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Bioelectrochemistry. 2002 May 15;56(1-2):113-5.

Amperometric urea biosensor based on urease and electropolymerized toluidine blue dye as a pH-sensitive redox probe.

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Department of Biochemical Technology, Faculty of Chemical and Food Technology, Slovak University of Technology, Radlinskeho 9, SK-812 37 Bratislava, Slovak Republic.


The electropolymerized toluidine blue film deposited on the glassy carbon electrode show amperometrically detectable pH sensitivity. This feature of polytoluidine blue (PTOB) film was used for a construction of an amperometric urea biosensor. We have observed a linear shift of the formal redox potential with increasing pH value between 4 and 8 giving the slope of 81 mV(Delta) pH(-1). Polytoluidine blue film has had a significantly increased stability and higher electrochemical activity compared to the adsorbed monomeric dye. The polytoluidine blue urea biosensor has been operating at a working potential of -200 mV vs. SCE. The sensitivity of the biosensor was 980 nA mM(-1) cm(-2). The biosensor showed linearity in concentration range up to 0.8 mM with the detection limit of 0.02 mM (S/N=3).

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