Background and objectives: The genus Johrenia belongs to Umbelliferae family and contains five species that are endemic to Iran. Johrenia paucijuga grows widely in the north-west, west and center of Iran. So far, there has been no research about phytochemistry of J. paucijuga. In the present study, phytochemicals of the plants have been isolated and their structures have been elucidated. Methods: The aerial parts were dried and cut into small pieces, then extracted with ethyl acetate and methanol using perculator apparatus at room temperature. The methanol extract was extracted again with, petroleum ether and butanol. The separation and isolation process was carried out using column (silica gel and Sephadex LH-20) and thin layer chromatographic (TLC) methods. Structure elucidation of the purified compounds were based on ¹H and ¹³C-NMR data, in comparison with those reported in the previous literatures. Results: The isolated compounds from the ethyl acetate and butanol extracts of J. paucijuga were identified as β-sitosterol, β- stigmasterol and quercetin. Quercetin is a bioactive flavonoid widely used as a health supplement. β- sitosterol and β-stigmasterol are phytosteroles (plants sterols) with chemical structure similar to that of cholesterol and are sometimes used in treating hypercholesterolemia. Conclusion: Regarding the valuable biological properties of the isolated compounds, different biological effects could be expected from the plant