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The purpose of the work was to investigate the qualitative and quantitative composition of Escherichia coli and Staphylococcus aureus lipids after exposure to disinfectants. Methods. Quantitative and Qualitative composition of total lipids and phospholipids were identified using method of thin layer chromatography. Disinfectants Barez® (active ingredients: essential oils, benzalkonium chloride, nanoparticles of argentum), Biochlor® (sodium hypochlorite), Geocide® (benzalkonium chloride; polyhexamethylene guanide hydrochloride, deltametrin), were used according to manufacturer’s recommendations. For this test, 0.5 ml of disinfectants (0.1 % Barez®, 0.1 % Biochlor®, 0.5 % Geocide®) were added into 3 ml of cell suspension (5×108 cell/ml). Controls were the cells nontreated with disinfectants. Cells suspensions were incubated in standard conditions for 1 hour. Results. The influence of disinfectants on the lipid composition of E. coli and S. aureus after the action of Barez®, Biochlor and Geocide was studied. Changes of quantitative and qualitative composition of the lipids containment were identified. Barez® had the most notable effect on quantitative and qualitative composition of total lipids and phospholipids. It was confirmed that reduction of total quantity of several phospholipids results to increased level of diglycerides. These data allow us to establish the sensitivity of microorganisms to the influence of disinfectants and to determine the optimal concentrations and exposure of disinfectants for high quality disinfection. Conclusions. It has been established that the disinfectants Barez®, Biochlor® and Geocide® cause significant changes in the lipid composition of E. coli and S. aureus cells, which leads to increased fluidity of cell membranes and loss of viability of bacteria. [ABSTRACT FROM AUTHOR]