期刊
ANALYTICA CHIMICA ACTA
卷 854, 期 -, 页码 86-94出版社
ELSEVIER
DOI: 10.1016/j.aca.2014.11.015
关键词
Short-chain fatty acid; Isotope labeling; Chemical derivatization; 3-Nitrophenylhydrazine; Ultrahigh performance liquid chromatography-multiple-reaction; monitoring mass spectrometry
资金
- Genome Canada - Metabolomics Innovation Centre (TMIC)
- Genome Canada and Genome British Columbia through the Science and Technology Innovation Centre (STIC)
Short-chain fatty acids (SCFAs) are produced by anaerobic gut microbiota in the large bowel. Qualitative and quantitative measurements of SCFAs in the intestinal tract and the fecal samples are important to understand the complex interplay between diet, gut microbiota and host metabolism homeostasis. To develop a new LC-MS/MS method for sensitive and reliable analysis of SCFAs in human fecal samples, 3-nitrophenylhydrazine (3NPH) was employed for pre-analytical derivatization to convert ten C-2-C-6 SCFAs to their 3-nitrophenylhydrazones under a single set of optimized reaction conditions and without the need of reaction quenching. The derivatives showed excellent in-solution chemical stability. They were separated on a reversed-phase C-18 column and quantitated by negative-ion electrospray ionization - multiplereaction monitoring (MRM)/MS. To achieve accurate quantitation, the stable isotope-labeled versions of the derivatives were synthesized in a single reaction vessel from C-13(6)-3NPH, and were used as internal standard to compensate for the matrix effects in ESI. Method validation showed on-column limits of detection and quantitation over the range from low to high femtomoles for the ten SCFAs, and the intra-day and inter-day precision for determination of nine of the ten SCFAs in human fecal samples was <= 8.8% (n = 6). The quantitation accuracy ranged from 93.1% to 108.4% (CVs <= 4.6%, n = 6). This method was used to determine the SCFA concentrations and compositions in six human fecal samples. One of the six samples, which was collected from a clinically diagnosed type 2 diabetes patient showed a significantly high molar ratio of branch-chain SCFAs to straight-chain SCFAs than the others. In summary, this work provides a new LC-MS/MS method for precise and accurate quantitation of SCFAs in human feces. (C) 2014 Elsevier B.V. All rights reserved.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据