Abstract: High altitude medicinal plants are highly demanding due to their therapeutic use but unfortunately mostly these plants are in threatened stage. So, there is an urgent need to conserve these species by applying appropriate agrotechnological practices. Experimental species was grown in polyhouse and open field conditions and after harvesting, the raw material was dried in different conditions. High Performance Liquid Chromatography (HPLC), was applied to separate and quantify iridoid glycosides (picrotin and picrotoxin) which are naturally present in Picrorhiza kurroa. The major active ingredients were quantitatively estimated in field and polyhouse grown plants. Good linear response over the range of 0.4 to 3.5% on the basis of dry weight was observed for each component. Both picrotin and picrotoxin were recorded in higher quantity respectively 3.1 and 1.2% of dry weight in polyhouse grown plants, While low 2.5 and 0.99% of dry weight of picrotin and picrotoxin, respectively in field grown plants. It was interesting to observe that the samples dried in room condition or in shade showed higher quantity of both the active contents in contrast to oven dried sample. It was also found that the broad leaf plants showed higher quantity of active ingredients in comparison to narrow leaf. On the basis of this study cultivation of the broad leaf variant inside the polyhouse and drying in room condition can be implemented for good quality and quantity of P. kurroa.