51], entrapped in polymers [52], carbon pastes [53] or zeolites [54]. Other catalytic biological systems such as micro-organisms, abzymes, organelles and tissue slices have also been combined with electrochemical transducers. 

Porphyrins are often employed in sensors on account of their ability to act as cation hosts and, with a suitable metal ion coordinated, as redox catalysts. Electropolymerised poly(metalloporphyrin)s have been used as potentiometric anion-selective electrodes [131] and as amperometric electrocatalytic sensors for many species including phenols [132], nitrous oxide [133] and oxygen [134]. Panasyuk et al. [135] have electropolymerised [nickel-(protoporphyrin IX)dimethylester] (Fig. 18.10) on glassy carbon in the presence of nitrobenzene in an attempt to prepare a nitrobenzene-selective amperometric sensor. Following extraction of the nitrobenzene the electrode was exposed to different species and cyclic voltammetric measurements made. A response was observed at the reduction potential of nitrobenzene (the polyporphyrin film acts only to accumulate the analyte and not in a catalytic fashion). Selectivity for nitrobenzene compared with w-nitroaniline and o-nitroto-luene was enhanced compared with an untreated electrode, while a glassy carbon-... 

Faradaic processes of electrode reactions, which are principle mechanism of obtaining analytical signal in amperometric sensors, significantly depend on working electrode material and state of its surface. The common working electrode materials include noble and seminoble metals, solid oxides of various elements and different kinds of carbon materials including carbon nanostructures. They are employed in conventional voltammetric measurements with various modes of electrode polarization, as amperometric chemical sensors, as well as for construction of amperometric biosensors. 

On the other hand, the molecular recognition by enzymes, which are also applied in the form of organelles, microorganisms and tissue slices, is accompanied by chemical conversion of the analyte to the respective products. Therefore this type of sensor is termed a metabolism sensor2. The initial state is usually reached when the analyte conversion is complete. With metabolism sensors, under certain conditions cosubstrates, effectors, and enzyme activities can be measured via substrate determination. Amperometric and potentiometric electrodes and thermistors are the preferred transducers, but in some cases optoelectronic sensors have also been used. With biomimetic sensors physical signals such as sound, stress, or light are measured through their ability to... 

In most amperometric cytochrome b2 electrodes the reaction is followed by anodic oxidation of ferrocyanide at a potential of +0.25 V or above. The first of such sensors was assembled by Williams et al. (1970), who immobilized the enzyme (from baker s yeast) physically at the tip of a platinum electrode within a nylon net of 0.15 mm thickness. The large layer thickness resulted in a response time of 3-10 min. Owing to the low specific enzyme activity used, the sensor was kinetically controlled. Therefore the linear measuring range extended only up to 0.1 Km-A similar sensor has been applied by Durliat et al. (1979) to continuous lactate analysis. The enzyme was contained in a reaction chamber of 1 pi volume in front of the electrode. This principle has also been employed in the first commercial lactate analyzer using an enzyme electrode (Roche LA 640, see Section 5.2.3.3X With a sensor stability of 30 days and a C V below 5%, 20-30 samples/h can be processed with this device. 

The substrate generation/tip collection (SG/TC) mode with an ampero-metric tip was historically the first SECM-type measurement performed (32). The aim of such experiments was to probe the diffusion layer generated by the large substrate electrode with a much smaller amperometric sensor. A simple approximate theory (32a,b) using the well-known c(z, t) function for a potentiostatic transient at a planar electrode (33) was developed to predict the evolution of the concentration profile following the substrate potential perturbation. A more complicated theory was based on the concept of the impulse response function (32c). While these theories have been successful in calculating concentration profiles, the prediction of the time-de-pendent tip current response is not straightforward because it is a complex function of the concentration distribution. Moreover, these theories do not account for distortions caused by interference of the tip and substrate diffusion layers and feedback effects. 

Coulometry is the name given to a group of other techniques that determine an analyte by measuring the amount of electricity consumed in a redox reaction. There are two categories referred as potentiostatic coulometry and amperostatic coulometry. The development of amperometric sensors, of which some are specific for chromatographic detection, open new areas of application for this battery of techniques. Combining coulometry with the well known Karl Fischer titration provides a reliable technique for the determination of low concentrations of water. 

Optodes provided with non-fluorescent esters of fluorophores have been used for the determination of external enzyme activities. The fluorophores are liberated by the enzymes and then seen by the optical Ober [214], As ecamples of p(02)-modulated optical biosensors, a glucose probe [213] and an ethanol probe [216] can be mentioned sensors based on glucose, alcohol, and other oxidases were reviewed by Opitz and Lttbbers [217]. The advantages of these 02-dependent optical biosensors are that, unlike corresponding amperometric sensors, they do not consume O2 and that they are strictly diffusion limited in their response. Fiber-optical devices are also available for the determination of substrates of dehydrogenases the NADH fluorescence produced by the immobilized enzyme is measured as a function of time [218, 219]. 

The direct measurement of analyte concentrations below approx. 10 mol/L with electrochemical sensors is so far only possible with the advanced microfabrication process of Ikarijama et al. [21-23]. Usually a chemical amplifier system has to be used for increased sensitivity. The only analytes which already provide this amplification system themselves are enzymes catalytically turning over their substrates. If the product of the enzymatic reaction can be assayed with a sensor, the activity of the respective enzyme can also be determined. As an example, an amperometric assay for glucose oxidase using benzoquinone as an oxidant has been published [37] ... 

When a voltammetric sensor operates with a small overpotential, the faradaic reaction rate is also small consequently, a high-precision instrument for the measurement is needed. An amperometric sensor is usually operated under limiting current or relatively small overpotential conditions. Amperometric sensors operate under an imposed fixed electrode potential. Under this condition, the cell current can be correlated with the bulk concentration of the detecting species (the solute). This operating mode is commonly classified as amperometric in most sensor work, but it is also referred to as the chronosupero-metric method, since time is involved. 

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