Objective: The present investigation was carried out to determine the possible bioactive components of stem, root, and leaf of Barleria courtallica Nees using a gas chromatography-mass spectrometry (GC-MS). Methods:The phytocomponents of the ethanol extracts of stem, root, and leaf of B. courtallica were investigated using PerkinElmer GC-MS, while the mass spectra of the compounds found in the extracts were matched with the National Institute of Standards and Technology version II library.Results: 25, 23, and 28 compounds were identified in the ethanol extracts of stem, root, and leaf of B. courtallica, respectively. The prevailing compounds of stem were β-sitosterol (20.27%), stigmasterol (17.07%), eicosane, 7-hexyl-(6.64%), 3,7,11,15-tetramethyl-2-hexadecan-1-ol (5.97%), and tetracosane, 11-decyl-(5.91%). The major constituents recorded from root extract of B. courtallica were β-sitosterol (22.94%), stigmasterol (20.17%), urs-12-en-28-oic acid, 3-hydroxy-, methyl ester, (3β)-(18.42%), and eiosane, 7-hexyl-(7.06%). The prevailing compounds of leaf were 3,7,11,15-tetramethyl-2-hexadecan-1-ol (34.42%), phytol (14.18%), β-sitosterol (12.71%), squalene (11.25%), stigmasterol (8.15%), phytol acetate (6.53%). Conclusions:From the results, it is evident that B. courtallica contains various bioactive compounds and is recommended as a plant of phytopharmaceutical importance.