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Reactive oxygen species (ROS) contributes to the oxidative stress associated with ageing and chronic diseases, highlighting the significance of natural antioxidants from traditional medicinal plants as a critical area of research. Gastrodia elata Blume (GE), a valuable traditional Chinese medicine mainly cultivated in Guizhou Province, China, is known for its medicinal properties including antioxidant activities. However, the metabolic profiles and underlying potential antioxidant mechanisms of GE , which can be influenced by environmental heterogeneity and cultivar variation in this region, remain poorly understood, limiting its full potential for comprehensive utilization. This study evaluated total contents of sugar, phenolic and flavonoid, antioxidant capacities and metabolic profiles of 11 GE samples from 8 counties in Guizhou using chemical assays and untargeted metabolomics. Network pharmacology and molecular docking revealed key antioxidant metabolites and their potential action mechanisms. Results showed that GE samples from high-altitude sites performed better in three functional parameters and antioxidant capacity. In addition, amino acid transformation and benzoate degradation were the main metabolic differences among these samples. Five key antioxidant metabolites, including isoreserpine and phytosphingosine, showed strong affinities with key targets such as GAPDH and EGFR through hydrogen and hydrophobic bonds, acting through pathways such as PI3K-Akt and IL-17 signaling. These findings provide insights into the cultivation and metabolic optimization of GE in Guizhou and enhance the understanding of its antioxidant properties, supporting its therapeutic and functional food applications. [Display omitted] • Quality and metabolite profiles of 11 GE from 7 counties in Guizhou were tested. • GE collected from high altitude showed better function and antioxidant activities. • Metabolic differences related to amino acid conversion and benzoate degradation. • 5 key antioxidant compounds showed good affinities with core antioxidant targets. [ABSTRACT FROM AUTHOR]

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