Introduction: Salvia macrosiphon Boiss. (Lamiaceae) is an aromatic perennial herb that its phytochemical analysis has shown presence of flavonoid and phenolic compounds. Material and Methods: Essential oil of the aerial parts of S. macrosiphon collected from south of Iran was obtained by hydrodistillation and analyzed using gas chromatography-mass spectrometry (GC/MS). Cytotoxic activity of the essential oil and the total extract of this herb were investigated by using MTT assay on three cell lines. Results: Twenty six components, representing 100% of the total oil, were identified. The main components of the essential oil were Linalool (19%), β-Cedrene (14.64%) and β-Elemene (13.33%). Both of the total extract and the essential oil showed remarkable cytotoxic activity against all tested cell lines. Moreover, the volatile oil exhibited significant antioxidant and anti-acetylcholinesterase activities; however the total extract did not exhibit any effect on inhibition of acetylcholinesterase. Conclusion: According to achieved results, this herb could be considered as a valuable source of antioxidant, cytotoxic and anti-AChE compounds.