(Decne.) Sch.Bip is a well known herbal medicine has been extensively used for many therapeutic purposes. In this article, we have accomplished targeted profiling of roots samples collected from different locations (Pangi, Lahaul, Barot and Kullu) of the western Himalayan region using NMR. This study allowed us to assigned four major sesquiterpenes of this plant, including costunolide (CL), dehydrocostus lactone (DCL), alantolactone (AL), isoalantolactone (IAL). Quantification of sesquiterpene lactones have also been accomplished using HNMR, and a wide diversity in the concentration of these active metabolites was observed. It has been observed that AL and IAL were major in the samples collected from Lahaul and Barot regions, and CL and DCL were major in the samples collected from Kullu and Pangi regions. This study concludes that roots possess wide chemical diversity, which may be due to various biotic and abiotic environmental related factors.