A quantitative method using ultra performance liquid chromatography with a photodiode array detector (UPLC-PDA) was established for the analysis of 2 major plant metabolites: β-asarone and α-asarone from Acorus gramineus, A. tatarinowii, A. calamus and Anemone altaica, and their contents are compared with other herbs of Acorus species. The method was validated according to the International Conference on harmonization (ICH) guideline for validation of analytical procedures with respect to precision, accuracy, and linearity. The average content of β-asarone in Acorus gramineus was significantly higher than that in others, with the second highest concentration observed in A. tatarinowii, and only a trace amounts found in A. calamus and Anemone altaica. In contrast, the average content of α-asarone in A. calamus was the highest, followed by that in Acorus gramineus and A. tatarinowii. principle component analysis (PCA) confirmed that β-asarone and α-asarone content differed among the species. These results suggest that this UPLC-PDA method can be considered as good quality control criteria for Acorus gramineus.