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. 2020 Nov 5;10:19158. doi: 10.1038/s41598-020-76119-w

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© The Author(s) 2020

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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Plots of significant features based upon the PCA analysis. (A) Loadings plot, where the spatial arrangement of the features (unique m/z-RT pairings) corresponds to the distribution of the samples (i.e., scores plot, Fig. 2). Speciofoline (3) appears to be a feature that is responsible for discriminating between the two sample groupings along the y-axis (PC2). (B) Volcano plot, highlighting significant features of the metabolites based on statistical testing (p value < 0.05, fold change > 2 or < 0.5) between the two groupings, with the further a feature’s position away from the origin (0, 0), the more significant the feature is. The speciofoline-rich group (red circles) yielded 120 significant features, while 19 features were significantly distinct in the non-speciofoline-rich grouping of commercial samples (purple circles). Speciofoline (3) was found to be a metabolite that differed significantly across sample groups, while mitragynine (1) and 7-hydroxymitragynine (2) were not significantly different between the two groups (orange circles). Other features predicted to be alkaloids based on mass defect (open circles). Labels on plot refer to the compound number and the type of ion observed (i.e.[3 + H]⁺ refers to the protonated molecule of speciofoline).