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. 2021 Jan 8;11:248. doi: 10.1038/s41598-020-80093-8

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Hierarchical clustering for 813 metabolites in maternal plasma. (a) Hierarchical Clustering on Principal Components. Percentage composition of microbial metabolites in the hierarchical clusters of the sample PCA loadings, using Ward’s method. Compounds were defined as microbial-derived (red), endogenous-derived (blue), and unknown identity (green). (b) Hierarchical clustering of the sample PCA loadings, plotted against the principle component score for Dimension 1. Plant phenolics were correlated in Cluster 1, and secondary bile acids in Cluster 5; Cluster 3 represented metabolites largely unaffected by alcohol. (c) Hierarchical Clustering on Spearman’s Correlation. The microbial-derived metabolites retained their relationships and plant-derived phenolics were correlated in cluster 4, while the secondary bile acids were in cluster 1. The correlation within Clusters 4 and 5 indicate metabolites having an immediate cellular process affected by the alcohol treatment, whereas the former clusters contain metabolites affected more distantly. For (b) and (c), a positive principal component score indicates the metabolite has increased abundance in response to alcohol; negative scores signify reduced abundance. Sample size is n = 9 control and n = 8 alcohol-exposed dams.