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SCFAs provide a valuable opportunity to study the human gut microbiome, which consists of trillions of microbes, is involved in numerous physiological processes, and is vital to the host's health. The study of SCFAs by GC-MS may be an innovative approach to discovering non-invasive biomarkers of disease. The complex matrices of fecal samples may reduce the recovery, accuracy, and reproducibility of the method, and lead to instrumental contamination in repetitive analysis. Thus, selective extraction and derivatization of SCFAs from fecal samples are required for clinical analysis of a biological sample.
The principle of GC relies on the carrier gas as the mobile phase, and the sample compounds are separated by differential interaction with the column stationary phase. Depending on their chemical nature, retention time upon reaching a detector as well as its response are used for a qualitative and quantitative evaluation of compounds. Feces pretreatment is crucial for the detection of SCFAs. Recently, solid-phase microextraction (SPME) combined with gas chromatography-mass spectrometry (GC-MS) has proved to be very useful for the analysis of fatty acids in aqueous extracts with low detection limits.
Fig.1 Overall analytical procedure for the determination of SCFAs in fecal samples by GC-MS/MS. (Park, 2017)
Samples should be prepared upon collection at 4℃ or stored promptly at -20℃ (short-term) or at -80℃ (long-term). To preserve the integrity of the samples, freeze-thaw cycles should be minimized. In addition, the recovery of SCFAs is decreased with each day the samples remain at 4℃ or higher. To reduce sample thawing time, all solutions should be prepared before starting. Ultrapure water should be used for all solutions.
GC-MS is the most commonly used analytical technique due to the chemical nature (i.e., volatile nature) of SCFAs and the advantage of detectors that can be associated with these instruments. The GC-MS system makes full use of the advantages of quantitative evaluation by the isotope dilution method. The method uses isotopes of the analyte as internal standards, providing specificity, accuracy, and robustness to the obtained quantitative data. GC-MS is widely used because of its high peak capacity and high detection sensitivity.
Creative Biolabs is a flexible and dynamic team dedicated to accelerating research projects through customized services. We have more than 10 years of experience in gut microbiota research. Our focus on scientific expertise, technology, innovation, and problem-solving has made us the partner of choice for many large pharmaceutical and biotech companies. If you are interested in our SCFAs analysis with GC technology, please feel free to contact us for more.
For Research Use Only. Not intended for use in food manufacturing or medical procedures (diagnostics or therapeutics). Do Not Use in Humans.
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