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Determination of short-chain fatty acids by N,N-dimethylethylenediamine derivatization combined with liquid chromatography/mass spectrometry and their implication in influenza virus infection

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Abstract

Short-chain fatty acids (SCFAs) are the end products of the fermentation of complex carbohydrates by the gut microbiota. Although SCFAs are recognized as important markers to elucidate the link between gut health and disease, it has been difficult to analyze SCFAs with mass spectrometry technologies due to their poor ionization efficiency and high volatility. Here, we present a novel and sensitive method for the quantification of SCFAs, including C2–C6 SCFAs and their hydroxy derivatives, by liquid chromatography/tandem mass spectrometry (LC–MS/MS) upon N,N-dimethylethylenediamine (DMED) derivatization with a run time of 10 min. Moreover, the quantification method of DMED-derivatized SCFAs in intestinal contents using isotope-labeled internal standards was also established. The method validation was performed by analyzing spiked intestinal samples; the limits of detection and quantification of SCFAs with this method were found to be 0.5 and 5 fmol, respectively; the recovery was greater than 80% and good linearity (0.9932 to 0.9979) of calibration curves was obtained over the range from 0.005 to 5000 pmol/μL; the intraday and interday precisions were achieved in the range of 1–5%. Furthermore, the validated method was applied to analyze SCFAs in the cecum and colon contents of mice infected with the influenza virus. The results showed that the concentration of most of the SCFAs tested here decreased significantly in a time-dependent manner after the infection, suggesting a possibility that SCFAs in intestinal samples could be used as severe disease markers. Overall, we here successfully developed a simple, fast, and sensitive method for SCFA analysis by LC–MS/MS combined with DMED derivatization. The method for the quantification of SCFAs will be a useful tool for both basic research and clinical studies.

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Funding

This work is supported by the Japanese Society for the Promotion of Science KAKENHI Grants (18K0743408, 19K0786109, 20K2335900, 21K1481201, and 21H02376) and by the Hokkaido Bureau of Economy, Trade, and Industry (strategic basic technology advancement support project).

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Author notes

  1. Divyavani Gowda and Yonghan Li contributed equally to this work.

Authors and Affiliations

  1. Faculty of Health Sciences, Hokkaido University, Kita-12 Nishi-5, Kita-ku, Sapporo, 060-0812, Japan

    Divyavani Gowda, Siddabasave Gowda B Gowda & Shu-Ping Hui

  2. Graduate School of Health Science, Hokkaido University, Kita-12, Nishi-5, Kita-ku, Sapporo, 060-0812, Japan

    Yonghan Li

  3. Graduate School of Global Food Resources, Hokkaido University, Kita-9 Nishi-9, Kita-ku, Sapporo, 060-0812, Japan

    Siddabasave Gowda B Gowda

  4. International Institute for Zoonosis Control, Hokkaido University, Kita-20, Nishi-10, Kita-ku, Sapporo, 001-0020, Japan

    Marumi Ohno

  5. Department of Nutrition, Sapporo University of Health Sciences, Nakanuma Nishi-4-3-1-15, Higashi-ku, Sapporo, 007-0894, Japan

    Hitoshi Chiba

Authors
  1. Divyavani Gowda
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  5. Hitoshi Chiba
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  6. Shu-Ping Hui
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Contributions

D.G., S.G.B.G.: conceptualization, methodology, supervision. Y.L., D.G., S.G.B.G.: formal analysis, visualization, writing—original draft. M.O., Y.L.: data curation, writing—review and editing. H.C., S.-P.H.: supervision, resources, writing—review and editing.

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Correspondence to Shu-Ping Hui.

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All the animal protocols were approved by the Animal Care and Use Committee of Hokkaido University (approval no. 17–0003).

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Gowda, D., Li, Y., B Gowda, S. et al. Determination of short-chain fatty acids by N,N-dimethylethylenediamine derivatization combined with liquid chromatography/mass spectrometry and their implication in influenza virus infection. Anal Bioanal Chem 414, 6419–6430 (2022). https://doi.org/10.1007/s00216-022-04217-x

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  • DOI: https://doi.org/10.1007/s00216-022-04217-x

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