Objective  To establish ultra performance liquid chromatography (UPLC) fingerprint for Wuyao decoction lyophilized powder, characterize the chemical components of its common peaks, and explore the material basis and potential mechanism of Wuyao decoction lyophilized powder in the treatment of primary dysmenorrhea using bioinformatics analysis techniques.

Methods  UPLC fingerprint for the lyophilized powder of Wuyao decoction was established, and the structures of the common peaks were characterized using liquid chromatography-mass spectrometry (LC-MS). Network pharmacology was applied to construct a "chemical component-target-pathway" relationship network. The potential active components and mechanisms of action for the treatment of primary dysmenorrhea were predicted by consulting literature, and finally verified through molecular docking using Autodock Vina.

Results  UPLC fingerprint for 10 batches of Wuyao decoction lyophilized powder reference samples was established, and the structures of 17 common peaks were identified. From these components, six key components, 12 core targets, and 10 signal pathways for Wuyao decoction in the treatment of primary dysmenorrhea were screened. KEGG pathway analysis showed high involvement of TNF, PI3K/Akt and MAPK signaling pathways. Molecular docking results indicated that chemical components such as diosmetin can stably bind to proteins such as the phosphoinositide-3-kinase catalytic subunit alpha peptide.

Conclusion  Diosmetin, desacetylmatricarin, ferulic acid, boldine, sophoradiol and pterodontic acid are likely to be the potential active ingredients in Wuyao decoction for alleviating primary dysmenorrhea, and act on core target proteins such as PIK3CA and ESR1. By regulating signaling pathways such as TNF, PI3K/Akt and MAPK, these components can jointly play a role in the treatment of primary dysmenorrhea. A preliminary exploration of the potential mechanism of Wuyao decoction in treating primary dysmenorrhea has been carried out, providing a multi-component, multi-target, and multi-pathway intervention strategy for relieving primary dysmenorrhea. The established UPLC fingerprint and its chemical composition analysis have laid a solid foundation for the in-depth study of subsequent pharmacodynamics.

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