Preparation of a potentiometric immobilized urease electrode and urea determination in serum

Polyvinylalcohol was activated with 2-fluoro-1-methylpyridiniumtoluene-4-sulphonate and urease (EC.3.5.1.5) was covalently linked to the activated matrix. PVA-urease was then immobilized on the surface of a pH glass electrode with gelatine gel and it was cross-linked using glutaraldehyde. This potentiometric membrane electrode provides a linearity to urea in the 8.910−5 to 1.110−3 M concentration range, but by changing the buffer concentration can be studied in the range of 10−4 to 10−2 M urea concentration. Reproducibility experiments (n:20) were carried out with the urease enzyme electrode and with photometric methods for pooled serum sample. Average values for the two methods were 5.96 and 5.86 mM, variation coefficients were 2.5 and 3.5% respectively.