Introduction. Due to the high prevalence and social significance of fungal infections, the search for new antifungal drugs is an actual direction t of modern pharmacology. One of the important components of preclinical microbiological studies is the culture media used in the process of a experiment.Aim. To validate the use Sabouraud liquid medium as an alternative to RPMI 1640 medium to optimize the experimental conditions for the determination of antifungal activity of new compounds by the

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... oserial dilution method.Materials and methods. The micromethod of two-fold serial dilutions to study the antifungal activity of new molecules was used. Antifungal activity was studied against the reference test strain Candida albicans NCTC 885-653. The evaluation of the validation parameters was carried out according to the principles set forth in the General Pharmacopoeia Monograph.1.1.0021.18 «Validation of microbiological methods». A new compound from the group of 4-(het)aryl-2,4-dioxobutanoic acid amide derivatives was used for test the validated method.Results and discussion. Results were obtained that meet the acceptance criteria for the studied validation parameters – trueness, precision, linearity, robustness and quantitation limit. The possibility of using Sabouraud's medium for the determination of antifungal activity was shown on the example of a new compound of the group of 4-(het)aryl-2,4-dioxobutanoic acid amide derivatives.Conclusion. The tested alternative liquid Sabouraud medium can be recommended for use in determining the antifungal activity of the newly synthesized compounds with the microdilutions method because the proven its better growth properties for the investigated yeast strain and the convergence of the results of determining antifungal activity using alternative and reference media.