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Updated: May 26, 2026
2D-HPLC-MS Technology Combined with Molecular Network for the Identification of Components in Tibetan Medicine Aconitum pendulum
Published on: December 8, 2023
An activity labelled molecular networking strategy-assisted discovery of antiplatelet aggregation-active components
Yan Xu1, Beibei Zhang2, Liangliang He3
1Department of Hematology, the First Affiliated Hospital of Zhengzhou University, Zhengzhou, China.
Introduction:
Allium chinense G. Don (ACGD) is a famous Chinese medicine that shares homology with medicine and food. However, the mechanism of ACGD-related components in antiplatelet aggregation remains unclear.
Methods:
This study identified steroidal saponins using a targeted molecular networking (TMN) approach and explored the mechanism of antiplatelet aggregation.
Results:
ACGD samples (1 mg/mL) extracted using different solvents (water, 30% methanol, 50% methanol, 70% methanol, and 100% methanol) all exhibited inhibitory effects on platelet aggregation induced by adenosine diphosphate, arachidonic acid, and collagen. The ACGD fraction extracted with 70% methanol (1 mg/mL) displayed the strongest inhibitory effects. Furthermore, a total of 36 steroidal saponins were visualized based on TMN analysis. These components were identified or tentatively characterized according to their retention behaviors, diagnostic ions, and fragmentation patterns. Among them, laxogenin-related furostanol and spirostanol saponins were the most abundant saponins in ACGD. A total of 15 saponins with antiplatelet aggregation activity, high content levels, different structural types, or authentic standards were selected to perform the network pharmacology analysis. The PI3K/Akt pathway is one of the most important signaling pathways in platelet activation. Moreover, based on CCK-8 assays in MEG-01 cells, compounds T3-12, T3-14, T3-15, T3-16, and T3-17 were non-toxic and did not induce proliferation at 5-20 μM. Moreover, compounds T3-15, T3-16, and T3-17 at 20 μM caused downregulation of the p-PI3K protein, whereas components T3-14, T3-16, and T3-17 at 20 μM decreased p-Akt in MEG-01 cells.
Discussion:
The TMN approach, integrated with diagnostic ions, was successfully applied to characterize target steroidal saponins in ACGD. Compounds T3-16 and T3-17 were confirmed to be active components in ACGD in antiplatelet aggregation through the PI3K/Akt pathway.
