A novel LCMSMS method for quantitative measurement of short-chain fatty acids in human stool derivatized with C-12- and C-13-labelled aniline

作者：Chan James Chun Yip; Kioh Dorinda Yan Qin; Yap Gaik Chin; Lee Bee Wah^*; Chan Eric Chun Yong^*

来源：Journal of Pharmaceutical and Biomedical Analysis, 2017, 138: 43-53.

DOI：10.1016/j.jpba.2017.01.044

• Short-chain fatty acids
• Butyric acid
• Human gut microbiome
• Human gut microbiota
• Human infant stool
• C-13-Aniline derivatization

A novel liquid chromatography tandem mass spectrometry (LCMSMS) method for the quantitative measurement of gut microbial-derived short-chain fatty acids (SCFAs) in human infant stool has been developed and validated. Baseline chromatographic resolution was achieved for 12 SCFAs (acetic, butyric, caproic, 2,2-dimethylbutyric, 2-ethylbutyric, isobutyric, isovaleric, 2-methylbutyric, 4-methylvaleric, propionic, pivalic and valeric acids) within an analysis time of 15 min. A novel sequential derivatization of endogenous and spiked SCFAs in stool via C-12- and C-13-aniline respectively, facilitated the accurate quantitation of C-12-aniline derivatized endogenous SCFAs based on calibration of exogenously C-13-derivatized SCFAs. Optimized quenching of derivatization agents prior to LCMSMS analysis further reduced to negligible levels the confounding chromatographic peak due to in-line derivatization of unquenched aniline with residual acetic acid present within the LCMS system. The effect of residual acetic acid, a common LCMS modifier, in analysis of SCFAs has not been addressed in previous SCFA assays. For the first time, a total of 9 SCFAs (acetic, butyric, caproic, isobutyric, isovaleric, 2-methylbutyric, 4-methylvaleric, propionic and valeric acids) were detected and quantitated in 107 healthy infant stool samples. The abundance and diversity of SCFAs in infant stool vary temporally from 3 weeks onwards and stabilize towards the end of 12 months. This in turn reflects the maturation of infant SCFA-producing gut microbiota community. In summary, this novel method is applicable to future studies that investigate the biological roles of SCFAs in paediatric health and diseases.

• 出版日期2017-5-10