Fast quantification of short-chain fatty acids in rat plasma by gas chromatography

Abstract: Short-chain fatty acids (SCFAs) are the main metabolites of the intestinal flora and play an important role in the interaction between the intestinal flora and host metabolism. Therefore, reliable methods are needed to accurately measure SCFAs concentrations. SCFAs are commonly analyzed by gas chromatography–mass spectrometry (GC-MS), which requires lengthy sample treatments and a long run time. This study aimed to develop a fast GC method with formic acid pretreatment for SCFAs quantification in the plasma of rat. Baseline chromatographic resolution was achieved for three SCFAs (acetic, propionic, and butyric) within an analysis time of 10.5 min. The method exhibited good recovery for a wide range of concentrations with a low limit of detection for each compound. The relative standard deviations (RSDs) of all targeted compounds showed good intra- and interday precision (<10%). We used our method to measure SCFAs levels in plasma samples from rats fed with a high fructose diet (HFD) to test the accuracy of the developed method. It was shown that SCFAs are indeed affected negatively by a HFD (60% fructose). This method was successfully employed to accurately determine SCFAs in the rat plasma with minimum sample preparation. Results showed potential damage of HFD, which produced lower SCFAs. Practical Application: Increasingly, microbiota and gut health research are being conducted by many food scientists to elucidate the relationships among the factors of food components, particularly the nondigestible carbohydrates, food processing conditions, and potential health impact. This research provides a useful, rapid, and accurate method that can save time in the analysis of short-chain fatty acids, which are commonly analyzed in gut health research.