Analytical factors for eight short-chain fatty acid analyses in mouse feces through headspace solid-phase microextraction–triple quadrupole gas chromatography tandem mass spectrometry

Abstract

This study developed a method for quantifying eight short-chain fatty acids (SCFAs) in mouse fecal samples using solid-phase microextraction (SPME) coupled with triple quadrupole gas chromatography tandem mass spectrometry. Furthermore, significant factors affecting SCFA analysis, including SPME fiber selection, pH, salting-out agent, and sample collection time, were investigated. Contrary to previous studies, we found that the CAR/PDMS fiber had the highest extraction efficiency for all SCFAs. The optimal extraction efficiency was observed at pH 2.0, particularly for low-molecular-weight SCFAs. NaH2PO4 showed a more effective extraction efficiency than NaCl, owing to its pH stability and less interference with the solvent matrix. Additionally, our results showed that the SCFA concentration increased over collection time. The composition ratio of the eight SCFAs was maintained for up to 24 h; thus, we concluded that samples should be collected within four hours to obtain reliable results. Our findings may improve laboratory methods for SCFA extraction and mouse fecal sample analysis. Graphical Abstract

A Chiral-LC-MS Method for the Simultaneous Quantification of Short-Chain Fatty Acids and D/L-Lactate in the Ruminal Fluid of Dairy Cows

“…For GC-based methods (including GC-FID and GC-MS), two types of protocols can be found for SCFA quantification, i.e., direct GC analysis and GC analysis after derivatisation [ 5 , 6 , 7 , 8 , 9 , 10 , 11 , 12 ]. While a sample pretreatment step (such as acidification, liquid–liquid extraction and solid-phase microextraction) is generally performed for direct GC injection, several derivatisation protocols (such as silylation, methylation and chloroformate derivatisation) have been reported for SCFA.…”