Abstract: To study the seasonal accumulation regularity of soluble sugar and gentiopicroside of Gentiana triflora, the contents of soluble sugar and gentiopicroside in different organs were determined with the method of anthrone colorimetry and the high performance liquid chromatography (HPLC), respectively. The results showed that the contents of soluble sugar and gentiopicroside in different organs of G. triflora presented fluctuation according to developmental stages. The differences of soluble sugar content in different organs were obvious. Soluble sugar contents in underground organs were the highest. During the whole developmental period, soluble sugar content increased at the beginning and then decreased. Meanwhile, soluble sugar content maintained at a higher level during its dormancy stage. Gentiopicroside content was the highest in the underground organs, followed by stem, flower, and leaf with the sequences from high to low. The two high accumulated peaks were found respectively in early July and middle of October in the underground organs. During the seasonal development period, the changes of soluble sugar and gentiopicroside in the underground organs of G. triflora showed a negative correlation, and the negative correlation was more obvious from plant germination to the early stage of fruit development. Therefore, the best harvest time of G. triflora would be in the early and middle of October. The contents of soluble sugar and gentiopicroside showed some correlations during entire development period.