AMPEROMETRIC BIOSENSORS

R. SAICHARAN

Types of Enzyme electrodes

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Saicharan/Unit III/ BS&BE/ DBT/SNIST

Introduction
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Amperometric biosensors function by the production

of a current when a potential is applied between two electrodes. They generally have response times, dynamic ranges and sensitivities similar to the potentiometric biosensors.

Saicharan/Unit III/ BS&BE/ DBT/SNIST

Introduction
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Because of the relatively large distance between the redox

center of enzymes and the transducer surface, it is difficult to achieve electron transfer between redox enzymes and electrodes. Alternately, the biocatalytic generation or consumption of electroactive species make oxidoreductase enzymes most suitable for amperometric biosensing. In particular, a large number of hydrogen peroxide generating oxidases and NAD+-dependent dehydrogenases The following general reactions:

Saicharan/Unit III/ BS&BE/ DBT/SNIST

Clark oxygen Electrode

The simplest amperometric biosensors in common usage
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involve the Clark oxygen electrode. This consists of a platinum cathode at which oxygen is reduced and a silver/silver chloride reference electrode. When a potential of -0.6 V, relative to the Ag/AgCl electrode is applied to the platinum cathode, a current proportional to the oxygen concentration is produced. Normally both electrodes are bathed in a solution of saturated potassium chloride and separated from the bulk solution by an oxygen-permeable plastic membrane (e.g. Teflon, polytetrafluoroethylene).

Saicharan/Unit III/ BS&BE/ DBT/SNIST

Glucose Biosensor
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Saicharan/Unit III/ BS&BE/ DBT/SNIST

Amperometric biosensors
Principles of biosensors: 1. The analyte is converted by an enzymatic process producing an easy reducible or oxidable species
2. This product is reduced or oxidized amperometric at constant potential I, A 0.6 V Whole blood E, V H2O2
Saicharan/Unit III/ BS&BE/ DBT/SNIST

Very selective process Only the analyte reacts Low potentials gives few interferences

GOx
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Glucose

Amperometric HPLC-detectors
Working electrode are placed in the effluent flow from the HPLC HPLC transport by constant flow flow
Side view:

Inert body

Working electrode

Inert spacer

Top view:

Saicharan/Unit III/ BS&BE/ DBT/SNIST

Summary for Amperometry

I(E) = k[Analyte]

Analysis based on electrolysis, but only partial (negligible) conversion

Analyte ze Products

The analytical signal is a steady-state current

There must be a constant transport of the analytes to the electrode. This transport can be by: Diffusion Amperometric sensors Solution flow Electrochemical HPLC-detectors
Amperometric HPLC-detectors are very sensitive, LD 0.01 1 ng better than UV, only slightly less than fluorescence
Saicharan/Unit III/ BS&BE/ DBT/SNIST

Selected oxidase- or dehydrogenase-based

electrodes
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Saicharan/Unit III/ BS&BE/ DBT/SNIST

Configurations
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Single-Use Sensors: The single use approach represents 99% of the commercial value.These are the glucose electrode strips available (in the USA) Intermittent-Use Sensors The second approach is to use a sensor intermittently. In this case, there is often a carrier stream and the samples are processed sequentially. Continuous-Use Sensors It is also possible to bring the electrode to the sample in the form of a sensor which is dipped into or implanted into the sample. The sensor puts out a continuous signal reflecting the analyte concentration as a function of time.

Saicharan/Unit III/ BS&BE/ DBT/SNIST

Comparison between Configuration

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Saicharan/Unit III/ BS&BE/ DBT/SNIST