Short-chain fatty acids (SCFA), also known as volatile fatty acids (VFAs), refer to organic fatty acids with 2-6 carbon atoms. SCFAs mainly include acetic acid, propionic acid, butyric acid, isobutyric acid, valeric acid, isovaleric acid, caproic acid and isohexanoic acid.
SCFAs can participate in the energy supply of intestinal epithelial cells, and gradually become signal molecules that regulate diet, intestinal microbes and the host. In the intestine, SCFAs can regulate intestinal immunity, glucose homeostasis, blood lipids, and reduce the risk of obesity and diabetes.
As the main energy substrate, SCFAs have anti-inflammatory and anti-cancer effects. SCFAs can affect chronic diseases through a variety of mechanisms, play an important role in maintaining the metabolic homeostasis of colonic epithelial cells, and protect the colon from external damage.
Creative Proteomics can provide you with one-stop high-sensitivity quantitative analysis of SCFAs, including SCFAs with C2-C12 carbon chain lengths of various biological materials (feces and serum, etc.).
Fig1. Gut microbial short-chain fatty acids and the risk of diabetes (Lau, W.L.;2019)
SCFA analysis provides valuable insights into the composition and activity of the gut microbiota. By quantifying the levels of different SCFAs, researchers can evaluate the microbial metabolic activity and identify alterations in the gut ecosystem associated with various diseases, such as inflammatory bowel disease, obesity, and diabetes.
SCFAs are strongly influenced by dietary factors, especially the consumption of dietary fiber. Analyzing SCFA profiles before and after dietary interventions helps in assessing the impact of different diets on gut microbial metabolism and overall health. This information can aid in the development of personalized dietary recommendations for individuals.
SCFAs have been proposed as potential biomarkers for several health conditions. By comparing SCFA profiles between healthy individuals and patients with specific diseases, researchers can identify unique patterns that may serve as diagnostic or prognostic markers. For example, altered levels of butyric acid have been associated with colorectal cancer and inflammatory bowel disease.
SCFA analysis is instrumental in evaluating the efficacy of probiotics and prebiotics. Probiotics are live bacteria that confer health benefits when consumed, while prebiotics are non-digestible substances that selectively promote the growth of beneficial gut bacteria. SCFA analysis helps determine if these interventions effectively modulate the gut microbiota and enhance SCFA production.
SCFA lipidomics analysis involves the comprehensive characterization and quantification of SCFAs in biological samples. The content of SCFA lipidomics analysis typically includes:
Identification and Quantification: SCFAs are identified and quantified using advanced analytical techniques such as gas chromatography-mass spectrometry (GC-MS) or liquid chromatography-mass spectrometry (LC-MS). These methods enable precise measurement of SCFA concentrations in complex biological samples.
Metabolite Profiling: In addition to SCFAs, lipidomics analysis can provide a broader perspective on the metabolic profile of the sample. It allows the identification and quantification of other metabolites, such as long-chain fatty acids, phospholipids, and sterols, which are relevant to gut health and metabolism.
Data Analysis: Sophisticated bioinformatics tools are employed to process and analyze the large datasets generated from SCFA lipidomics analysis. Statistical methods, pathway analysis, and multivariate analysis help uncover significant patterns and associations within the data.
GC-FID is a widely recognized and reliable technique for SCFA analysis, known for its exceptional sensitivity and selectivity. This method involves the separation of SCFAs in a gaseous phase using a capillary column, followed by the detection of the compounds using flame ionization. The combustion of SCFAs in the flame generates ions, which are then measured by the FID detector for quantification.
For GC-FID analysis of SCFAs, several instrumental setups are commonly employed, including the Agilent 7890A GC-FID, Shimadzu GC-2010 Plus, and Thermo Scientific TRACE 1310 GC systems.
LC-MS is another powerful analytical technique used for SCFA analysis, particularly for the detection of polar and non-volatile SCFAs. This approach offers high sensitivity, a wide dynamic range, and the capability to simultaneously analyze a broad range of metabolites alongside SCFAs. In LC-MS, SCFAs are separated using a liquid chromatography system, and their detection and quantification are performed using mass spectrometry.
Commonly employed LC-MS instrumentations for SCFA analysis include the Agilent 1260 Infinity II LC system, Shimadzu Nexera X2, and Thermo Scientific Vanquish Flex, coupled with mass spectrometers such as the Agilent 6460 Triple Quadrupole, Thermo Scientific Q Exactive Plus, and Waters Xevo TQ-S.
By utilizing these cutting-edge platforms and instruments, such as the GC-FID models mentioned above and LC-MS setups featuring the indicated systems, we have successfully conducted high-quality SCFA analysis. These instruments offer precise separation, identification, and quantification of SCFAs in complex samples, ensuring accurate and reliable results for a wide range of research applications.
Determination of short-chain fatty acids by GC-MS (Wang et al., 2020)
Quick-freeze in liquid nitrogen and store at -80°C, and send it on dry ice.
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