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Highly Specific and Rapid Method for the Determination of Uric Acid in Erythrocytes
Authors: Žáková Pavla | Kanďár Roman | Malíková Jana | Čegan Alexander | Ventura Karel
Year: 2003
Type of publication: ostatní - přednáška nebo poster
Name of source: Clinical Chemistry and Laboratory Medicine Special Supplement
Publisher name: Walter de Gruyter GmbH & Co. KG
Place: Berlin
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Language Name Abstract Keywords
cze Vysoce specifická a rychlá metoda pro stanovení kyseliny močové v erytrocytech uric acid, erythrocytes, antioxidant, HPLC, electrochemical detection, patients with chronic renal failure
eng Highly Specific and Rapid Method for the Determination of Uric Acid in Erythrocytes Uric acid is antioxidant in humans and protects erythrocyte ghosts against lipid peroxidation. Aim: To develop and to evaluate a method for the determination of uric acid in erythrocytes by HPLC with electrochemical detection. Erythrocytes were obtained from patients with chronic renal failure before hemodialysis (n = 30, 15 women and 15 men) and blood donors (n = 30, 15 women and 15 men). The erythrocytes were washed threetimes, each with an equal volume of isotonic solution and hemolysed with cold water. To precipitate proteins was used 10% trichloroacetic acid/10 mM oxalic acid. The supernatants were extracted with 3 vol of water-saturated ethyl ether to remove the trichloroacetic acid and oxalic acid. Twenty microl extracted supernatant was injected onto a Discovery C18, 150 x 4.0 mm, 5 microm column. The mobile phase consisted of 0.05 M sodium phosphate, 0.05 M sodium acetate, 189 microM dodecyltrimethylammonium chloride, and 3.66 microM tetraoctylammonium bromide in 30/70 methanol/water (v/v), pH 4.80. The flow rate was 0.5 mL/min. Electrochemical detector ESA Coulochem Model 5100A settings were as follows: E1 = 30 mV and E2 = 280 mV. We found the significant differences in uric acid concentrations between the patients and the control group (109.6 +- 24.9 nmol/g Hb vs. 76.5 +- 24.3 nmol/g Hb; p < 0.001, Student's t-test). Conclusion: We have developed and evaluated a HPLC method for the determination of uric acid in erythrocytes. Supported by grants MSMT 253 100 002, GACR 203/02/0023. uric acid, erythrocytes, antioxidant, HPLC, electrochemical detection, patients with chronic renal failure

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