A candidate reference method for determination of serum uric acid by reversed-phase liquid chromatography with ultraviolet detection

Abstract

We established a method for measuring uric acid in serum by reversed-phase liquid chromatography with ultraviolet detection.
Serum (0.1ml) was mixed with 1.0 ml of 0.3 mol/l perchloric acid. After vortexmixing for 10 sec, the samples were centrifuged at 3,000 × rpm for 10 min. The supernatant diluted two-fold with 0.2 mol/l disodium hydrogen phosphate was injected onto a reversed-phase ODS column. The mobile phase was a 75 m mol/l pH 2.2sodium phosphate buffer, containing 2 % methanol. The eluent was monitored at 284nm.
Analytic recovery of added uric acid was 100.6%±0.4% (× ±S. D. ) and the C. V. was 0.7∼0.9 %. The values obtained by this method correlated with those obtained by the uricase-catalase method of Kageyama (r=0.996).