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Clinical Chemistry and Laboratory Medicine (CCLM)

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Allantoin as a marker of oxidative stress in human erythrocytes

Roman Kand'ár1 / Pavla Žáková2

1Department of Biological and Biochemical Sciences, Faculty of Chemical Technology, University of Pardubice, Pardubice, Czech Republic

2Department of Biological and Biochemical Sciences, Faculty of Chemical Technology, University of Pardubice, Pardubice, Czech Republic

Corresponding author: Mgr. Roman Kand'ár, PhD, Department of Biological and Biochemical Sciences, Faculty of Chemical Technology, University of Pardubice, Strossova 239, 530 03 Pardubice, Czech Republic Phone: +420-466037715, Fax: +420-466037068,

Citation Information: Clinical Chemistry and Laboratory Medicine. Volume 46, Issue 9, Pages 1270–1274, ISSN (Online) 1437-4331, ISSN (Print) 1434-6621, DOI: https://doi.org/10.1515/CCLM.2008.244, July 2008

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Background: Uric acid is the final product of purine metabolism in humans. It was determined that this compound has important antioxidative properties and it may be oxidized to allantoin by various reactive oxygen species. Therefore, the measurement of allantoin may be useful for the determination of oxidative stress in humans.

Methods: We measured allantoin and uric acid in human plasma and erythrocytes obtained from patients with chronic renal failure before hemodialysis (n=30) and blood donors (n=30). We used a method based on selective isolation of allantoin from deproteinized plasma and erythrocyte lysate samples on AG 1-X8 resin and its derivatization to glyoxylate-2, 4-dinitrophenylhydrazone. Separation of glyoxylate-2, 4-dinitrophenylhydrazone from interfering substances was achieved on reversed phase HPLC with gradient elution and UV/VIS detection at 360 nm. Uric acid was determined by reversed phase HPLC with UV/VIS detection at 292 nm.

Results: We found significant differences in allantoin and uric acid concentration between the patients with chronic renal failure and the control group both in plasma (20.5±6.5 μmol/L and 323.9±62.9 μmol/L vs. 2.1±1.1 μmol/L and 270.1±62.3 μmol/L, p<0.05) and erythrocytes [82.8±39.1 nmol/g hemoglobin (Hb) and 110.7±28.8 nmol/g Hb vs. 20.1±6.1 nmol/g Hb and 82.1±23.7 nmol/g Hb, p<0.05].

Conclusions: Significant higher levels of allantoin in both plasma and erythrocytes of patients with chronic renal failure indicate that allantoin may be used as a good marker of oxidative stress.

Clin Chem Lab Med 2008;46:1270–4.

Keywords: allantoin; antioxidants; human erythrocytes; oxidative stress; uric acid

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