期刊
CLINICAL BIOCHEMISTRY
卷 46, 期 7-8, 页码 665-669出版社
PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/j.clinbiochem.2013.01.006
关键词
Ascorbic acid; Uric acid; Plasma antioxidants; HPLC-UV; Ion-exclusion; Atherosclerosis
资金
- project 'Search for nuclear and mitochondrial biomarkers of atherosclerosis in isolated populations from Azores Islands' [M2.1.2/F/009/2007]
- Regional Government of the Azores
- Fundacao para a Ciencia e a Tecnologia [SFRH/BD/34322/2007]
- Fundação para a Ciência e a Tecnologia [SFRH/BD/34322/2007] Funding Source: FCT
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. (C) 2013 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.
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