A Facile Profiling Method of Short Chain Fatty Acids Using Liquid Chromatography-Mass Spectrometry

Abstract

Short chain fatty acids (SCFAs) are the main products of dietary fibers that are not digested by the human body, and they have been shown to affect human metabolism and inflammation. The amount of SCFAs in the body is related to many human diseases, and studies have focused on elucidating their roles and target molecules in both metabolic and immune responses. Thus, the quantitation of SCFAs in biological samples becomes crucial in understanding their important roles in the human body. Herein, a facile profiling method of SCFAs using liquid chromatography-tandem mass spectrometry (LC-MS/MS) was developed and then applied to biological samples. C2-C6 SCFAs were derivatized while using 4-acetamido-7-mercapto-2,1,3-benzoxadiazole for 5 min. at room temperature prior to LC-MS/MS analysis, and characteristic fragmentation patterns and increased hydrophobicity after chemical derivatization enabled specific discrimination among 12 SCFAs. Derivatization was fast and reliable, and the reaction products were stable for a week at 4 °C. The developed method was applied to measure SCFAs in mouse feces, plasma, and human exhaled breath condensates. This fast and simple method can save labor and effort to profile SCFAs from various biological samples.

High-fiber diets attenuate emphysema development via modulation of gut microbiota and metabolism

“…Standard metabolites and internal standards were purchased from Sigma-Aldrich (St. Louis, MO, USA), and all solvents were purchased from J. T. Baker (Center Valley, PA, USA). The SCFA metabolites in feces and BA metabolites were measured using a previously described procedure 70 , 71 . Mouse serum (20 µL) was mixed well with 500 μL of cold acetonitrile and 50 μL of internal standard solution (50 μL of 1 μM cholic acid-d 5 solution).…”

Sensitive and quantitative determination of short-chain fatty acids in human serum using liquid chromatography mass spectrometry

“…Han et al (2015) measured the 3-NPH derivatives of the SCFAs in faeces, so this work demonstrates that the derivatising reagent is also sensitive enough for serum applications, where the concentrations of the SCFAs are several orders of magnitude lower [ 9 ]. Song et al (2019) selected 4-acetamido-7-mercapto-2,1,3-benzoxadiazole as the derivatising reagent to monitor SCFAs in mouse serum, but with LOQs typically 3–10 times higher than reported here, minor SCFAs such as valeric and isovaleric acid would not be quantifiable in human serum [ 13 ]. Wei et al (2020) quantified the SCFAs in mouse serum after derivatisation with d 0 /d 6 -N, N-dimethyl-6,7-dihydro-5H-pyrrolo [3,4-d] pyrimidine-2-amine using HRAM-MS operating in PRM mode and reported LOQs in the range 0.5–4 fg on column (for a 10-μL sample injection) [ 11 ].…”

Plasma Short-Chain Fatty Acids and Their Derivatives in Women with Gestational Diabetes Mellitus

“…Determination of SCFA in biological materials allows a better understanding of their role, including outside of diabetes mellitus. Multiple analytical approaches for the detection of SCFA have been developed covering advanced chromatographic, mass spectrometric, electromigration and spectroscopic methods [16,[30][31][32][33][34][35][36][37][38][39]. The most common methods combine gas chromatography (GC), or liquid chromatography (LC) coupled with mass spectrometry (MS), most often with a derivatization step, which is time consuming but provides better chromatographic behavior.…”

“…In particular, GC-MS offers high sensitivity [31][32][33]. In recent years, LC-MS approaches that allow fast and efficient sample preparation and analysis were introduced [16,30,34]. Despite developments in SCFA analysis, the methods remain laborious and, therefore, SCFA determination is still not utilized in routine clinical practice.…”