The study aims to identify quality markers (Q-Markers) for the evaluation of Hunan Aurantii Fructus (HAF) germplasm resources based on the "spectrum-efficacy" correlation by the establishment of chromatographic quantitative analysis of multiple components,combining network pharmacology,molecular docking technology,and multivariate statistical analysis.High performance liquid chromatography and headspace solid-phase microextraction-gas chromatography were used to perform content determination of germplasm resource samples from counties and cities in Hunan.Target collection and network pharmacology analysis were carried out on multi-components to construct a "component-target-pathway" network and predict candidate Q-Markers and core targets.Molecular docking technology was used to theoretically simulate the binding activity of ligands and receptors to identify key Q-Markers.Principal component loading factor analysis and cluster analysis were utilized to excavate Q-Markers that distinguish HAF germplasm resource samples.The results showed that the germplasm resource samples had the highest contents of naringin,neohesperidin,and D-limonene,with significant fluctuations in the content differences of each target compound. Seven candidate Q-Markers,including tangeretin,exerted antioxidant and anti-inflammatory effects through seven core targets such as PIK3CD in the PI3K/AKT and RAS/MAPK pathways,demonstrating good molecular docking activity among them.Multivariate statistical analysis indicated that five Q-Markers,including limonin,could be used to differentiate germplasm resource samples.These markers were group-specific,while the differences between samples were unrelated to geography.This method can evaluate the quality of HAF germplasm resources and furnishing a valuable tool for variety selection and breeding initiatives.

"> Hunan Aurantii Fructus germplasm resources, quality markers, chromatography, network pharmacology, molecular docking, multivariate statistical analysis