Rapid, sensitive and simultaneous determination of ascorbic and uric acids in human plasma by ion-exclusion HPLC-UV

Abstract

This article appeared in a journal published by Elsevier. The attached copy is furnished to the author for internal non-commercial research and education use, including for instruction at the authors institution and sharing with colleagues.Other uses, including reproduction and distribution, or selling or licensing copies, or posting to personal, institutional or third party websites are prohibited. Objectives: Ascorbic (AA) and uric (UA) acids act as antioxidants and are capable to react with biologically relevant oxidants. We aimed to developed a simple, rapid, sensitive, and accurate ion-exclusion HPLC-UV methodology for the simultaneously determination of AA and UA in human plasma.Methods: Analytical pre-requisites, such as the use of heparin as an anticoagulant and meta-phosphoric acid as a stabilizer were added for accurate and reliable measurements. Chromatographic separation was achieved by an isocratic elution on a HEMA-BIO 1000 SB analytical column using a phosphate buffer, pH 2.4, as a mobile phase.Results: Results indicated an excellent linearity with correlation coefficients (r 2 ) ≥ 0.999. The LOD of AA and UA was 1.02 and 1.42 nmol/mL, respectively, while LOQ ranged from 0.306 to 0.426 nmol/mL. A great repeatability for both antioxidants was found, where the CV (%) values for intra-day were lower than 1.8% and under 6.5% for the inter-day assay. The recovery of AA ranged from 92% to 96% and from 99% to 100% for UA.Conclusion: This validated method allows the determination of both antioxidants within 10 min, and is well suited to routine measurements and/or high-throughput clinical analysis. The methodology was applied to assess the antioxidant status of a group of Azorean subjects.

A sensitive and selective chemosensor for ascorbic acid based on a fluorescent nitroxide switch

“…1b inset illustrated that this chemosensor exhibited a linear response toward negative logarithm of AsA concentration from 80 nM to 50 μM, the linear equation was y¼2915-381.4x and the linear relative coefficient was R 2 ¼ 0.993. The detection limit of AsA was estimated to 20 nM, which is lower than the AsA concentration detected by previous researches [15,28] in physiological sample. We also compared our technique with other methods (Table S1), and the results showed that our chemosensor provided lower detection limit and wider line range for the detection of AsA.…”

“…A number of specific assays for reductants including ascorbic acid, uric acid, tocopherol etc. have been developed based on high performance liquid chromatography (HPLC) [15,16]. These techniques require several complicated preliminary steps.…”

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

“…For example, the course of typical gout passes through four stages: asymptomatic hyperuricemia, acute gouty inflammation, intermittent gout and chronic tophaceous gout (Guggi et al, 2002;Liu et al, 2011). It is mainly excreted in the urine (Huda et al, 2014;Ferin et al, 2013). Because it is possible to prevent hyperuricemia by making lifestyle improvements, early detection and treatment are important (Matsuura, 2007).…”

Comparison of Uric Acid Quantity with Different Food in Human Urine by Flow Injection Chemiluminescence Analysis

“…Consequently, monitoring the concentration of UA in bodily fluids is essential in diagnosis and treatment of the corresponding diseases. Many methods have been reported for the determination of UA, such as capillary electrophoresis (CE) [7], enzymatic assay (EA) [8], high-performance liquid chromatography (HPLC) [9, 10], differential pulse voltammetric (DPV) [11, 12], and UV-Vis spectroscopy [13, 14]. Chemiluminescence (CL) has also been developed for UA determination [15–17].…”