Quadram Institute Bioscience, Norwich Research Park, Norwich NR4 7UQ, UK.
National Research Centre, Chemistry of Flavour and Aroma Department, Dokki, Cairo 12622, Egypt.
Molecules. 2021 Oct 26;26(21):6444. doi: 10.3390/molecules26216444.
The gut microbiota is critical to the maintenance of physiological homeostasis and as such is implicated in a range of diseases such as colon cancer, ulcerative colitis, diabetes, cardiovascular diseases, and neurodegenerative diseases. Short chain fatty acids (SCFAs) are key metabolites produced by the gut microbiota from the fermentation of dietary fibre. Here we present a novel, sensitive, and direct LC-MS/MS technique using isotopically labelled internal standards without derivatisation for the analysis of SCFAs in different biological matrices. The technique has significant advantages over the current widely used techniques based on sample derivatization and GC-MS analysis, including fast and simple sample preparation and short LC runtime (10 min). The technique is specific and sensitive for the quantification of acetate, butyrate, isobutyrate, isovalerate, lactate, propionate and valerate. The limits of detection were all 0.001 mM except for acetate which was 0.003 mM. The calibration curves for all the analytes were linear with correlation coefficients r > 0.998. The intra- and inter-day precisions in three levels of known concentrations were <12% and <20%, respectively. The quantification accuracy ranged from 92% to 120%. The technique reported here offers a valuable analytical tool for use in studies of SCFA production in the gut and their distribution to host tissues.
肠道微生物群对于维持生理内稳态至关重要，因此与多种疾病有关，如结肠癌、溃疡性结肠炎、糖尿病、心血管疾病和神经退行性疾病。短链脂肪酸 (SCFA) 是肠道微生物群从膳食纤维发酵产生的关键代谢物。在这里，我们提出了一种新颖、灵敏、直接的 LC-MS/MS 技术，使用同位素标记的内标，无需衍生化，可分析不同生物基质中的 SCFA。该技术与目前广泛使用的基于样品衍生化和 GC-MS 分析的技术相比具有显著优势，包括快速简单的样品制备和短的 LC 运行时间（10 分钟）。该技术对乙酸盐、丁酸盐、异丁酸、异戊酸盐、乳酸盐、丙酸盐和戊酸盐的定量具有特异性和灵敏度。除乙酸盐外，所有分析物的检测限均为 0.001 mM，乙酸盐的检测限为 0.003 mM。所有分析物的校准曲线均具有相关性系数 r > 0.998。在三个已知浓度水平下，日内和日间精密度均<12%和<20%。定量准确性范围为 92%至 120%。这里报道的技术为研究肠道中 SCFA 的产生及其向宿主组织的分布提供了一种有价值的分析工具。
