| Summary: | In the current study, the total content from two <i>Apocynum</i> species leaves (<i>Apocynum venetum</i> and <i>Apocynum hendersonii</i>) collected from the Ili River Valley Region were extracted, and their bioactivities were investigated. The results showed a significant variation in the total flavonoid contents in the leaf samples collected at different periods (June, July, August, and September), with the highest content in August (60.11 ± 0.38 mg RE/g DW for <i>A. venetum</i> and 56.56 ± 0.24 mg RE/g DW for <i>A. hendersonii</i>), and the lowest in June (22.36 ± 0.05 mg RE/g DW for <i>A. venetum</i> and 20.79 ± 0.02 mg RE/g DW for <i>A. hendersonii</i>). The total flavonoid content was comparably higher in <i>A. venetum</i> than in <i>A. hendersonii</i>. Leaves extracts from the two species demonstrated strong bioactivity, which positively correlated with the total flavonoid contents. The anti-oxidative activity of <i>A. venetum</i> was higher than that of <i>A. hendersonii</i> in tandem with its higher flavonoid contents; the antibacterial activity, however, was conversely opposite. Furthermore, a total of 83 flavonoid metabolites were identified in the two species based on UPLC-ESI-MS/MS, out of which 24 metabolites were differentially accumulated. The variability in these metabolites might be the reason for the different bioactivities displayed by the two species. The present study provides insight into the optimal harvest time for <i>Apocynum</i> species planted in the major distribution area of the Ili River Valley and the specific utilization of <i>A. venetum</i> and <i>A. hendersonii</i>.
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