Amperometric multidetection with composite enzyme electrodes

The use of composite biosensors for multianalyte detection strategies is discussed. Graphite–Teflon rigid composite biosensors offer the possibility of coimmobilization of several enzymes by simple physical inclusion in the bulk of the electrode matrix with no covalent linkages. A novel trienzyme graphite–Teflon–glucose oxidase (GOD)–alcohol oxidase (AOD)–peroxidase (HRP)–ferrocene bisosensor yielded amperometric steady-state currents similar to those obtained with graphite–Teflon–GOD–HRP–ferrocene and graphite–Teflon–AOD–HRP–ferrocene electrodes for the same concentration of glucose and ethanol, respectively. The performance of the trienzyme biosensor for multianalyte detection was evaluated with the simultaneous determination of glucose and ethanol after separation by HPLC, in samples of sweet wine. The simultaneous analysis of several analytes in the same sample should imply that, with an adequate dilution, the concentration levels of the analytes can be included within the ranges of linearity of the corresponding calibration plots. The use of two composite biosensors in a parallel configuration, so that different analytes can be simultaneously detected with no need of chromatographic separation, is also discussed. The usefulness of this approach was evaluated by the simultaneous analysis of glucose and ethanol in sweet wine, and of glucose and lactic acid in red wine.     

Amperometric enzyme electrodes 3. Alcohol oxidase

A rapid, simple method for the measurement of ethanol or methanol has been developed by coupling amperometric hydrogen peroxide monitoring to the alcohol oxidase enzyme system. Alcohols can be measured within several seconds at concentrations as low as 0.5 mg/100 ml. The only reagent required is alcohol oxidase in phosphate buffer.     

Amperometric enzyme electrodes: Part I. Theory

The theory of the steady state operation of an amperometric enzyme is derived. The reaction scheme includes diffusion of substrate and product through a membrane, the kinetics of the enzyme substrate reaction and the electrochemical regeneration of the enzyme. A simple diagnostic plot is derived which allows the rate limiting process to be identified, and the balance between the transport of substrate through the membrane and the enzyme and electrochemical kinetics to be determined. The effects of inhibition by the accumulation of product behind the membrane are also considered.     

Amperometric enzyme electrodes II. Amino acid oxidase

An amperometric method based on an L-amino acid oxidase electrode has been developed for the determination of several L-amino acids. The time of measurements is less than 12 s if a kinetic method is used, and 1 min if a steady-state method is used. The only reagent required is a phosphate buffer solution.     

Amperometric enzyme electrodes for the determination of l-glutamate

Electrodes for amperometric measurement of l-glutamate were prepared by immobilization of l-glutamate oxidase on an Immobilon-AV Affinity membrane and attachment to an oxygen/hydrogen peroxide sensor. The response of the hydrogen peroxide sensor was linear over the concentration range 5.0 x 10(-8)-5.0 x 10(-4)Ml-glutamate, with a limit of detection of 35nM. Attachment of a size-exclusion membrane (cut-off for molecular weight > 100) or of a hydrophobic oxygen membrane eliminated electro-oxidizable interferences, but the response was attenuated by a factor of 2-3. The response may be amplified 10-fold by co-immobilizing l-glutamate dehydrogenase with the l-glutamate oxidase. The electrode initially lost 25% of its activity but was then stable for more than 320 days and at least 200 assays. The electrode was successfully used to assay glutamate in a protein tablet and in several food products. A flow-injection system was assembled for the continuous assay of l-glutamate.     

Amperometric enzyme sensor for glucose based on graphite paste-modified electrodes

Amperometric enzyme electrode for glucose is described based on the incorporation of glucose oxidase (GOD) into graphite paste modified with tetracyanoquinodimethane (TCNQ). The incorporated enzyme exhibits high activity and long-term stability over the earlier TCNQ-based glucose sensor (1). The sensor provides a linear response to glucose over a wide concentration range. The response time of the sensor is 15-50 sec, and the detection limit is 0.5 mM. Stable response to the substrate was obtained during a period of 35 d. Application of the sensor in the plasma analysis is reported.     

Preparation and analytical testing of mediator containing photolithographically patterned enzyme membrane electrodes

The application of mediators for measurements with amperometric enzyme sensors have been investigated to improve the behavior of sensors with respect to interfering substances or for working under anaerobic conditions. The aim of this investigation is to develop photolithographically patterned enzyme membranes containing mediators, which facilitate the inexpensive technological preparation of patterned sensors. Thin layer platinum electrodes were coated with the enzyme membranes and cross-linked by means of UV light. Measurements were made in a wall-jet configuration using flow injection techniques with or without oxygen in the solutions. Optimum properties can be obtained with glucose oxidase containing membranes using tetrathiafulvalenes. The interfering substances ascorbic acid, uric acid and acetaminophenol showed no influence on the glucose measurements in the range of physiological concentrations. The membrane served as a diffusion barrier; a decrease in the applied potential to 300 mV vs. SCE also improved the ratio of the glucose response to the interference response.     

Amperometric enzyme electrodes for the determination of volatile alcohols in the headspace above fruit and vegetable juices

The surface of a glassy carbon electrode (GCE) was modified by electropolymerization of acridine red followed by drop-coating of graphene. The morphology was characterized by scanning electron microscopy. Uric acid (UA) is effectively accumulated on the surface of the modified electrode and generates a sensitive anodic peak in solutions of pH 6.5. Differential pulse voltammetry was used to evaluate the electrochemical response of the modified GCE to UA. Compared to the bare GCE, the GCE modified with acridine red, and to the graphene modified electrode, the new GCE displays high electrochemical activity in giving an oxidation peak current that is proportional to the concentration of UA in the range from 0.8 to 150?μM, with a detection limit of 0.3?μM (at an S/N of 3). The modified electrode displays excellent selectivity, sensitivity, and a wide linear range. It has been applied to the determination of UA in real samples with satisfactory results.

Nucleation in analytical chemistry

Precipitation involves two processes, nucleation and subsequent crystal growth. The nucleation process is of extreme importance in determining the number and size of the final crystalline particles. The significance of experimental studies of nucleation is discussed and the need for further research indicated.     

Chemiluminescence in analytical chemistry

Analytical applications based on chemiluminescence are reviewed. Analyses in the gas phase for atmospheric pollutants such as sulphur compounds, ozone and oxides of nitrogen are described. The commonest chemiluminescent systems used in the liquid phase are then discussed. Their applications as indicators in different types of titration are outlined. Determinations of organic and inorganic substances are classified according to their action as oxidant, catalyst or inhibitor. Special applications are described in fields such as forensic science, microbiology, polymer technology, radiation chemistry and flow mechanics.     

Quality in analytical chemistry

Summary In order to enhance the quality of analyticochemical statements it is common practice, to optimize the analytical method. But furthermore it is also necessary to fit the design of the study including sampling procedures and calibrations to the aims of the investigation and its consequences as close as possible. The presentation of results should mention all premises which were not empirically tested. To prevent misinterpretation of the results, their respective field of application should be specified. As regards the characterization of methods, it must be explained whether it is to be valid for a specific analysis series, for a measuring system or for the method as such. Thus, the quality in analytical chemistry is measurable in terms of the scientific nature of the statements; that is in terms of their degree of objective verifiability and in terms of their deduction by recognized methods of all disciplines involved, including statistics.     

Markers in analytical chemistry

The growing use of markers in analytical literature in the 10 years, 1991-2000, is presented and discussed because of their relevance in modern analytical chemistry. The complementary and contradictory aspects of markers and others related words, such as tracer, indicator, index, labelling compound, etc., are clarified. To offer a general overview, several classifications of markers are outlined. The main distinction between markers is their internal or external fitness for purpose. Selected examples are assessed on this basis.     

Ultrasound in analytical chemistry

Ultrasound is a type of energy which can help analytical chemists in almost all their laboratory tasks, from cleaning to detection. A generic view of the different steps which can be assisted by ultrasound is given here. These steps include preliminary operations usually not considered in most analytical methods (e.g. cleaning, degassing, and atomization), sample preparation being the main area of application. In sample preparation ultrasound is used to assist solid-sample treatment (e.g. digestion, leaching, slurry formation) and liquid-sample preparation (e.g. liquid–liquid extraction, emulsification, homogenization) or to promote heterogeneous sample treatment (e.g. filtration, aggregation, dissolution of solids, crystallization, precipitation, defoaming, degassing). Detection techniques based on use of ultrasonic radiation, the principles on which they are based, responses, and the quantities measured are also discussed.     

Sensors in analytical chemistry

Chemical sensorics is an independent domain of modern analytical chemistry. In Germany and Japan, multivolume encyclopedias dedicated to sensors were published. The publication of the nine-volume encyclopedia [1] in Germany was immediately followed by the appearance of the annual publication [2] incorporating additional data available from US. The preparations to publication of the next encyclopedia of sensors are under way now. It san be said with a good reason that chemical sensorics is a well-established sphere which is still under active development. This is specifically a testing ground for novel ideas and novel materials. This issue of Rossiiskii Khimicheskii Zhurnal (Zhurnal Rossiiskogo Khimicheskogo O-va im. D. I. Mendeleeva) is dedicated to chemical and biochemical sensors. It makes readers acquainted with the current state of the art in this sphere and covers various types of sensors. Here, we attempt to outline the general situation in this domain of analytical chemistry.