Objective: Russian Oregano (Origanum vulgare L.) and Turkish oregano (Origanum onites L.) are pharmacopoeial plants that are used for producing medicinal plant raw materials (crude herbal drugs). The aim of the study was to determine the composition and content of biologically active substances in Origanum crude herbal drugs. Materials and methods: Complex of modern physicochemical methods was used in pharmacognostical research. Thin-layer chromatography, gas-liquid chromatography was applied for the determination of crude herbal drugs composition. The content of total flavonoids in terms of cynaroside was determined by differential UV spectrophotometry, total tannins content – by redox titration, essential oil – by distillation. Microscopic analysis was performed on a Biomed-C2 microscope with 5×, 10× eyepieces, 4×, 10× and 40× objectives, photographies – were taken with a Canon Power Short SX 210 IS digital camera. Results: Thymol is a dominant component in essential oil. Two species contain a cynaroside (flavonoid). The content of essential oil in O. vulgare and O. onites herb was 0.12±0.03% and 2.60±0.02% respectively; the total flavonoids in terms of cinaroside – 0.98±0.02% and 1.25±0.01%; total polyphenols – 11.54±0.35% and 19.43± 0.72% (condensed polyphenols precipitated by gelatin – 4.94±0.15% and 13.62±0.21%); ascorbic acid – 0.35± 0.05% and 0.40±0.06%. Existing ones were modified and a number of new unified procedures were developed, allowing qualitative and quantitative analysis of phenolic compounds in O. vulgare and O. onites herb. A complex of anatomical and diagnostic features (the structure of the stomatal complex, simple hairs, ethereal-oil glands) was established to carry out CHD identification. Conclusion: Composition and content of the polyphenolic complex and hydrophilic biologically active compounds of Origanum vulgare L. and Origanum onites L. crude herbal drugs (CHD) were determined. The obtained data will be used to improve the existing regulatory documentation for Origanum crude herbal drugs standardization. © RJPT All right reserved.