Background: Menopause associated asthma impacts a subset of women and tends to be more severe and is less responsive to current treatments. We recently developed a model of menopause associated asthma using a combination of 4-Vinylcyclohexene Diepoxide (VCD) and House Dust Mite (HDM) exposures. The goal of this study was to uncover potential biomarkers and drivers in menopausal asthma by assessing serum and BALF samples from mice with and without menopause with and without asthma by large-scale targeted metabolomics. Methods: Four groups of mice were assessed: menopausal asthma (VCD/HDM), asthma (HDM), menopause (VCD) and control (Vehicle). Lung function during a methacholine challenge was assessed by Flexivent. Serum and BALF samples were collected liquid chromatography tandem mass spectrometer was used to examine ~300 metabolites from >25 metabolic pathways. Results: Over 50 metabolites, impacting multiple metabolic pathways, were significantly different across the 4 study groups. Particularly, Glutamate, GABA, phosphocreatine, and pyroglutamic acid, which are involved in glutamate and glutamine metabolism, glutathione metabolism, and arginine and proline metabolism, were significantly impacted in the menopausal asthmatic mice compared to non-menopausal asthmatic mice. Glutamic Acid, Histamine, Uridine, Cytosine, Cytidine, and Acetamide has significant correlations with total airways resistance. Conclusions: Using metabolic profiling of serum and bronchoalveolar lavage fluid from mouse models we identified metabolites and metabolic pathways that may aid in discriminating potential biomarkers of menopause-associated asthma.