Determination of gouty arthritis' biomarkers in human urine using reversed-phase high-performance liquid chromatography

Abstract

Creatinine, uric acid, hypoxanthine and xanthine are important diagnostic biomarkers in human urine for gouty arthritis or renal disease diacrisis. A simple method for simultaneous determination of these biomarkers in urine based on reversed-phase high-performance liquid chromatography (RP-HPLC) with ultraviolet (UV) detector was proposed. After pretreatment by dilution, centrifugation and filtration, the biomarkers in urine samples were separated by ODS-BP column by elution with methanol/50 mM NaH2PO4 buffer solution at pH 5.26 (5:95). Good linearity between peak areas and concentrations of standards was obtained for the biomarkers with correlation coefficients in the range of 0.9957–0.9993. The proposed analytical method has satisfactory repeatability (the recovery of data in a range of creatinine, uric acid, hypoxanthine and xanthine was 93.49–97.90%, 95.38–96.45%, 112.46–115.78% and 90.82–97.13% with standard deviation of <5%, respectively) and the limits of detection (LODs, S/N≥3) for creatinine, uric acid, hypoxanthine, and xanthine were 0.010, 0.025, 0.050 and 0.025 mg/L, respectively. The established method was proved to be simple, accurate, sensitive and reliable for the quantitation of gouty arthritis' biomarkers in human urine samples. The ratio of creatinine to uric acid was found to be a possible factor for assessment of gouty arthritis.

Simultaneous Detection of Dopamine and Uric Acid Using a Poly(l-lysine)/Graphene Oxide Modified Electrode

“…Thus, the detection of UA has the vital significance in the field of clinical diagnosis. Several methods, such as enzymatic [12], voltammetric [13], high performance liquid chromatographic [14], and phospho-tungstic acid deoxidizing methods, have been utilized for the detection of UA. Because DA and UA always coexist in human biological fluids, it is highly desirable to propose a selective, sensitive and simultaneous detection technique for analytical research and diagnostic applications Electrochemical methods with advantage of sensitivity, rapidity and low cost have been widely adopted for the simultaneous detection of UA and DA [15,16,17].…”

Simultaneous determination of renal function biomarkers in urine using a validated paper-based microfluidic analytical device

“…In this test, alkaline picrate reacts with creatinine to form an orange-red product that is measured spectrophotometrically. Other current methods include HPLC-MS/MS [2], HPLC-DAD [3,4], spectrophotometry [5], spectrophotometry using a portable microfluidic system [6], and electrophoresis with electrochemical [7], UV [8], and colorimetric detection [9].…”

“…Analytical procedures for the measurement of uric acid include HPLC-DAD [3,4], electrophoresis with UV detection [8], spectrophotometry with chemometric tools [12], electrochemical methods [13], colorimetric detection with multiple indicators [14], enzymatic reaction [15], and gold nanoparticles [16]. Only methods that involve separation techniques, such as chromatography [3,4] and electrophoresis [8], are able to determine both analytes simultaneously. However, these techniques involve the use of large volumes of toxic organic solvents, produce substantial amounts of waste that may be dangerous to the operator and the environment [17], and require laborious clean-up steps.…”

Uric acid quantification in fingernail of gout patients and healthy volunteers using HPLC‐UV

“…It is mainly excreted in the urine (Huda et al, 2014;Ferin et al, 2013). Urine, serum and saliva samples have been extensively investigated for the UA assay in biological specimens (Shibasaki et al, 2012;Punzi and So, 2013;Xiang et al, 2014). In recent years, several methods have been presented by different researchers for the determination of UA in biological samples.…”

“…Furthermore, it is well known that UA is present in saliva (Stibůrková et al, 2014;Chauhan et al, 2014). Urine, serum and saliva samples have been extensively investigated for the UA assay in biological specimens (Shibasaki et al, 2012;Punzi and So, 2013;Xiang et al, 2014). However, to the best of our knowledge, the study of gout based on the determination of UA in human fingernails has not yet been reported.…”