Direct determination of uric acid in serum by a fluorometric-enzymatic method based on uricase

Abstract

The present paper describes a method for the fluorometric determination of uric acid in blood serum by its reaction with uricase (UOx). The procedure is based on the changes in fluorescence that take place during the enzymatic reaction of UOx with uric acid when the solution is excited at 287 nm and the emission is measured at 330 nm. A mathematical model which relates the analytical signal to the analyte concentration was developed and the model also served to obtain some of the thermodynamic constants of the system (the Michaelis constant and the turnover number). The optimum reaction conditions and its analytical characteristics were studied, linear response range (3×10 −5–6×10 −4 M) and reproducibility (4%, n=7). The method was applied to the determination of uric acid in three blood serum samples. The results were compared with those obtained by a commercial clinical analyzer and no systematic errors were observed.