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      저자 : Ashleigh B. Theberge (검색결과 1 건)
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        Chemical derivatization-based LC–MS/MS method for quantitation of gut microbial short-chain fatty acids

        Won-Suk Song,Han-Gyu Park,Seong-Min Kim,Sung-Hyun Jo,Byung-Gee Kim,Ashleigh B. Theberge,Yun-Gon Kim 한국공업화학회 2020 Journal of Industrial and Engineering Chemistry Vol.83 No.-

        Short chain fatty acids (SCFAs) are end products of fermentation by anaerobic gut microbiota. They can beused as beneficial metabolites to regulate the host’s physiological processes. Despite their importance,SCFAs are difficult to analyze with mass spectrometric technologies due to their poor ionizationefficiency and susceptibility to water loss during ionization of low molecular weight organic acids. Here,we developed a sensitive and reliable method to quantify SCFAs by liquid chromatography tandem massspectrometry (LC–MS/MS) in multiple reaction monitoring (MRM) mode. SCFAs were chemicallyderivatized by Girard’s reagent T (GT), providing a permanent cationic charge. This techniquedemonstrated an excellent quantitative capacity, showing good linearity (R2> 0.99) and limit ofquantification (femtomole levels) forfive SCFAs (i.e., acetate, propionate, butyrate, valerate, andcaproate). Next, we applied this derivatization method to quantitate SCFAs from a small volume of totalextracellular metabolites produced by Eubacterium rectale (E. rectale), one of main butyrate-producinggut bacteria. GT-labeled SCFAs were quantitated well in small volumes of culture medium (5, 10, 15, and20 mL), with the amount of SCFA measured being proportional to the volume of culture medium, asexpected. We also investigated plant-derived polysaccharides as prebiotics that could enhance theproduction of butyrate by E. rectale. Finally, the production of butyrate was successfully monitored in aco-culture system for E. rectale and Bifidobacterium longum (B. longum) by analyzing GT-labeled butyrate. Taken together, our results suggest that this highly sensitive method would be useful for quantifyingSCFAs extracted from stool in an aqueous solution to monitor gut health.

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