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HS-GC with A Simple Pretreatment Protocol to Determine Short-chain Fatty Acids in Mouse Feces and the Effect of Polysaccharide from Acanthopanax trifoliatus (L.) Merr on Their Distribution in Diabetic Mice

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Abstract

Increasing numbers of studies suggest that short-chain fatty acids (SCFAs) are closely related to diabetes. However, the complexity of fecal pretreatment makes the detection of SCFAs in feces relatively difficult. In this study, we established a sample pretreatment protocol that involved no tedious steps, requiring only the grinding of appropriate amounts of feces into powder in liquid nitrogen. The SCFA content in feces was then detected using the headspace-gas chromatography (HS-GC) method. The method achieved good linearity (R2 ≥ 0.9940), and the limits of quantification (LOQs) and detection (LODs) of all investigated SCFAs ranged from 0.15 to 0.45 mM and 0.05 to 0.15 mM, respectively. The recoveries of the SCFAs ranged from 91.55 to 103.96% in mouse fecal samples. The HS-GC method was then applied to evaluate the effects of a neutral polysaccharide from Acanthopanax trifoliatus (ATP1-1) on the fecal SCFA content in streptozotocin (STZ)-induced diabetic mice. After 6 weeks of treatment with 140 mg/kg/d ATP1-1, the expression levels of acetic acid, propionic acid, butyric acid, isobutyric acid, isovaleric acid and valeric acid were increased by 215.4, 88.8, 95.3, 300.8, 260.7 and 158.0%, respectively, which indicated that the hypoglycemic effect of ATP1-1 may be related to its impact on SCFAs. Our simplified pretreatment of fecal samples is suitable for the analysis and detection of SCFAs in mouse feces by HS-GC.

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Acknowledgements

This work was supported by the “Special funds of key disciplines construction from Guangdong and Zhongshan cooperating”. We would like to give special thanks for the generous support of Xinyu Peng, director of the Pharmaceutical Research Office of the Institute of Animal Health, Guangdong Academy of Agricultural Sciences. We sincerely thank Dr. Qing Zhou from the School of Pharmacy of Guangdong Pharmaceutical University for his guidance in writing the paper.

Funding

This study was funded by the “Special funds of key disciplines construction from Guangdong and Zhongshan cooperating”.

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Authors and Affiliations

  1. School of Pharmacy, Guangdong Pharmaceutical University, Guangzhou, 510006, China

    Yue Liu, Yuzi Lin, Ping Li, Huiwen Yang & Yufang Pan

  2. Zhongshan Institute for Drug Discovery, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Zhongshan, 528400, China

    Yue Liu

  3. Department of Pharmacy, Xiamen Children’s Hospital, Xiamen, 361006, China

    Ping Li

  4. School of Life Sciences and Biopharmaceuticals, Guangdong Pharmaceutical University, Guangzhou, 510006, China

    Qingsong Huang

  5. Guangdong Provincial Key Laboratory of Advanced Drug Delivery Systems, Guangdong Pharmaceutical University, Guangzhou, 510006, China

    Yufang Pan

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  1. Yue Liu
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Correspondence to Huiwen Yang or Yufang Pan.

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All animal experiments were carried out according to protocols approved by the Institutional Animal Ethics Committee of Guangdong Pharmaceutical University.

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Liu, Y., Lin, Y., Li, P. et al. HS-GC with A Simple Pretreatment Protocol to Determine Short-chain Fatty Acids in Mouse Feces and the Effect of Polysaccharide from Acanthopanax trifoliatus (L.) Merr on Their Distribution in Diabetic Mice. Chromatographia 85, 809–816 (2022). https://doi.org/10.1007/s10337-022-04187-3

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