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Gentiana straminea Maxim. is a well-known Tibetan traditional herb, which has been used to treat rheumatic arthritis, iceteric hepatitis, and other diseases for thousands years. However, there is still lack of comprehensive active constituents profiling of this species throughout the Qinghai-Tibet Plateau (QTP). This study was designed to provide a comprehensive quality map of G.straminea germplasm based on twelve active constituents (loganic acid, gentiopicroside, swertiamarin, sweroside, 6-O-β-D-glucosylgentiopicroside, oleanic acid, morroniside, trilobatin, isoorientin, isovite, Shanzhisidemethylester and quercetin) on the QTP. G.straminea root samples collected throughout QTP in the flowering period were analyzed by the LC-MS/MS. Statistics analysis methods PCA, clustering and ecological regions analysis for G.straminea constituents differentiation was demonstrated. The active constituents varied greatly across the QTP; the majority of constituents were secoiridoid derivatives, with gentiopicroside being the most abundant compound. Most constituents were significantly affected by the latitudes and altitudes other than longitudes. PCA and hierarchical clustering analysis showed that all samples could be separated into six distinct groups, and 15 populations showed the highest constituent abundances. Further, geographical region analysis showed that the highest quality populations mainly located near the source region of Yellow River, especially in the Qinghai and Sichuan areas. Additionally, correlation analysis showed that there were relationships among genetiopicroside, loganic acid, and other compounds, which might be related to the enzymatic pathways involved in the metabolism of these constituents. Conclusion : LC-MS/MS method allowed separation of quality profiling of G.straminea on the QTP, 15 populations showed the highest constituent abundances. In six geographical groups, the highest quality populations mainly located near the source region of Yellow River, especially in the Qinghai and Sichuan areas, which may be due to the climate caused by the westerlies and Indian Ocean monsoons in the QTP. [ABSTRACT FROM AUTHOR]