To determine the content of esculetin in the different parts of Viola philippica from various habitats by HPLC to provide the basis for the reasonable application and the quality assessment of the herb. Methods:The content of esculetin in roots, leaf stems and leaves of Viola philippica was determined by HPLC, and the proportion of various parts in the total grass was calculated, and the content of esculetin in Viola philippica from different habitats was compared. Results:The index ingredient esculetin was de-tected out in the roots, leaf stems and leaves, the content in leaf blades was the highest followed by leaf stalks, and that in root was the lowest. There were notable differences in the quality of Viola philippica from different habitats. Conclusion:The content of esculetin in the leaves of Viola philippica is the highest proportion, and that in Viola philippica from Hebei is the higher. The difference among the other habitats is relatively small. The results can provide reference for the harvest and reasonable application of Viola philippica.