Enhanced Detection of Short‐Chain Fatty Acids Using Gas Chromatography Mass Spectrometry

Abstract

Short‐chain fatty acids (SCFAs) are produced mainly by intestinal microbiota and play an important role in many host biological processes such as immune system development, glucose and energy homeostasis, and regulation of immune response and inflammation. In addition, they participate in the regulation of anorectic hormones, which have a role in appetite control, tumor suppression, and regulating the central and peripheral nervous systems. As such, there is great interest in monitoring levels of SCFAs in various biological samples. Due to the highly hydrophilic and volatile characteristics of SCFAs, optimizing extraction and sample preparation procedures is often a central component to further improve SCFA quantification. Here, we describe a rapid and highly sensitive analytical method for measuring SCFAs in human serum and feces. Briefly, SCFAs are protected by adding sodium hydroxide, followed by a one‐step extraction (pH > 7). Then, SCFAs are quantified by gas chromatography coupled to mass spectrometry (GC‐MS) after derivatization with N‐tert‐butyldimethylsilyl‐N‐methyltrifluoroacetamide (MTBSTFA). This method demonstrates excellent sensitivity, linearity, and derivatization efficiency for simultaneous determination of 14 different SCFAs. Further, this validated method can be successfully applied to quantify SCFAs in micro‐scale biological samples. In summary, we describe efficient and advanced sample preparation and detection procedures that are critically needed for monitoring SCFA concentrations in human biological samples. © 2021 Wiley Periodicals LLC. Basic Protocol: SCFA extraction and detection from fecal and serum samples with gas chromatography−mass spectrometry

Microbiome and metabolome profiles of high screen time in a cohort of healthy college students

“…GC–MS conditions used here were adopted from a previously published protocol 74 . Briefly, GC–MS experiments were performed on an Agilent 7820A GC-5977B MSD system (Santa Clara, CA) by injecting 1 µL of prepared samples.…”

Metabolic Profiling of Neocortical Tissue Discriminates Alzheimer’s Disease from Mild Cognitive Impairment, High Pathology Controls, and Normal Controls

“…Following storage at −20 °C for 20 min and centrifugation at 14 000 rpm for 10 min, 450 μL of supernatant were collected and sample pH was adjusted to 10 by adding 30 μL of NaOH/H 2 O (1:4, v/v). Samples were then dried, and they were measured using the same protocol as that outlined for GC–MS untargeted profiling …”

“…Samples were then dried, and they were measured using the same protocol as that outlined for GC−MS untargeted profiling. 53…”

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.…”

“…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].…”