Amperometric, screen-printed, glucose biosensor for analysis of human plasma samples using a biocomposite water-based carbon ink incorporating glucose oxidase.

This paper describes the optimisation of a screen-printing water-based carbon ink containing cobalt phthalocyanine (CoPC) and glucose oxidase (GOD) for the fabrication of a glucose biosensor. To optimise the performance of the biosensor, the loadings of the electrocatalyst (CoPC) and enzyme (GOD) were varied. It was found that the maximum linear range was achieved with a CoPC loading of 20% (m/m, relative to the mass of carbon) and a GOD loading of 628 U per gram of carbon. In our studies we chose to employ chronoamperometry, as this technique is commonly used for commercial devices. The optimum operating applied potential was found to be +0.5 V, following an incubation period of 60 s. The optimum supporting electrolyte was found to be 0.05 M phosphate buffer at pH 8.0, which resulted in a linear range of 0.2-5 mM, the former represents the detection limit. The sensitivity was 1.12 microA mM(-1). The effect of temperature was also investigated, and it was found that 40 degrees C gave optimal performance. The resulting amperometric biosensors were evaluated by measuring the glucose concentrations for 10 different human plasma samples containing endogenous glucose and also added glucose. The same samples were analysed by a standard spectrophotometric method, and the results obtained by the two different methods were compared. A good correlation coefficient (R(2) = 0.95) and slope (0.98) were calculated from the experimental data, indicating that the new devices hold promise for biomedical studies.

A novel, disposable, screen-printed amperometric biosensor for glucose in serum fabricated using a water-based carbon ink.

Screen-printed amperometric glucose biosensors have been fabricated using a water-based carbon ink. The enzyme glucose oxidase (GOD) and the electro-catalyst cobalt phthalocyanine were mixed with the carbon ink prior to the screen-printing process; therefore, biosensors are prepared in a one-step fabrication procedure. Optimisation of the biosensor performance was achieved by studying the effects of pH, buffer strength, and applied potential on the analytical response. Calibration studies were performed under optimum conditions, using amperometry in stirred solution, with an operating potential of +500 mV versus SCE. The sensitivity was found to be 1170 nA mM(-1), with a linear range of 0.025-2 mM; the former represents the detection limit. The disposable amperometric biosensor was evaluated by carrying out replicate determinations on a sample of bovine serum. This was achieved by the method of multiple standard additions and included a correction for background currents arising from oxidizable serum components. The mean serum concentration was calculated to be 8.63 mM and compared well with the supplier's value of 8.3 mM; the coefficient of variation was calculated to be 3.3% (n=6).

Rapid electrochemical detection and identification of catalase positive micro-organisms.

The rapid detection and identification of bacteria has application in a number of fields, e.g. the food industry, environmental monitoring and biomedicine. While in biomedicine the number of organisms present during infection is multiples of millions in the other fields it is the detection of low numbers of organisms that is important, e.g. an infective dose of Escherichia coli O157:H7 from contaminated food is less than 100 organisms. A rapid and sensitive technique has been developed to detect low numbers of the model organism E. coli O55, combining Lateral Flow Immunoassay (LFI) for capture and amperometry for sensitive detection. Nitrocellulose membranes were used as the solid phase for selective capture of the bacteria using antibodies to E. coli O55. Different concentrations of E. coli O55 in Ringers solution were applied to LFI strips and allowed to flow through the membrane to an absorbent pad. The capture region of the LFI strip was placed in close contact with the electrodes of a Clarke cell poised at +0.7 V for the detection of hydrogen peroxide. Earlier research identified that the consumption of hydrogen peroxide by bacterial catalase provided a sensitive indicator of aerobic and facultative anaerobic microorganisms numbers. Modification and application of this technique to the LFI strips demonstrated that the consumption of 8 mM hydrogen peroxide was correlated with the number of microorganisms presented to the LFI strips in the range of 2 x 10(1)-2 x 10(7) colony forming units (cfu). Capture efficiency was dependent on the number of organisms applied and varied from 71% at 2 x 10(2) cfu to 25% at 2 x 10(7) cfu. The procedure was completed in less than 10 min and could detect less than 10 cfu captured from a 200 microl sample applied to the LFI strip. The approached adopted provides proof of principle for the basis of a new technological approach to the rapid, quantitative and sensitive detection of bacteria that express catalase activity.

Voltammetric behaviour and trace determination of copper at a mercury-free screen-printed carbon electrode.

Screen-printed carbon electrodes (SPCEs), without chemical modification, have been investigated as disposable sensors for the measurement of trace levels of Cu(2+). Cyclic voltammetry was employed to elucidate the electrochemical behaviour of Cu(2+) at these electrodes in a variety of supporting electrolytes. For all of the electrolytes studied the anodic peaks, obtained on the reverse scans, showed that the Cu(2+) had been deposited as a thin layer on the surface of the SPCE. The anodic peak of greatest magnitude was obtained in 0.1 M malonic acid. The possibility of determining Cu(2+) at trace levels using this medium was examined by differential pulse anodic stripping voltammetry (DPASV). The effect of Bi(3+), Cd(2+), Fe(3+), Hg(2)(2+), Pb(2+), Sb(3+) and Zn(2+) on the Cu stripping peak was examined and under the conditions employed, only Hg(2)(2+) was found to significantly effect the response gained. The sensors were evaluated by carrying out Cu(2+) determinations on spiked and unspiked serum and water samples. The mean recovery was found in all cases to be >90% and the performance characteristics indicated the method holds promise for trace Cu(2+) levels by employment of Hg-free SPCEs using DPASV.