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. 2019 Dec 13;9:19075. doi: 10.1038/s41598-019-55600-1

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© The Author(s) 2019

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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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GF-to-SPF ratios of propanoic acid and butyric acid determined by the 2PA derivatization LC-MS/MS method (Absolute) and by the CIL-LC-MS/MS method (CIL). Bars indicate means ± SD. Gray bars indicate the ratio was obtained from absolute quantitation method; Three independent plasma samples were prepared for each group mice and separately labeled by 2PA. GF/SPF ratios of propanoic acid and butyric acid were calculated by mean of the absolute concentrations. White bars indicate the ratio was obtained from CIL-LC-MS/MS method; The plasma samples were obtained by combining equal volume (n = 3) of each group mice. The pooled plasma was labeled with 2PA (for SPF) and 2PA-d6 (for GF), respectively. Then labeled samples were mixed (1:1) and subjected to the differential analysis. GF/SPF ratios of propanoic acid and butyric acid were calculated by the mean of ion peak area in triplicate measurements.