An Improved Uplc Method For Determining Uric Acid In Rat Serum And Comparison Study With Commercial Colorimetric Kits

Uric acid (UA) is the final product of purine metabolism in humans. Elevated serum UA levels lead to the development of hyperuricemia, gout, kidney diseases, and metabolic syndrome. Accurate determination of UA plays a critical role in clinical diagnosis and laboratory investigation. An ultra-performance liquid chromatography (UPLC) with ultraviolet detection method has been developed and validated for UA analysis. Separation was achieved by a Waters ethylene bridged hybrid (BEH) Amide column (50 mm x 2.1 mm i.d., 1.7 mu m) with acetonitrile and 0.1% acetic acid in deionized water in the proportion of 90 to 10 (v/v) as the mobile phase. The limit of detection and limit of quantification were 0.09 and 0.18 mu mol/L, respectively. The method was validated by evaluating recovery (98.37-104.20%), accuracy (0.47-0.90%), and precision (1.24-1.81% for intra-batch and 1.76-3.98% for inter-batch). This method was then applied to UA determination in rat serum of hyperuricemia model. The results from UPLC, high-performance liquid chromatography (HPLC), and uric acid kits (phosphor-tungstic acid-based kit and uricase-based kit) were compared. The UPLC results were in very good agreement with HPLC. The developed method could be employed as a useful tool for the determination of UA in biofluids.