Bi-enzymatic amperometric biosensor for oxalate

The development of a new amperometric biosensor for oxalate utilising two enzymes, oxalate oxidase (OXO) and horseradish peroxidase (HRP), incorporated into carbon paste electrode modified with silica gel coated with titanium oxide containing toluidine blue is described. OXO has been immobilised on silica gel modified with titanium oxide surface using glutaraldehyde for crosslinking. HRP has been immobilised with covalent binding with carbodiimide on graphite powder. The biosensor showed a good performance with a linear response range between 0.1 and 2.0 mmol l−1 of oxalate, fit by the equation i=0.33(±0.04)+2.29(±0.04) [oxalate], where i is the current in μA and [oxalate] is the oxalate concentration in mmol l−1 with a correlation coefficient of 0.998 for n=20. The biosensor could be used for 80 determinations when stored in a succinate buffer at pH 3.8 in a refrigerator. The response time was about 0.5 s. The detection limit, considering three times the noise, was 0.09 mmol l−1 for oxalate. The time for oxalate determination in spinach samples decreased by 3 days when this biosensor was used, compared to the AOAC method.