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Abstract

In this study, two new amperometric carbon paste enzyme electrodes for determination of uric acid were developed. The carbon paste was prepared by mixing uricase enzyme, 1,4-benzoquinone or poly(vinylferrocene) (PVF) as a mediator, graphite powder, paraffin oil and then the paste was placed into cavity of a teflon electrode body. Determination of uric acid was performed by oxidation of enzymatically generated H₂O₂. The effects of enzyme loading, mediator amount, buffer type, pH, buffer concentration, working potential and temperature were investigated for both electrodes. The working range of the 1,4-benzoquinone modified enzyme electrode was 1.9 × 10^–8–2.7 × 10^–3 M, detection limit 1.9 × 10^–8 M and response time 150 s. Optimum buffer type, pH, buffer concentration, working potential, temperature and amounts of enzyme and mediator for 1,4-benzoquinone modified enzyme electrode were found to be Tris, 8.0, 0.20 M, +0.25 V, 30 °C, 2.0 Unit and 13%, respectively. The working range of the PVF modified enzyme electrode was 7.4 × 10^–8–7.0 × 10^–3 M, detection limit 7.4 × 10^–8 M and response time 120 s. Optimum buffer type, pH, buffer concentration, working potential, temperature and amounts of enzyme and mediator for PVF modified enzyme electrode were found to be phosphate, 8.0, 0.05 M, +0.70 and +0.30 V, 40 °C, 2.0 Unit and 10.9%, respectively. The repeatability, storage stability of the enzyme electrodes and interference effects were also investigated. Enzyme electrodes were used for determination of uric acid in serum samples and the results were in a good agreement with those obtained by commercial enzymatic kits.

Keywords: Amperometry; 1,4-Benzoquinone; Carbon paste; Enzyme electrode; Poly(vinylferrocene); Uric acid; Biosensors; Carbon; Enzymes.

References: 41 live references.