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Quantitative analysis of short-chain fatty acids in human plasma and serum by GC–MS

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Abstract

Short-chain fatty acids (SCFAs) are volatile fatty acids produced by gut microbial fermentation of dietary nondigestible carbohydrates. Acetate, propionate, and butyrate SCFA measures are important to clinical and nutritional studies for their established roles in promoting healthy immune and gut function. Additionally, circulating SCFAs may influence the metabolism and allied function of additional tissues and organs. The accurate quantification of SCFAs in plasma/serum is critical to understanding the biological role of SCFAs. The low concentrations of circulating SCFAs and their volatile nature present challenges for quantitative analysis. Herein, we report a sensitive method for SCFA quantification via extraction with methyl tert-butyl ether after plasma/serum acidification. The organic extract of SCFAs is injected directly with separation and detection using a polar GC column coupled to mass spectrometry. The solvent-to-sample ratio, plasma volume, and amount of HCl needed for SCFA protonation were optimized. Method validation shows good within-day and inter-day repeatability. The limit of detection was 0.3–0.6 µg/mL for acetate and 0.03–0.12 µg/mL for propionate and butyrate. Successful application of this method on clinical plasma and serum samples was demonstrated in six datasets. By simplifying the sample preparation procedure, the present method reduces the risk of contamination, lowers the cost of analysis, increases throughput, and offers the potential for automated sample preparation.

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All data generated or analyzed during this study are included in this published article.

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Acknowledgements

We thank the UCHealth Garth Englund Blood Center (Fort Collins, CO, USA) for providing control plasma samples, and Jane Andales for suggestions of quality assurance for this assay.

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Authors and Affiliations

  1. Analytical Resources Core–Bioanalysis and Omics, Colorado State University, Fort Collins, CO, 80523, USA

    Linxing Yao & Corey D. Broeckling

  2. Department of Environmental Health Sciences, Yale School of Public Health, Yale University, New Haven, CT, 06510, USA

    Emily A. Davidson

  3. Department of Cellular & Molecular Physiology, Yale School of Medicine, Yale University, New Haven, CT, 06510, USA

    Emily A. Davidson

  4. Rush University Medical Center and Rush University, Chicago, IL, 60612, USA

    Maliha W. Shaikh & Christopher B. Forsyth

  5. Department of Horticulture and Landscape Architecture, Colorado State University, Fort Collins, CO, 80523, USA

    Jessica E. Prenni

Authors
  1. Linxing Yao
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  3. Maliha W. Shaikh
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  4. Christopher B. Forsyth
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  6. Corey D. Broeckling
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Contributions

L. Yao and C.D. Broeckling designed experiments, L. Yao and E.A. Davidson conducted the experiments and acquired the data, L. Yao conducted data analysis, L. Yao, E.A. Davidson, M.W. Shaikh, C.B. Forsyth, J. E. Prenni, C.D. Broeckling contributed to manuscript writing.

Corresponding author

Correspondence to Linxing Yao.

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Plasma obtained from local blood bank has an approval from the Institutional Review Board of Colorado State University (IRB#16-063B).

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The authors declare no competing interests.

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Published in the topical collection Analytical Methods and Applications in the Materials and Life Sciences with guest editors Ute Resch-Genger, Matthias Koch, Björn Meermann, and Michael G. Weller.

This manuscript is dedicated to the 150th anniversary of BAM.

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Yao, L., Davidson, E.A., Shaikh, M.W. et al. Quantitative analysis of short-chain fatty acids in human plasma and serum by GC–MS. Anal Bioanal Chem 414, 4391–4399 (2022). https://doi.org/10.1007/s00216-021-03785-8

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  • DOI: https://doi.org/10.1007/s00216-021-03785-8

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