Introduction: This study was designed to evaluate the phytocomponents present in aerial part and in vitro induced callus methanol extracts of Dipterygium glaucum by Gas Chromatography-Mass Spectrometry (GC-MS) technique. Methods: GC-MS analysis of aerial part sample and callus produced from leaf of D. glaucum extracted in methanol solvents was performed using GC-MS QP 2010 Plus (Shimadzu, Japan) system comprising an auto sampler (AOC-20i) and a gas chromatograph interfaced to a mass spectrometer. Results: This study was carried out to identified and comparative analysis of bioactive phytochemicals from aerial part extract and callus extract of D. glaucum. This analysis revealed that both the extracts have 69 different types of phytochemical components in varying quantities. Some of the important phytochemical compounds were Stigmasterol, β-sitosterol, γ-sitosterol, Campesterol, Squalene, n-Hexadecanoic acid, Stearic acid, Myristic acid, Quinazoline, Linalyl acetate etc. These chemical compounds have anticancer, antitumor, anti-inflammatory, antidiabetic and antioxidants properties. Conclusion: This study represents the detection and identification of different phytochemical compounds from aerial part and callus extract of D. glaucum. Thus, due to the presence of various important bioactive phytocomponents this plant is recommended as a pharmaceutically important plant.