Untargeted metabolomics reveals that multiple reproductive toxicants are present at the endometrium

Recent epidemiologic research shows many environmental chemicals exhibit endocrine disrupting effects on the female reproductive system. Few studies have examined exposure at reproductive organs. Our aim was to perform a preliminary untargeted metabolomic characterization of menstrual blood, a novel biofluid, to identify environmental toxins present in the endometrium and evaluate the suitability of this sample type for exposome research. Whole blood menstrual samples were collected from four women using a menstrual cup. Samples were analyzed for small molecules that include both environmental chemicals and endogenous metabolites using untargeted liquid chromatography with high-resolution mass spectrometry (LC-HRMS). Principal component analysis (PCA) and ANOVA was used to identify differences within and between individuals' menstrual blood metabolomic profiles, and the influence of the sample processing method. To assess the presence of environmental exposures, LC-HRMS chemical profiles were matched to the ToxCast chemical database, which includes 4557 commonly used commercial chemicals. Select compounds were confirmed by comparison to reference standards. PCA of metabolome profiles showed analysis of menstrual blood samples were highly reproducible, with high variability in detected metabolites between participants and low variability between analytical replicates of an individual's sample. Endogenous metabolites detected in menstrual blood samples achieved good coverage of the human blood metabolome. We found 1748 annotations for environmental chemicals, including suspected reproductive toxicants such as phenols, parabens, phthalates, and organochlorines. Storage temperature for the first 24 h did not significantly influence global metabolomic profiles. Our results show chemical exposures linked to reproductive toxicity and endocrine disruption are present in menstrual blood, a sampling medium for the endometrium.