Development and Validation of a Simple High Performance Liquid Chromatography/UV Method for Simultaneous Determination of Urinary Uric Acid, Hypoxanthine, and Creatinine in Human Urine

Abstract

Uric acid and hypoxanthine are produced in the catabolism of purine. Abnormal urinary levels of these products are associated with many diseases and therefore it is necessary to have a simple and rapid method to detect them. Hence, we report a simple reverse phase high performance liquid chromatography (HPLC/UV) technique, developed and validated for simultaneous analysis of uric acid, hypoxanthine, and creatinine in human urine. Urine was diluted appropriately and eluted with C-18 column 100 mm × 4.6 mm with a C-18 precolumn 25 mm × 4.6 mm in series. Potassium phosphate buffer (20 mM, pH 7.25) at a flow rate of 0.40 mL/min was employed as the solvent and peaks were detected at 235 nm. Tyrosine was used as the internal standard. The experimental conditions offered a good separation of analytes without interference of endogenous substances. The calibration curves were linear for all test compounds with a regression coefficient, r2 > 0.99. Uric acid, creatinine, tyrosine, and hypoxanthine were eluted at 5.2, 6.1, 7.2, and 8.3 min, respectively. Intraday and interday variability were less than 4.6% for all the analytes investigated and the recovery ranged from 98 to 102%. The proposed HPLC procedure is a simple, rapid, and low cost method with high accuracy with minimum use of organic solvents. This method was successfully applied for the determination of creatinine, hypoxanthine, and uric acid in human urine.

A Nanozyme‐Based Artificial Peroxisome Ameliorates Hyperuricemia and Ischemic Stroke

“…The UOD oxidizes uric acid (UA) to allantoin and has been used therapeutically for the prevention and treatment of hyperuricemia. [ 42 ] By measuring the specific absorbance of UA at 293 nm, [ 43 ] we observed that pero ‐nanozysome effectively degraded UA in vitro. With the UA as the substrate, the pero ‐nanozysome showed almost fivefold higher degradation rate for UA compared with N ‐nanozyme (Figure S5a, Supporting Information).…”

An Advanced Statistical Approach Using Weighted Linear Regression in Electroanalytical Method Development for Epinephrine, Uric Acid and Ascorbic Acid Determination

“…Moreover, the reported LOD and LOQ for AA using HPLC were 1.2 × 10 −3 g/L and 3.4 × 10 −3 g/L, respectively [ 38 ]. Furthermore, the reported LOD and LOQ for UA using HPLC were 0.30 μg/mL and 0.89 μg/mL, respectively [ 39 ]. Therefore, the electroanalytical methods presented in this work are competitive with HPLC for the analysis of EP, AA, and UA in terms of LOD and LOQ.…”

Enzymeless Electrocatalytic Detection of Uric Acid Using Polydopamine/Polypyrrole Copolymeric film

“…In a healthy body system, UA concentration ranges from 0.1‐4 mM and 1.5‐4.4 mM in blood and urine respectively . Elevated concentration of UA in human body has been implicated with diseases like gout, Lesch‐Nyan disease, obesity, diabetes, heart diseases and kidney‐related diseases . Hence, its prompt detection and monitoring is vital for human health protection.…”

“…Several analytical procedures have been developed for detection of uric acid in the past. These include isotopic dilution mass spectroscopy and liquid chromatography, chemiluminescence and electrochemical biosensors . etc.…”