| 摘要 |
FIELD: clinical biochemistry, in particular evaluation of fibrinogen oxidative modification degree under oxidative stress action. ^ SUBSTANCE: claimed method includes treatment of tested sample with 20 % trichloroacetic acid (TCA) solution, addition of 1 ml of 0.1 M 2,4-dinitrophenyl hydrazine in 2 M HCl to precipitate; incubation at room temperature for 1 h; centrifugation at 3000 g for 20 min; three-stage precipitate washing with 1 ml 1:1 ethanol/acetate solution; precipitate drying; dissolving thereof in 2.5 ml of 8 M urea followed by evaluation of protein oxidative modification based on level of formed dinitrophenyl hydrazones using spectrophotometrical detection of optical density, wherein 0.5 ml of 20 mM CaCl2 solution is added in 1 ml of citrate blood plasma, mixture is incubated on water bath at 37°C for 10 min, fibrin clot is isolated, dried on paper filter, weighted, converted to fibrinogen concentration (mg/ml), and before treatment with 20 % TCA solution 0.5 ml of 0.9 % NaCl solution is added to fibrinogen clot and further TCA solution is added in ratio of 1:1. ^ EFFECT: accelerated method of high information content. |
