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- Title
A rapid and convenient derivatization method for quantitation of short-chain fatty acids in human feces by ultra-performance liquid chromatography/tandem mass spectrometry.
- Authors
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
- Abstract
Rationale: 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. Methods: 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). Results: 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. Conclusions: 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.
- Subjects
TANDEM mass spectrometry; SHORT-chain fatty acids; LIQUID chromatography; FECES; GUT microbiome; DERIVATIZATION
- Publication
Rapid Communications in Mass Spectrometry: RCM, 2020, Vol 34, Issue 9, p1
- ISSN
0951-4198
- Publication type
Article
- DOI
10.1002/rcm.8730