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A planar microchip-based creatinine biosensor employing an oxidizing layer (e.g., a PbO2 film), where interfering redox-active substances are broken (i.e., oxidized) to redox-inactive products, was developed to facilitate the microfabrication of the sensor and to provide improved, reliable determination of creatinine in physiological samples. The feasibility of using hydrophilic polyurethanes in permselective barrier membranes for creatinine biosensors and the effect of adding a silanizing agent (adhesion promoter) on the sensor performance (e.g., sensitivity, stability, and lifetime) are described. The proposed creatinine microsensor with a three-layer configuration, i.e., enzyme, protecting, and oxidizing layers, exhibits good electrochemical performance in terms of response time (t95% = 98 s at 100-->200 microM creatinine change), linearity (1-1000 microM, r = 0.9997), detection limit (0.8 microM), and lifetime (approximately 35 days). The creatinine biosensor devised in a differential sensing arrangement that compensates the erroneous results from creatine is considered to be suitable for assay of serum specimens.

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