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Radiotherapy is one of the most promising and in-demand methods used for treatment of oncological diseases, especially in case of brain, head, neck tumors, and tumors of some other localization. However, in 50% - 90% cases even the application of the optimized methods of radiotherapy, providing significant increase in patient’s life expectancy, leads to radiation damage accompanied by progressive disorders of such cognitive functions as memory, attention, thinking etc. Nowadays there are no analytical systems providing evaluation of the risk of development of radiation damage of brain, approaches to its early diagnostics, methods of medicated prevention and therapy. The aim of this investigation was to develop the multiplex fluorescent procedures for prognostics and diagnostics of cognitive complications in the result of radiotherapy using the appearance and content of neuromediator exchange markers as a control test. We proposed a bio-sensing system for sensitive determination of catecholamines (dopamine, epinephrine, norepinephrine), and their metabolites based on producing highly fluorescent derivatives with benzylamine (BA) or 1,2-diphenylethylenediamine (DPhEDA) in the presence of horseradish peroxidase (HRP). A 96-well polystyrene plate modified by bio-recognizing film {chitosan-DPhEDA/BA-HRP} was applied as a template for fluorescence measurements. The spectra of the solutions were recorded in the interval 400 – 500 nm (λex = 305 – 360 nm). The bio-sensing system for the determination of nanomolar concentrations of the markers is selective enough for the analysis of blood plasma. The reproducibility (RSD, %) of a response of the biosensing system does not exceed 10% (n = 5). The developed techniques were tested for rapid determination of the markers in blood plasma of mice for a comprehensive assessment of the general state of their health before and after radiation therapy.