Background. Promising drugs for the treatment of a number of infectious diseases are most often represented by multicomponent injectable dosage forms, which include a set of inorganic salts, antibacterial and antiviral substances, as well as auxiliary substances that act as prolongators and preservatives. The shelf life of the drug and its pharmacokinetic characteristics depend on the amount of these substances and their ratio in the composition. The aim of the study. To develop and validate the procedure for joint determination of β-cyclodextran and polyvinylpyrrolidone in multicomponent preparations by high-performance liquid chromatography (HPLC). Materials and methods. In this study, the optimal column and sorbent was selected for quantitative determination of PVP separately from β-cyclodextran with the method of exclusion chromatography and the validation of the technique was performed using correctness, convergence (precision), linearity, and range of application. In this work, we used a LC-20 block liquid chromatograph (Shimadzu Co. Japan) and a chromatography method with simultaneous application of two columns. Results. Optimal columns were chosen for quantitative determination of PVP separately from β-cyclodextran by the method of exclusion chromatography. The following columns were chosen: the main one – 250 mm x 0.46 mm, filled with the Equisil ODS sorbent, particle size 5 μm, (manufactured by Dr. Maisch, Germany); Protective – size 20 mm x 4.0 mm Discovery HS C18, particle size 5 μm (Supelco, USA). Conclusion. The method of quantitative determination and establishment of the authenticity of β-cyclodextran corresponds to all metrological requirements to the technique characterized by sufficient convergence and accuracy. The systematic error of the technique for each determined component satisfies the requirements of practical insignificance.