[Aim]  To investigate metabolic differences and potential regulatory mechanisms related to colony strength in Apis cerana cerana. [Methods]  Metabolites in strong and weak colonies were screened using non-targeted metabolomics and liquid chromatography-mass spectrometry (LC-MS), after which KEGG enrichment analysis was performed. Glutathione S-transferase (GST), superoxide dismutase (SOD) and cytochrome P450 (CYP450), activity were determined using biochemical methods. [Results]  In positive and negative ion modes, 616 and 212 distinct metabolites were identified in workers from strong and weak colonies. After annotation, betaine, flavonoids, lysoPA (0:0/18:2(9Z,12Z)), and other metabolites related to antioxidant, anti-inflammatory, antibacterial and anti-apoptosis, were identified. Differential metabolites were significantly enriched in six metabolic pathways, including the metabolism of xenobiotics by cytochrome P450, the AMPK signaling pathway, the glutathione metabolism, arginine biosynthesis, thermogenesis and folate biosynthesis. In addition, CYP450, SOD and GST activity was higher in strong colonies than in weak ones. [Conclusion]  There were significant differences in the metabolites of strong and weak A. c. cerana colonies. Worker bees in strong colonies had a stronger energy metabolism, and were better able to resist oxidative stress, extreme temperatures, exposure to harmful substances, inflammation and bacterial infection.