A novel LCMSMS method for quantitative measurement of short-chain fatty acids in human stool derivatized with 12C- and 13C-labelled aniline
A novel LCMSMS method for quantitative measurement of short-chain fatty acids in human stool derivatized with 12C- and 13C-labelled aniline
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DOI：
10.1016/j.jpba.2017.01.044
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发表时间：
2017-05-10
影响因子：
3.4
通讯作者：
Chan, Eric Chun Yong
中科院分区：
文献类型：
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作者：
Chan, James Chun Yip;Kioh, Dorinda Yan Qin;Chan, Eric Chun Yong
建立了一种新的液相色谱串联质谱（LCMSMS）方法，用于定量测定人类婴儿粪便中肠道微生物衍生的短链脂肪酸（SCFAs）。在15分钟的分析时间内，12种scfa（乙酸、丁酸、己酸、2,2-二甲基丁酸、2-乙基丁酸、异丁酸、异戊酸、2-甲基丁酸、4-甲基戊酸、丙酸、戊酸和戊酸）达到了基线色谱分辨率。通过C-12-苯胺和c -13-苯胺分别对粪便中内源性和加标的SCFAs进行了新的顺序衍生化，在对外源性c -13-苯胺衍生的SCFAs进行校准的基础上，促进了C-12-苯胺衍生的内源性SCFAs的准确定量。在LCMSMS分析之前，衍生化剂的优化淬火进一步降低了由于在LCMS系统中存在残留乙酸的未淬灭苯胺的在线衍生化而导致的混淆色谱峰的水平。残留乙酸（一种常见的LCMS修饰剂）在SCFA分析中的作用在以前的SCFA分析中没有得到解决。本文首次在107例健康婴儿粪便样本中检测并定量了9种SCFAs（乙酸、丁酸、己酸、异丁酸、异戊酸、2-甲基丁酸、4-甲基戊酸、丙酸和戊酸）。婴儿粪便中SCFAs的丰度和多样性从3周开始暂时变化，并在12个月结束时保持稳定。这反过来反映了婴儿产生scfa的肠道微生物群落的成熟。总之，这种新方法适用于未来研究SCFAs在儿科健康和疾病中的生物学作用。(C) 2017 Elsevier B.V.版权所有
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. (C) 2017 Elsevier B.V. All rights reserved.