Total alkaloids content of both Orobanche aegyptiaca and Orobanche ramose Was detected using High-performance liquid chromatography (HPLC). The different kinds and concentration of alkaloids were also studied. Anti-oxidant effectiveness of both species was investigated using 2, 2-Diphenyl-1-picrylhydrazyl (DPPH). In O. ramose five alkaloids were identified (Lupanin, Sparteine, Oxoretamine, Isospartein and Oxosparteine), in case of O. aegyptiaca only four alkaloids were identified (Lupanine, Spartein, Isosparteine and Oxosparteine). The highest level of alkaloids was found in O. aegyptiaca (138.93 µg/gm) and the rate inhibition of free radicals was (85.31%).