METHOD FOR DETERMINING ANTIRADICAL ACTIVITY OF BIOLOGICAL FLUIDS

摘要

FIELD: medicine. SUBSTANCE: method involves applying 2,2-azo-bis-iso-butyronitryl preliminary dissolved in dimethylsulfoxide to 50 mM concentration. Luminol is first dissolved in dimethylsulfoxide and its concentration is brought to 253 mM by adding Henks solution. Then, 800 mcl of the produced 2,2-azo-bis-iso- butyronitryl solution is mixed with 200 mcl of 5 mM Na₂HPO₄ solution and 50 mcl of luminol solution at T+37 C. The mixture is stirred at 800 rpm speed during 15 min and then, stationary luminescence level is recorded during 1 min in the course of radical formation process (I₀). Next to it, 15 mcl of biological fluid under study is added and new luminescence level (I) is recorded on the stationary kinetic plot segment. Antiradical activity of biological fluid is calculated from formula APA = k₇xconst = (i^-1/2-i^1/2)/(2x[In], where I₀ is the initial chemiluminescence intensity, I is the chemiluminescence intensity on stationary segment after adding biological fluid under study, k₇ is the link rupture speed constant in reactions with In participating, [In} is the inhibitor concentration in the sample, i-I/I₀. EFFECT: high accuracy of diagnosis; simplified method. 2 tbl