Objective To rapidly and accurately analyze and identify the complex chemical components in Honghua Qinggan Shisanwei pills by high performance liquid chromatography-Q-Exactive mass spectrometry (HPLC-Q-Exactive MS), clarify the active ingredients for the alleviation of non-alcoholic fatty liver disease, and explore the potential mechanism of action of Honghua Qinggan Shisanwei pills by network pharmacology and molecular docking.

Methods  Gradient elution was performed using methanol and aqueous solution containing 0.1% formic acid as mobile phases. The scan was switched by alternating positive and negative ion scanning modes with the scanning range of m/z 110-1 200. The compounds in the formula were identified by comparing them with reference substance, references, and self-constructed databases. Based on liquid chromatography-mass spectrometry data, active compounds were further screened using the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) database. Possible targets of Honghua Qinggan Shisanwei pills were predicted and analyzed using online databases such as SwissTargetPrediction and GeneCards, followed by KEGG and GO enrichment analyses. The core targets were identified using the protein-protein interaction network. Finally, molecular docking technology was employed to verify the binding viability of the active compounds with the targets.

Results  A total of 75 compounds were identified from Honghua Qinggan Shisanwei pills, including 31 flavonoids, 17 organic acids, 9 tannins, 8 cyclic enol ether terpenes, 3 lignans and 7 others. 5 core targets were identified using network pharmacological methods: serine/threonine kinase 1 (AKT1), epidermal growth factor receptor (EGFR), human epidermal growth factor receptor 2/tyrosine kinase receptor 2 (HER2/ERBB2), phosphatidylinositol 3-kinase regulatory subunit 1 (PI3KR1) and non-receptor tyrosine kinase Src. The KEGG enrichment pathways mainly included the PI3K-AKT signaling pathway, the EGFR tyrosine kinase inhibitor resistance pathway and the ERBB signaling pathway.

Conclusion  The complex chemical components of Honghua Qinggan Shisanwei pills were rapidly and accurately identified. Additionally, the potential mechanisms for the treatment of non-alcoholic fatty liver disease were investigated, providing a scientific reference for the quality control of Honghua Qinggan Shisanwei pills and subsequent research.

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