A rapid and convenient derivatization method for quantitation of short-chain fatty acids in human feces by ultra-performance liquid chromatography/tandem mass spectrometry.

Huan Fu, Qing-Li Zhang, Xiao-Wu Huang, Zheng-Hua Ma, Xiao-Li Zheng, Shi-Lin Li, Hao-Nan Duan, Xiao-Chu Sun, Fei-Fei Lin, Li-Juan Zhao, Guo-Sheng Teng, Jia Liu

Rapid communications in mass spectrometry : RCM 2020 May 15

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Short-chain fatty acids (SCFAs) are associated with intestinal microbiota and diseases in humans. SCFAs have a low response in mass spectrometry, and in order to increase sensitivity, reduce sample consumption, shorten analysis time, and simplify sample preparation steps, a derivatization method was developed. We converted seven SCFAs into amide derivatives with 4-aminomethylquinoline. The reaction occurred for 20 min at room temperature. The analytes were separated on a reversed-phase C18 column and quantitated in the positive ion electrospray ionization mode using multiple reaction monitoring. Acetic acid-d4 was used as the stable-isotope-labeled surrogate analyte for acetic acid in the working solutions, while the other stable-isotope-labeled standards were used as internal standards (ISs). Method validation showed that the intra-day and inter-day precision of quantitation for the seven SCFAs over the whole concentration range was ≤3.8% (n = 6). The quantitation accuracy ranged from 85.5% to 104.3% (n = 6). Most important, the collected feces were vortexed immediately with ethanol. This study provides a new derivatization method for a precise, accurate, and rapid quantitation of SCFAs in human feces using ultra-performance liquid chromatography/tandem mass spectrometry. This method successfully determined the concentration of SCFAs in human feces and could assist in the exploration of intestinal microbiota and diseases. © 2020 John Wiley & Sons, Ltd.

Citation

Huan Fu, Qing-Li Zhang, Xiao-Wu Huang, Zheng-Hua Ma, Xiao-Li Zheng, Shi-Lin Li, Hao-Nan Duan, Xiao-Chu Sun, Fei-Fei Lin, Li-Juan Zhao, Guo-Sheng Teng, Jia Liu. A rapid and convenient derivatization method for quantitation of short-chain fatty acids in human feces by ultra-performance liquid chromatography/tandem mass spectrometry. Rapid communications in mass spectrometry : RCM. 2020 May 15;34(9):e8730