The construction of sensors is based on the approach for operation of oxidase based biosensors by low potential selective detection of H₂O₂ on electrodes modified by electrochemically deposited Prussian Blue (ferric hexacyanoferrate). The approach was first demonstrated by our team in 1994 [1,2] and had been developed in order to achieve the most advantageous H₂O₂ transducer. Electrochemical biosensor for lactate has been made by immobilization of the enzyme lactate oxidase on the top of H₂O₂ transducer (planar sensor for hydrogen peroxide). Immobilization protocol included a formation of the enzyme containing Nafion polyelectrolyte membrane according to procedure developed earlier [2]. Exposure of lactate oxidase to water-organic mixtures with a high (85%) content of the organic sol­vent, resulted in stabilization of the enzyme by membrane-forming polyelectrolyte. The different approaches affecting the immobilization procedure have been studied. Combining the attractive performance of a Prussian-Blue based hydrogen peroxide transducer and optimal immobilization protocol, we have been developing low cost and mass-productive planar sensors for H₂O₂ and biosensors for glucose and lactate based on screen-printed electrodes modified with chemically deposited nano-layers of Prussian Blue. The best results were obtained for lactate oxidase immobilized in siloxans membranes. The corresponding biosensor has following analytical characteristics: improved sensitivity (about 70 mA/M cm²) and signal-to-noise ratio, low detection limit to compare with known analogs. Lactate may be analyzed in range of 1.10^-6-1.10^-3 M in FIA regime. The biosensor retained the sensitivity within 3 days (for 500 measurements) and could be used for multiuse measurements.

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1. Electroanalytical system based on lactate biosensor with preliminary concentration for non-invasive diagnostics