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. 2021 Apr 14;11:8133. doi: 10.1038/s41598-021-86980-y

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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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Combined protocol of fibres and SCFA analysis. (A) A schematic workflow for the combined analysis of a very small gastrointestinal sample volume. HPAEC chromatograms of GOS faecal fermentation samples (B, C) and chicory FOS faecal fermentation samples (D, E). The black lines represent fibres present in the intact sample supernatant, and the red lines represent fibres present in the aqueous phase after the SCFA extraction procedure with HCl (B, D) or without HCl (C, E). The numbers in the chromatograms represent the degree of polymerization, 1 = monomers, 2 = dimers, ≥ 3 = oligomers, and * are components present in the quenching reagent and in the medium. The calibration curves of concentrations of acetate (F), propionate (G) and butyrate (H) with and without the addition of HCl in the extraction protocol, and SCFA concentrations (I) in the same faecal sample with and without the addition of HCl (mean ± SEM, n = 3–5) . GC–MS gas chromatography–mass spectrometry, HPAEC-PAD high performance anion exchange chromatography with pulsed amperometric detection.