Determination of Short-Chain Fatty Acids in Mouse Feces by High-Performance Liquid Chromatography Using 2-Nitrophenylhydrazine as a Labeling Reagent

It has been suggested that imbalances in gut microbiota are related to diseases associated with metabolism, the central nervous system, etc. Therefore, analysis of short-chain fatty acids (SCFAs) produced by gut microbiota is very important as an indicator of causation, demonstrating the effects on...

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Bibliographic Details
Published in:Biological & pharmaceutical bulletin 2019/05/01, Vol.42(5), pp.845-849
Main Authors: Inoue, Hirofumi, Takayama, Kento, Takahara, Chiho, Tabuchi, Norihiko, Okamura, Nobuyuki, Narahara, Naoko, Kojima, Eijiro, Date, Yuuko, Tsuruta, Yasuto
Format: Article
Language:English
Subjects:
2-nitrophenylhydrazine
Acetonitrile
Causation
Central nervous system
Elution
Fatty acids
Feces
High performance liquid chromatography
HPLC
Intestinal microflora
Liquid chromatography
Metabolism
Microbiota
Phosphoric acid
short-chain fatty acid
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Summary:It has been suggested that imbalances in gut microbiota are related to diseases associated with metabolism, the central nervous system, etc. Therefore, analysis of short-chain fatty acids (SCFAs) produced by gut microbiota is very important as an indicator of causation, demonstrating the effects on the host due to changes in the gut microbiota. We developed a HPLC method for the determination of SCFAs in mouse feces. After homogenization, the SCFAs in mouse feces and 2-ethylbutyric acid (internal standard) were derivatized with 2-nitrophenylhydrazine (2-NPH) in the presence of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide. The 2-NPH derivatives of SCFAs and the internal standard were separated on a reversed-phase column (octadecyl silyl column) by gradient elution using phosphoric acid (pH 2.5)–acetonitrile at 50°C and detected by absorbance measurement at 400 nm. The recovery of the method was 90–115%, with a precision (relative standard deviation) of 1.3–7.7%. The determination of SCFAs by the present method can provide useful information for biological and clinical research.
ISSN:0918-6158
1347-5215
DOI:10.1248/bpb.b18-01017