This study aimed to investigate the chemical constituents of   Cotula cinerea at three different habitats (Al-Arish, Saint Catherine and Matruh) during growth season of 2008. Phytochemical screening of the aerial plant parts revealed the presence of sterols (except Saint Catherine habitat) volatile oils, tannins, saponins, flavonoids, alkaloids, carbohydrates and/or glycosides, sulphates and chlorides. Determination of constants and other constituents revealed some differences in these values between the three different habitats. The percentages of total flavonoids and saponins reached to their maximum values in the plants at Matruh habitat, while the highest values of alkaloid and tannins were detected in the plants at Al-Arish habitat. The free and combined sugars were detected by using GLC. The free amino acids were detected by using paper liquid chromatography, while the protein amino acids were detected by using the amino acid analyzer.  The fundamental chemical properties of lipids of Cotula cinerea collected from the three different habitats were determined. The obtained GLC results showed that the unsaponifiable matter of the lipids contained squalene and cholesterol at the three habitats, while stigmasterol and β-sitosterol were present only in Al-Arish and Saint Catherine habitats. GLC chromatograms of the fatty acids content showed that the plants of Matruh habitat contained eleven fatty acids, while the plants of Al-Arish and Saint Catherine contained only eight fatty acids. The higher percentage was that of oleic acid in the three studied habitats.