Mass changes and dissolution of platinum during electrical stimulation in artificial perilymph solution.

The electrochemistry of platinum electrodes in artificial perilymph solution subjected to cyclic and steady-state potentials was studied by a quartz crystal electrochemical microbalance and by analysis of electrolyte for dissolved platinum. The effect of amino acid adsorption appears to be one of blocking sites for platinum oxidation and oxide reduction, a process in which the amino acid competes with chloride from phosphate-buffered saline. For amino acids such as cysteine, which are known to interact strongly with platinum, the voltammograms became nearly featureless and only a small change in mass was observed during cycling of the potential. There were no mass changes of an electrode in solution containing human serum albumen, but its presence did inhibit dissolution of platinum. The overall magnitudes of dissolved platinum found in the buffered solutions were low, remaining below 5 ppb in most cases. Dissolution was greatest in solutions containing high concentrations of cysteine. Extrapolation of the results to implanted auditory prosthesis electrodes indicated that platinum dissolution would not lead to toxic concentrations in the body.

Parameters important in tuning the response of monolayer enzyme electrodes fabricated using self-assembled monolayers of alkanethiols.

Enzyme electrodes were observed experimentally to have a broad dynamic range, high sensitivity and excellent reproducibility. The theoretically predicted response of the monolayer enzyme electrodes was in good agreement with that observed experimentally over the broad range of experimental conditions tested. The response is limited by the rate of enzyme turnover by a mediating species rather than mass transport. As a consequence of this limitation, the response was very sensitive to the enzyme loading and the concentration of mediator in the sample solution but insensitive to mass transport variables such as solution stirring or the diffusion coefficients of the substrate or cosubstrate.

Electrodeposited polytyramine as an immobilisation matrix for enzyme biosensors.

The application of an electrodeposited polytyramine film as an immobilisation matrix for the construction of enzyme biosensors is described. Glucose oxidase (used as a model enzyme) is covalently attached to free amine groups on the polytyramine film using the coupling reagents 1-ethyl-3(3-dimethylaminopropyl) carbodiimide hydrochloride and N-hydroxysuccinimide. The resultant recognition interface consisted of multilayers of GOx immobilised onto the polymer surface. This method of constructing enzyme biosensors is shown to produce a highly reproducible and stable device. The biosensor showed no loss in electrode response after four months of dry storage and exhibited only minor loss in response after 20 days of repeated use. The resultant biosensor had a linear range of 0.1-28 mM glucose and a detection limit of 0.01 mM.

Sub-ppt detection limits for copper ions with Gly-Gly-His modified electrodes.

An electrochemical metal ion sensor has been developed with a detection limit of less than 0.2 ppt by the covalent attachment of the tripeptide Gly-Gly-His as a recognition element to a 3-mercaptopropionic acid modified gold electrode.

Linearity and the limitations of least squares calibration.

The magnitude of errors that can arise in practice from the limitations of the least squares method of calibration is estimated. Data generated from y = xn (0.7 < or = n < or = 1.3 and 1 < or = x < or = 30, or < or = 60) was analysed by least squares regression. Each y-value was then presented to the linear model and an x-value predicted. The relative errors on small x-values reached 70% of the concentration value even when r2 exceeded 0.999. Estimates of the errors on each predicted x-value, determined from the standard errors of the slope and intercept failed to reveal large errors at small x-values. Reducing the range over which linear regression is performed improved the errors. Other data sets with a heteroscedastic error distribution show that linear regression by least squares can also lead to the rejection of methods that performed sufficiently well for their application. Heteroscedastic data may be treated by repeated measurements at the lower end of the range. Data from a validation of an HPLC method for isoflavones in legumes is used to show the errors in recovery when a check sample is presented to the instrument using a calibration which satisfies the linearity tests. It is recommended that both y- and relative x-residuals are inspected. It is proposed that over-reliance on linear calibration supported by r2 may make a major contribution to large, hitherto unexplained, inter-laboratory errors.

Structure and solution equilibria of D-glucose and D-mannose sulfite adducts.

An X-ray crystallographic study has confirmed that the potassium bisulfite adducts of D-glucose and D-mannose have open-chain structures with R and S configurations respectively at C-1. NMR studies have shown that each sugar gives rise to two bisulfite compounds, and solution-state structures and conformations of these isomers have been deduced from analysis of (1)H NMR spectra. (13)C NMR data for the four adducts are given. Furanose forms of the D-glucose and D-mannose have been detected in the equilibrium solutions.

Amperometric detection of amines using cobalt electrodes after separation by ion-moderated partition chromatography.

A simple and low-cost amperometric sensor for amines has been developed using a cobalt wire electrode working in alkaline solution. The sensor may be used as a detector for high-performance liquid chromatography (HPLC) that avoids the need for derivatization or post-column reaction. Experimental conditions for flow injection analysis (FIA) and HPLC separation, including the applied potential, pH and concentrations of organic modifier and carrier solution, were optimized. A cobalt wire electrode, in the constant potential amperometric mode, gives an excellent response toward amines in ion-exclusion chromatography in unbuffered solution. The sensitivities of the detection and separation of amines on the column are affected by flow rate, the concentration of the mobile phase and the concentration of organic solvent in the mobile phase, whereas the applied potential only affects the sensitivity of the detector. A cobalt electrode is more sensitive than a copper electrode, and comparable in sensitivity to a UV detector for most amines tested. The detection sensitivity is comparable to that obtained with GC methods, but the procedures are far simpler. The detection limits of the order of nanomoles obtained under the chromatographic conditions used offer an alternative for the determination of amines in a variety of matrices, such as in environmental, biomedical and pharmaceutical samples.

A photo-cured coated-wire calcium ion selective electrode for use in flow injection potentiometry.

A multi-sensor cell containing a new photo-cured calcium ion selective electrode sensor is reported. Four membranes containing different components are prepared to determine the one with optimum selectivity and sensitivity. This is shown to consist of the N,N,N',N'-tetracyclohexyl-3-oxapentanediamide ligand (ETH 129) as the ionophore, 2-nitrophenyl octyl ether as the plasticiser and tetradodecyl ammonium tetrakis(4-chlorophenyl) borate as the lipophilic additive. The photo-curing process is applied after coating a thin membrane on a silver wire as substrate transducer to produce the calcium sensor. The curing process is demonstrated to be faster (1 min) than previous methods and does not require a nitrogen atmosphere for reproducible production of membrane response characteristics. Four sensors constructed with the identical optimum membrane are shown to function reproducibly in a multi-sensor flow-through cell using the steady-state mode of flow measurement, and an average calibration slope of 28.5+/-0.4 mV change per activity decade is observed over a log-linear concentration range between 0.01 and 10 mM. The sensor is also shown to respond to changes in pH. Hence, in the flow injection potentiometric mode, a constant carrier buffer composition of pH 8.3 is required for accurate potentiometric calcium determinations. The sensor is used to determine calcium in water samples by flow injection potentiometry. The accuracy of the electrode determination relative to atomic absorption spectroscopy was in the range 5-9% for three different water samples.

Continuous flow analysis of lead (II) and mercury (II) with substituted diazacrown ionophore membrane electrodes.

A novel flow cell for use with ion-selective membrane electrodes is reported in which the carrier stream is drawn through a tube that suppresses the pump noise. PVC membrane electrodes based on 7,16-dithenoyl-1,4,10,13-tetraoxa-7,16-diazacyclooctadecane (DTODC), and 7,16-di-(2-thiopheneacetyl)-1,4,10,13-tetraoxa-7,16-diazacyclooctadecane (DTAODC) for lead (II), and 7,16-dithenyl-1,4,10,13-tetraoxa-7,16-diazacyclooctadecane (DTDC) and 7,16-di-(2-methylquinolyl)-1,4,10,13-tertraoxa-7,16-diazacyclootadecane (DQDC), for mercury (II) were prepared and evaluated. The linear ranges were pPb: 5.5-3.0 (DTODC) and 6.0-2.0 (DTAODC); pHg: 5.5-3.0 (DTDC) and 4.5-2.5 (DQDC). With flow rate of 3 ml min(-1) the repeatability of measurements was less than 5% RSD (n = 3). The system was applied to the determination of lead (II) and mercury (II) in spiked natural water samples.

Flow injection potentiometry for enzymatic assay of cholesterol with a tungsten electrode sensor.

Flow injection potentiometry (FIP) for the enzymatic determination of cholesterol is reported. The assay utilises a combination of three enzymes: cholesterol esterase (CE), cholesterol oxidase (COD) and peroxidase (POD). The method is developed by the use of a tungsten wire electrode as a sensor vs. Ag/AgCl in conjunction with a redox mediator ferrocyanide. CE converts esterified cholesterol to free cholesterol, which is then oxidised by COD with hydrogen peroxide as product. Ferrocyanide is converted to ferricyanide by hydrogen peroxide, catalysed by POD, and the tungsten electrode responds to the ratio of ferricyanide to ferrocyanide. Flow injection potentiometry gave well-defined peaks for cholesterol samples with a fast response (30 s). Linear calibration was obtained from 0.05 to 3.0 mM cholesterol, with a slope of 60.2 mV/decade change in cholesterol concentration, and detection limit 0.01 mM cholesterol (S/N=3). Repeatability was 3% (CV). Interferences from commonly found species were shown to be negligible. The sensor cell is simple to construct, and it was free from surface contamination problems over long periods of use. The application of the sensor for the determination of serum cholesterol was demonstrated.

Field-portable flow-injection analysers for monitoring of air and water pollution.

There has been a rapid growth in the development of field-portable analytical instrumentation capable of in-situ and real-time feedback of data from remote sites. Advances have been made in applications for many technologies aided by developments in electronics, computing and telecommunications systems. This report presents a brief review of these developments and particularly of portable flow-injection systems applied in both the liquid and gas phase modes of operation with potentiometric sensors and gas sensors of the tin-oxide semiconductor types.

Parameters important in fabricating enzyme electrodes using self-assembled monolayers of alkanethiols.

The fabrication of enzyme electrodes using self-assembled monolayers (SAMs) has attracted considerable interest because of the spatial control over the enzyme immobilization. A model system of glucose oxidase covalently bound to a gold electrode modified with a SAM of 3-mercaptopropionic acid was investigated with regard to the effect of fabrication variables such as the surface topography of the underlying gold electrode, the conditions during covalent attachment of the enzyme and the buffer used. The resultant monolayer enzyme electrodes have excellent sensitivity and dynamic range which can easily be adjusted by controlling the amount of enzyme immobilized. The major drawback of such electrodes is the response which is limited by the kinetics of the enzyme rather than mass transport of substrates. Approaches to bringing such enzyme electrodes into the mass transport limiting regime by exploiting direct electron transfer between the enzyme and the electrode are outlined.

Amperometric detection of organic thiols at a tungsten wire electrode following their separation by liquid chromatography.

The amperometric detection of thiols following their separation by reversed-phase chromatography and reaction into a post-column mercury carrier stream is shown to be a sensitive method when using a tungsten wire sensor as the working electrode in a three-electrode flow cell. Five organic thiols (cysteine, homocysteine, reduced glutathione, D,L-penicillamine and 2-mercaptopropionic acid) and thiourea could be separated within approx. 18 min. The analytical performance is comparable, and stability superior, to chemically modified electrodes previously reported.

The electrogenerated chemiluminescent behavior of hemin and its catalytic activity for the electrogenerated chemiluminescence of lucigenin.

The electrogenerated chemiluminescent (ECL) behavior of hemin at a platinum electrode in the alkaline solution has been investigated in detail. Under the optimum conditions the linear response range of hemin is 1.0 x 10(-5)-1.0 x 10(-8) g ml(-1), the detection limit was 1.0 x 10(-8) g ml(-1), and the relative standard derivation for 1 x 10(-7) g ml(-1) hemin was 2.8%. It has been also found that hemin would catalyze the ECL of lucigenin at a platinum electrode in a neutral solution in the presence of hydrogen peroxide, the catalytic ECL intensity was linear with the concentration of hemin in the range of 1.0 x 10(-14)-1.0 x 10(-10) g ml(-1). IgG labeled with hemin was used to examine the ECL catalytic activity of hemin after conjugating to protein, and the results showed that hemin retained ECL catalytic activity when conjugated to protein.