Introduction. Due to the increasing requirements for the quality of pharmaceutical products, the requirements for standardization of herbal medicines are also growing, which in turn necessitates the development of approaches to quality control of medicinal plant raw materials. The use of such an advanced method as high-performance thin-layer chromatography (HPTLC) will improve the quality of drug standardization, which is associated with the advantages of this method: minimized sample preparation, parallel determination of small volumes, elution time, high sensitivity, efficiency and speed. Objective. Quantitative determination of rutin by high-performance thin-layer chromatography, as well as comparison of the results of quantitative determination of the amount of flavonoids in terms of rutin by the spectrophotometric method. Material and methods. Medicinal plant raw materials – violet herb – were subjected to extraction using a developed method with 70 and 96% ethanol. Quantitative determination of rutin in extracts was carried out by HPTLC and spectrophotometry. Results. The developed method for the quantitative determination of rutin was validated in terms of linearity, specificity and accuracy. The difference in the extractive ability of the solvents used was shown. The percentage of rutin determined using the HPTLC method in an ethanol (96%) extract (3 µl) is 3.0826%, in an ethanol (70%) extract (3 µl) – 1.5911%. The percentage of rutin determined using the HPTLC method in an ethanol (96%) extract (5 µl) is 3.0894%, in an ethanol (70%) extract (5 µl) – 1.6064%. The percentage of total flavonoids in terms of rutin determined using the spectrophotometric method in the ethanol (96%) extract is 3.4289%, in the ethanol (70%) extract – 2.4176%. Conclusion. As a result of the study, a method for the quantitative determination of rutin was developed and validated, which can be used in the standardization of violet herb.