The essential oil of Hedychium spicatum rhizome is used in perfumery and medicines. The rhizome is reported to be stomachic, carminative, a bronchodilator stimulant and a tonic. For studying seasonal variation of the composition of its volatiles, rhizomes of H. spicatum were collected from the Council of Scientific and Industrial ResearchâInstitute of Himalayan Bioresource Technology (CSIR-IHBT), Palampur, Himachal Pradesh, India. To study the spatial diversity in essential oil content and composition, the rhizomes were collected from five different locations of Himachal Pradesh. Essential oils obtained by hydro-distillation were evaluated for their chemical composition by gas chromatography (GC) and GC/mass spectrometry (GC/MS) analyses. Twenty-two compounds were identified from the rhizome oil with major components 1,8-cineole, Î²-eudesmol, Î²-pinene and 10-epiâÎ³-eudesmol. All the essential oil samples were evaluated for their antioxidant activity by 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging activity. The essential oil extracted from the rhizomes collected from Biling had highest antioxidant activity; hence, it was also tested for cytotoxic activity against C-6, A549 and SiHa cell lines.