Quantitative Determination of Uric Acid in Serum by Reversed-Phase Liquid Chromatography Using an Internal Standard

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Abstract
A rapid and sensitive high-performance liquid chromatography procedure is reported for the analysis of uric acid in serum. The method employs a single treatment of serum with cyclohexane and an excess of ammonium sulfate to eliminate endogenous serum components which easily contaminate the chromatographic system. Reversed-phase chromatography is performed at room temperature using a binary-solvent gradient, monitoring the effluent at 290 nm. Quantitation is based on peak-height ratio of uric acid to internal standard (β-hydroxyethyltheophylline). A linear response is obtained to 160 mg/1. Within-day and between-day precisions for the mean of this range varied from 0.59 to 1.1% and from 0.71 to 1.2%, respectively. Numerous potential interfering substances tested do not interfere.